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Two subfamilies of land planarians (Geoplanidae) are endemic to the Neotropical region, namely Geoplaninae (with 29 genera and 346 nominal species, most of which are from Brazil) and its sister-group Timyminae, with only two Chilean species. The systematics of these groups through morphology and molecular data (COI and 28S rDNA genes), including nine new Chilean species, is re-assessed in this study. The great morphological diversity of the Chilean species is congruent with the molecular trees and, accordingly, five new genera (Adinoplana, Harana, Myoplana, Sarcoplana and Transandiplana) are proposed, each characterized by putative synapomorphies. Seven new tribes are also erected (Adinoplanini, Gusanini, Haranini, Inakayaliini, Myoplanini, Polycladini and Sarcoplanini), each one monogeneric, except Geoplanini (which includes all genera under the current concept of Geoplaninae plus the Chilean Transandiplana) and Sarcoplanini (with Sarcoplana and the already known Mapuplana, Pichidamas and Wallamapuplana). Re-diagnoses of Geoplaninae, Timymini, Gusana, Inakayalia, Polycladus and Pichidamas are proposed and biogeographic remarks on Transandiplana are provided

Scientific reports 12, 16571 (2022). doi:10.1038/s41598-022-20170-2, Published by Macmillan Publishers Limited, part of Springer Nature, [London]

The objective of this study was to evaluate the accuracy of the SureTune3 postoperative imaging software in determining the location of a deep brain stimulation (DBS) electrode based on clinical outcomes and the adverse effects (AEs) observed.Twenty-six consecutive patients with Parkinson disease (n = 17), essential tremor (n = 8), and dystonia (n = 1) who underwent bilateral DBS surgery (52 electrodes) were included in this study. Presurgical assessments were performed in all patients prior to surgery and at 3 and 6 months after surgery, using quality-of-life and clinical scales in each case. The SureTune3 software was used to evaluate the anatomical positioning of the DBS electrodes.Following DBS surgery, motor and quality-of-life improvement was observed in all patients. Different AEs were detected in 12 patients, in 10 of whom (83.3%) SureTune3 related the symptoms to the positioning of an electrode. A clinical association was observed with SureTune3 for 48 of 52 (92.3%) electrodes, whereas no association was found between the AEs or clinical outcomes and the SureTune3 reconstructions for 4 of 52 electrodes (7.7%) from 4 different patients. In 2 patients, the contact chosen was modified based on the SureTune3 data, and in 2 cases, the software helped determine that second electrode replacement surgery was necessary.The anatomical position of electrodes analyzed with SureTune3 software was strongly correlated with both the AEs and clinical outcomes. Thus, SureTune3 may be useful in clinical practice, and it could help improve stimulation parameters and influence decisions to undertake electrode replacement surgery.

Injury : international journal of the care of the injured 53(8), 2774-2782 (2022). doi:10.1016/j.injury.2022.05.009, Published by Elsevier Science, Amsterdam [u.a.]

Grau, José Horacio, Almeida, Ana Laura, Sluys, Ronald, Carbayo, Fernando (2022): A new genus and two new species of land planarians (Platyhelminthes: Tricladida: Geoplanidae) from Southern Chile. Journal of Natural History 56 (13-16): 947-967, DOI: 10.1080/00222933.2022.2097137, URL: http://dx.doi.org/10.1080/00222933.2022.2097137

There is currently a degree of divergence among the main clinical practice guidelines on the management of risk factors for peripheral arterial disease (PAD). This project aims to gain understanding of the management of PAD risk factors in clinical practice and to reach a multidisciplinary consensus on the strategies to be followed in order to optimize its identification, treatment, and follow-up.A multidisciplinary consensus following the Delphi methodology.Professionals (n = 130) with extensive experience in PAD participated in this consultation. The results suggest that in order to optimize the control of risk factors, efforts should be aimed at: (1) promoting the involvement and awareness of all specialists in the identification of and screening for the disease; (2) guaranteeing the possibility of evaluating the ankle-brachial index (ABI) in all the medical specialties involved; (3) promoting strategies for patients to quit smoking through the use of drugs, programs, or referrals to specialized units; (4) promoting an appropriate Mediterranean-based diet and the prescription of daily exercise; (5) raising awareness of the importance of ensuring LDL cholesterol values below 70 mg/dL, especially in symptomatic but also in asymptomatic patients (55 mg/dL following the publication of the ESC/EAS guide); (6) recommending the use of antiplatelet therapy in asymptomatic patients with diabetes mellitus (DM) and/or a pathological ABI; and (7) protocolizing the annual evaluation of ABI in high-risk patients.This document presents the 22 agreed-upon strategies which are intended to help professionals optimize multidisciplinary management of PAD risk factors.

Introduction: This study aimed to develop an individualized artificial intelligence model to help radiologists assess the severity of COVID-19’s effects on patients’ lung health.Methods: Data was collected from medical records of 1103 patients diagnosed with COVID-19 using RT- qPCR between March and June 2020, in Hospital Madrid-Group (HM-Group, Spain). By using Convolutional Neural Networks, we determine the effects of COVID-19 in terms of lung area, opacities, and pulmonary air density. We then combine these variables with age and sex in a regression model to assess the severity of these conditions with respect to fatality risk (death or ICU).Results: Our model can predict high effect with an AUC of 0.736. Finally, we compare the performance of the model with respect to six physicians’ diagnosis, and test for improvements on physicians’ performance when using the prediction algorithm.Discussion: We find that the algorithm outperforms physicians (39.5% less error), and thus, physicians can significantly benefit from the information provided by the algorithm by reducing error by almost 30%.

Tuberculosis (TB) is still considered a leading cause of death and a substantial threat to global child health. Both TB infection and disease are curable using antibiotics. However, most children who die of TB are never diagnosed or treated. In clinical practice, experienced physicians assess TB by examining chest X-rays (CXR). Pediatric CXR has specific challenges compared to adult CXR, which makes TB diagnosis in children more difficult. Computer-aided diagnosis systems supported by Artificial Intelligence have shown performance comparable to experienced radiologist TB readings, which could ease mass TB screening and reduce clinical burden. We propose a multi-view deep learning-based solution which, by following a proposed template, aims to automatically regionalize and extract lung and mediastinal regions of interest from pediatric CXR images where key TB findings may be present. Experimental results have shown accurate region extraction, which can be used for further analysis to confirm TB finding presence and severity assessment. Code publicly available at https://github.com/dani-capellan/pTB_LungRegionExtractor., This work has been accepted at the SPIE Medical Imaging 2023, Image Processing conference

PDF file - 12K, Ascribed functions of 10-biomarker diagnostic panel.

XLSX file - 12K, Summary of cubic-root transformed^ biomarker data by institute.

Supplementary Data from Identification of PMF1 Methylation in Association with Bladder Cancer Progression

Purpose: Polyamines are important regulators of cell growth and death. The polyamine modulated factor-1 (PMF-1) is involved in polyamine homeostasis. After identifying an enriched CpG island encompassing the PMF1 promoter, we aimed at evaluating the clinical relevance of PMF1 methylation in bladder cancer.Experimental Design: The epigenetic silencing of PMF1 by hypermethylation was tested in bladder cancer cells (n = 11) after azacytidine treatment. PMF1 methylation status was evaluated in 507 bladder tumors and 118 urinary specimens of bladder cancer patients and controls. PMF1 protein expression was analyzed by immunohistochemistry on tissue arrays containing bladder tumors for which PMF1 methylation was assessed (n = 218).Results: PMF1 hypermethylation was associated with gene expression loss, being restored in vitro by a demethylating agent. An initial set of 101 primary frozen bladder tumors served to identify PMF1 hypermethylation in 88.1% of the cases. An independent set of 406 paraffin-embedded tumors also revealed a high PMF1 methylation rate (77.6%). PMF1 methylation was significantly associated with increasing stage (P = 0.025). Immunohistochemical analyses revealed that PMF1 methylation was associated with cytoplasmic PMF1 expression loss (P = 0.032). PMF1 protein expression patterns were significantly associated with stage (P < 0.001), grade (P < 0.001), and poor overall survival using univariate (P < 0.001) and multivariate (P = 0.011) analyses. Moreover, PMF1 methylation in urinary specimens distinguished bladder cancer patients from controls (area under the curve = 0.800).Conclusion: PMF1 was identified to be epigenetically modified in bladder cancer. The association of PMF1 methylation with tumor progression and its diagnostic ability using urinary specimens support including PMF1 assessment for the clinical management of bladder cancer patients.

PDF file - 672K, Figure S1. High eIF3b mRNA expression is associated with human bladder cancer. Figure S2. eIF3b antibody specificity Figure S3. High eIF3b mRNA expression is associated with aggressive phenotypes in human prostate cancer Figure S4. Time course of eIF3b depletion. Figure S5. Depletion of eIF3b inhibits prostate cancer cell growth. Figure S6. Depletion of eIF3b affects the cell cycle in cancer cells. Figure S7. Depletion of eIF3b does not induce apoptosis in UMUC3 cells. Figure S8. Depletion of eIF3b decreases cell spreading and adhesion through integrin alpha5 in PC3 cells. Figure S9. Depletion of eIF3c inhibits cell growth, protein synthesis and disrupts actin cytoskeleton organization and focal adhesion.

XLSX file - 8K, Comparison of biomarker concentration data across 3 independent cohort studies (n = 755).

PDF file - 136K, Translation initiation factor eIF3b expression in human cancer and its role in tumor growth and lung colonization Table S1. Clinicopathologic characteristics of bladder patient samples in microarray datasets Table S2. Clinicopathologic characteristics of prostate patient samples in microarray datasets Table S3. eIF3 subunits mRNA expression in normal bladder vs. bladder tumor Table S4. Half-life of selected proteins involved in current study.

Supplementary Figures 1-5 from Src and Caveolin-1 Reciprocally Regulate Metastasis via a Common Downstream Signaling Pathway in Bladder Cancer

Supplementary Methods, Table 1, Figure Legends 1-5 from Src and Caveolin-1 Reciprocally Regulate Metastasis via a Common Downstream Signaling Pathway in Bladder Cancer

Background Identifying predictive non-invasive biomarkers of immunotherapy response is crucial to avoid premature treatment interruptions or ineffective prolongation. Our aim was to develop a non-invasive biomarker for predicting immunotherapy clinical durable benefit, based on the integration of radiomics and clinical data monitored through early anti-PD-1/PD-L1 monoclonal antibodies treatment in patients with advanced non-small cell lung cancer (NSCLC). Methods In this study, 264 patients with pathologically confirmed stage IV NSCLC treated with immunotherapy were retrospectively collected from two institutions. The cohort was randomly divided into a training (n = 221) and an independent test set (n = 43), ensuring the balanced availability of baseline and follow-up data for each patient. Clinical data corresponding to the start of treatment was retrieved from electronic patient records, and blood test variables after the first and third cycles of immunotherapy were also collected. Additionally, traditional radiomics and deep-radiomics features were extracted from the primary tumors of the computed tomography (CT) scans before treatment and during patient follow-up. Random Forest was used to implementing baseline and longitudinal models using clinical and radiomics data separately, and then an ensemble model was built integrating both sources of information. Results The integration of longitudinal clinical and deep-radiomics data significantly improved clinical durable benefit prediction at 6 and 9 months after treatment in the independent test set, achieving an area under the receiver operating characteristic curve of 0.824 (95% CI: [0.658,0.953]) and 0.753 (95% CI: [0.549,0.931]). The Kaplan-Meier survival analysis showed that, for both endpoints, the signatures significantly stratified high- and low-risk patients (p-value< 0.05) and were significantly correlated with progression-free survival (PFS6 model: C-index 0.723, p-value = 0.004; PFS9 model: C-index 0.685, p-value = 0.030) and overall survival (PFS6 models: C-index 0.768, p-value = 0.002; PFS9 model: C-index 0.736, p-value = 0.023). Conclusions Integrating multidimensional and longitudinal data improved clinical durable benefit prediction to immunotherapy treatment of advanced non-small cell lung cancer patients. The selection of effective treatment and the appropriate evaluation of clinical benefit are important for better managing cancer patients with prolonged survival and preserving quality of life.

Purpose: Studies have shown increased invasive Group A Streptococcus (GAS) disease, including bloodstream infections (GAS-BSI). However, the epidemiological data of GAS-BSI are limited in children. We aimed to describe GAS-BSI in Spanish children over 13 years (2005-2017). Methods: Multicenter retrospective cohort study from 16 Spanish hospitals. Epidemiology, symptomatology, laboratory, treatment, and outcome of GAS-BSI in children ≤16 years were analyzed. Results: 109 cases of GAS-SBI were included, with incidence rate of 4.3 episodes/100,000 children attended at the emergency department/year. We compared incidence between two periods (P1:2005-June 2011 vs P2:July 2011-2017) and observed a non-significant increase along the study period (APC:+6.0% [95%CI:-2.7,+15.4]; p=0.163). Median age was 24.1 months (IQR:14.0–53.7), peaking during the first four years of life (89/109 cases;81.6%). Primary BSI (46.8%), skin and soft tissue (21.1%), and osteoarticular infections (18.3%) were the most common syndromes. We compared children with primary BSI with those with a known source and observed that the former had shorter hospital stay (7vs.13 days; p=0.003) and received intravenous antibiotics less frequently (72.5%vs.94.8%; p=0.001) and for shorter periods (10vs.21 days; p=0.001). 22% of cases required PICU admission. Factors associated with severity were respiratory distress, pneumonia, thrombocytopenia, and surgery, but in multivariate analysis, only respiratory distress remained significant (adjusted OR:9.23 [95%CI:2.16-29.41]). Two children (1.8%) died. Conclusion: We observed a trend in increased incidence of GAS-BSI within the study period. Younger children were more frequently involved, and primary BSI was the most common and less severe syndrome. PICU admission was frequent, being respiratory distress the main risk factor.

The Chilean land planarian genus Gusana (Platyhelminthes, Tricladida, Geoplaninae) currently comprises three species that were described in the 19th century. Four new species of the genus are described herein, namely G. hualpensis Carbayo, sp. nov., G. lujanae Almeida & Carbayo, sp. nov., G. melipeucensis Almeida & Carbayo, sp. nov. and G. purensis Bolonhezi, Almeida & Carbayo, sp. nov. An integrative taxonomy approach was adopted by combining morphological (anatomy, histology) and molecular (COI and 28S genes) information. Additionally, the monophyletic status of Gusana is investigated and the species are delimited by a recently introduced molecular delimitation method based on pairwise genetic distances. All Gusana species are very similar in attributes such as body shape, dorsal colour pattern and internal organs, namely the pharynx and copulatory apparatus and only differ in the details. Gusana is retrieved as monophyletic and the molecular delimitation method recovered the same species recognised morphologically. A re-diagnosis of the genus and replacement of G. lata into Pseudogeoplana are also proposed. A new role of the penis papilla is also suggested based on the spermatophore found in Gusana lujanae.

Breast cancer is the most common cancer in women worldwide. Screening programs and imaging improvements have increased the detection of clinically occult non-palpable lesions requiring preoperative localization. Wire guided localization (WGL) is the current standard of care for the excision of non-palpable carcinomas during breast conserving surgery. Due to the current limitations of intraoperative tumor localization approaches, the integration of multimodal imaging information may be especially relevant in surgical planning. This research proposes a novel method for performing preoperative image-to-surgical surface data alignment to determine the position of the tumor at the time of surgery and aid preoperative planning. First, the volume of the breast in the surgical position is reconstructed and a set of surface correspondences is defined. Then, the preoperative (prone) and intraoperative (supine) volumes are co-registered using landmark driven non-rigid registration methods. We compared the performances of diffeomorphic and Bspline based registration methods. Finally, our method was validated using clinical data from 67 patients considering as target registration error (TRE) the distance between the estimated tumor position and the reference surgical position. The proposed method achieved a TRE of 16.21 ± 8.18 mm and it could potentially assist the surgery planning and guidance of breast cancer treatment in the clinical practice. This work was supported in part by the Spanish Ministry of Science and Innovation under Project RTI2018-098682-B-I00 (MCIU/AEI/FEDER,UE), Project PI18/01625 (Instituto de Salud Carlos III) and Grant BGP18/00178 under Beatriz Galindo Programme; in part by the European Union's European Regional Development Fund (ERDF); and in part by the Madrid Government (Comunidad de Madrid-Spain) under the Multiannual Agreement with Universidad Politécnica de Madrid in the line Support for Research and Development Projects for Beatriz Galindo researchers, in the context of the V Plan Regional de Investigación Científíca e Innovación Tecnológica (PRICIT).

Why some animals can regenerate while many others cannot remains a fascinating question. Even amongst planarian flatworms, well-known for their ability to regenerate complete animals from small body fragments, species exist that have restricted regeneration abilities or are entirely regeneration incompetent. Towards the goal of probing the evolutionary dynamics of regeneration, we have assembled a diverse live collection of planarian species from around the world. The combined quantification of species-specific head regeneration abilities and comprehensive transcriptome-based phylogeny reconstructions reveals multiple independent transitions between robust whole-body regeneration and restricted regeneration in the freshwater species. Our demonstration that theRNAi-mediated inhibition of canonical Wnt signalling can nevertheless bypass all experimentally tractable head regeneration defects in the current collection indicates that the pathway may represent a hot spot in the evolution of planarian regeneration defects. Combined with our finding that Wnt signalling has multiple roles in the reproductive system of the model speciesS. mediterranea, this raises the possibility of a trade-off between egg-laying and asexual reproduction by fission/regeneration as a driver of regenerative trait evolution. Although initial quantitative comparisons of Wnt signalling levels, reproductive investment, and regenerative abilities across the collection confirm some of the model’s predictions, they also highlight the diversification of molecular mechanisms amongst the divergent planarian lineages. Overall, our study establishes a framework for the mechanistic evolution of regenerative abilities and planarians as model taxon for comparative regeneration research.

The arrhythmic substrate of ventricular tachycardias in many structural heart diseases is located in the epicardium, often resulting in poor outcomes with currently available therapies. Cardiosphere-derived cells (CDCs) have been shown to modify myocardial scarring. A total of 19 Large White pigs were infarcted by occlusion of the mid-left anterior descending coronary artery for 150 min. Baseline cardiac magnetic resonance (CMR) imaging with late gadolinium enhancement sequences was obtained 4 weeks post-infarction and pigs were randomized to a treatment group (intrapericardial administration of 300,000 allogeneic CDCs/kg), (n = 10) and to a control group (n = 9). A second CMR and high-density endocardial electroanatomical mapping were performed at 16 weeks post-infarction. After the electrophysiological study, pigs were sacrificed and epicardial optical mapping and histological studies of the heterogeneous tissue of the endocardial and epicardial scars were performed. In comparison with control conditions, intrapericardial CDCs reduced the growth of epicardial dense scar and epicardial electrical heterogeneity. The relative differences in conduction velocity and action potential duration between healthy myocardium and heterogeneous tissue were significantly smaller in the CDC-treated group than in the control group. The lower electrical heterogeneity coincides with heterogeneous tissue with less fibrosis, better cardiomyocyte viability, and a greater quantity and better polarity of connexin 43. At the endocardial level, no differences were detected between groups. Intrapericardial CDCs produce anatomical and functional changes in the epicardial arrhythmic substrate, which could have an anti-arrhythmic effect. This study was supported by the Instituto de Salud Carlos III, Madrid, Spain (PI18/01895 and DTS21/00064); Red de Terapia Celular from the Instituto de Salud Carlos III, Madrid, Spain (RD16/0011/0029); Ricors-Red de Investigación Cooperativa Orientada a Resultados en Salud-RICORS TERAV (RD21.0017.0002), European Union's H2020 Program under grant agreement No. 874827 (BRAVE), and the Sociedad Española de Cardiología, Madrid, Spain.

MYOPLANA JOAOPAULOI ALMEIDA & CARBAYO SP. NOV. (FIGS 24–27) Zoobank registration: urn: lsid: zoobank. org:act: A4D8AE61-85C5-49A4-A57E-E2B5AD577454 Holotype: MNHNCL PLAT-15050 (Field code, F4875). Parque Nacional Nahuelbuta, Región de Purén, Chile (37°49′′39.2′′S, 073°00′′35.0′′W), coll. F. Carbayo et al., 9 December 2010. Cephalic region: transverse sections on 13 slides; ovarian region: horizontal sections on 27 slides; pre-pharyngeal region: transverse sections on 28 slides; pharynx and copulatory apparatus: sagittal sections on 32 slides. Type locality: Parque Nacional Nahuelbuta, Purén, Malleco Province, Región La Araucanía, Chile. The species is only known from this locality. Etymology: The specific name pays homage to João Paulo Gonzaga de Paula, a teacher in the public school E. M. E. F. Henrique Souza Filho Henfil, and most influential in the humanistic formation of children. Diagnosis: Species of Myoplana with an inconspicuous dorsal light mid stripe. Short, distal portion of the sperm ducts runs anteriorly. Penis papilla projects from the ventro-anterior region of the male atrium. Description External aspect: The specimen was not measured alive. Preserved, it was 32 mm in length, 5.5 mm in width, and 1.7 in height. The body margins are parallel; the anterior tip is pointed and the posterior tip is rounded. The dorsum is slightly convex; the ventral surface is flat (Fig. 24). The dorsal colour of the live specimen is black-brown (RAL 8022), passing into beige-brown (RAL 8024) in the anterior extremity. A thin inconspicuous light stripe runs medially (Fig. 24C). The colour of the body margins is cream (RAL 9001). The ventral surface is light-grey (RAL 7035), mottled with brownish dots in the extremities of the body (Fig. 28B, C). The eyes are of a single-cup type measuring 40 µm in diameter. They are placed in haloes and are distributed in a row contouring the anterior 5 mm of the body. Backward they form one to two marginal rows until the posterior tip. The sensory pits are approximately 45 µm deep. They contour the anterior region of the body and extend backward ventromarginally along a portion the body equivalent to 28% of its length. The mouth is positioned at a distance from the anterior extremity equal to 75% of the body length; the gonopore, 87.5%. Internal morphology: The creeping sole occupies the entire ventral surface. The entire epidermis is pierced by the necks of abundant rhabditogen cells and by three types of cells producing erythrophil amorphous secretion, xanthophil amorphous secretion and erythrophil granules, respectively. Ventrally, erythrophil granules are much more abundant, while the xanthophil secretion is less abundant than dorsally. The narrow glandular margin consists of two types of gland cells producing erythrophil and xanthophil granules, respectively. The main nervous system is organized in a 220 µm thick plate, representing approximately 13.5% of the body height (Fig. 25A). The cutaneous musculature comprises three layers, namely, a subepidermal layer of circular fibres (5 µm thick), followed by a double layer (20 µm) with diagonal fibres and then a well-developed, innermost layer of longitudinal fibres (55–145 µm thick, dorsally and ventrally, respectively). The latter layer is divided into a subepithelial portion and a portion sunken into the parenchyma representing 78% of the total thickness of the ventral layer (Fig. 25). The cutaneous musculature thickness relative to the body height in pre-pharyngeal region is 15%. Four parenchymal muscle layers are present, namely, a dorsal layer of decussate fibres, a supraintestinal layer of transverse muscle, a subintestinal layer of transverse muscle and a transneural layer of diagonal fibres. The muscle fibres of the transneural layers are located among the components of the main nerve plate and extend until the inner cutaneous nerve plexus. Oblique muscle fibres run from the dorsal to the lateroventral epidermis. The mouth is situated at a distance from the root of the pharynx, equivalent to one-third of pharyngeal pouch length (Fig. 26A). The pharyngeal pouch is close to the prostatic vesicle. An oesophagus is present. The oesophagus to pharynx length ratio is 42%. The pharynx is bell-shaped (Fig. 26A). Three types of gland cells discharge their xanthophil, erythrophil and cyanophil granules, respectively, through the covering epithelium of the distal portion of the pharynx. The outer pharyngeal musculature consists of a subepithelial layer of longitudinal muscle (20 µm thick), followed by a layer of circular muscle (67.5 µm thick) and a layer of longitudinal muscle (40 µm thick). The inner pharyngeal musculature consists of a subepithelial layer (10– 12 µm thick) of longitudinal fibres, followed by a layer (40 µm) of diagonal fibres, a circular muscle (200 µm) and an innermost longitudinal muscle (72 µm). Numerous radial muscle fibres run between the outer and inner pharyngeal epithelia. The testes are approximately 330 µm in diameter and are distributed into two to three rows at each side of the body. They are dorsally located between the supra-intestinal parenchymal musculature and the intestine (Fig. 25A). The anteriormost testes are located at a distance from the anterior tip of the body equivalent to 15.5% of the body length; the posteriormost testes, the equivalent to 70%, i.e. they are lateral to the pharyngeal root. The sperm ducts run posteriorly until a position lateral to the male atrium. Subsequently, they bend anteriorly before opening laterally into the proximal region of the prostatic vesicle (Fig. 26C). These ducts are lined with a squamous epithelium and are filled with sperm, except in their distal section. The prostatic vesicle is extrabulbar and is attached to the anterior side of the penis bulb. The pear-shaped, anterior-half of this vesicle runs dorsoposteriorly, subsequently penetrates the anterior region of the penis bulb and continues with the sinuous ductalhalf, which communicates with the ejaculatory duct. This vesicle is lined with an epithelium varying from squamous to columnar. Cilia covering the epithelium are only present in the ductal portion. The epithelium of the prostatic vesicle is pierced by the necks of three types of gland cells producing erythrophil, xanthophil and cyanophil granules, respectively. This epithelium of the extrabulbar portion is surrounded by a single layer (25 µm thick) of decussate fibres, while that of the intrabulbar portion is surrounded by a layer (7.5 µm) of circular fibres. The ejaculatory duct opens at the tip of the penis papilla. This duct is lined with a cuboidal, ciliated epithelium, which is traversed by the necks of gland cells producing cyanophil granules. A circular muscle (2 µm thick) surrounds this duct. The small penis papilla is conical and lies horizontally. This papilla occupies approximately the anterior-tenth of the male atrium and is placed in the ventral region of the penis bulb (Figs 26C, 27B). The penis papilla is lined with a columnar epithelium, which is pierced the necks of gland cells producing fine erythrophil granules. This epithelium is underlain by a muscle of scattered circular fibres (4 µm thick). Narrow anteriorly, the male atrium widens progressively to subsequently be narrowed by a distal, traverse fold, the dorsal section of which is continued with a lateral fold of the female atrium (Figs 26C, 27D). The male atrium is lined with a columnar epithelium, which is rugged in some sections. This epithelium is pierced by the necks of gland cells producing fine erythrophil granules; additionally, the necks of glands producing cyanophil granules pierce the dorsoanterior-half of the atrium. The atrial epithelium is underlain by a layer of circular muscle (5 µm thick), followed by a layer of longitudinal muscle (10 µm thick), which is only present in the anteriormost and posteriormost sections of the atrium. The ovoid ovaries have a maximum diameter of 280 µm and are located at a distance from the anterior tip of the body, equivalent to 8.5% of the body length. The ovovitelline ducts emerge from the dorsal portion of the ovaries and run ventrally above the nerve plate. Anteriorly to the gonopore, these ducts run dorsoposteriorly to join the common glandular ovovitelline duct. This common duct is located dorsad to the female atrium and runs posteroventrally to communicate with the conspicuous female genital canal. This canal is C-shaped in lateral view and projects from the posterior region of the female atrium. The female atrium is funnel-shaped and presents a lateral fold that continues from the male atrium (Figs 26C, 27E). The female genital canal and female atrium are lined with a columnar (60 µm high) epithelium, the cells of which are stained reddish apically. This epithelium is pierced by the necks of gland cells producing erythrophil granules and is underlain by a circular muscle (25 µm thick) with longitudinal fibres interspersed. Toward the gonopore canal, these two types of muscle fibres are separated into two layers, each 7 µm thick. Additionally, an ectal reinforcement of longitudinal fibres is located posterior to the female genital canal (Fig. 27E)., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 868-872, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977

HARANA HARAI ALMEIDA & CARBAYO SP. NOV. (FIGS 17–19) Zoobank registration: urn: lsid: zoobank. org:act: AC02BCF9-C45D-41AD-BBBC-D1589201E570 Holotype: MNHNCL PLAT-15042 (Field code, F4738): Parque Nacional Bosque Fray Jorge, Región de Coquimbo, Chile (30°39′′45.0′′S, 071°40′′57.4′′W). coll. F. Carbayo et al., 4 December 2010. Cephalic region: transverse-to-horizontal sections on two slides; a portion behind the cephalic region: horizontal sections on three slides; pharynx and copulatory apparatus: sagittal sections on eight slides. Type locality: Parque Nacional Bosques de Fray Jorge, Chile. Species only known from this locality. Etymology: The specific epithet pays homage to Prof. Marcos Ryotaro Hara (University of São Paulo). Description External aspect: The live specimen measured approximately 8 mm long and 1 mm wide when creeping. The length of the preserved specimen was 6.5 mm long, while its width was 1.2 mm. The body is elongated and subcylindrical, with the anterior tip rounded and the posterior tip pointed (Fig. 17A). The creeping sole is 78% of body width in the prepharyngeal region, as measured from sagittal sections. The mouth is positioned at a distance from the anterior extremity of the body equivalent to 83% of the body length; the gonopore 92%. The dorsal colour of the live specimen consists of numerous grey-brown (RAL 8019) dots mottling the pearl-beige (RAL 1035) ground colour (Fig. 17A). The ventral side is pearl-beige, darker in the anterior tip. The eyes are of a single pigmented cup measuring 25 µm in diameter. Clear haloes around the eyes are absent. The eyes are distributed in a single row that encircles the anterior tip of the body and extends marginally until the posterior tip. The sensory pits are 25 µm deep and are distributed in a single row ventrolateral along a body portion with about 8% of the body length. Internal morphology: Numerous rhabditogen cells and two types of gland cells producing erytrophil and cyanophil granules, respectively, pierce the dorsal epidermis of the pre-pharyngeal region. Other gland cells discharge their fine erythrophil granules through the ventral epidermis. A glandular margin is absent. All of these gland cells are scarce in the cephalic region. The cutaneous musculature consists of a thin subepithelial layer of circular muscle, followed by a thin layer of decussate fibres and an innermost layer of longitudinal muscle comprising bundles of two to four fibres each. The longitudinal layer is 4 µm thick dorsally and 8 µm ventrally. The thickness of cutaneous muscle relative to the body height is 2.3%. The cutaneous musculature in the cephalic region is thinner. The parenchymal musculature is weak. A parenchymal layer of transverse subintestinal fibres is relatively well developed (Fig. 18B), while other muscle layers are lacking. Instead, dorsal diagonal fibres and transverse supraintestinal fibres are scattered. The ventral nerve plate is poorly defined. A straight tube (Fig. 18A, B) runs medially among the fibres of the subintestinal parenchymal layer of transverse muscle. The tube runs from the near anterior tip of the body and is at least 1 mm long. The body portion behind the cephalic region was denatured for DNA extraction, and eventual communication of this tube with other organs could not be observed. The tube is 25 µm in diameter and is lined with a weakly stained cuboidal epithelium. A thin longitudinal muscle underlies the lining epithelium of the tube. The pharyngeal pouch extends over the copulatory apparatus and extends 750 µm behind it (Fig. 19B, C). The pharyngeal pouch is approximately twice as long as the pharynx. The anteriormost portion of the pharynx was denatured, but its general appearance is that of a cylindrical type (Fig. 18C). The posterior portion of the pharynx lies over the prostatic vesicle. The outer pharyngeal epithelium is underlain by a layer of longitudinal muscle (5 µm thick), followed by a layer of circular fibres (17 µm) and an innermost layer of longitudinal muscle (8 µm); the inner pharyngeal epithelium is underlain by a layer of circular muscle (12 µm), followed by a layer of longitudinal muscle (5 µm) (Fig. 19A). Testes were not found in the sections. The sperm ducts are located over the main nervous system and contain sperm in their distal portion. These ducts communicate laterally with the respective branch of the prostatic vesicle (Fig. 19B). The extrabulbar prostatic vesicle is tubular. The prostatic vesicle has the shape of an inverted J in lateral view and its distal portion penetrates the anterior region of the welldeveloped penis bulb. The prostatic vesicle is lined with ciliated, cuboidal epithelium. This epithelium is crossed by gland cells producing erythrophil granules and is surrounded by a circular muscle (10 µm thick). The ejaculatory duct is horizontal and slightly sinuous and opens at the tip of the penis papilla. The ejaculatory duct is lined with ciliated, cuboidal epithelium and is surrounded by a 3 µm thick circular muscle. The penis papilla is cylindrical, having a distal enlargement that makes the papilla resemble a club (Fig. 19B–E). The papilla lies horizontally and occupies the entire male atrium. The proximal two-thirds of the penis papilla are lined with a columnar epithelium, while the epithelium of the enlarged, distal-third is cuboidal. Three types of gland cells discharge their erythrophil, cyanophil and light cyanophil granules, respectively, through the covering epithelium of the penis papilla. The erythrophil type is particularly abundant along the proximal two-thirds of the papilla. The cyanophil type is restricted to the dorsoproximal region of the penis papilla, while the light cyanophil type is found only at the tip of the penis papilla. The lining epithelium of this organ is underlain by a layer (5 µm thick) of circular muscle, followed by a layer of longitudinal fibres (5 µm). The male atrium is smooth, except for some small folds close to the insertion of the penis papilla. The communication of the male atrium with the female atrium is narrowed by a thin fold located dorsally to the level of the gonopore (Fig. 19B–E). The male atrium is lined with a squamous epithelium dorsally and with a columnar epithelium ventrally. This ventral epithelium is pierced by the necks of gland cells producing erythrophil granules. The atrial epithelium is underlain by a layer (2 µm thick) of circular muscle, followed by a layer (3 µm thick) of longitudinal muscle, the latter underlying only the ventral epithelium. The ovaries were not found in the sections available. Vitellaria are abundant around the intestine. The ovovitelline ducts run backward above the ventral nerve plate. Posterior to the gonopore canal, these ducts ascend medially inclined to unite with the common glandular ovovitelline duct below the female atrium (Fig. 19B, C, E). This common duct ascends posterior to the female atrium to join the female genital canal. This canal projects posteroventrally from the posterodorsal portion of the female atrium (Fig. 19B). The female atrium to male atrium length ratio is 2.5: 4.0. The female atrium is irregular in shape and is inclined toward the gonopore.This atrium is lined with a columnar, 100 µm high epithelium and the apical portion of its cells is erythrophil (Fig. 19E). Toward the gonopore canal, the columnar epithelium passes gradually into a cuboidal type. Two types of gland cells producing fine cyanophil and scarce erythrophil granules, respectively, pierce the epithelium of the female atrium. The atrial epithelium is underlain by a longitudinal muscle (2 µm thick), followed by a circular muscle (8 µm). A weak common muscle coat of longitudinal fibres wraps the male and the female atria. Remarks on the neae tribe Haranini and its genus: H a r a n a i s a l way s r e t r i e v e d a s a n in g r o u p o f Geoplaninae sister to Timymini. The extraordinarily long pharyngeal pouch of Harana harai precludes it from being fitted in any of the tribes except Timymini. Moreover, H. harai and Timyma share a sister-group relationship in all analyses, and it is reasonable to ponder that the long extension of the pharyngeal pouch in both taxa is homologous. Such a pharyngeal pouch extending posteriorly over the copulatory apparatus is only known in these two species among all the land planarians. In addition to the long pharyngeal pouch, Haranini and Timymini are morphologically different from each other in that the cephalic region is semi-lunate in Timymini (vs. regular in Haranini), and the sensory pits are intercalated with sensory papillae (vs. sensory papillae absent in Haranini). The ventral position of the testes in Timymini, a condition which is an exception within Geoplanini, unfortunately could not be checked in H. harai since the body region seemingly housing the testes of this small animal was removed for DNA extraction. This same constraint impeded tracking the complete course of the straight tube level with the subintestinal transverse parenchymal muscle. On the other hand, the tube is a feature which is only found in Haranini., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 861-863, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977

TRANSANDIPLANA GRAUI ALMEIDA & CARBAYO SP. NOV. (FIGS 13–16) Zoobank registration: urn: lsid: zoobank. org:act: 9247667A-A4EE-4722-B935-7332CAA3F792 Holotype: MNHNCL PLAT-15049 (Field code, F4696). Huasco, Región de Atacama, Chile (28°27′′57.4′′S, 071°11′′09.5′′W), coll. F. Carbayo et al., 2 December 2010. Cephalic region: transverse sections on 13 slides; ovarian region: horizontal sections on 4 slides; prepharyngeal region: transverse sections on seven slides; pharynx and copulatory apparatus: sagittal sections on 13 slides. Type locality: Huasco, Región de Atacama, Chile. The species is only known from this locality. Etymology: The specific epithet pays homage to Dr José Horácio Grau, for his friendship and contribution to the knowledge of the Chilean planarians. Description External aspect: The live holotype (Fig. 13A) is about 25 mm in length and 2 mm in width. The body margins are parallel. The anterior extremity of the body is rounded, while the posterior is pointed. The dorsal side is convex; the ventral one is flat. The dorsal colour is graphite-grey (RAL 7024) – slightly clearer on the body margins– and adorned with a median grey-white (RAL 9002) stripe (Fig. 13A). The ventral surface is pure white (RAL 9010) – darker in the cephalic region – and exhibits a median, thin, whitish stripe throughout the body length (Fig. 13B). The preserved holotype measured 25 mm long, 2.5 mm wide and 1 mm high. The eyes are of a single-cup type measuring 35–38 µm in diameter. The anterior 2 mm of the body are encircled with a uniserial row of eyes. Behind this region, the eyes spread on to the dorsal surface to the extent of a band 40% of the body width on either side. This band is gradually thinner toward the posterior tip of the body, where the eyes are only marginally located. Scarce sensory depressions are located ventromarginally in the anterior region of the body (Fig. 13C–E). The depth of these depressions is equivalent to two-thirds of the height of the surrounding epidermal cells. The suboptimal quality of the sections of the cephalic region, which lacks part of the ventral portion, hindered a detailed description of their distribution. There are no sensory pits. The mouth is positioned at a distance from the anterior extremity equal to 67.6% of the body length; the gonopore is at 78.8%. Internal morphology: The creeping sole occupies 85% of the ventral surface of the body. Rhabditogen cells, gland cells producing erythrophil granules and gland cells producing xanthophil amorphous secretion discharge their secretion through the dorsal epidermis of the pre-pharyngeal region. Gland cells producing cyanophil granules and gland cells producing amorphous erythrophil secretion discharge their secretion through the ventral epidermis. A glandular margin is absent. The gland cells exhibit the same distribution in the cephalic region, although they are scarcer. The main nervous system is organized in approximately 24 longitudinal nerve cords.The shape of each cord resembles a necklace of beads (Fig. 14A, B, D). Cerebral ganglia could not be discerned. The cutaneous musculature comprises three layers, namely, a subepidermal layer of circular muscle (2–3 µm thick), followed by a double layer with diagonal fibres (4 µm) and a third layer of longitudinal muscle (20–25 µm dorsally, 17–20 µm ventrally). Muscle fibres of the longitudinal muscle layer are arranged into bundles with six to 15 fibres each (Fig. 14E, F). The cutaneous musculature thickness relative to body height at the pre-pharyngeal region is 6.7%. The musculature in the cephalic region maintains the organization observed in the pre-pharyngeal region. The parenchymal musculature comprises three layers of scattered fibres, namely, a dorsal double layer (12–15 µm thick) of decussate fibres, a supraintestinal layer of transverse muscle (40–50 µm) and a subintestinal layer of transverse muscle (35–50 µm) (Fig. 14C, D). The mouth is situated at the end of the pharyngeal pouch (Fig. 15A). A distinct oesophagus is present. The oesophagus to pharynx length ratio is 24%. The pharynx is cylindrical (Fig. 15A). The oesophagic musculature consists of a subepithelial layer of circular muscle (40 µm thick), followed by a layer of longitudinal muscle (25 µm thick). Two types of gland cells discharge their erythrophil and cyanophil granules, respectively, through the covering epithelium of the distal portion of the pharynx. The outer pharyngeal musculature consists of a subepithelial layer of longitudinal muscle (2.5 µm thick), followed by a layer of circular muscle (7.5 µm thick). The inner pharyngeal musculature consists of a single muscle of circular and longitudinal fibres interwoven. The testes range between club- and pear-shaped and are 180–230 µm in diameter. The testes are surrounded by a pigmented covering (Fig. 14C, E, F), and are distributed into one to two rows at each side of the body. They are located dorsally, beneath the supraintestinal parenchymal muscle, and between the intestinal branches. The anteriormost testes lie at a distance from the anterior tip of the body equivalent approximately to 10% of the body length; the posteriormost testes are located 2 mm (8% of body length) anterior to the pharynx, i.e. at a distance from the anterior body tip equivalent to 52.4% of the body length. The sperm ducts are located dorsally to the ovovitelline ducts. The distal portion of the sperm ducts is bent dorsally and medially to open into the mid-dorsal region of the prostatic vesicle (Fig. 15C). The distal portion of one of the ducts is dilated to form a spermiducal vesicle filled with sperm. The anteriorhalf of the prostatic vesicle is extrabulbar, dilated and with a folded wall. The posterior-half is narrow and located within the penis bulb, whose dorso-anterior region is traversed by the prostatic vesicle (Fig. 15B– D). The ejaculatory duct runs within the penis papilla to open at its tip (Fig. 15C, D). The prostatic vesicle is lined with a columnar epithelium in its proximal region; otherwise, the lining epithelium is cuboidal. Abundant gland cells discharge cyanophil granules into the prostatic vesicle (Fig. 15B, D). The dilated portion of the prostatic vesicle is surrounded by a single muscle (10–65 µm thick) of intermingled fibres, while the canalicular portion is surrounded by a single circular muscle (15 µm thick). The ejaculatory duct is lined with a cuboidal, ciliated epithelium, through which two types of gland cells discharge cyanophil and erythrophil granules, respectively. Muscle fibres surrounding this duct are not apparent. The penis papilla is conical, with the dorsal insertion shifted backward and its basis somewhat bulged. This papilla occupies the entire length of the male atrium (Fig. 15C, D). The epithelium of the penis papilla is cuboidal-to-columnar and is pierced by the openings of two types of numerous gland cells producing erythrophil and cyanophil granules, respectively. The epithelium is underlain by a single circular muscle (10 µm thick). Multiple radial and longitudinal muscle fibres are located in the stroma of the penis papilla. The male atrium is smooth (Fig. 15C, D) and is lined with a low epithelium (10 µm high) proximally and tall (30 µm thick) in the other regions. Two types of gland cells discharge their cyanophil and erythrophil granules, respectively, through the atrial epithelium. Cyanophil granules are particularly abundant in the proximal region of the male atrium. The atrial epithelium is underlain by a subepithelial layer of circular muscle (7.5 µm thick), followed by a layer of longitudinal muscle (5 µm thick). The ovaries are ovoid, with 190 µm in maximum diameter (Fig. 16A) and are located at a distance from the anterior body tip equivalent to about 11% of the body length. The ovovitelline ducts emerge from the dorsoposterior portion of the ovaries (Fig. 16A). Laterally to the gonopore and the female atrium, the ovovitelline ducts ascend dorsoposteriorly, to subsequently bending anteriorly to communicate with the common ovovitelline duct. This unpaired duct runs dorso-anteriorly to join the posterodorsal region of the female atrium. In this joining point, a short diverticulum of projects anteriorly (Fig. 15C). The female atrium is spherical-to-ovoid. A dorsal fold located above the gonopore narrows the communication of the male and female atria. The female atrium is approximately as long as the male atrium and is lined with a columnar (150 µm high) epithelium provided with an irregular surface and stratified appearance. This epithelium has scattered and unevenly distributed cilia, presenting some spaces with a vacuolar aspect filled with cyanophil secretion (Fig. 16B, C). This atrial epithelium is pierced by the necks of two types of gland cells producing erythrophil and cyanophil granules, respectively. The lining epithelium of the female atrium is underlain by a subepithelial layer of longitudinal muscle (2.5 µm thick), followed by a layer of circular muscle (7.5 µm thick); ectally to this muscle is a longitudinal loose muscle (20 µm) coating the atrium (Fig. 16B). Remarks on Geoplanini and the neae genus Transandiplana: Transandiplana is nested in Geoplanini in all molecular trees. This tribe also includes Amaga Ogren & Kawakatsu, 1990, Bogga Grau & Sluys, 2012, Barreirana Ogren & Kawakatsu, 1990, Cephaloflexa Carbayo & Leal-Zanchet, 2003, Choeradoplana Graff, 1896, Cratera Carbayo et al., 2013, Difroehlichia Leal-Zanchet & Marques, 2018, Geobia Diesing, 1862, Geoplana Schultze, 1856, Imbira Carbayo et al., 2013, Issoca Froehlich, 1954, Luteostriata Carbayo, 2010, Matuxia Carbayo et al., 2013, Notogynaphallia Ogren & Kawakatsu, 1990, Obama Carbayo et al., 2013, Paraba Carbayo et al., 2013, Pasipha Ogren & Kawakatsu, 1990, Piima Carbayo, 2020, Pseudogeoplana (collective genus) Ogren & Kawakatsu, 1990, Supramontana Carbayo & Leal-Zanchet, 2003, Winsoria Negrete et al., 2020 and Xerapoa Froehlich, 1955. Geoplanini differ from the remaining tribes by a unique combination of characters, namely, dorsum convex (vs. carinate in Adinoplanini), dorsal eyes (vs. only marginal in Haranini, Inakayaliini, Polycladini, Sarcoplanini and Timymini), dorsal longitudinal cutaneous muscle not sunken into the parenchyma (vs. sunken in Gusanini; the Geoplanini Choeradoplana gladismariae Carbayo & Froehlich, 2012 is an exception), pharyngeal pouch anterior to the copulatory apparatus (vs. extending posteriorly over the copulatory apparatus in Haranini and Timymini), female atrium without musculoglandular organs (vs. with it in Adinoplanini and Sarcoplanini) and female genital duct not dilated (vs. dilated in Inakayaliini). Thus, Geoplanini lack exclusive morphological characteristics. However, the eye distribution pattern might help to recognize most members of the tribe. Although some subcylindrical species of Geoplanini exhibit dorsal eyes, Geoplanini with a flattened body also present dorsal eyes. Exceptions to this pattern are some Geoplanini taxa such as Bogga and Amaga, which, while being flattened, have only marginal eyes. Nonetheless, the phylogenetic position of these two genera has not yet been assessed. The new Geoplanini genus Transandiplana is represented by one single individual. It is retrieved as an ingroup of Geoplanini in all analyses, having an unstable position, either sister to Geoplana Stimpson, 1858 (Fig. 2; Supporting Information, Fig. S4), to Paraba multicolor (Graff, 1899) (Supporting Information, Fig. S3) or it is a branch of a polytomy (Supporting Information, Figs S2, S 5, S 6). Transandiplana graui largely resembles some Geoplanini genera in the general shape of the copulatory apparatus, including the lack of structures such as musculoglandular organs. However, while it is similar to other Geoplanini, it also exhibits some features uncommon in Geoplanini, namely, sensory depressions, the main nervous system consisting of multiple longitudinal cords, testes provided with a pigmented covering, and the relatively large distance between the testes and the pharynx. With this combination of features, T. graui does not fit well in any of the Geoplaninae tribes. Sensory depressions are known from Sarcoplanini. The main nervous system in Geoplanini consists of either two longitudinal cords (usually in small and thin species such as Xerapoa) or it exhibits the aspect of an even plate (especially in large and flat organisms such as Obama). Transandiplana graui is an exception, as it presents both a slender body and a main nervous system consisting of multiple cords. These two latter features (dark spots covering testes and relative position of posteriormost testes) are underreported in most old species descriptions. Testes covered with dark spots were only reported for Obama ladislaƲii (Graff, 1899) (in: Álvarez-Presas et al., 2015). Given the general morphological resemblance of T. graui with other Geoplanini taxa and the phylogenetic position as an ingroup of Geoplanini, we tentatively place the species and the genus within Geoplanini. Transandiplana graui resembles ‘ Geoplana ’ caleta E. M. Froehlich, 1978 in the general aspect of the body and internal organs. However, important diagnostic traits (ventrolateral sensory depressions; main nervous system comprising multiple longitudinal cords; testes surrounded by a pigmented covering) and molecular data of G. caleta remain unknown. Thus, a systematic replacement would be speculative. Interestingly, both species occur in an adverse climatic region, namely, the Huasco river valley, in the arid Norte Chico zone (Atacama) of the Chilean Andes. Annual rainfall in the region is 42 mm /year, and the relative humidity ranges between 66–74% (see: Juliá et al., 2008).

POLYCLADUS BLANCHARD, 1845 Type species: Polycladus gayi Blanchard, 1845, fixed by monotypy by Ogren & Kawakatsu (1990). Neae diagnosis: Polycladini with mouth and gonopore in the posterior-quarter of the body. Copulatory apparatus with a well-developed penis papilla. Female genital canal with dorsal entrance into the genital antrum. Distribution: As for that of the tribe. Remarks on the neae tribe Polycladini: In the molecular phylogenies, Polycladus is represented by an unidentified specimen. This specimen is retrieved as a sister to Gusana in all analyses. Apart from the unidentified Polycladus sp. (in: Carbayo et al., 2013), only Polycladus gayi is formally described. The diagnosis of the genus was revised by Graff (1896) and Ogren & Kawakatsu (1990). The most remarkable features of Polycladus are the large size of the body and a body width of 40% of the body length. Only some species of Obama (Geoplanini) resemble Polycladus in this aspect, but their bodies do not reach such a width. Therefore, Polycladus does not fit into any tribe, except Polycladini. Polycladus also presents two additional exclusive traits, as discussed below, namely, a transverse subneural parenchymal muscle and a longitudinal transneural parenchymal muscle. Several works have dealt with Polycladus gayi (see: Barahona-Segovia et al., 2020 and references therein), but actually only Graff (1899) and Schmidt (1902) have reported details of its internal morphology. The most conspicuous characteristic of this genus is the extraordinarily wide (2.4 times as long as wide in P. gayi) and flattened body (Graff, 1899), along with marginal eyes only (Graff, 1899; Schmidt, 1902). The musculature of the species is not clearly understood. Graff (1899) detailed illustrations of the cutaneous and parenchymal muscles (Graff, 1899: plate 30, figs 3, 4). The cutaneous musculature is described as comprising three muscle layers, namely, a layer of circular muscle, a layer of diagonal fibres and a third innermost layer of longitudinal muscle. The parenchymal musculature shown in Graff’s drawing of a transverse section of the body (plate 30, fig. 3) is depicted as being comprised of ‘obere Transversalmuskeln’, ‘mittlere Transversalmuskeln’ and ‘ ventrale Transversalmuskeln’. The two latter muscles are crossed by fibres of ‘ventrale Longitudinalmuskeln’. In another drawing of a sagittal section (plate 30, fig. 4), only ‘dorsale Longitudinalmuskeln’ and ‘dorsoventrale Muskeln’ are depicted. The ‘dorsale Longitudinalmuskeln’ are represented with dashed lines, suggesting that they might not be longitudinal but diagonal. Although Graff (1899) stated that the parenchymal musculature in land planarians consists of longitudinal, transverse and dorsoventral fibres, all Geoplaninae (except Timyma) present a dorsal parenchymal layer of diagonal muscle. In our experience, the orientation of the fibres of the cutaneous and parenchymal muscle layers in Geoplaninae is best discerned, if not only, on horizontal sections (for an example, see Figs 14C, 18B, 25B, C of this paper). Summing up, apart from the dorsoventral muscle fibres, the parenchymal musculature in Polycladus gayi comprises a dorsal layer of decussate fibres (‘dorsale Longitudinalmuskeln’), a transverse supraintestinal muscle (‘obere Transversalmuskeln’), a transverse subintestinal muscle (‘mittlere Transversalmuskeln’), a transverse subneural muscle (‘ ventrale Transversalmuskeln’) and a longitudinal transneural muscle (‘ventrale Longitudinalmuskeln’), which is intermingledwithfibresofthesubintestinalandsubneural muscles. The transverse subneural parenchymal muscle and the parenchymal transneural layer of longitudinal muscle are unique among the Geoplaninae., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 876-877, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Blanchard E. 1845. Recherches sur l'organisation des vers. Annales des Sciences Naturelles, Zoologie, 3 e serie 8: 119 - 149.","Carbayo F, Alvarez-Presas M, Olivares CT, Marques FPL, Froehlich EM, Riutort M. 2013. Molecular phylogeny of Geoplaninae (Platyhelminthes) challenges current classification: proposal of taxonomic actions. Zoologica Scripta 42: 508 - 528.","Graff LV. 1896. Uber das System und die geographische Verbreitung der Landplanarien. Verhandlungen der Deutsche Zoologischen Gesellschaft 6: 75 - 93.","Barahona-Segovia RM, Araya JF, Paninao-Monsalvez L. 2020. New records of the giant planarian Polycladus gayi Blanchard, 1845 (Platyhelminthes: Geoplanidae) with notes on its conservation biology. Zootaxa 4822: 595 - 600.","Graff LV. 1899. Monographie der Turbellarien II. Tricladida Terricola (Landplanarien). Atlas Von Achtundfunfzig Tafeln zur Monographie der Turbellarien II. Tricladida Terricola (Landplanarien), pls I - LVIII. Leipzig: Wilhelm Engelmann.","Schmidt AT. 1902. Zur Kenntnis der Tricladenaugen und der Anatomie von Polycladus gayi. Zeitschrift fur Wissenschaftliche Zoologie 72: 545 - 564."]}

HARANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: F3527621-B8FF-482B-9814-07855F164AE7 Type genus: Harana Almeida & Carbayo gen. nov. Diagnosis: Small-sized Geoplaninae with a longitudinal tube located among the fibres of the subintestinal transverse parenchymal muscle. Long pharyngeal pouch, extending behind the copulatory apparatus. Haranini comprises only the genus Harana. Etymology: The name of the tribe is based on the name of its type genus. Distribution: Only known from the type locality of the type species., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 859, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977

HARANA HARAI ALMEIDA & CARBAYO SP. NOV. (FIGS 17–19) Zoobank registration: urn: lsid: zoobank. org:act: AC02BCF9-C45D-41AD-BBBC-D1589201E570 Holotype: MNHNCL PLAT-15042 (Field code, F4738): Parque Nacional Bosque Fray Jorge, Región de Coquimbo, Chile (30°39′′45.0′′S, 071°40′′57.4′′W). coll. F. Carbayo et al., 4 December 2010. Cephalic region: transverse-to-horizontal sections on two slides; a portion behind the cephalic region: horizontal sections on three slides; pharynx and copulatory apparatus: sagittal sections on eight slides. Type locality: Parque Nacional Bosques de Fray Jorge, Chile. Species only known from this locality. Etymology: The specific epithet pays homage to Prof. Marcos Ryotaro Hara (University of São Paulo). Description External aspect: The live specimen measured approximately 8 mm long and 1 mm wide when creeping. The length of the preserved specimen was 6.5 mm long, while its width was 1.2 mm. The body is elongated and subcylindrical, with the anterior tip rounded and the posterior tip pointed (Fig. 17A). The creeping sole is 78% of body width in the prepharyngeal region, as measured from sagittal sections. The mouth is positioned at a distance from the anterior extremity of the body equivalent to 83% of the body length; the gonopore 92%. The dorsal colour of the live specimen consists of numerous grey-brown (RAL 8019) dots mottling the pearl-beige (RAL 1035) ground colour (Fig. 17A). The ventral side is pearl-beige, darker in the anterior tip. The eyes are of a single pigmented cup measuring 25 µm in diameter. Clear haloes around the eyes are absent. The eyes are distributed in a single row that encircles the anterior tip of the body and extends marginally until the posterior tip. The sensory pits are 25 µm deep and are distributed in a single row ventrolateral along a body portion with about 8% of the body length. Internal morphology: Numerous rhabditogen cells and two types of gland cells producing erytrophil and cyanophil granules, respectively, pierce the dorsal epidermis of the pre-pharyngeal region. Other gland cells discharge their fine erythrophil granules through the ventral epidermis. A glandular margin is absent. All of these gland cells are scarce in the cephalic region. The cutaneous musculature consists of a thin subepithelial layer of circular muscle, followed by a thin layer of decussate fibres and an innermost layer of longitudinal muscle comprising bundles of two to four fibres each. The longitudinal layer is 4 µm thick dorsally and 8 µm ventrally. The thickness of cutaneous muscle relative to the body height is 2.3%. The cutaneous musculature in the cephalic region is thinner. The parenchymal musculature is weak. A parenchymal layer of transverse subintestinal fibres is relatively well developed (Fig. 18B), while other muscle layers are lacking. Instead, dorsal diagonal fibres and transverse supraintestinal fibres are scattered. The ventral nerve plate is poorly defined. A straight tube (Fig. 18A, B) runs medially among the fibres of the subintestinal parenchymal layer of transverse muscle. The tube runs from the near anterior tip of the body and is at least 1 mm long. The body portion behind the cephalic region was denatured for DNA extraction, and eventual communication of this tube with other organs could not be observed. The tube is 25 µm in diameter and is lined with a weakly stained cuboidal epithelium. A thin longitudinal muscle underlies the lining epithelium of the tube. The pharyngeal pouch extends over the copulatory apparatus and extends 750 µm behind it (Fig. 19B, C). The pharyngeal pouch is approximately twice as long as the pharynx. The anteriormost portion of the pharynx was denatured, but its general appearance is that of a cylindrical type (Fig. 18C). The posterior portion of the pharynx lies over the prostatic vesicle. The outer pharyngeal epithelium is underlain by a layer of longitudinal muscle (5 µm thick), followed by a layer of circular fibres (17 µm) and an innermost layer of longitudinal muscle (8 µm); the inner pharyngeal epithelium is underlain by a layer of circular muscle (12 µm), followed by a layer of longitudinal muscle (5 µm) (Fig. 19A). Testes were not found in the sections. The sperm ducts are located over the main nervous system and contain sperm in their distal portion. These ducts communicate laterally with the respective branch of the prostatic vesicle (Fig. 19B). The extrabulbar prostatic vesicle is tubular. The prostatic vesicle has the shape of an inverted J in lateral view and its distal portion penetrates the anterior region of the welldeveloped penis bulb. The prostatic vesicle is lined with ciliated, cuboidal epithelium. This epithelium is crossed by gland cells producing erythrophil granules and is surrounded by a circular muscle (10 µm thick). The ejaculatory duct is horizontal and slightly sinuous and opens at the tip of the penis papilla. The ejaculatory duct is lined with ciliated, cuboidal epithelium and is surrounded by a 3 µm thick circular muscle. The penis papilla is cylindrical, having a distal enlargement that makes the papilla resemble a club (Fig. 19B–E). The papilla lies horizontally and occupies the entire male atrium. The proximal two-thirds of the penis papilla are lined with a columnar epithelium, while the epithelium of the enlarged, distal-third is cuboidal. Three types of gland cells discharge their erythrophil, cyanophil and light cyanophil granules, respectively, through the covering epithelium of the penis papilla. The erythrophil type is particularly abundant along the proximal two-thirds of the papilla. The cyanophil type is restricted to the dorsoproximal region of the penis papilla, while the light cyanophil type is found only at the tip of the penis papilla. The lining epithelium of this organ is underlain by a layer (5 µm thick) of circular muscle, followed by a layer of longitudinal fibres (5 µm). The male atrium is smooth, except for some small folds close to the insertion of the penis papilla. The communication of the male atrium with the female atrium is narrowed by a thin fold located dorsally to the level of the gonopore (Fig. 19B–E). The male atrium is lined with a squamous epithelium dorsally and with a columnar epithelium ventrally. This ventral epithelium is pierced by the necks of gland cells producing erythrophil granules. The atrial epithelium is underlain by a layer (2 µm thick) of circular muscle, followed by a layer (3 µm thick) of longitudinal muscle, the latter underlying only the ventral epithelium. The ovaries were not found in the sections available. Vitellaria are abundant around the intestine. The ovovitelline ducts run backward above the ventral nerve plate. Posterior to the gonopore canal, these ducts ascend medially inclined to unite with the common glandular ovovitelline duct below the female atrium (Fig. 19B, C, E). This common duct ascends posterior to the female atrium to join the female genital canal. This canal projects posteroventrally from the posterodorsal portion of the female atrium (Fig. 19B). The female atrium to male atrium length ratio is 2.5: 4.0. The female atrium is irregular in shape and is inclined toward the gonopore.This atrium is lined with a columnar, 100 µm high epithelium and the apical portion of its cells is erythrophil (Fig. 19E). Toward the gonopore canal, the columnar epithelium passes gradually into a cuboidal type. Two types of gland cells producing fine cyanophil and scarce erythrophil granules, respectively, pierce the epithelium of the female atrium. The atrial epithelium is underlain by a longitudinal muscle (2 µm thick), followed by a circular muscle (8 µm). A weak common muscle coat of longitudinal fibres wraps the male and the female atria. Remarks on the neae tribe Haranini and its genus: H a r a n a i s a l way s r e t r i e v e d a s a n in g r o u p o f Geoplaninae sister to Timymini. The extraordinarily long pharyngeal pouch of Harana harai precludes it from being fitted in any of the tribes except Timymini. Moreover, H. harai and Timyma share a sister-group relationship in all analyses, and it is reasonable to ponder that the long extension of the pharyngeal pouch in both taxa is homologous. Such a pharyngeal pouch extending posteriorly over the copulatory apparatus is only known in these two species among all the land planarians. In addition to the long pharyngeal pouch, Haranini and Timymini are morphologically different from each other in that the cephalic region is semi-lunate in Timymini (vs. regular in Haranini), and the sensory pits are intercalated with sensory papillae (vs. sensory papillae absent in Haranini). The ventral position of the testes in Timymini, a condition which is an exception within Geoplanini, unfortunately could not be checked in H. harai since the body region seemingly housing the testes of this small animal was removed for DNA extraction. This same constraint impeded tracking the complete course of the straight tube level with the subintestinal transverse parenchymal muscle. On the other hand, the tube is a feature which is only found in Haranini.

INAKAYALIA NEGRETE ET AL., 2020 Type species: Inakayalia ƲaldiƲiana (Grau & Carbayo, 2011), designated by Negrete et al. (2020). Neae diagnosis: Inakayaliini with medium-sized slender body with nearly parallel margins; dorsal surface convex and ventral body surface flat or slightly concave; monolobulate eyes extending dorsally along the body with large clear haloes; bell-shaped pharynx; extrabulbar, voluminous, horizontal prostatic vesicle; penis papilla nearly dome-shaped; common ovovitelline duct dorsal to female atrium; short, anterodorsally flexed female genital canal, ascending from the posterodorsal region of female atrium; female atrium with narrow lumen. Distribution: As for that of the tribe., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 863-865, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Negrete L, Alvarez-Presas M, Riutort M, Brusa F. 2020. Integrative taxonomy of land planarians (Platyhelminthes: Geoplanidae) from the Andean-Patagonian Forests from Argentina and Chile, with the erection of two new genera. Journal of Zoological Systematics and EVolutionary Research 59: 588 - 612."]}

GUSANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: 9E0939FB-0E21-4209-B514-A52634BD1ED0 Type genus: Gusana E.M. Froehlich, 1978. Diagnosis: Geoplaninae with a broad and foliaceus body, tapering abruptly to the anterior tip. Anterior tip triangular. Dorsal and ventral longitudinal cutaneous muscle partly sunken into the parenchyma. Sensory pits as a simple invagination or obliquely elongated and internally branched. Gusanini comprise only the genus Gusana. Etymology: The name of the tribe is based on the name of its type genus. Distribution: Regions Biobío and La Araucanía (Chile)., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 859, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Froehlich EM. 1978. On a collection of Chilean land planarians. Boletins de Zoologia da UniVersidade de Sao Paulo 3: 7 - 80."]}

TIMYMA E.M. FROEHLICH, 1978 Type species: Timyma juliae E.M. Froehlich, 1978, designated by E. M. Froehlich (1978). Diagnosis: Timymini with mouth approximately in midbody. Without parenchymatic longitudinal muscle. Pharyngeal pouch extends posteriorly beyond copulatory apparatus. Outer pharyngeal musculature tripartite, with an outer longitudinal muscle layer, a midcircular and an inner longitudinal muscle layer. Prostatic vesicle extrabulbar. Penis papilla absent. Distal section of male atrium narrowed. Ovovitelline ducts emerge from the ventral aspect of ovaries and subsequently ascend laterally to the female atrium to join dorsally to it. Female genital canal dilated to form an ootype projected from the dorsoposterior aspect of the female atrium. Without adhesive musculoglandular organs. Copulatory apparatus without adenodactyls (as re-diagnosed by Almeida et al., 2021). Distribution: Regions Coquimbo and Valparaiso, Chile., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 892, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Froehlich EM. 1978. On a collection of Chilean land planarians. Boletins de Zoologia da UniVersidade de Sao Paulo 3: 7 - 80.","Almeida AL, Francoy TM, Alvarez-Presas M, Carbayo F. 2021. Convergent evolution: a new subfamily for bipaliinlike Chilean land planarians (Platyhelminthes). Zoologica Scripta 50: 500 - 508."]}

INAKAYALIA CYANEA ALMEIDA & CARBAYO SP. NOV. (FIGS 20–23) Zoobank registration: urn: lsid: zoobank. org:act: 3CF36CAC-C035-45EC-B439-79C0EE4DC8E3 Material examined: All specimens collected in Parque Nacional Nahuelbuta, Chile (37°48′′00.0′′S, 073°00′′00.0′′W), coll. F. Carbayo et al., 13 December 2010. Holotype: MNHNCL PLAT-15043 (Field code, F4912). Cephalic region: transverse sections on 14 slides; ovarian region: horizontal sections on 100 slides; prepharyngeal region: transverse sections on 30 slides; copulatory apparatus: sagittal sections on 122 slides. Paratypes: MZUSP PL 2283 (Field code, F4914). Cephalic region: transverse sections on ten slides; ovarian region: horizontal sections on 13 slides; prepharyngeal region: transverse sections on ten slides; pharynx and copulatory apparatus: sagittal sections on 73 slides. MZUSP PL2284 (Field code, F4917). Cephalic region: transverse sections on 15 slides; ovarian region: horizontal sections on 12 slides; prepharyngeal region: transverse sections on 20 slides; pharynx: sagittal sections on 22 slides; copulatory apparatus: sagittal sections on seven slides. Type locality: Parque Nacional Nahuelbuta, Región de la Araucanía, Chile. The species is only known from this locality. Etymology: The specific epithet derives from the Greek Κ&upsi;ανό&sigmav; meaning blue, alluding to the colour of the body. Diagnosis: Species of Inakayalia with a long and widened prostatic vesicle, branched proximally, and provided with a long posterodorsal diverticulum; long unpaired and dilated common ovovitelline duct. Description External aspect: The three specimens (Fig. 20) are mature and measured between 23 and 27 mm in length and 6–8.3 mm in width at rest. Preserved, they measured 28.5–44.5 mm long, 5–6 mm wide and 1.2–1.3 mm high. At rest, the body is lanceolate with undulating margins (Fig.20D, E). The cephalic region narrows to the rounded tip; the posterior narrows abruptly to the pointed tip. The dorsum is flattened except for the convex median region. The ventral side is flat. With approximately one-eighth of the body length, the cephalic region exhibits two black-blue (RAL 5004) bands, separated from each other by a median pure white (RAL 9010) line (Fig. 20A–C). These two bands are completely (Fig. 20A, B) or incompletely (Fig. 20C) interrupted by a zigzagged, pure white or beige transverse band. Behind the transverse band, the dorsum is black-blue (RAL 5004), darker along the median and paramedian zones to form bands, each with 10–16% of the body width. Additionally, large blue-grey (RAL 7031) haloes mottle the dorsum behind the transverse band except for the median zone. The ground colour of the ventral side is cream (RAL 9001) (Fig. 20D, E). Numerous small black-blue pigment dots are in the cephalic region. A whitish transverse band continued from the dorsal side separates the cephalic region from the remaining ventral surface, which is covered with numerous dots, either graphitegrey (RAL 7024) or ochre-brown (RAL 8001). The eyes are of a single-cup type measuring 32–38 µm in diameter. They are organized in a singleto-biserial row around the anterior eighth of the body; behind this body region, they spread onto the dorsal surface and are encircled by the blue-grey haloes. The sensory pits are simple invaginations 50–57 µm deep and are located ventromarginally in a single row running along an anterior region with 18% of body length. The mouth is positioned at a distance from the anterior extremity equal to 63.4–66.3% of the body length; the gonopore, 83.6–86.7%. Internal morphology: The creeping sole has 95% of the body width. Abundant gland cells producing coarse (1 µm) xanthophil granules and scarce gland cells producing erythrophil granules discharge their secretion through the entire epidermis of the prepharyngeal region. The cell necks of the xanthophil type are 10–15 µm in diameter and become more abundant toward the body margins of the dorsal side. The glandular margin consists of xanthophil gland cells (Fig. 21A). Rhabdites are discharged through the dorsal epidermis. As the creeping sole narrows toward the anterior extremity of the body, the xanthophil glands become scarce dorsally and abundant ventrally. The cutaneous musculature comprises three layers, namely, a subepithelial layer of circular muscle (5 µm thick), followed by a double layer (15–40 µm) with decussate fibres and then a well-developed, innermost layer of longitudinal fibres (30–105 µm thick, both dorsally and ventrally) (Fig. 21B–C). The cutaneous musculature thickness relative to the body height is 13–15%. Toward the anterior region of the body, these muscle layers become thinner until they disappear. There are three strong parenchymal muscle layers, namely, a dense dorsal layer of decussate fibres (30 µm thick), a supra-intestinal layer of transverse fibres (100 µm thick) and a denser subintestinal layer of transverse fibres (100 µm thick; Fig. 21B–C). These muscle layers are thinner in the anterior region of the body. Two of the three main branches of the intestine, namely, the paired ones, may connect to each other at the level of the prostatic vesicle. The oesophagus to pharynx length ratio is 31–33%. The mouth is situated at a distance from the root of the pharynx equivalent to 41–54% of the pharyngeal pouch length (Fig. 21D). The distal portion of the pharyngeal pouch is close to the prostatic vesicle. The pharynx is bell-shaped, with its dorsal insertion slightly anterior to the mouth. The epithelium of the distal portion of the pharynx is pierced by the necks of four types of gland cells, producing xanthophil granules, erythrophil granules, cyanophil granules and amorphous secretion, respectively. The outer pharyngeal musculature consists of a subepithelial layer of longitudinal muscle (5 µm thick), followed by a layer of circular muscle (15 µm thick) and an innermost layer of longitudinal muscle (5 µm thick). The inner pharyngeal musculature consists of a subepithelial circular muscle (75 µm thick), followed by a longitudinal muscle (10 µm thick; Fig. 21E). The rounded-to-irregular testes measure 325– 450 µm in diameter. They are organized into two to four rows in three vertical levels at each side of the body, between the supra-intestinal and subintestinal parenchymal muscles (Fig. 21A). The anteriormost testes lie at a distance from the anterior tip of the body equivalent to 17.3% of the body length; the posteriormost ones, the equivalent to 65% of the body length, i.e. they are lateral to the pharyngeal root. The sperm ducts run immediately above the subintestinal parenchymal muscle. Laterally to the pharyngeal pouch, each duct opens into the anteroventral region of the respective lateral, short branch of the prostatic vesicle (Fig. 22A). The rest of the prostatic vesicle is unpaired and large, measuring up to 1.3 mm in length. Approximately the anterior-half of the vesicle occupies two-thirds of the body height and exhibits numerous folds filling its lumen (Figs 22B, C, 23A–C). The posterior-half consists of a dorsal, blind diverticulum (200–250 µm long) and a ventral, widened duct (600 µm long), both running posteriorly. The latter penetrates the anterior section of the penis bulb to communicate with the ejaculatory duct. The prostatic vesicle is lined with a ciliated epithelium, being cuboidal in the paired portion and columnar in the unpaired one. Abundant gland cells discharge erythrophil granules through the lining epithelium of the prostatic vesicle. The epithelium of this vesicle is underlain by a muscle layer (25–100 µm thick) of decussate fibres. The ejaculatory duct is wide and opens at the tip of the penis papilla (Fig. 22A). This duct is lined with a cuboidal-to-columnar, ciliated epithelium, surrounded by a circular muscle (50 µm thick). The penis papilla is cylindrical, with a rounded tip and is horizontally located (Figs 22A–C, 23B). This papilla is covered with a columnar epithelium, which is pierced by the necks of two types of gland cells producing erythrophil and cyanophil granules, respectively. This epithelium is underlain by a 15 µm thick muscle with interwoven circular and longitudinal fibres. The male atrium is relatively short and not folded. It is lined with a cuboidal-to-columnar epithelium, which is crossed by two types of gland cells producing erythrophil and cyanophil granules, respectively. This epithelium is underlain by a layer of circular muscle, followed by a layer of longitudinal fibres, each layer being 5 µm thick in the proximal region of the atrium and 30 µm in the distal. The ovaries are ovoid and have a maximum diameter of 400 µm in the longitudinal body axis. These ovaries are located immediately above the ventral nerve plate (Fig. 21C) and at a distance from the anterior tip of the body corresponding to 9.6% of the body length. The ovovitelline ducts emerge from the lateral aspect of the ovaries and run ventrally above the main nerve plate. Close to the mid-region of the prostatic vesicle, each ovovitelline duct opens laterally into the long, dilated common ovovitelline duct (Figs 22A, C, 23B–E). This long duct is six times wider than the ovovitelline ducts and exhibits longitudinal folds. The common ovovitelline duct ascends gradually to communicate with the common glandular ovovitelline duct. This glandular duct runs posteriorly over the male and female atria to join the female genital canal, which projects dorso-anteriorly from the dorsoposterior region of the female atrium. The female atrium is elongated to funnel-shaped and is slightly shorter than the male atrium. The common ovovitelline duct is lined with a columnar epithelium, which is crossed by three types of gland cells producing xanthophil, erythrophil and cyanophil granules, respectively (Fig. 23E). This duct is surrounded by a single muscle layer (50 µm thick) comprising circular, diagonal and longitudinal thin fibres. The female genital canal and the female atrium are lined with a 75 µm high columnar, non-ciliated epithelium and the apical side of its cells contains xanthophil granules. Additionally, gland cells discharge erythrophil granules through the epithelium of the female genital canal and that of the female atrium. These epithelia are underlain by a 50–120 µm thick muscle consisting of intermingled longitudinal and circular fibres. The common muscle coat consists of sparse longitudinal and circular muscle fibres. Remarks on the neae tribe Inakayaliini and its genus: The phylogenetic position of Inakayaliini is unstable. It was recovered as sister to Myoplanini (Fig. 2; Supporting Information, Figs S1, S 2) or to Adinoplanini (Supporting Information, Figs S3, S 4). Inakayaliini is monogeneric and currently houses four species. Two of them are represented in our phylogenetic trees, namely, I. ƲaldiƲiana and I. cyanea. These two species are sister to each other. Inakayalia cyanea matches all diagnostic features of the genus, except that the wall of its penis papilla is not irregular but smooth, and the dilated portion of the female ducts does not correspond to ovovitelline ducts, but the common ovovitelline duct. Therefore, the genus is re-diagnosed by omitting the mention of the irregular wall of the penis papilla and the dilation of the female genital ducts. This latter feature is transferred to the diagnosis of the new tribe. Inakayalia cyanaea is readily distinguished from the other three species in the genus in that it presents a long prostatic vesicle (vs. shorter) and a dilated common ovovitelline duct (vs. paired ovovitelline ducts dilated). Inakayalia cyanea is also the only species in the genus with the testes extending vertically between the supraintestinal and the subintestinal parenchymal muscle (see Fig. 21A).

TRANSANDIPLANA GRAUI ALMEIDA & CARBAYO SP. NOV. (FIGS 13–16) Zoobank registration: urn: lsid: zoobank. org:act: 9247667A-A4EE-4722-B935-7332CAA3F792 Holotype: MNHNCL PLAT-15049 (Field code, F4696). Huasco, Región de Atacama, Chile (28°27′′57.4′′S, 071°11′′09.5′′W), coll. F. Carbayo et al., 2 December 2010. Cephalic region: transverse sections on 13 slides; ovarian region: horizontal sections on 4 slides; prepharyngeal region: transverse sections on seven slides; pharynx and copulatory apparatus: sagittal sections on 13 slides. Type locality: Huasco, Región de Atacama, Chile. The species is only known from this locality. Etymology: The specific epithet pays homage to Dr José Horácio Grau, for his friendship and contribution to the knowledge of the Chilean planarians. Description External aspect: The live holotype (Fig. 13A) is about 25 mm in length and 2 mm in width. The body margins are parallel. The anterior extremity of the body is rounded, while the posterior is pointed. The dorsal side is convex; the ventral one is flat. The dorsal colour is graphite-grey (RAL 7024) – slightly clearer on the body margins– and adorned with a median grey-white (RAL 9002) stripe (Fig. 13A). The ventral surface is pure white (RAL 9010) – darker in the cephalic region – and exhibits a median, thin, whitish stripe throughout the body length (Fig. 13B). The preserved holotype measured 25 mm long, 2.5 mm wide and 1 mm high. The eyes are of a single-cup type measuring 35–38 µm in diameter. The anterior 2 mm of the body are encircled with a uniserial row of eyes. Behind this region, the eyes spread on to the dorsal surface to the extent of a band 40% of the body width on either side. This band is gradually thinner toward the posterior tip of the body, where the eyes are only marginally located. Scarce sensory depressions are located ventromarginally in the anterior region of the body (Fig. 13C–E). The depth of these depressions is equivalent to two-thirds of the height of the surrounding epidermal cells. The suboptimal quality of the sections of the cephalic region, which lacks part of the ventral portion, hindered a detailed description of their distribution. There are no sensory pits. The mouth is positioned at a distance from the anterior extremity equal to 67.6% of the body length; the gonopore is at 78.8%. Internal morphology: The creeping sole occupies 85% of the ventral surface of the body. Rhabditogen cells, gland cells producing erythrophil granules and gland cells producing xanthophil amorphous secretion discharge their secretion through the dorsal epidermis of the pre-pharyngeal region. Gland cells producing cyanophil granules and gland cells producing amorphous erythrophil secretion discharge their secretion through the ventral epidermis. A glandular margin is absent. The gland cells exhibit the same distribution in the cephalic region, although they are scarcer. The main nervous system is organized in approximately 24 longitudinal nerve cords.The shape of each cord resembles a necklace of beads (Fig. 14A, B, D). Cerebral ganglia could not be discerned. The cutaneous musculature comprises three layers, namely, a subepidermal layer of circular muscle (2–3 µm thick), followed by a double layer with diagonal fibres (4 µm) and a third layer of longitudinal muscle (20–25 µm dorsally, 17–20 µm ventrally). Muscle fibres of the longitudinal muscle layer are arranged into bundles with six to 15 fibres each (Fig. 14E, F). The cutaneous musculature thickness relative to body height at the pre-pharyngeal region is 6.7%. The musculature in the cephalic region maintains the organization observed in the pre-pharyngeal region. The parenchymal musculature comprises three layers of scattered fibres, namely, a dorsal double layer (12–15 µm thick) of decussate fibres, a supraintestinal layer of transverse muscle (40–50 µm) and a subintestinal layer of transverse muscle (35–50 µm) (Fig. 14C, D). The mouth is situated at the end of the pharyngeal pouch (Fig. 15A). A distinct oesophagus is present. The oesophagus to pharynx length ratio is 24%. The pharynx is cylindrical (Fig. 15A). The oesophagic musculature consists of a subepithelial layer of circular muscle (40 µm thick), followed by a layer of longitudinal muscle (25 µm thick). Two types of gland cells discharge their erythrophil and cyanophil granules, respectively, through the covering epithelium of the distal portion of the pharynx. The outer pharyngeal musculature consists of a subepithelial layer of longitudinal muscle (2.5 µm thick), followed by a layer of circular muscle (7.5 µm thick). The inner pharyngeal musculature consists of a single muscle of circular and longitudinal fibres interwoven. The testes range between club- and pear-shaped and are 180–230 µm in diameter. The testes are surrounded by a pigmented covering (Fig. 14C, E, F), and are distributed into one to two rows at each side of the body. They are located dorsally, beneath the supraintestinal parenchymal muscle, and between the intestinal branches. The anteriormost testes lie at a distance from the anterior tip of the body equivalent approximately to 10% of the body length; the posteriormost testes are located 2 mm (8% of body length) anterior to the pharynx, i.e. at a distance from the anterior body tip equivalent to 52.4% of the body length. The sperm ducts are located dorsally to the ovovitelline ducts. The distal portion of the sperm ducts is bent dorsally and medially to open into the mid-dorsal region of the prostatic vesicle (Fig. 15C). The distal portion of one of the ducts is dilated to form a spermiducal vesicle filled with sperm. The anteriorhalf of the prostatic vesicle is extrabulbar, dilated and with a folded wall. The posterior-half is narrow and located within the penis bulb, whose dorso-anterior region is traversed by the prostatic vesicle (Fig. 15B– D). The ejaculatory duct runs within the penis papilla to open at its tip (Fig. 15C, D). The prostatic vesicle is lined with a columnar epithelium in its proximal region; otherwise, the lining epithelium is cuboidal. Abundant gland cells discharge cyanophil granules into the prostatic vesicle (Fig. 15B, D). The dilated portion of the prostatic vesicle is surrounded by a single muscle (10–65 µm thick) of intermingled fibres, while the canalicular portion is surrounded by a single circular muscle (15 µm thick). The ejaculatory duct is lined with a cuboidal, ciliated epithelium, through which two types of gland cells discharge cyanophil and erythrophil granules, respectively. Muscle fibres surrounding this duct are not apparent. The penis papilla is conical, with the dorsal insertion shifted backward and its basis somewhat bulged. This papilla occupies the entire length of the male atrium (Fig. 15C, D). The epithelium of the penis papilla is cuboidal-to-columnar and is pierced by the openings of two types of numerous gland cells producing erythrophil and cyanophil granules, respectively. The epithelium is underlain by a single circular muscle (10 µm thick). Multiple radial and longitudinal muscle fibres are located in the stroma of the penis papilla. The male atrium is smooth (Fig. 15C, D) and is lined with a low epithelium (10 µm high) proximally and tall (30 µm thick) in the other regions. Two types of gland cells discharge their cyanophil and erythrophil granules, respectively, through the atrial epithelium. Cyanophil granules are particularly abundant in the proximal region of the male atrium. The atrial epithelium is underlain by a subepithelial layer of circular muscle (7.5 µm thick), followed by a layer of longitudinal muscle (5 µm thick). The ovaries are ovoid, with 190 µm in maximum diameter (Fig. 16A) and are located at a distance from the anterior body tip equivalent to about 11% of the body length. The ovovitelline ducts emerge from the dorsoposterior portion of the ovaries (Fig. 16A). Laterally to the gonopore and the female atrium, the ovovitelline ducts ascend dorsoposteriorly, to subsequently bending anteriorly to communicate with the common ovovitelline duct. This unpaired duct runs dorso-anteriorly to join the posterodorsal region of the female atrium. In this joining point, a short diverticulum of projects anteriorly (Fig. 15C). The female atrium is spherical-to-ovoid. A dorsal fold located above the gonopore narrows the communication of the male and female atria. The female atrium is approximately as long as the male atrium and is lined with a columnar (150 µm high) epithelium provided with an irregular surface and stratified appearance. This epithelium has scattered and unevenly distributed cilia, presenting some spaces with a vacuolar aspect filled with cyanophil secretion (Fig. 16B, C). This atrial epithelium is pierced by the necks of two types of gland cells producing erythrophil and cyanophil granules, respectively. The lining epithelium of the female atrium is underlain by a subepithelial layer of longitudinal muscle (2.5 µm thick), followed by a layer of circular muscle (7.5 µm thick); ectally to this muscle is a longitudinal loose muscle (20 µm) coating the atrium (Fig. 16B). Remarks on Geoplanini and the neae genus Transandiplana: Transandiplana is nested in Geoplanini in all molecular trees. This tribe also includes Amaga Ogren & Kawakatsu, 1990, Bogga Grau & Sluys, 2012, Barreirana Ogren & Kawakatsu, 1990, Cephaloflexa Carbayo & Leal-Zanchet, 2003, Choeradoplana Graff, 1896, Cratera Carbayo et al., 2013, Difroehlichia Leal-Zanchet & Marques, 2018, Geobia Diesing, 1862, Geoplana Schultze, 1856, Imbira Carbayo et al., 2013, Issoca Froehlich, 1954, Luteostriata Carbayo, 2010, Matuxia Carbayo et al., 2013, Notogynaphallia Ogren & Kawakatsu, 1990, Obama Carbayo et al., 2013, Paraba Carbayo et al., 2013, Pasipha Ogren & Kawakatsu, 1990, Piima Carbayo, 2020, Pseudogeoplana (collective genus) Ogren & Kawakatsu, 1990, Supramontana Carbayo & Leal-Zanchet, 2003, Winsoria Negrete et al., 2020 and Xerapoa Froehlich, 1955. Geoplanini differ from the remaining tribes by a unique combination of characters, namely, dorsum convex (vs. carinate in Adinoplanini), dorsal eyes (vs. only marginal in Haranini, Inakayaliini, Polycladini, Sarcoplanini and Timymini), dorsal longitudinal cutaneous muscle not sunken into the parenchyma (vs. sunken in Gusanini; the Geoplanini Choeradoplana gladismariae Carbayo & Froehlich, 2012 is an exception), pharyngeal pouch anterior to the copulatory apparatus (vs. extending posteriorly over the copulatory apparatus in Haranini and Timymini), female atrium without musculoglandular organs (vs. with it in Adinoplanini and Sarcoplanini) and female genital duct not dilated (vs. dilated in Inakayaliini). Thus, Geoplanini lack exclusive morphological characteristics. However, the eye distribution pattern might help to recognize most members of the tribe. Although some subcylindrical species of Geoplanini exhibit dorsal eyes, Geoplanini with a flattened body also present dorsal eyes. Exceptions to this pattern are some Geoplanini taxa such as Bogga and Amaga, which, while being flattened, have only marginal eyes. Nonetheless, the phylogenetic position of these two genera has not yet been assessed. The new Geoplanini genus Transandiplana is represented by one single individual. It is retrieved as an ingroup of Geoplanini in all analyses, having an unstable position, either sister to Geoplana Stimpson, 1858 (Fig. 2; Supporting Information, Fig. S4), to Paraba multicolor (Graff, 1899) (Supporting Information, Fig. S3) or it is a branch of a polytomy (Supporting Information, Figs S2, S 5, S 6). Transandiplana graui largely resembles some Geoplanini genera in the general shape of the copulatory apparatus, including the lack of structures such as musculoglandular organs. However, while it is similar to other Geoplanini, it also exhibits some features uncommon in Geoplanini, namely, sensory depressions, the main nervous system consisting of multiple longitudinal cords, testes provided with a pigmented covering, and the relatively large distance between the testes and the pharynx. With this combination of features, T. graui does not fit well in any of the Geoplaninae tribes. Sensory depressions are known from Sarcoplanini. The main nervous system in Geoplanini consists of either two longitudinal cords (usually in small and thin species such as Xerapoa) or it exhibits the aspect of an even plate (especially in large and flat organisms such as Obama). Transandiplana graui is an exception, as it presents both a slender body and a main nervous system consisting of multiple cords. These two latter features (dark spots covering testes and relative position of posteriormost testes) are underreported in most old species descriptions. Testes covered with dark spots were only reported for Obama ladislaƲii (Graff, 1899) (in: Álvarez-Presas et al., 2015). Given the general morphological resemblance of T. graui with other Geoplanini taxa and the phylogenetic position as an ingroup of Geoplanini, we tentatively place the species and the genus within Geoplanini. Transandiplana graui resembles ‘ Geoplana ’ caleta E. M. Froehlich, 1978 in the general aspect of the body and internal organs. However, important diagnostic traits (ventrolateral sensory depressions; main nervous system comprising multiple longitudinal cords; testes surrounded by a pigmented covering) and molecular data of G. caleta remain unknown. Thus, a systematic replacement would be speculative. Interestingly, both species occur in an adverse climatic region, namely, the Huasco river valley, in the arid Norte Chico zone (Atacama) of the Chilean Andes. Annual rainfall in the region is 42 mm /year, and the relative humidity ranges between 66–74% (see: Juliá et al., 2008)., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 854-859, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Graff LV. 1896. Uber das System und die geographische Verbreitung der Landplanarien. Verhandlungen der Deutsche Zoologischen Gesellschaft 6: 75 - 93.","Carbayo F, Alvarez-Presas M, Olivares CT, Marques FPL, Froehlich EM, Riutort M. 2013. Molecular phylogeny of Geoplaninae (Platyhelminthes) challenges current classification: proposal of taxonomic actions. Zoologica Scripta 42: 508 - 528.","Schultze M. 1856. Beitrage zur Kenntnis der Landplanarien nach Mittheilungen des Dr. Fritz Muller in Brasilien und nach eigenen Untersuchungen. Abhandlungen der Naturforschenden Gesellschaft zu Halle 4: 19 - 38.","Negrete L, Alvarez-Presas M, Riutort M, Brusa F. 2020. Integrative taxonomy of land planarians (Platyhelminthes: Geoplanidae) from the Andean-Patagonian Forests from Argentina and Chile, with the erection of two new genera. Journal of Zoological Systematics and EVolutionary Research 59: 588 - 612.","Stimpson W. 1858. Prodromus descriptionis animalium evertebratorum quae in expeditione ad Oceanum Pacificum Septentrionalem, Johanne Rodgers Duce a Republica Federate missa, observavit et descripsit. Proceedings of the Academy of Natural Sciences of Philadelphia 9: 19 - 31.","Graff LV. 1899. Monographie der Turbellarien II. Tricladida Terricola (Landplanarien). Atlas Von Achtundfunfzig Tafeln zur Monographie der Turbellarien II. Tricladida Terricola (Landplanarien), pls I - LVIII. Leipzig: Wilhelm Engelmann.","Alvarez-Presas M, Amaral SV, Carbayo F, LealZanchet AM, Riutort M. 2015. Focus on the details: morphological evidence supports new cryptic land flatworm (Platyhelminthes) species revealed with molecules. Organism DiVersity and EVolution 15: 379 - 403.","Froehlich EM. 1978. On a collection of Chilean land planarians. Boletins de Zoologia da UniVersidade de Sao Paulo 3: 7 - 80.","Julia C, Montecinos S, Maldonado A. 2008. Caracteristicas climaticas de la region de Atacama. In: Squeo FA, Arancio G, Gutierrez JR, eds. Libro Rojo de la Flora NatiVa y de los Sitios Prioritarios para su ConserVacion: Region de Atacama, Vol. 3. La Serena: Ediciones Universidad de La Serena, 25 - 42."]}

SARCOPLANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: D16FF0B2-C7D9-4082-9C98-09D81831A7AF Diagnosis: Geoplaninae with narrow-to-wide creeping sole, ranging between 51 and 83% of the body width. Eyes marginal. With sensory depressions. Subneural parenchymal decussate muscle present. Prostatic vesicle extrabulbar. Generally provided with a cephalic retractor muscle, branched glands associated with the prostatic vesicle and genital musculoglandular organs. Sarcoplanini include the genera Liana E.M. Froehlich, 1978, Mapuplana Grau et al., 2022, Pichidamas Bulnes et al., 2018, Sarcoplana and Wallmapuplana Negrete et al., 2020. Type genus: Sarcoplana Almeida & Carbayo gen. nov. Distribution: Regiones Maule, La Araucanía, Los Lagos, and Aisén (Chile); Provinces Neuquén, Rio Negro and Chubut (Argentina)., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 877, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Froehlich EM. 1978. On a collection of Chilean land planarians. Boletins de Zoologia da UniVersidade de Sao Paulo 3: 7 - 80.","Grau JH, Almeida AL, Sluys R, Carbayo F. 2022. A new genus and two new species of land planarians (Platyhelminthes, Tricladida, Geoplanidae) from Southern Chile, including the Chonos archipelago. Journal of Natural History 56: 947 - 967.","Bulnes VN, Grau JH, Carbayo F. 2018. A new Chilean genus and species of land planarian (Platyhelminthes: Tricladida, Geoplaninae) with cephalic retractor muscle and adenodactyl. Journal of Natural History 52: 2553 - 2566.","Negrete L, Alvarez-Presas M, Riutort M, Brusa F. 2020. Integrative taxonomy of land planarians (Platyhelminthes: Geoplanidae) from the Andean-Patagonian Forests from Argentina and Chile, with the erection of two new genera. Journal of Zoological Systematics and EVolutionary Research 59: 588 - 612."]}

GUSANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: 9E0939FB-0E21-4209-B514-A52634BD1ED0 Type genus: Gusana E.M. Froehlich, 1978. Diagnosis: Geoplaninae with a broad and foliaceus body, tapering abruptly to the anterior tip. Anterior tip triangular. Dorsal and ventral longitudinal cutaneous muscle partly sunken into the parenchyma. Sensory pits as a simple invagination or obliquely elongated and internally branched. Gusanini comprise only the genus Gusana. Etymology: The name of the tribe is based on the name of its type genus. Distribution: Regions Biobío and La Araucanía (Chile).

HARANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: F3527621-B8FF-482B-9814-07855F164AE7 Type genus: Harana Almeida & Carbayo gen. nov. Diagnosis: Small-sized Geoplaninae with a longitudinal tube located among the fibres of the subintestinal transverse parenchymal muscle. Long pharyngeal pouch, extending behind the copulatory apparatus. Haranini comprises only the genus Harana. Etymology: The name of the tribe is based on the name of its type genus. Distribution: Only known from the type locality of the type species.

PICHIDAMAS BULNES ET AL., 2018 Type species: Pichidamas piru Bulnes et al., 2018, designated by Bulnes et al., (2018). Neae diagnosis: Sarcoplanini with a small- to mediumsized body, subcylindrical, slender with nearly parallel margins and anterior extremity rounded. Eyes monolobulate, distributed along the body margins. Cephalic retractor muscle present, derived from ventral longitudinal cutaneous musculature. Five parenchymal muscle layers, the ventralmost layer consisting of decussate fibres and located to the inside of the peripheral nerve plexus. Distal portion of male atrium with an adenodactyl. Penis eversible. Ovovitelline ducts ventral, opening to the common ovovitelline duct from posteroventral. Distribution: Regions Maule and Los Lagos, Chile. PICHIDAMAS GNYTHOS ALMEIDA & CARBAYO, Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 877, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Bulnes VN, Grau JH, Carbayo F. 2018. A new Chilean genus and species of land planarian (Platyhelminthes: Tricladida, Geoplaninae) with cephalic retractor muscle and adenodactyl. Journal of Natural History 52: 2553 - 2566."]}

MYOPLANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: 92DB3E7F-3F9C-47FE-B68A-83FDADA3A19A Diagnosis: Geoplaninae with the ventral peripheral nervous plexus divided into two plexuses. With a transneural parenchymal muscle, this consisting of diagonal fibres. Inner pharyngeal musculature consisting of four muscle layers. Type genus: Myoplana Almeida & Carbayo gen. nov. Myoplanini comprises only the genus Myoplana. Etymology: The name of the tribe is based on the name of its type genus. Distribution: Región de la Araucanía, Chile.

SARCOPLANA MUSCULOSA ALMEIDA & CARBAYO SP. NOV. (FIGS 40–44) Zoobank registration: urn: lsid: zoobank. org:act: 67BAB3EC-6FCC-4799-89E9-48133F0A40D1 Holotype: MNHNCL PLAT-15047 (Field code, F4886). Parque Nacional Nahuelbuta, Purén, Región de La Araucanía, Chile (37°49′′37.2′′S, 073°00′′32.4′′W), coll. F. Carbayo et al., 11 December 2010. Cephalic region: transverse sections on 11 slides; portion immediately behind the cephalic region: horizontal sections on six slides; pre-pharyngeal region: transverse sections on 17 slides; pharynx and copulatory apparatus: sagittal sections on four slides. Type locality: Parque Nacional Nahuelbuta, Región de La Araucanía, Chile. The species only known from this locality. Etymology: The specific epithet is from the Latin adjective musculosus, muscular, alluding to the thick cutaneous musculature. Description External aspect: At rest, the live specimen measured approximately 18 mm long and 3 mm wide (Fig. 40A, B). The body length may double when crawling (Fig. 40C). The body margins are parallel. The anterior tip is rounded, while the posterior is pointed. The dorsum is convex and the ventral side is flat. The preserved specimen measured 17.5 mm long, 2.5 mm wide and approximately 1.5 mm high. The dorsum displays a pure orange (RAL 2004) median stripe 21% of the body width, which is divided longitudinally by a thin carmine-red (RAL 3002) midline (2.3% of the body width; Fig. 40A, C). The median stripe is absent in both extremities of the body. External to the median stripe is a black-red (RAL 3007) band 37% of the body width, the margins of which are darker. Some pure orange spots occur in the bands. These bands converge in the cephalic region. The ventromarginal sensory border is a line with beige-grey colour (Fig. 40B). The body margins are pure orange. The ground colour of the ventral side is pure orange, provided with a pair of bands, each with 26% of body width, consisting of brown-red (RAL 3011) dots. The inner and outer margins of the bands are darker (Fig. 40B). The monolobate eyes measure 4 5–5 0 µm in diameter. They are distributed in an irregular row contouring the cephalic region and extending marginally to the posterior tip of the body. Sensory pits are absent. Instead, the ventromarginal epithelium of the cephalic region possesses sensory depressions. These depressions reach the underlying basal lamina and are provided with cilia (Fig. 41A). The sensory depressions are absent at the anterior tip of the body. The mouth is positioned at a distance from the anterior extremity of the body equal to 66% of the body length; the gonopore 77%. Internal morphology: The epidermis is ciliated only on the creeping sole, this occupying 83% of the body width. Gland cells producing erythrophil granules and cells producing rhabdites discharge through the entire epidermis. The erythrophil type is more abundant in the body margin, while the rhabditogen cells are more numerous in the ventral surface of the cephalic region (Fig. 41A). Gland cells producing weakly cyanophil granules also discharge through the ventral and marginal epidermis. A glandular margin is absent. The cutaneous musculature comprises three layers, namely, a subepidermal, layer (2 µm thick) of circular muscle, followed by a double layer (13 µm) with decussate fibres and an innermost layer of longitudinal muscle, the fibres of which are gathered into bundles (Fig. 41B–F). This latter layer is 70 µm thick dorsally and 160 µm ventrally. It is thinner than the body margins, where it remains conspicuous (Fig. 41B, F). The ventral layer of the longitudinal muscle is divided into a thin outer muscle and a thick inner muscle. These outer and inner ventral portions of the longitudinal muscle are separated by a secondary peripheral nerve net (Fig. 41C). The relative thickness of the cutaneous musculature is 19.5% of body height. The parenchymal musculature comprises four layers along the entire body: a dorsal layer (30 µm thick) of decussate fibres located to the inside of the peripheral nervous net; a dense layer of supraintestinal transverse muscle (40 µm); a dense layer of subintestinal transverse muscle (75 µm); and a subneural layer of decussate fibres (40 µm) (Fig. 41B, C, E, F). Additionally, abundant oblique muscle fibres run in transverse body planes along the body. The muscular organization changes in the anterior region of the body. At 1.9 mm from the anterior tip of the body, the longitudinal cutaneous muscle is 40 µm thick dorsally and 180 µm ventrally. In this region, the relative thickness of the cutaneous musculature is 21.6% of the body height (Fig. 41F). The cutaneous and parenchymal muscles are thinner at 1.35 mm from the body tip. At 0.6 mm, the inner ventral cutaneous longitudinal muscle concentrates medially, so that one-quarter of the body width at each side of the body lacks this muscle. In this region, a cephalic retractor muscle is flat lenticular in cross-sections (Fig. 42A). At 0.4 mm from the anterior tip, the secondary peripheral cutaneous nerve net is inconspicuous so that the ventral cutaneous muscle is no longer divided into an outer and an inner layer. Here, the longitudinal muscle is roughly lenticular in cross-section (Fig. 42B). Toward the anterior tip of the body, the retractor muscle becomes progressively smaller as its muscle fibres progressively detach from it to run obliquely to the dorsum and body margins (Fig. 42C, D). The mouth is located at a distance from the anterior region of the pharyngeal pouch, equivalent to 65% of the pouch length. The pharynx is cylindrical (Fig. 43A, B). The ventro-anterior portion of the pharynx was cut off for DNA extraction, and thus the presence of an oesophagus could not be ascertained. The outer pharyngeal musculature consists of a subepithelial layer (5 µm thick) of longitudinal muscle, followed by a layer (8 µm) of circular fibres. The inner pharyngeal musculature consists of a single subepithelial layer of circular muscle, with longitudinal fibres interspersed (40 µm). The stroma of the pharynx has circular and longitudinal fibres. The testes are pear-shaped and measure approximately 400 µm high. They are dorsally located beneath the transverse supraintestinal parenchymal muscle and between the intestinal branches (Fig. 41B). They are distributed in a row of one to two testes at each side of the body. The anteriormost testes are placed at a distance from the anterior tip of the body equivalent to approximately 35% of the body length, that is, 1.2 mm behind the ovaries; the posteriormost testes lie at a distance from the anterior tip equivalent to 44% of body length, that is, 100 µm anterior to the pharynx. The sperm ducts run above the subintestinal parenchymal muscle and more or less dorsally to the ovovitelline ducts. The distal portion of the sperm ducts bends dorsally to the sagittal plane to open into the proximal section of the respective branch of the prostatic vesicle (Fig. 43C). The prostatic vesicle is a sinuous tube roughly C-shaped in lateral view. Its proximal portion is bifurcate. This vesicle penetrates the anterior region of the penis bulb to join the ejaculatory duct. The penis bulb is well developed and is mainly constituted of longitudinal fibres. Most of the ejaculatory duct is sinuous and located within the penis bulb, while its distal section is straight and opens at the tip of the penis papilla. The penis papilla is 300 µm long and lies horizontally. This papilla is conical and presents some folds (Figs 43C, 44A). The prostatic vesicle is lined with a cuboidal, apparently non-ciliated epithelium. This epithelium is pierced by the necks of gland cells producing fine (0.5 µm) erythrophil granules and is surrounded by a circular muscle (10 µm thick). The ejaculatory duct is lined with a cuboidal ciliated epithelium. The basal-half of the penis papilla is lined with a columnar epithelium traversed by the necks of numerous openings of gland cells producing erythrophil granules. The distal-half of the papilla is lined with a cuboidal epithelium through which some gland cells of the same type discharge. The epithelium of the penis papilla is underlain by some longitudinal muscle fibres. The male atrium is narrow, elongated and roughly smooth (Fig. 43C). This atrium is lined with a cuboidal-to-columnar epithelium, the apical surface of which is erythrophil. Numerous gland cells discharge erythrophil granules through the atrial epithelium, which is underlain by a layer (10 µm thick) of circular muscle, followed by a layer (10 µm) of longitudinal fibres. The atrial wall dorsal to the gonoduct presents the openings of two different musculoglandular organs, one located behind another (Fig. 43C). The anterior organ (named mg 1 in the figures) consists of a 310 µm long and 30 µm wide, bowed and vertical blind duct embedded into the parenchyma and surrounded by muscle fibres. The duct of this musculoglandular organ is lined with a 10 µm high columnar epithelium, and the cells of this epithelium contain fine erythrophil granules (0.5 µm in diameter) produced by gland cells located outside the organ. The epithelium of the duct is underlain by a layer (10 µm thick) of circular muscle, followed by a layer (50 µm) of muscle fibres variously oriented, most circular. Beneath the epithelium of the innermost portion of the duct is a cyanophil, granular mass. The lumen of the canal contains some erythrophil granules. The posterior musculoglandular organ (named mg 2 in the figures) is ampulla-shaped. It consists of a 130 µm long duct leading to a deeper, enlarged portion with 120 µm in diameter (Figs 43B, C, 44A). The duct is lined with a cuboidal, strongly erythrophil epithelium. A 30 µm thick longitudinal muscle underlies this epithelium. The cells of the lining epithelium of the enlarged portion are not discernible. Abundant gland cells with their bodies outside the organ discharge fine cyanophil granules into the lumen of the enlarged portion of the organ. Surrounding this enlarged portion of the musculoglandular organ is a 30 µm thick muscle net, followed by a layer (30 µm thick) of longitudinal fibres. The ovaries are rounded-to-ovoid and approximately 100 µm in diameter. They are incompletely developed. These ovaries are located at a distance from the anterior tip of the body corresponding to 28% of the body length and 1.2 mm anterior to the anteriormost testes. The ovaries lie immediately above the ventral nerve plate. The ovovitelline ducts emerge laterally from the dorsal side of the ovaries. Subsequently, these ducts run posteriorly above the nervous plate and immediately underneath the transverse subintestinal parenchymal muscle (Fig. 41C). Just behind the level of the gonopore, one ovovitelline duct ascends gradually to enter the common ovovitelline duct behind the female atrium. This duct is short and oriented dorsally and communicates with the female genital canal. This female canal projects posteroventrally from the posterior wall of the female atrium (Fig. 43C). The suboptimal quality of the sections did not allow examination of the second ovovitelline duct nor the type of epithelium lining the common ovovitelline duct and the female genital canal. The female atrium is elongated and narrow. The dorsal wall of this atrium is more or less smooth, whereas the ventral wall is provided with three shallow recesses, each 100–200 µm in size (Fig. 44C, D). The female atrium is lined with a columnar, 35–45 µm high epithelium. The free surface of this epithelium is erythrophil and resembles the bristles of a brush. Gland cells producing fine erythrophil granules pierce this epithelium. The recesses are lined with a low epithelium. The female atrium contains clumps of xanthophil granules. The lining epithelium of the female atrium is surrounded by a 5 µm thick layer of longitudinal fibres, followed by a 10 µm thick layer of circular fibres. The male atrium to female atrium ratio is 84%. A common muscle coat wraps the distal-half of the prostatic vesicle and the male and female atria. This coat is comprised of abundant longitudinal muscle fibres. Remarks on the neae tribe Sarcoplanini and its genera: The molecular phylogenies retrieved Sarcoplanini as a monophyletic group comprising Mapuplana, Pichidamas, Sarcoplana and Wallmapuplana. The intergeneric relationships are unstable. The monotypic genus Liana can be included in this tribe based on the morphological similarity of L. guasa Froehlich, 1978 with Sarcoplanini members, as shown below. The species of Sarcoplanini share three unique characteristics among the Geoplaninae, namely, sensory depressions, a cephalic retractor muscle with a particular fibre organization (possibly secondarily lost in Wallmapuplana) and a subneural parenchymal decussate muscle (but the fibre orientation of this muscle is unknown in Wallmapuplana). These characteristics readily distinguish the Sarcoplanini members from the remaining Geoplaninae. Furthermore, branched glands associated with the prostatic vesicle are present in three of the four genera (Mapuplana, Pichidamas and Wallmapuplana). Additional traits shared by all species in Sarcoplanini, and which probably evolved convergently in other lineages of Geoplanidae, are marginal distribution of the eyes [also present in Adinoplanini, Myoplanini, Haranini, Caenoplanini (Rhynchodeminae) and some Geoplanini)], a small penis papilla (e.g. Amaga, Gusana, but it is large in Liana), a copulatory apparatus provided with musculoglandular organs [possibly secondarily lost in Mapuplana; also present in Australasian taxa, such as some Bipalium, Artioposthia Graff, 1896 (see: Fyfe, 1947), Coleocephalus Fyfe, 1953 (see: Winsor, 1998)] and a female genital canal with the postflex condition (i.e. the canal approaching the female atrium from behind as in Pasipha, Gigantea, and Gusana, among others). The genera of Sarcoplanini differ from each other by several structures. Sarcoplana stands apart from the remaining Sarcoplanini genera from the presence of a secondary peripheral nerve net located in the ventral side of the body (convergent in Myoplana). Mapuplana and Liana are the only genera of Sarcoplanini having part of the ventral longitudinal cutaneous muscle sunken into the parenchyma. These two genera differ in that the penis papilla is small in Mapuplana (vs. large in Liana). Wallmapuplana is the only genus of Sarcoplanini lacking a cephalic retractor muscle, while Pichidamas is the only genus having a large musculoglandular organ of adenodactyl type. The genus Liana deserves a detailed discussion. This monotypic genus was proposed for L. guasa (Froehlich, 1978). The species was described from incompletely mature individuals. The main diagnostic features of the genus are: elongated body, broad creeping sole, sensory depressions (‘minute sensory pits’ in the original description), longitudinal ventral cutaneous muscle partially sunken into the parenchyma, cutaneous muscle thickness relative to the body height is 10%, testes are dorsal; copulatory apparatus without adenodactyls; penis papilla short and blunt; female canal approaches from horizontal or ventral aspect (Froehlich, 1978). This species also has a subneural layer of decussate parenchymal muscle (‘a layer of fibres obliquely oriented to the right and to the left’ in Froehlich, 1978: 21) interwoven with fibres of the sunken longitudinal cutaneous muscle. The relative thickness of the cutaneous musculature increases to 21% when the sunken muscle portion is also considered (see: Froehlich, 1978: fig. 24). There are no gene sequences available of this species. Among the Geoplanini tribes, Liana fits well Sarcoplanini:the creeping sole is wide; the eyes marginal; sensory depressions and subneural parenchymal decussate muscle are present. The original description of L. guasa does not mention a cephalic retractor muscle but a modification of the musculature organization in the cephalic region, which is compatible with a retractor organ (‘ At the anterior end, the dorsal longitudinal [Ʋentrally?] cutaneous fibers bend dorsally to end on the basement membrane. Laterally, toaeards the Ʋentral sensory border the cutaneous musculature progressiƲely loses height becoming minimal if not absent. Ventrally it regains height toaeards the median line, attaining a little more than half the height of the dorsal portion. […] The Ʋentral longitudinal parenchymal [cutaneous?] musculature progressiƲely disappears toaeards the anterior extremity. At the same time it appears there a layer of diagonal fibers interspersed aeith rarer and rarer longitudinal fibers. Presumably the longitudinal fibers change direction anteriorly but it cannot be discerned’, E. M. Froehlich, 1978, p. 22). The presence of two taxonomically relevant diagnostic features of Sarcoplanini, namely, branched glands associated with the prostatic vesicle and genital musculoglandular organs, could not be verified since the individuals are only partially mature. Liana does not fit in any of the remaining tribes because it lacks the following features: carinate dorsal side, musculoglandular organs in the female atrium (Adinoplanini); body leaf-like with dorsal eyes (Geoplanini); anterior region of the body triangular, ventral and dorsal longitudinal cutaneous muscle sunken into the parenchyma, sensory pits (Gusanini); long pharyngeal pouch (Haranini and Timymini); dilated female genital ducts (Inakayaliini); transneural parenchymal muscle of diagonal fibres (Myoplanini); extraordinarily wide and flattened body, marginal eyes, subneural parenchymal of transverse muscle and a transneural parenchymal layer of longitudinal muscle (Polycladini); semi-lunate head plate (Timymini). Therefore, we place the genus Liana in Sarcoplanini.

GUSANA E.M. FROEHLICH, 1978 Ty p e s p e c i e s: G e o p l a n a c r u c i a t a G r a f f, 1 8 9 9, designated by E. M. Froehlich (1978). Neae diagnosis: Gusanini with body colour pattern with cross-banding; creeping sole broad, with more than half the body width. Sensory border wide, around the anterior tip. Cutaneous musculature thickness relative to body height at the pre-pharyngeal region ranges between 16 and 24%. Testes dorsally located. Male atrium large. Penis papilla of small intra-antral type. Female canal enters ventrally. Without adhesive musculoglandular organs and sensory papillae. Copulatory apparatus without adenodactyls. Distribution: As for that of the tribe. Remarks on Gusanini: This monogeneric tribe includes six species of Gusana (Almeida et al., 2022). This genus was recovered as a clade in Geoplaninae by Almeida et al. (2022), and also in this paper. Gusana was erected by E. M. Froehlich (1978) and re-diagnosed by Ogren & Kawakatusu (1990) and Almeida et al. (2022). The tribe is diagnosed herein by a set of diagnostic features of Gusana herein elevated to the tribe level. Therefore, the diagnosis of Gusana is shortened., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 859, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Froehlich EM. 1978. On a collection of Chilean land planarians. Boletins de Zoologia da UniVersidade de Sao Paulo 3: 7 - 80.","Almeida AL, Alvarez-Presas M, Bolonhezi L, Carbayo F. 2022. Integrative taxonomy increases biodiversity knowledge of Gusana (Platyhelminthes, Tricladida, Geoplanidae) with the description of four new Chilean species. InVertebrate Systematics 36: 533 - 556."]}

MYOPLANINI ALMEIDA & CARBAYO TRIB. NOV. Zoobank registration: urn: lsid: zoobank. org:act: 92DB3E7F-3F9C-47FE-B68A-83FDADA3A19A Diagnosis: Geoplaninae with the ventral peripheral nervous plexus divided into two plexuses. With a transneural parenchymal muscle, this consisting of diagonal fibres. Inner pharyngeal musculature consisting of four muscle layers. Type genus: Myoplana Almeida & Carbayo gen. nov. Myoplanini comprises only the genus Myoplana. Etymology: The name of the tribe is based on the name of its type genus. Distribution: Región de la Araucanía, Chile., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on page 867, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977

INAKAYALIA CYANEA ALMEIDA & CARBAYO SP. NOV. (FIGS 20–23) Zoobank registration: urn: lsid: zoobank. org:act: 3CF36CAC-C035-45EC-B439-79C0EE4DC8E3 Material examined: All specimens collected in Parque Nacional Nahuelbuta, Chile (37°48′′00.0′′S, 073°00′′00.0′′W), coll. F. Carbayo et al., 13 December 2010. Holotype: MNHNCL PLAT-15043 (Field code, F4912). Cephalic region: transverse sections on 14 slides; ovarian region: horizontal sections on 100 slides; prepharyngeal region: transverse sections on 30 slides; copulatory apparatus: sagittal sections on 122 slides. Paratypes: MZUSP PL 2283 (Field code, F4914). Cephalic region: transverse sections on ten slides; ovarian region: horizontal sections on 13 slides; prepharyngeal region: transverse sections on ten slides; pharynx and copulatory apparatus: sagittal sections on 73 slides. MZUSP PL2284 (Field code, F4917). Cephalic region: transverse sections on 15 slides; ovarian region: horizontal sections on 12 slides; prepharyngeal region: transverse sections on 20 slides; pharynx: sagittal sections on 22 slides; copulatory apparatus: sagittal sections on seven slides. Type locality: Parque Nacional Nahuelbuta, Región de la Araucanía, Chile. The species is only known from this locality. Etymology: The specific epithet derives from the Greek Κ&upsi;ανό&sigmav; meaning blue, alluding to the colour of the body. Diagnosis: Species of Inakayalia with a long and widened prostatic vesicle, branched proximally, and provided with a long posterodorsal diverticulum; long unpaired and dilated common ovovitelline duct. Description External aspect: The three specimens (Fig. 20) are mature and measured between 23 and 27 mm in length and 6–8.3 mm in width at rest. Preserved, they measured 28.5–44.5 mm long, 5–6 mm wide and 1.2–1.3 mm high. At rest, the body is lanceolate with undulating margins (Fig.20D, E). The cephalic region narrows to the rounded tip; the posterior narrows abruptly to the pointed tip. The dorsum is flattened except for the convex median region. The ventral side is flat. With approximately one-eighth of the body length, the cephalic region exhibits two black-blue (RAL 5004) bands, separated from each other by a median pure white (RAL 9010) line (Fig. 20A–C). These two bands are completely (Fig. 20A, B) or incompletely (Fig. 20C) interrupted by a zigzagged, pure white or beige transverse band. Behind the transverse band, the dorsum is black-blue (RAL 5004), darker along the median and paramedian zones to form bands, each with 10–16% of the body width. Additionally, large blue-grey (RAL 7031) haloes mottle the dorsum behind the transverse band except for the median zone. The ground colour of the ventral side is cream (RAL 9001) (Fig. 20D, E). Numerous small black-blue pigment dots are in the cephalic region. A whitish transverse band continued from the dorsal side separates the cephalic region from the remaining ventral surface, which is covered with numerous dots, either graphitegrey (RAL 7024) or ochre-brown (RAL 8001). The eyes are of a single-cup type measuring 32–38 µm in diameter. They are organized in a singleto-biserial row around the anterior eighth of the body; behind this body region, they spread onto the dorsal surface and are encircled by the blue-grey haloes. The sensory pits are simple invaginations 50–57 µm deep and are located ventromarginally in a single row running along an anterior region with 18% of body length. The mouth is positioned at a distance from the anterior extremity equal to 63.4–66.3% of the body length; the gonopore, 83.6–86.7%. Internal morphology: The creeping sole has 95% of the body width. Abundant gland cells producing coarse (1 µm) xanthophil granules and scarce gland cells producing erythrophil granules discharge their secretion through the entire epidermis of the prepharyngeal region. The cell necks of the xanthophil type are 10–15 µm in diameter and become more abundant toward the body margins of the dorsal side. The glandular margin consists of xanthophil gland cells (Fig. 21A). Rhabdites are discharged through the dorsal epidermis. As the creeping sole narrows toward the anterior extremity of the body, the xanthophil glands become scarce dorsally and abundant ventrally. The cutaneous musculature comprises three layers, namely, a subepithelial layer of circular muscle (5 µm thick), followed by a double layer (15–40 µm) with decussate fibres and then a well-developed, innermost layer of longitudinal fibres (30–105 µm thick, both dorsally and ventrally) (Fig. 21B–C). The cutaneous musculature thickness relative to the body height is 13–15%. Toward the anterior region of the body, these muscle layers become thinner until they disappear. There are three strong parenchymal muscle layers, namely, a dense dorsal layer of decussate fibres (30 µm thick), a supra-intestinal layer of transverse fibres (100 µm thick) and a denser subintestinal layer of transverse fibres (100 µm thick; Fig. 21B–C). These muscle layers are thinner in the anterior region of the body. Two of the three main branches of the intestine, namely, the paired ones, may connect to each other at the level of the prostatic vesicle. The oesophagus to pharynx length ratio is 31–33%. The mouth is situated at a distance from the root of the pharynx equivalent to 41–54% of the pharyngeal pouch length (Fig. 21D). The distal portion of the pharyngeal pouch is close to the prostatic vesicle. The pharynx is bell-shaped, with its dorsal insertion slightly anterior to the mouth. The epithelium of the distal portion of the pharynx is pierced by the necks of four types of gland cells, producing xanthophil granules, erythrophil granules, cyanophil granules and amorphous secretion, respectively. The outer pharyngeal musculature consists of a subepithelial layer of longitudinal muscle (5 µm thick), followed by a layer of circular muscle (15 µm thick) and an innermost layer of longitudinal muscle (5 µm thick). The inner pharyngeal musculature consists of a subepithelial circular muscle (75 µm thick), followed by a longitudinal muscle (10 µm thick; Fig. 21E). The rounded-to-irregular testes measure 325– 450 µm in diameter. They are organized into two to four rows in three vertical levels at each side of the body, between the supra-intestinal and subintestinal parenchymal muscles (Fig. 21A). The anteriormost testes lie at a distance from the anterior tip of the body equivalent to 17.3% of the body length; the posteriormost ones, the equivalent to 65% of the body length, i.e. they are lateral to the pharyngeal root. The sperm ducts run immediately above the subintestinal parenchymal muscle. Laterally to the pharyngeal pouch, each duct opens into the anteroventral region of the respective lateral, short branch of the prostatic vesicle (Fig. 22A). The rest of the prostatic vesicle is unpaired and large, measuring up to 1.3 mm in length. Approximately the anterior-half of the vesicle occupies two-thirds of the body height and exhibits numerous folds filling its lumen (Figs 22B, C, 23A–C). The posterior-half consists of a dorsal, blind diverticulum (200–250 µm long) and a ventral, widened duct (600 µm long), both running posteriorly. The latter penetrates the anterior section of the penis bulb to communicate with the ejaculatory duct. The prostatic vesicle is lined with a ciliated epithelium, being cuboidal in the paired portion and columnar in the unpaired one. Abundant gland cells discharge erythrophil granules through the lining epithelium of the prostatic vesicle. The epithelium of this vesicle is underlain by a muscle layer (25–100 µm thick) of decussate fibres. The ejaculatory duct is wide and opens at the tip of the penis papilla (Fig. 22A). This duct is lined with a cuboidal-to-columnar, ciliated epithelium, surrounded by a circular muscle (50 µm thick). The penis papilla is cylindrical, with a rounded tip and is horizontally located (Figs 22A–C, 23B). This papilla is covered with a columnar epithelium, which is pierced by the necks of two types of gland cells producing erythrophil and cyanophil granules, respectively. This epithelium is underlain by a 15 µm thick muscle with interwoven circular and longitudinal fibres. The male atrium is relatively short and not folded. It is lined with a cuboidal-to-columnar epithelium, which is crossed by two types of gland cells producing erythrophil and cyanophil granules, respectively. This epithelium is underlain by a layer of circular muscle, followed by a layer of longitudinal fibres, each layer being 5 µm thick in the proximal region of the atrium and 30 µm in the distal. The ovaries are ovoid and have a maximum diameter of 400 µm in the longitudinal body axis. These ovaries are located immediately above the ventral nerve plate (Fig. 21C) and at a distance from the anterior tip of the body corresponding to 9.6% of the body length. The ovovitelline ducts emerge from the lateral aspect of the ovaries and run ventrally above the main nerve plate. Close to the mid-region of the prostatic vesicle, each ovovitelline duct opens laterally into the long, dilated common ovovitelline duct (Figs 22A, C, 23B–E). This long duct is six times wider than the ovovitelline ducts and exhibits longitudinal folds. The common ovovitelline duct ascends gradually to communicate with the common glandular ovovitelline duct. This glandular duct runs posteriorly over the male and female atria to join the female genital canal, which projects dorso-anteriorly from the dorsoposterior region of the female atrium. The female atrium is elongated to funnel-shaped and is slightly shorter than the male atrium. The common ovovitelline duct is lined with a columnar epithelium, which is crossed by three types of gland cells producing xanthophil, erythrophil and cyanophil granules, respectively (Fig. 23E). This duct is surrounded by a single muscle layer (50 µm thick) comprising circular, diagonal and longitudinal thin fibres. The female genital canal and the female atrium are lined with a 75 µm high columnar, non-ciliated epithelium and the apical side of its cells contains xanthophil granules. Additionally, gland cells discharge erythrophil granules through the epithelium of the female genital canal and that of the female atrium. These epithelia are underlain by a 50–120 µm thick muscle consisting of intermingled longitudinal and circular fibres. The common muscle coat consists of sparse longitudinal and circular muscle fibres. Remarks on the neae tribe Inakayaliini and its genus: The phylogenetic position of Inakayaliini is unstable. It was recovered as sister to Myoplanini (Fig. 2; Supporting Information, Figs S1, S 2) or to Adinoplanini (Supporting Information, Figs S3, S 4). Inakayaliini is monogeneric and currently houses four species. Two of them are represented in our phylogenetic trees, namely, I. ƲaldiƲiana and I. cyanea. These two species are sister to each other. Inakayalia cyanea matches all diagnostic features of the genus, except that the wall of its penis papilla is not irregular but smooth, and the dilated portion of the female ducts does not correspond to ovovitelline ducts, but the common ovovitelline duct. Therefore, the genus is re-diagnosed by omitting the mention of the irregular wall of the penis papilla and the dilation of the female genital ducts. This latter feature is transferred to the diagnosis of the new tribe. Inakayalia cyanaea is readily distinguished from the other three species in the genus in that it presents a long prostatic vesicle (vs. shorter) and a dilated common ovovitelline duct (vs. paired ovovitelline ducts dilated). Inakayalia cyanea is also the only species in the genus with the testes extending vertically between the supraintestinal and the subintestinal parenchymal muscle (see Fig. 21A)., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 865-867, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977

SARCOPLANA MUSCULOSA ALMEIDA & CARBAYO SP. NOV. (FIGS 40–44) Zoobank registration: urn: lsid: zoobank. org:act: 67BAB3EC-6FCC-4799-89E9-48133F0A40D1 Holotype: MNHNCL PLAT-15047 (Field code, F4886). Parque Nacional Nahuelbuta, Purén, Región de La Araucanía, Chile (37°49′′37.2′′S, 073°00′′32.4′′W), coll. F. Carbayo et al., 11 December 2010. Cephalic region: transverse sections on 11 slides; portion immediately behind the cephalic region: horizontal sections on six slides; pre-pharyngeal region: transverse sections on 17 slides; pharynx and copulatory apparatus: sagittal sections on four slides. Type locality: Parque Nacional Nahuelbuta, Región de La Araucanía, Chile. The species only known from this locality. Etymology: The specific epithet is from the Latin adjective musculosus, muscular, alluding to the thick cutaneous musculature. Description External aspect: At rest, the live specimen measured approximately 18 mm long and 3 mm wide (Fig. 40A, B). The body length may double when crawling (Fig. 40C). The body margins are parallel. The anterior tip is rounded, while the posterior is pointed. The dorsum is convex and the ventral side is flat. The preserved specimen measured 17.5 mm long, 2.5 mm wide and approximately 1.5 mm high. The dorsum displays a pure orange (RAL 2004) median stripe 21% of the body width, which is divided longitudinally by a thin carmine-red (RAL 3002) midline (2.3% of the body width; Fig. 40A, C). The median stripe is absent in both extremities of the body. External to the median stripe is a black-red (RAL 3007) band 37% of the body width, the margins of which are darker. Some pure orange spots occur in the bands. These bands converge in the cephalic region. The ventromarginal sensory border is a line with beige-grey colour (Fig. 40B). The body margins are pure orange. The ground colour of the ventral side is pure orange, provided with a pair of bands, each with 26% of body width, consisting of brown-red (RAL 3011) dots. The inner and outer margins of the bands are darker (Fig. 40B). The monolobate eyes measure 4 5–5 0 µm in diameter. They are distributed in an irregular row contouring the cephalic region and extending marginally to the posterior tip of the body. Sensory pits are absent. Instead, the ventromarginal epithelium of the cephalic region possesses sensory depressions. These depressions reach the underlying basal lamina and are provided with cilia (Fig. 41A). The sensory depressions are absent at the anterior tip of the body. The mouth is positioned at a distance from the anterior extremity of the body equal to 66% of the body length; the gonopore 77%. Internal morphology: The epidermis is ciliated only on the creeping sole, this occupying 83% of the body width. Gland cells producing erythrophil granules and cells producing rhabdites discharge through the entire epidermis. The erythrophil type is more abundant in the body margin, while the rhabditogen cells are more numerous in the ventral surface of the cephalic region (Fig. 41A). Gland cells producing weakly cyanophil granules also discharge through the ventral and marginal epidermis. A glandular margin is absent. The cutaneous musculature comprises three layers, namely, a subepidermal, layer (2 µm thick) of circular muscle, followed by a double layer (13 µm) with decussate fibres and an innermost layer of longitudinal muscle, the fibres of which are gathered into bundles (Fig. 41B–F). This latter layer is 70 µm thick dorsally and 160 µm ventrally. It is thinner than the body margins, where it remains conspicuous (Fig. 41B, F). The ventral layer of the longitudinal muscle is divided into a thin outer muscle and a thick inner muscle. These outer and inner ventral portions of the longitudinal muscle are separated by a secondary peripheral nerve net (Fig. 41C). The relative thickness of the cutaneous musculature is 19.5% of body height. The parenchymal musculature comprises four layers along the entire body: a dorsal layer (30 µm thick) of decussate fibres located to the inside of the peripheral nervous net; a dense layer of supraintestinal transverse muscle (40 µm); a dense layer of subintestinal transverse muscle (75 µm); and a subneural layer of decussate fibres (40 µm) (Fig. 41B, C, E, F). Additionally, abundant oblique muscle fibres run in transverse body planes along the body. The muscular organization changes in the anterior region of the body. At 1.9 mm from the anterior tip of the body, the longitudinal cutaneous muscle is 40 µm thick dorsally and 180 µm ventrally. In this region, the relative thickness of the cutaneous musculature is 21.6% of the body height (Fig. 41F). The cutaneous and parenchymal muscles are thinner at 1.35 mm from the body tip. At 0.6 mm, the inner ventral cutaneous longitudinal muscle concentrates medially, so that one-quarter of the body width at each side of the body lacks this muscle. In this region, a cephalic retractor muscle is flat lenticular in cross-sections (Fig. 42A). At 0.4 mm from the anterior tip, the secondary peripheral cutaneous nerve net is inconspicuous so that the ventral cutaneous muscle is no longer divided into an outer and an inner layer. Here, the longitudinal muscle is roughly lenticular in cross-section (Fig. 42B). Toward the anterior tip of the body, the retractor muscle becomes progressively smaller as its muscle fibres progressively detach from it to run obliquely to the dorsum and body margins (Fig. 42C, D). The mouth is located at a distance from the anterior region of the pharyngeal pouch, equivalent to 65% of the pouch length. The pharynx is cylindrical (Fig. 43A, B). The ventro-anterior portion of the pharynx was cut off for DNA extraction, and thus the presence of an oesophagus could not be ascertained. The outer pharyngeal musculature consists of a subepithelial layer (5 µm thick) of longitudinal muscle, followed by a layer (8 µm) of circular fibres. The inner pharyngeal musculature consists of a single subepithelial layer of circular muscle, with longitudinal fibres interspersed (40 µm). The stroma of the pharynx has circular and longitudinal fibres. The testes are pear-shaped and measure approximately 400 µm high. They are dorsally located beneath the transverse supraintestinal parenchymal muscle and between the intestinal branches (Fig. 41B). They are distributed in a row of one to two testes at each side of the body. The anteriormost testes are placed at a distance from the anterior tip of the body equivalent to approximately 35% of the body length, that is, 1.2 mm behind the ovaries; the posteriormost testes lie at a distance from the anterior tip equivalent to 44% of body length, that is, 100 µm anterior to the pharynx. The sperm ducts run above the subintestinal parenchymal muscle and more or less dorsally to the ovovitelline ducts. The distal portion of the sperm ducts bends dorsally to the sagittal plane to open into the proximal section of the respective branch of the prostatic vesicle (Fig. 43C). The prostatic vesicle is a sinuous tube roughly C-shaped in lateral view. Its proximal portion is bifurcate. This vesicle penetrates the anterior region of the penis bulb to join the ejaculatory duct. The penis bulb is well developed and is mainly constituted of longitudinal fibres. Most of the ejaculatory duct is sinuous and located within the penis bulb, while its distal section is straight and opens at the tip of the penis papilla. The penis papilla is 300 µm long and lies horizontally. This papilla is conical and presents some folds (Figs 43C, 44A). The prostatic vesicle is lined with a cuboidal, apparently non-ciliated epithelium. This epithelium is pierced by the necks of gland cells producing fine (0.5 µm) erythrophil granules and is surrounded by a circular muscle (10 µm thick). The ejaculatory duct is lined with a cuboidal ciliated epithelium. The basal-half of the penis papilla is lined with a columnar epithelium traversed by the necks of numerous openings of gland cells producing erythrophil granules. The distal-half of the papilla is lined with a cuboidal epithelium through which some gland cells of the same type discharge. The epithelium of the penis papilla is underlain by some longitudinal muscle fibres. The male atrium is narrow, elongated and roughly smooth (Fig. 43C). This atrium is lined with a cuboidal-to-columnar epithelium, the apical surface of which is erythrophil. Numerous gland cells discharge erythrophil granules through the atrial epithelium, which is underlain by a layer (10 µm thick) of circular muscle, followed by a layer (10 µm) of longitudinal fibres. The atrial wall dorsal to the gonoduct presents the openings of two different musculoglandular organs, one located behind another (Fig. 43C). The anterior organ (named mg 1 in the figures) consists of a 310 µm long and 30 µm wide, bowed and vertical blind duct embedded into the parenchyma and surrounded by muscle fibres. The duct of this musculoglandular organ is lined with a 10 µm high columnar epithelium, and the cells of this epithelium contain fine erythrophil granules (0.5 µm in diameter) produced by gland cells located outside the organ. The epithelium of the duct is underlain by a layer (10 µm thick) of circular muscle, followed by a layer (50 µm) of muscle fibres variously oriented, most circular. Beneath the epithelium of the innermost portion of the duct is a cyanophil, granular mass. The lumen of the canal contains some erythrophil granules. The posterior musculoglandular organ (named mg 2 in the figures) is ampulla-shaped. It consists of a 130 µm long duct leading to a deeper, enlarged portion with 120 µm in diameter (Figs 43B, C, 44A). The duct is lined with a cuboidal, strongly erythrophil epithelium. A 30 µm thick longitudinal muscle underlies this epithelium. The cells of the lining epithelium of the enlarged portion are not discernible. Abundant gland cells with their bodies outside the organ discharge fine cyanophil granules into the lumen of the enlarged portion of the organ. Surrounding this enlarged portion of the musculoglandular organ is a 30 µm thick muscle net, followed by a layer (30 µm thick) of longitudinal fibres. The ovaries are rounded-to-ovoid and approximately 100 µm in diameter. They are incompletely developed. These ovaries are located at a distance from the anterior tip of the body corresponding to 28% of the body length and 1.2 mm anterior to the anteriormost testes. The ovaries lie immediately above the ventral nerve plate. The ovovitelline ducts emerge laterally from the dorsal side of the ovaries. Subsequently, these ducts run posteriorly above the nervous plate and immediately underneath the transverse subintestinal parenchymal muscle (Fig. 41C). Just behind the level of the gonopore, one ovovitelline duct ascends gradually to enter the common ovovitelline duct behind the female atrium. This duct is short and oriented dorsally and communicates with the female genital canal. This female canal projects posteroventrally from the posterior wall of the female atrium (Fig. 43C). The suboptimal quality of the sections did not allow examination of the second ovovitelline duct nor the type of epithelium lining the common ovovitelline duct and the female genital canal. The female atrium is elongated and narrow. The dorsal wall of this atrium is more or less smooth, whereas the ventral wall is provided with three shallow recesses, each 100–200 µm in size (Fig. 44C, D). The female atrium is lined with a columnar, 35–45 µm high epithelium. The free surface of this epithelium is erythrophil and resembles the bristles of a brush. Gland cells producing fine erythrophil granules pierce this epithelium. The recesses are lined with a low epithelium. The female atrium contains clumps of xanthophil granules. The lining epithelium of the female atrium is surrounded by a 5 µm thick layer of longitudinal fibres, followed by a 10 µm thick layer of circular fibres. The male atrium to female atrium ratio is 84%. A common muscle coat wraps the distal-half of the prostatic vesicle and the male and female atria. This coat is comprised of abundant longitudinal muscle fibres. Remarks on the neae tribe Sarcoplanini and its genera: The molecular phylogenies retrieved Sarcoplanini as a monophyletic group comprising Mapuplana, Pichidamas, Sarcoplana and Wallmapuplana. The intergeneric relationships are unstable. The monotypic genus Liana can be included in this tribe based on the morphological similarity of L. guasa Froehlich, 1978 with Sarcoplanini members, as shown below. The species of Sarcoplanini share three unique characteristics among the Geoplaninae, namely, sensory depressions, a cephalic retractor muscle with a particular fibre organization (possibly secondarily lost in Wallmapuplana) and a subneural parenchymal decussate muscle (but the fibre orientation of this muscle is unknown in Wallmapuplana). These characteristics readily distinguish the Sarcoplanini members from the remaining Geoplaninae. Furthermore, branched glands associated with the prostatic vesicle are present in three of the four genera (Mapuplana, Pichidamas and Wallmapuplana). Additional traits shared by all species in Sarcoplanini, and which probably evolved convergently in other lineages of Geoplanidae, are marginal distribution of the eyes [also present in Adinoplanini, Myoplanini, Haranini, Caenoplanini (Rhynchodeminae) and some Geoplanini)], a small penis papilla (e.g. Amaga, Gusana, but it is large in Liana), a copulatory apparatus provided with musculoglandular organs [possibly secondarily lost in Mapuplana; also present in Australasian taxa, such as some Bipalium, Artioposthia Graff, 1896 (see: Fyfe, 1947), Coleocephalus Fyfe, 1953 (see: Winsor, 1998)] and a female genital canal with the postflex condition (i.e. the canal approaching the female atrium from behind as in Pasipha, Gigantea, and Gusana, among others). The genera of Sarcoplanini differ from each other by several structures. Sarcoplana stands apart from the remaining Sarcoplanini genera from the presence of a secondary peripheral nerve net located in the ventral side of the body (convergent in Myoplana). Mapuplana and Liana are the only genera of Sarcoplanini having part of the ventral longitudinal cutaneous muscle sunken into the parenchyma. These two genera differ in that the penis papilla is small in Mapuplana (vs. large in Liana). Wallmapuplana is the only genus of Sarcoplanini lacking a cephalic retractor muscle, while Pichidamas is the only genus having a large musculoglandular organ of adenodactyl type. The genus Liana deserves a detailed discussion. This monotypic genus was proposed for L. guasa (Froehlich, 1978). The species was described from incompletely mature individuals. The main diagnostic features of the genus are: elongated body, broad creeping sole, sensory depressions (‘minute sensory pits’ in the original description), longitudinal ventral cutaneous muscle partially sunken into the parenchyma, cutaneous muscle thickness relative to the body height is 10%, testes are dorsal; copulatory apparatus without adenodactyls; penis papilla short and blunt; female canal approaches from horizontal or ventral aspect (Froehlich, 1978). This species also has a subneural layer of decussate parenchymal muscle (‘a layer of fibres obliquely oriented to the right and to the left’ in Froehlich, 1978: 21) interwoven with fibres of the sunken longitudinal cutaneous muscle. The relative thickness of the cutaneous musculature increases to 21% when the sunken muscle portion is also considered (see: Froehlich, 1978: fig. 24). There are no gene sequences available of this species. Among the Geoplanini tribes, Liana fits well Sarcoplanini:the creeping sole is wide; the eyes marginal; sensory depressions and subneural parenchymal decussate muscle are present. The original description of L. guasa does not mention a cephalic retractor muscle but a modification of the musculature organization in the cephalic region, which is compatible with a retractor organ (‘ At the anterior end, the dorsal longitudinal [Ʋentrally?] cutaneous fibers bend dorsally to end on the basement membrane. Laterally, toaeards the Ʋentral sensory border the cutaneous musculature progressiƲely loses height becoming minimal if not absent. Ventrally it regains height toaeards the median line, attaining a little more than half the height of the dorsal portion. […] The Ʋentral longitudinal parenchymal [cutaneous?] musculature progressiƲely disappears toaeards the anterior extremity. At the same time it appears there a layer of diagonal fibers interspersed aeith rarer and rarer longitudinal fibers. Presumably the longitudinal fibers change direction anteriorly but it cannot be discerned’, E. M. Froehlich, 1978, p. 22). The presence of two taxonomically relevant diagnostic features of Sarcoplanini, namely, branched glands associated with the prostatic vesicle and genital musculoglandular organs, could not be verified since the individuals are only partially mature. Liana does not fit in any of the remaining tribes because it lacks the following features: carinate dorsal side, musculoglandular organs in the female atrium (Adinoplanini); body leaf-like with dorsal eyes (Geoplanini); anterior region of the body triangular, ventral and dorsal longitudinal cutaneous muscle sunken into the parenchyma, sensory pits (Gusanini); long pharyngeal pouch (Haranini and Timymini); dilated female genital ducts (Inakayaliini); transneural parenchymal muscle of diagonal fibres (Myoplanini); extraordinarily wide and flattened body, marginal eyes, subneural parenchymal of transverse muscle and a transneural parenchymal layer of longitudinal muscle (Polycladini); semi-lunate head plate (Timymini). Therefore, we place the genus Liana in Sarcoplanini., Published as part of Almeida, Ana Laura, Álvarez-Presas, Marta & Carbayo, Fernando, 2023, The discovery of new Chilean taxa revolutionizes the systematics of Geoplaninae Neotropical land planarians (Platyhelminthes: Tricladida), pp. 837-898 in Zoological Journal of the Linnean Society 197 (4) on pages 885-892, DOI: 10.1093/zoolinnean/zlac072, http://zenodo.org/record/7813977, {"references":["Froehlich EM. 1978. On a collection of Chilean land planarians. Boletins de Zoologia da UniVersidade de Sao Paulo 3: 7 - 80.","Graff LV. 1896. Uber das System und die geographische Verbreitung der Landplanarien. Verhandlungen der Deutsche Zoologischen Gesellschaft 6: 75 - 93.","Fyfe M. 1947. The classification and reproductive organs of New Zealand land planarians, Part III. Artioposthia mariae (Dendy), A. australis (Dendy), Geoplana iris Dendy, and A. garVeyi (Dendy). Transactions of the Royal Society of Neae Zealand 76: 517 - 523."]}