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Background: Liver flukes from the genus Fasciola are the causative agents for human and livestock fascioliasis. Accurate identification of Fasciola spp. is essential to understanding the epidemiology of fascioliasis. This study aimed to determine the morphometric and molecular characterization of Fasciola spp. in livestock from Northwestern provinces of Iran. Methods: Five hundred adult Fasciola flukes were obtained from different definitive hosts (cattle, sheep, goats, and buffaloes) in four local abattoirs in the northwestern provinces of Iran (West‐Azerbaijan, East‐Azerbaijan, Ardabil, and Zanjan) from September 2021 to August 2022. All samples were examined by morphometric criteria; then, 49 samples were identified using PCR‐RFLP based on ITS1 region and 23 sequence of isolates analyzed by cox1 marker. PCR‐RFLP methods compared morphometric results, and cox1 gene sequences were used to confirm PCR‐RFLP results and phylogenetic analysis. Results: The differences between the body length, body width, cephalic cone length, cephalic cone width, body area, and distance between the ventral sucker and posterior end of the body in two species were significant (p < 0.05). Based on the morphometric criteria, 139 samples (27.8%) were identified as Fasciola gigantica and 361 (72.2%) as Fasciola hepatica. Similarly, PCR‐RFLP analysis of ITS1 region confirmed morphometric results. No hybrid forms of Fasciola were detected. Partial sequences of cox1 showed 13 variable sites with eight haplotypes in F. hepatica and 12 variable sites with five haplotypes in F. gigantica. Conclusion: Based on the results of this study, the PCR‐RFLP method can be used to confirm the morphological method of Fasciola species, but it is insufficient to study their genetic diversity. Also, sequences of cox1 results of the present study showed that F. hepatica and F. gigantica species in the Northern provinces of Iran have different genetic structures and haplotypes. [ABSTRACT FROM AUTHOR]

Clonorchis sinensis is an important zoonotic parasite that is mainly prevalent in China, Korea, Vietnam and the Russian Far East. To explore the genetic variation and population structure of C. sinensis, an in silico analysis was conducted based on mitochondrial cytochrome c oxidase subunit 1 (COX1), ribosomal internal transcribed spacer 1 (ITS1) and ribosomal internal transcribed spacer 2 (ITS2) sequences. The sequences obtained from NCBI were truncated for further analyses, including haplotype network, phylogenetic, gene flow, diversity and neutrality analyses. The results showed that there were 20, 11 and 4 haplotypes for COX1, ITS1 and ITS2, respectively. The results of both the haplotype network and phylogenetic analyses indicated that the haplotypes for each type of sequence from the same country were not all clustered together. Haplotype diversity values were all lower than 0.5. Values of nucleotide diversity were higher than 0.005, except for ITS2. Tajima's D and Fu's Fs values were all negative, and p-values showed significant differences, indicating that the population of C. sinensis is growing. Fst values were all lower than 0.05. In conclusion, this study found that there are specific variations of C. sinensis in different countries, and the population of this parasite is growing with less genetic variation. The findings provide a crucial foundation for understanding the molecular epidemiology and population dynamics of C. sinensis. [ABSTRACT FROM AUTHOR]

Background: Scabies, an infestation of the mite Sarcoptes scabiei, has seen an increase in clinical diagnoses in the Netherlands since 2011. This study aimed to analyse PCR-positive S. scabiei skin samples through partial genome sequencing and to link findings to patient epidemiological characteristics. Methods: Skin samples were collected from individuals in the Netherlands between January 2016 and January 2023. On the PCR-positive S. scabiei skin samples, partial mitochondrial cytochrome c oxidase subunit 1 gene (cox1) sequencing was performed to assess genetic variability. Epidemiological information was collected through interviews. We examined associations between cox1 subtypes, epidemiological factors and treatment outcomes. Results: Sequencing results were obtained from 128 patients, with epidemiological information available for 55 (43%) of these patients. Fifteen distinct cox1 subtypes were identified. Subtype 01 was most prevalent (45%) and present across all age groups and social settings. The remaining subtypes were less common and not consistently found in all contexts. Five clusters were identified, each with identical cox1 subtypes. Comparative analysis with GenBank sequences revealed genetic similarities with strains from Australia, the USA and China, suggesting the global distribution and transmission of specific subtypes. A substantial proportion (73%) of patients with scabies required multiple treatments to eradicate the infestation, with no subtype-related differences. Conclusions: This is the first study linking S. scabiei sequencing results to patient epidemiological data. Several subtypes clustered in specific geographic regions and social contexts, underscoring localised transmission patterns. Further research with larger sample sizes is needed to enhance our understanding of the transmission of this mite. This study provides valuable insights that will strengthen scabies control efforts. [ABSTRACT FROM AUTHOR]

The trematode family Cyathocotylidae infects various hosts worldwide, including birds, mammals, reptiles, and fish. However, the lack of molecular data from adult worms hinders phylogenetic, epidemiological, and host association studies. This study aims to identify the common cyathocotylid trematodes infecting African catfish in Egypt using morphological and molecular evidence. Out of 142 Clarias gariepinus, 123 fish (86.6%) harbored cyathocotylid metacercariae, with a mean metacercarial intensity of 201 ± 38.5/g. Cyathocotylid metacercariae prevalence gradually rose as host size increased. Although there was no significant difference between groups, larger fish had a higher mean metacercarial intensity. The prevalence and intensity were unrelated to the fish gender. Histopathological examination of metacercariae-infected catfish revealed varying degrees of degenerative changes, including intermuscular edema leading to muscle fiber dispersion and atrophy, involving 11% to over 81% of muscle sections. We identified three cyathocotylid metacercariae and eight cyathocotylid adult species from experimental infection using morphometric and molecular data, including internal transcribed spacer (ITS) and/or mitochondrial cytochrome c oxidase subunit 1 (cox1) sequences. We determined the phylogenetic position of these cyathocotylid samples. The ITS sequence analysis linked the isolated Cyathocotylidae sp. 1 and 2 metacercariae to Prohemistomum vivax adults. Mesostephanus appendiculatoides and Paracoenogonimus ovatus were reported for the first time in Egypt. These findings may provide valuable genetic data for future molecular epidemiological and phylogenetic studies of cyathocotylid trematodes. [ABSTRACT FROM AUTHOR]

Abstract Background Scabies, an infestation of the mite Sarcoptes scabiei, has seen an increase in clinical diagnoses in the Netherlands since 2011. This study aimed to analyse PCR-positive S. scabiei skin samples through partial genome sequencing and to link findings to patient epidemiological characteristics. Methods Skin samples were collected from individuals in the Netherlands between January 2016 and January 2023. On the PCR-positive S. scabiei skin samples, partial mitochondrial cytochrome c oxidase subunit 1 gene (cox1) sequencing was performed to assess genetic variability. Epidemiological information was collected through interviews. We examined associations between cox1 subtypes, epidemiological factors and treatment outcomes. Results Sequencing results were obtained from 128 patients, with epidemiological information available for 55 (43%) of these patients. Fifteen distinct cox1 subtypes were identified. Subtype 01 was most prevalent (45%) and present across all age groups and social settings. The remaining subtypes were less common and not consistently found in all contexts. Five clusters were identified, each with identical cox1 subtypes. Comparative analysis with GenBank sequences revealed genetic similarities with strains from Australia, the USA and China, suggesting the global distribution and transmission of specific subtypes. A substantial proportion (73%) of patients with scabies required multiple treatments to eradicate the infestation, with no subtype-related differences. Conclusions This is the first study linking S. scabiei sequencing results to patient epidemiological data. Several subtypes clustered in specific geographic regions and social contexts, underscoring localised transmission patterns. Further research with larger sample sizes is needed to enhance our understanding of the transmission of this mite. This study provides valuable insights that will strengthen scabies control efforts. Graphical Abstract

There have been report of possible food poisoning in Lithuania following the consumption of raw steaks from a roe deer (Capreolus capreolus). Two people experienced symptoms of abdominal pain, watery diarrhea, loss of appetite, and nausea. The parasitological examination of the roe deer revealed macrocysts of Sarcocystis sp. to the naked eye. Microscopic sarcocysts of Sarcocystis spp. were detected in different muscles, and a high sarcocyst load was observed in the roe deer muscles by a light microscope. Based on molecular results, S. capreolicanis, S. gracilis, S. linearis, S. oviformis, and S. silva were identified in meat samples of the roe deer. This is the first report of a possible link between venison poisoning and Sarcocystis spp. infection in Lithuania. [ABSTRACT FROM AUTHOR]

Sarcocystis species are prevalent in Iran and have economic and public health consequences on ani-mals and humans. The present study aimed at molecular identification of Sarcocystis spp. isolated from Iranian buffaloes in Guilan province based on 18s ribosomal RNA (18s rRNA) and cytochrome oxidase subunit I (Cox1) sequences. For this purpose, a total of 100 buffalo carcasses were sampled from four tissues (oesophagus, diaphragm, shoulder, thigh), and assessed microscopically for sarco-cysts. Samples were suspected of being infected with Sarcocystis spp. according to morphological evaluation. Amplification of 18s rRNA and Cox1 sequences were performed from DNA extracted from any detected Sarcocystis. The phylogenetic tree was plotted by Neighbor-Jinning (NJ) phylo-grams. Phylogenetic analysis of 18s rRNA and Cox1 genes showed no differences between Sarcocys-tis isolates of the same species and a low level of genetic variability was found among isolates and other Sarcocystis cruzi sequences. S. cruzi was identified as the species of Iranian buffalo in the Guilan province of Iran. This is the first study of the Cox1 gene and 18s rRNA sequences to confirm the presence of S. cruzi in Iranian buffaloes.

Abstract Background Southeast Asia is regarded as a hotspot for the diversity of ixodid ticks. In this geographical region, Vietnam extends through both temperate and tropical climate zones and therefore has a broad range of tick habitats. However, molecular-phylogenetic studies on ixodid tick species have not been reported from this country. Methods In this study, 1788 ixodid ticks were collected from cattle, buffalos and a dog at 10 locations in three provinces of northern Vietnam. Tick species were identified morphologically, and representative specimens were molecularly analyzed based on the cytochrome c oxidase subunit I (cox1) and 16S rRNA genes. Fifty-nine tick species that are indigenous in Vietnam were also reviewed in the context of their typical hosts in the region. Results Most ticks removed from cattle and buffalos were identified as Rhipicephalus microplus, including all developmental stages. Larvae and nymphs were found between January and July but adults until December. Further species identified from cattle were Rhipicephalus linnaei, Rhipicephalus haemaphysaloides, Amblyomma integrum and Haemaphysalis cornigera. Interestingly, the latter three species were represented only by adults, collected in one province: Son La. The dog was infested with nymphs and adults of R. linnaei in July. Phylogenetically, R. microplus from Vietnam belonged to clade A of this species, and R. haemaphysaloides clustered separately from ticks identified under this name in China, Taiwan and Pakistan. Amblyomma integrum from Vietnam belonged to the phylogenetic group of haplotypes of an Amblyomma sp. reported from Myanmar. The separate clustering of H. cornigera from Haemaphysalis shimoga received moderate support. Conclusions Three tick species (R. linnaei, A. integrum and H. cornigera) are reported here for the first time in Vietnam, thus increasing the number of indigenous tick species to 62. Clade A of R. microplus and at least R. linnaei from the group of Rhipicephalus sanguineus sensu lato occur in the country. There is multiple phylogenetic evidence that different species might exist among the ticks that are reported under the name R. haemaphysaloides in South and East Asia. This is the first report of A. integrum in Southeastern Asia. Graphical Abstract

Bertiella spp. is a mite-borne cestode parasite that inhabits the small intestine of wide range of mammals, including non-human primates. In the present study, the morphological and molecular analysis of Bertiella studeri recovered from the small intestine of a bonnet macaque (Macaca radiata) from Wayanad, Kerala (South India) was performed. Acetic alum carmine staining identified the cestode morphologically based on the characters like broader proglottids, which contain irregularly alternating genital pores, single set of reproductive organs, 280 testes and a tubular transverse uterus. Molecular characterization was done using 18SrRNA, ITS1-5.8S and COX1 genes. Phylogenetic trees were constructed using MEGA X based on the Maximum Likelihood (ML) method (Hasegawa-Kishino-Yano (HKY) model). Cytochrome oxidase I gene could detect the existence of genetic variation in the parasite from two different hosts viz., monkey (Kerala, Argentina, and Kenya) and human (Sri Lanka). A minimum spanning network of haplotypes was generated by the haplotype networking with the above sequences using the popARTv1.7. Haplotype analysis based on COX1 revealed that the parasite haplotype was different in each country with highest population frequency in Sri Lanka.

Epiphytic algae are important components of marine ecosystems. Despite the importance of epiphytic algae in marine ecosystems, our knowledge of these taxa is still limited. The marine red algal genus Reinboldiella, which is distributed mainly in the northwestern Pacific region, is characterized by tiny thalli that are epiphytic on various other red algae. Six species of the genus have been reported in Japan, Taiwan, and Korea. In this study, 50 Korean Reinboldiella specimens, previously known as Reinboldiella schmitziana, were collected from 11 localities and reexamined to verify their taxo-nomic status. We describe a new species, Reinboldiella koreana sp. nov., based on rbcL phylogeny and morphological observations. A total of four grateloupiacean species (Pachymeniopsis lanceolata, P. elliptica, Grateloupia turuturu, and G. asiatica) were identified as host species of R. koreana sp. nov. Due to the restricted host range of R. koreana sp. nov., a phylogenetic tree was reconstructed using the cox1 marker to confirm the co-phylogenetic relationships between the epiphytic algae and their grateloupiacean hosts. Two of the four cox1 haplotypes of R. koreana were found in different host species, regardless of which haplotype they were. It indicates that there are no co-phylogenetic relationships between R. koreana and host species. [ABSTRACT FROM AUTHOR]

• Low-level laser light suppresses oxidative stress and enhances antioxidant defenses. • Sargassum latifolium showed antiatherosclerotic effects in combination with low-level laser irradiation in rats. • The synergistic effect of each algal dry biomass concentration with low-level laser irradiation on rats. • Sargassum latifolium showed 74.8 % of FRAB scavenging power and, 76.08 % Cox1 inhibition. Atherosclerosis is considered the most common underlying cause of most cardiovascular diseases. Most conventional therapies focus on lipid levels but fail to reduce oxidative stress and inflammation. In contrast, seaweeds can solve such problems, with their antihypercholesterolemic properties. The current study aims to evaluate the antiatherosclerotic effects of the brown alga Sargassum latifolium and low-level laser irradiation in rats fed on a hypercholesterolemic diet, individually or in combination. Two doses of S. latifolium dry biomass (5 and 10 %) were orally gavaged to rats with and without low-level laser irradiation (650 nm laser 100 mW) compared with Atorvastatine Ca (Lipitor). The alga phytochemical contents were estimated with (GC–MS) analysis, evaluating its antioxidant, anti-inflammatory, and antihypercholesterolemic activities. S. latifolium showed 74.8 % of FRAP scavenging power and 76.08 % Cox1 inhibition. In addition to suppressing weight gain and blood viscosity in hypercholesterolemic rats, the algal laser treatment demonstrated a significant improvement in serum HDL-cholesterol and a reduction in serum total lipids, triglycerides, total cholesterol, LDL-cholesterol, and VLDL-cholesterol concentrations. Additionally, there was a significant improvement in serum HDL-cholesterol and a significant reduction in elevated atherogenic index, risk factor, inflammatory marker (CRP), creatine kinase, lactate dehydrogenase activities, and serum liver and kidney function biomarkers. The best results were achieved by treatment of hypercholesterolemic rats with a high dose of S. latifolium with laser irradiation. In conclusion, the synergistic effect of S. latifolium seaweed (especially high dose) and low-level laser has great potential, which exceeds that of anticholesterol drugs for preventing and/or therapy of hypercholesterolemia and maintaining overall health. Thus, this synergistic effect is a potential source of a new therapy that provides antihypercholesterolemic and antiatherosclerotic effects similar to, or better than, those of more traditional pharmaceutical drugs. [ABSTRACT FROM AUTHOR]

Sarcocystis species are prevalent in Iran and have economic and public health consequences on animals and humans. The present study aimed at molecular identification of Sarcocystis spp. isolated from Iranian buffaloes in Guilan province based on 18s ribosomal RNA (18s rRNA) and cytochrome oxidase subunit I (Cox1) sequences. For this purpose, a total of 100 buffalo carcasses were sampled from four tissues (oesophagus, diaphragm, shoulder, thigh), and assessed microscopically for sarcocysts. Samples were suspected of being infected with Sarcocystis spp. according to morphological evaluation. Amplification of 18s rRNA and Cox1 sequences were performed from DNA extracted from any detected Sarcocystis. The phylogenetic tree was plotted by Neighbor-Jinning (NJ) phylograms. Phylogenetic analysis of 18s rRNA and Cox1 genes showed no differences between Sarcocystis isolates of the same species and a low level of genetic variability was found among isolates and other Sarcocystis cruzi sequences. S. cruzi was identified as the species of Iranian buffalo in the Guilan province of Iran. This is the first study of the Cox1 gene and 18s rRNA sequences to confirm the presence of S. cruzi in Iranian buffaloes. [ABSTRACT FROM AUTHOR]

The phylum Nematoda remains very poorly sampled for mtDNA, with a strong bias toward parasitic, economically important or model species of the Chromadoria lineage. Most chromadorian mitogenomes share a specific order of genes encoded on one mtDNA strand. However, the few sequenced representatives of the Dorylaimia lineage exhibit a variable order of mtDNA genes encoded on both strands. While the ancestral arrangement of nematode mitogenome remains undefined, no evidence has been reported for Enoplia, the phylum's third early divergent major lineage. We describe the first mitogenome of an enoplian nematode, Campydora demonstrans, and contend that the complete 37‐gene repertoire and both‐strand gene encoding are ancestral states preserved in Enoplia and Dorylaimia versus the derived mitogenome arrangement in some Chromadoria. The C. demonstrans mitogenome is 17,018 bp in size and contains a noncoding perfect inverted repeat with 2013 bp‐long arms, subdividing the mitogenome into two coding regions. This mtDNA arrangement is very rare among animals and instead resembles that of chloroplast genomes in land plants. Our report broadens mtDNA taxonomic sampling of the phylum Nematoda and adds support to the applicability of cox1 gene as a phylogenetic marker for establishing nematode relationships within higher taxa. Research Highlights: Genes are encoded on both strands in the mitochondrial genome of enoplid nematode Campydora demonstrans.The coding portion of C. demonstrans mitochondrial genome is subdivided in two regions by two arms of a long noncoding inverted repeat.cox1 gene can serve as a suitable phylogenetic marker for resolving within‐higher‐taxon relationships among nematodes. [ABSTRACT FROM AUTHOR]

Purpose: Human ophthalmomyiasis is a rare ocular parasitosis that results from the accidental infestation of dipteran larvae of several species, including Oestrus ovis (Linnaeus, 1758). This study aims to present the fourth documented human case of ophthalmomyiasis in Mexico, identifying the etiological agent through morphological and molecular analyses. Additionally, we investigated the phylogenetic position and genetic distances among different specimens globally characterized based on mitochondrial Cox1 sequences. Methods: A total of five larval specimens were extracted from the patient's eye, with two specimens allocated for identification based on morphological features using a stereomicroscope, and the remaining three preserved in absolute ethanol, one of them used for subsequent analysis using molecular methods. The mitochondrial Cox1 region was amplified and sequenced using automated Sanger sequencing. The resulting sequence was deposited in GenBank under accession number OR440699 and subjected to BlastN analysis against 35 other Cox1 sequences of O. ovis from GenBank. The identity and phylogenetic position of the strains were further explored using parsimony and maximum likelihood phylogenetic methods. Results: Morphological examination of the larval specimens extracted from the patient's eye unequivocally identified them as O. ovis species. BlastN analysis and comprehensive phylogenetic investigations involving a total of 36 Cox1 sequences confirmed the taxonomic identity of the larvae. Notably, our sequence was positioned within the cluster formed by the Brazilian and two Iranian samples. This finding underscores a shared genetic ancestry among these distinct geographical isolates and provides valuable insights into the evolutionary relationships within O. ovis populations. Conclusion: The presence of O. ovis infestation in Mexico City suggests potential shifts in environmental conditions favoring fly proliferation, highlighting the need for vigilance in urban healthcare settings. [ABSTRACT FROM AUTHOR]

[Display omitted] • Previous population genetic analysis of Opisthorchis viverrini eggs revealed two populations. • Cat- and human-specific populations use different intermediate fish hosts. • There are distinct morphologies in egg, metacercaria, and adult worm stages of O. viverrini. • Our study confirms distinct human- and cat-specific O. viverrini populations. Infection by the zoonotic fish-borne trematode, Opisthorchis viverrini, remains a crucial health issue in Thailand and neighboring countries. Recently, molecular analysis revealed two populations of putative O. viverrini : one found primarily in human hosts ("human-specific" population) and the other primarily in cats ("cat-specific" population). It is unclear how the infective stages (metacercariae) of these different populations circulate among definitive and reservoir hosts in nature. To gain an insight into this, mitochondrial cox 1 and nad 1 gene sequences of metacercariae from fish intermediate hosts were examined. None of 192 metacercariae from cyprinid fish in Lao PDR and Thailand had sequences typical of "cat-specific" O. viverrini , suggesting that cyprinid fish are not the main second intermediate hosts of this population. Interestingly, all 20 O. viverrini -like metacercariae from snakehead fish (Channa striata) shared 99.51–100% sequence identity with eggs from cats naturally infected in a previous study. Hence, we propose a modification of the known transmission dynamics of O. viverrini : consumption of metacercariae within snakehead fish provides another pathway for cats and (occasionally) humans to acquire infection. We also performed morphological comparisons of eggs, metacercariae, and adult flukes (raised in hamsters) of both Opisthorchis populations. The "cat-specific" population has eggs that are narrower and adults that are shorter and wider than in the human-specific population. The metacercaria of the "cat-specific" population is elliptical, while that of the "human-specific" population is oval, occasionally rounded. Our results confirmed that O. viverrini -like metacercariae from snakehead fish are the infective stages of the "cat-specific" fluke. This provides a new insight into the dissemination and transmission of each population in the second intermediate host. The identity of the cat-specific population is discussed. [ABSTRACT FROM AUTHOR]

Eight Eimeria spp. (Apicomplexa: Eimeriidae) have been isolated from the ring-necked pheasant (Phasianus colchicus Linnaeus), native to the temperate zone of Asia and eastern regions of Europe. Enteric coccidiosis has become a major issue associated with the breeding of farmed pheasants for game bird release or meat production. In this study, 35 fecal samples were collected from two-to-three-month-old ring-necked pheasants from four pheasant-rearing farms in Ehime Prefecture, Japan. Microscopic examination using a saturated sugar solution technique detected numerous subspherical oocysts from the samples of one farm and ellipsoidal Eimeria phasiani Tyzzer, 1929 oocysts from the three other farms. The subspherical oocysts were artificially sporulated and measured 18.6 µm by 15.7 µm with a 1.18 shape index (n = 150). Each oocyst contained four 10.7 µm × 5.8 µm sporocysts (n = 30) and one coarse refractile polar granule; no micropyle or residua were detected. Each sporocysts contained two sporozoites with one large and one small refractile body and sparsely distributed residua. The complete, 1,443-bp cytochrome c oxidase subunit I gene (cox1) of this isolate exhibited low sequence identity with published Eimeria spp. sequences including E. phasiani that was previously recorded in the same area. Meanwhile, the oocyst morphology most closely resembled that of Eimeria tetartooimia Wacha, 1973, but with distinct refractile polar granules and sporocyst residua. The available GenBank cox1 sequence of E. tetartooimia exhibited a sequence identity of < 94.5% with the study isolate. Here, the coccidian isolate identified in this study represents a new Eimeria iyoensisn. sp. capable of infecting ring-necked pheasant. [ABSTRACT FROM AUTHOR]

Background: Southeast Asia is regarded as a hotspot for the diversity of ixodid ticks. In this geographical region, Vietnam extends through both temperate and tropical climate zones and therefore has a broad range of tick habitats. However, molecular-phylogenetic studies on ixodid tick species have not been reported from this country. Methods: In this study, 1788 ixodid ticks were collected from cattle, buffalos and a dog at 10 locations in three provinces of northern Vietnam. Tick species were identified morphologically, and representative specimens were molecularly analyzed based on the cytochrome c oxidase subunit I (cox1) and 16S rRNA genes. Fifty-nine tick species that are indigenous in Vietnam were also reviewed in the context of their typical hosts in the region. Results: Most ticks removed from cattle and buffalos were identified as Rhipicephalus microplus, including all developmental stages. Larvae and nymphs were found between January and July but adults until December. Further species identified from cattle were Rhipicephalus linnaei, Rhipicephalus haemaphysaloides, Amblyomma integrum and Haemaphysalis cornigera. Interestingly, the latter three species were represented only by adults, collected in one province: Son La. The dog was infested with nymphs and adults of R. linnaei in July. Phylogenetically, R. microplus from Vietnam belonged to clade A of this species, and R. haemaphysaloides clustered separately from ticks identified under this name in China, Taiwan and Pakistan. Amblyomma integrum from Vietnam belonged to the phylogenetic group of haplotypes of an Amblyomma sp. reported from Myanmar. The separate clustering of H. cornigera from Haemaphysalis shimoga received moderate support. Conclusions: Three tick species (R. linnaei, A. integrum and H. cornigera) are reported here for the first time in Vietnam, thus increasing the number of indigenous tick species to 62. Clade A of R. microplus and at least R. linnaei from the group of Rhipicephalus sanguineus sensu lato occur in the country. There is multiple phylogenetic evidence that different species might exist among the ticks that are reported under the name R. haemaphysaloides in South and East Asia. This is the first report of A. integrum in Southeastern Asia. [ABSTRACT FROM AUTHOR]

Background: A molecular approach for the identification of unknown species by the using mitochondrial cox1 gene is an effective and reliable as compared with morphological-based identification. Hirudinaria manillensis referred to as Asian Buffalo Leech, is found in South Asia and traditionally used as medicine owing to its medicinal properties. Methods and results: The study aimed to isolate and identify the leech species using cox1 gene sequencing and their phylogenetic relationships. The nucleotide sequences of cytochrome c oxidase subunit I (cox1) mitochondrial genes were analyzed for species identification and the phylogenetic relationship of crucial therapeutic leech Hirudinaria manillensis. The isolated DNA from the leech sample was amplified with cox1 gene-specific primers. BLAST results with the H. manillensis sequence showed 89.24% homology with H. manillensis and phylogenetic tree analysis revealed the genetic relationship with other GenBank submitted sequences. Conclusion: The present study concluded that the cox1 gene could be an effective way to identify the leech H. manillensis and provided sufficient phylogenetic information to distinguish H. manillensis indicating a significant mtDNA-based approach to species identification. [ABSTRACT FROM AUTHOR]

Simple Summary: This research analyzed for the first time the diversity in the Czech population of the Western Honey Bee. It used two mitochondrial DNA markers to identify haplotypes. Sequencing revealed both common and novel haplotypes. The cox1 sequences showed more diversity than the tRNAleu-cox2 sequences. Most haplotypes belong to the C lineage. Some haplotypes showed links to the bees of African origin. No indigenous M lineage haplotype was found. Thus, the native population of the Dark Honey Bee, Apis mellifera mellifera, has been completely replaced by the C lineage. The haplotype networks and phylogenetic trees showed that some A and C lineage haplotypes were distinct. The haplotype diversity data indicate that there is ongoing population expansion and introgression. The Czech honeybee population is diverse, with different haplotypes in both tRNAleu-cox2 and cox1 sequences. The study aimed to analyze the genetic diversity in the Czech population of Apis mellifera using mitochondrial DNA markers, tRNAleu-cox2 intergenic region and cox1 gene. A total of 308 samples of bees were collected from the entire Czech Republic (from colonies and flowers in 13 different regions). Following sequencing, several polymorphisms and haplotypes were identified. Analysis of tRNAleu-cox2 sequences revealed three DraI haplotypes (C, A1, and A4). The tRNAleu-cox2 region yielded 10 C lineage haplotypes, one of which is a newly described variant. Three A lineage haplotypes were identified, two of which were novel. A similar analysis of cox1 sequences yielded 16 distinct haplotypes (7 new) within the population. The most prevalent tRNAleu-cox2 haplotype identified was C1a, followed by C2a, C2c, C2l, and C2d. For the cox1 locus, the most frequent haplotypes were HpB02, HpB01, HpB03, and HpB04. The haplotype and nucleotide diversity indices were high in both loci, in tRNAleu-cox2 with values of 0.682 and 0.00172, respectively, and in cox1 0.789 and 0.00203, respectively. The Tajima's D values were negative and lower in tRNAleu-cox2 than in cox1. The most frequent haplotypes were uniformly distributed across all regions of the Czech Republic. No haplotype of the indigenous M lineage was identified. High diversity and the occurrence of rare haplotypes indicate population expansion and continuous import of tribal material of the C lineage. [ABSTRACT FROM AUTHOR]

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Identifying marine trematode parasites in host tissue can be complicated when there is limited morphological differentiation between species infecting the same host species. This poses a challenge for regular surveys of the parasite communities in species of socio-economic and ecological importance. Our study focused on identifying digenean trematode species infecting the marine bivalve Cerastoderma edule across Europe by comparing morphological and molecular species identification methods. Cockles were sampled from ten locations to observe the trematode parasites under a stereomicroscope (morphological identification) and to isolate individuals for phylogenetic analyses using two gene markers, the small sub-unit ribosomal (18S) RNA gene (SSU rDNA) and the mitochondrial cytochrome c oxidase subunit 1 (cox1). For the first time, we compared both morphological identification and phylogenetic analyses for each of the 13 originally identified species. First, we identified a group of five species for which morphological identification matched molecular results (Bucephalus minimus, Monorchis parvus, Renicola parvicaudatus, Psilostomum brevicolle, Himasthla interrupta). Second, we identified a group of six species for which molecular results revealed either misidentifications or cryptic diversity (Gymnophallus choledochus, Diphterostomum brusinae, Curtuteria arguinae, Himasthla quissetensis, H. elongata, H. continua). Third, our analyses showed that all sequences of two expected species, Gymnophallus minutus and G. fossarum, matched between the two, strongly suggesting that only G. minutus is present in the studied area. Our study clearly demonstrates that molecular tools are necessary to validate the trematode species composition. However, with 17 distinct genetic lineages detected, some of which are not fully identified, future studies are needed to clarify the identity and status (regular vs. accidental infection) of some of these cryptic trematode species.

ABSTRACT Background Liver flukes from the genus Fasciola are the causative agents for human and livestock fascioliasis. Accurate identification of Fasciola spp. is essential to understanding the epidemiology of fascioliasis. This study aimed to determine the morphometric and molecular characterization of Fasciola spp. in livestock from Northwestern provinces of Iran. Methods Five hundred adult Fasciola flukes were obtained from different definitive hosts (cattle, sheep, goats, and buffaloes) in four local abattoirs in the northwestern provinces of Iran (West‐Azerbaijan, East‐Azerbaijan, Ardabil, and Zanjan) from September 2021 to August 2022. All samples were examined by morphometric criteria; then, 49 samples were identified using PCR‐RFLP based on ITS1 region and 23 sequence of isolates analyzed by cox1 marker. PCR‐RFLP methods compared morphometric results, and cox1 gene sequences were used to confirm PCR‐RFLP results and phylogenetic analysis. Results The differences between the body length, body width, cephalic cone length, cephalic cone width, body area, and distance between the ventral sucker and posterior end of the body in two species were significant (p < 0.05). Based on the morphometric criteria, 139 samples (27.8%) were identified as Fasciola gigantica and 361 (72.2%) as Fasciola hepatica. Similarly, PCR‐RFLP analysis of ITS1 region confirmed morphometric results. No hybrid forms of Fasciola were detected. Partial sequences of cox1 showed 13 variable sites with eight haplotypes in F. hepatica and 12 variable sites with five haplotypes in F. gigantica. Conclusion Based on the results of this study, the PCR‐RFLP method can be used to confirm the morphological method of Fasciola species, but it is insufficient to study their genetic diversity. Also, sequences of cox1 results of the present study showed that F. hepatica and F. gigantica species in the Northern provinces of Iran have different genetic structures and haplotypes.

Taxonomic identification of alien bivalve species in the Mediterranean Sea can be challenging because of high morphological variability and the occurrence of cryptic species complexes, as in the case of pearl oysters. While the presence of the Lessepsian species Pinctada radiata is well-established in the eastern Mediterranean Sea, the presence of P. fucata was recently suggested based on morphological data. In this study we performed an integrative assessment based on morphological and genetic data on pearl oysters collected across the Mediterranean Basin. Molecular species delimitation and phylogenetic analyses allowed a straightforward taxonomic assignment of all collected specimens to P. radiata. These specimens show the entire suite of morphological character states considered diagnostic of either P. fucata or P. radiata by previous studies. This finding clearly demonstrates that these morphological characters have no taxonomic value, and their variability observed in specimens from the Mediterranean Sea represents (part of) the intraspecific variability of P. radiata. While no evidence has been found for the presence of P. fucata, the earlier occurrences of P. radiata from the eastern and southern Mediterranean Sea are complemented with verified occurrence in the western and northern regions, demonstrating a further spreading of this non-native species throughout the Mediterranean Sea. This study clarifies the taxonomic identification and geographical distribution of pearl oysters in the Mediterranean Sea and substantiates the importance of molecular identification of alien bivalves characterized by extensive variation in shell characters.

Objectives: Alveolar echinococcosis is caused by Echinococcus multilocularis, a parasite of zoo¬notic significance with a wide range of intermediate and final hosts, and the parasite survives suc¬cessfully in diversified conditions. Plentiful studies have been done to study the genetic structure of the population of the parasite and the level of intimate kinship using mitochondrial (mt) DNA. The present study was conducted to investigate the population structure, genetic variation, and phylogenetic relationship of various isolates of E. multiocularis submitted to GenBank worldwide. Sequences of mt genes (mt-cytochrome c oxidase (cox1), mt-NADH dehydrogenase (nad1)) of E. multilocularis were analyzed to achieve the set goals. Materials and Methods: A total of 275 and 124 gene sequences of mt-cox1 and mt-nad1 belong¬ing to E. multilocularis, respectively, were retrieved from the National Center for Biotechnology Information GenBank. The retrieved sequences were subjected to alignment with respective reference sequences using MEGA software. The PopArt software was used to establish medi¬an-joining networks, while DnaSp was used to calculate neutrality and diversity indices. MrBayes software was used to investigate the phylogenetic association between haplotypes based on Bayesian phylogeny. Results: Approximately 13 and 20 distinctive haplotypes of nad1 and cox1 genes, respectively, were observed in the present study. In both of the mt genes, diversity indices indicated low haplo¬type (mt-cox1 = 0.140; mt-nad1 = 0.374) and nucleotide (mt-cox1 = 0.00111; mt-nad1 = 0.00287) diversities. The values of Tajima's D and Fu Fs for a population of both of the genes under study were found to be negative. Conclusion: This study is a maiden attempt to provide insights into the population structure and genetic variation of E. multilocularis on a global scale. However, it is suggested that to better understand the population structure and genetic diversity of E. multilocularis, more geographical locations and amplifications of full-length gene sequences should be considered, which could be helpful in widening the insights into the genetic diversity of E. multilocularis. [J Adv Vet Anim Res 2024; 11(2.000): 264-274]

The monogeneric family Calomyscidae includes the brush-tailed mice, genus Calomyscus , which have a Palearctic distribution ranging from the Hindu Kush Mountains in western Pakistan to the eastern Mediterranean region. Zagros Mountains—stretching from northwest to south of Iran—was assumed for long as the range of a single species, Calomyscus bailwardi ; however, recent studies revealed considerable heterogeneities among the geographical populations presumably referring to the persistence of undescribed diversity. One such group from the western Zagros Mountains has recently been recognized as C. behzadi Akbarirad, Dezhman, Aliabadian, Siahsarvie, Shafaeipour, and Mirshamsi, 2021. Using molecular, morphometric, geometric morphometric, and karyotypic data, we examine the divergence of 2 additional groups from western Iran and name these as new species. The first of these species is distributed in the northern Zagros and western Alborz Mountains in northwestern Iran was recovered as the sister species to C. urartensis from the South Caucuses. The second species from a single locality in the central Zagros Mountains was clustered closely with C. grandis from the western Alborz Mountains. Morphologic analyses, karyological features, and genetic distances disentangled these 2 new species from their sister species. [ABSTRACT FROM AUTHOR]

In the present study, tissue samples (tongue, esophagus and heart) were investigated from dromedary camels of India for identification and characterization of Sarcocystis spp. using histopathology, PCR and gene sequencing. Genomic DNA extracted from these tissue samples was used for PCR amplification of the cytochrome c oxidase subunit I gene (cox1) of Sarcocystis spp. and the partial sequence of small subunit ribosomal RNA (18S rRNA) gene of the S. cameli. The PCR products were purified, sequenced and analyzed using bioinformatics tools. Based on phylogenetic analysis of the cox1 gene, the sequences of the present study clustered with those of S. cameli, hosted by dromedary camels of Iraq and a close association was observed with S. masoni hosted by dogs and alpacas of China. Until now, there are no 18S rRNA sequences of S. cameli available in GenBank and this is the first study recording 18S rRNA sequences of S. cameli which were grouped with S. masoni from alpaca of China and guanaco and llama of Argentina in phylogenetic analysis. These findings could be useful for further studies on the characterization through molecular epidemiology, genetic diversity and host specificity of S. cameli. [ABSTRACT FROM AUTHOR]

Sparganosis has been a neglected parasitic zoonosis for a long time. The accurate identification of Spirometra tapeworms in clinical practice is poorly understood. A case of breast sparganosis was reported in Henan Province of central China. One plerocercoid approximately 3.5 cm in length was collected from the patient. The clinical isolate was identified as Spirometra mansoni based on the barcoding sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1). Finally, the epidemiology of sparganosis in central China was reviewed. Comprehensive public health education should be carried out, and the risky habit of eating live tadpoles must be discouraged in Henan Province. [ABSTRACT FROM AUTHOR]

Objectives: Alveolar echinococcosis is caused by Echinococcus multilocularis, a parasite of zoonotic significance with a wide range of intermediate and final hosts, and the parasite survives successfully in diversified conditions. Plentiful studies have been done to study the genetic structure of the population of the parasite and the level of intimate kinship using mitochondrial (mt) DNA. The present study was conducted to investigate the population structure, genetic variation, and phylogenetic relationship of various isolates of E. multiocularis submitted to GenBank worldwide. Sequences of mt genes (mt-cytochrome c oxidase (cox1), mt-NADH dehydrogenase (nad1)) of E. multilocularis were analyzed to achieve the set goals. Materials and Methods: A total of 275 and 124 gene sequences of mt-cox1 and mt-nad1 belonging to E. multilocularis, respectively, were retrieved from the National Center for Biotechnology Information GenBank. The retrieved sequences were subjected to alignment with respective reference sequences using MEGA software. The PopArt software was used to establish median-joining networks, while DnaSp was used to calculate neutrality and diversity indices. MrBayes software was used to investigate the phylogenetic association between haplotypes based on Bayesian phylogeny. Results: Approximately 13 and 20 distinctive haplotypes of nad1 and cox1 genes, respectively, were observed in the present study. In both of the mt genes, diversity indices indicated low haplotype (mt-cox1 = 0.140; mt-nad1 = 0.374) and nucleotide (mt-cox1 = 0.00111; mt-nad1 = 0.00287) diversities. The values of Tajima's D and Fu Fs for a population of both of the genes under study were found to be negative. Conclusion: This study is a maiden attempt to provide insights into the population structure and genetic variation of E. multilocularis on a global scale. However, it is suggested that to better understand the population structure and genetic diversity of E. multilocularis, more geographical locations and amplifications of full-length gene sequences should be considered, which could be helpful in widening the insights into the genetic diversity of E. multilocularis. [ABSTRACT FROM AUTHOR]

Three species of Geosesarma : G. celebense , G. leprosum and G. clavicrure have been previously described from the Indonesian island of Sulawesi. To evaluate their identities, we examined type material of the three species, and they are re-described and illustrated. A short segment of 229 bp of COX1 gene from the Sulawesian Geosesarma was obtained (when available) and used for genetic comparisons. Our record of G. celebense from Selatan, South Sulawesi, and the genetic affinity between the type from North Sulawesi and the southern material indicates that G. celebense is well distributed throughout Sulawesi. The present study also describes a new species of Geosesarma from central Sulawesi. Geosesarma schubarti new species differs significantly from other Sulawesian Geosesarma in the morphology of the carapace, which is distinctly squarish in shape with a clear second anterolateral tooth. Genetic distances, using the segment of COX1 marker, showed all Sulawesian Geosesarma species, including the new one, are distant from each other. Our morphological comparisons also show that G. clavicrure is actually a junior subjective synonym of Bresedium brevipes ; and the sequences of G. clavicrure grouped stably with B. brevipes in the 16S phylogenetic tree, and G. clavicrure therefore needs to be removed from the genus Geosesarma. As a consequence, with the new species, there remain three known species for Sulawesi (G. celebense , G. leprosum , G. schubarti), but based on currently available evidence it is considered likely that there exist additional species from this genus on the island. [ABSTRACT FROM AUTHOR]

Furcocercariae, of the trematodes from the family Diplostomidae, were found in freshwater snails—Radix auricularia, which were collected in a reservoir located on Popov Island (Peter the Great Bay, Sea of Japan). The life cycle was experimentally reproduced for the first time, while morphometric data for the development stages were studied and described for the newly discovered trematode. Moreover, molecular data for nuclear and mitochondrial markers were also obtained. It was determined that the morphometric characteristics of the trematode coincided with the species Diplostomum cf. vanelli, the molecular data analysis validates the species independence. Furthermore, the study highlights the issue of species identification in the Diplostomum genus. [ABSTRACT FROM AUTHOR]

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Hydatid cyst disease, caused by Echinococcus granulosus sensu lato (s.l.), is linked to endemic areas in the Levant region, such as Jordan. There is a scarcity of epidemiological data about this disease. Our goal was to pinpoint the particular genotypes accountable for sheep and goat infection in unexplored regions of Southern Jordan. Partial sequences of cytochrome C oxidase (Cox1, 450 bp) and NADH dehydrogenase subunit 2 (Nad2, 820 bp) of the E. granulosus from Jordan were analysed and compared with E. granulosus s.l. reference genomes, to infer the phylogeny of the species. This analysis demonstrated a significant degree of intraspecific similarity and identified the presence of E. granulosus sensu stricto (G1, G3). The G1 genotype was the most prevalent (Cox1, n = 17/17); (Nad2, n = 13/18). The study examined partial Cox1 sequences from different countries (Jordan, Lebanon, Iran, India) and regions (South America) to analyze the genetic architecture and phylogeography of the G1 genotype at regional and global levels. One primary haplotype (Hap_1) was present in the research locations. We hypothesized that a certain branch of G1, known as Hap_1, was prevalent among regions that were not geographically connected. Future research on the biology and distribution of E. granulosus s.s. will be enhanced by this information. [ABSTRACT FROM AUTHOR]

Up to the present study, only 8.5% of the 522 macroalgal species reported at the Azores have sequences deposited in GenBank and BOLD public repositories. The sequences of four genetic markers (cox1, rbcL, UPA, tufA) were obtained for recently collected samples from two Azorean islands. DNA barcode-assisted identification was conducted on newly generated and unpublished sequences from public repositories. A literature review of recently published studies, including the molecular identifications of Azorean macroalgae, was also performed. The results confirm the occurrence of 51 species (including subspecific ranks) and provide four new records, namely, three cryptogenic species (Olokunia boudouresquei, Padina gymnospora, and Ulva lacinulata) and one introduced species (Ulva australis). This study contributes 23 DNA barcodes generated for the first time to the Azores, which now has 10.5% of its marine flora represented in public repositories. Additionally, UPA sequences were generated for the first time for the five taxa.

Clonorchis sinensis is an important zoonotic parasite that is mainly prevalent in China, Korea, Vietnam and the Russian Far East. To explore the genetic variation and population structure of C. sinensis, an in silico analysis was conducted based on mitochondrial cytochrome c oxidase subunit 1 (COX1), ribosomal internal transcribed spacer 1 (ITS1) and ribosomal internal transcribed spacer 2 (ITS2) sequences. The sequences obtained from NCBI were truncated for further analyses, including haplotype network, phylogenetic, gene flow, diversity and neutrality analyses. The results showed that there were 20, 11 and 4 haplotypes for COX1, ITS1 and ITS2, respectively. The results of both the haplotype network and phylogenetic analyses indicated that the haplotypes for each type of sequence from the same country were not all clustered together. Haplotype diversity values were all lower than 0.5. Values of nucleotide diversity were higher than 0.005, except for ITS2. Tajima’s D and Fu’s Fs values were all negative, and p-values showed significant differences, indicating that the population of C. sinensis is growing. Fst values were all lower than 0.05. In conclusion, this study found that there are specific variations of C. sinensis in different countries, and the population of this parasite is growing with less genetic variation. The findings provide a crucial foundation for understanding the molecular epidemiology and population dynamics of C. sinensis.

The genus Sarcocystis is an abundant group of Apicomplexa parasites found in mammals, birds, and reptiles. These parasites are characterised by the formation of sarcocysts in the muscles of intermediate hosts and the development of sporocysts in the intestines of definitive hosts. The identification of Sarcocystis spp. is usually carried out in carcasses of animals, while there is a lack of studies on the detection of Sarcocystis species in blood samples. In the current study, blood samples of 214 yellow-necked mice (Apodemus flavicollis) and 143 bank voles (Clethrionomys glareolus) from Lithuania were examined for Sarcocystis. The molecular identification of Sarcocystis was carried out using nested PCR of cox1 and 28S rRNA and subsequent sequencing. Sarcocystis spp. were statistically (p < 0.01) more frequently detected in the bank vole (6.3%) than in yellow-necked mice (0.9%). The analysed parasites were observed in four different habitats, such as mature deciduous forest, bog, natural meadow, and arable land. Three species, Sarcocystis funereus, Sarcocystis myodes, and Sarcocystis cf. glareoli were confirmed in the bank vole, whereas only Sarcocystis myodes were found in yellow-necked mice. The obtained results are important in the development of molecular identification of Sarcocystis parasites in live animals. [ABSTRACT FROM AUTHOR]

The yellow-legged hornet (Vespa velutina) is a social Hymenoptera native from Asia and an invasive species in Europe, where it was first detected in France in 2004. Since then, the species has spread across the continent invading mainland Spain and Mallorca island (Balearic archipelago, Western Mediterranean) in 2010 and 2015, respectively. Yellow-legged hornets cause severe damage to ecosystem by predating over a wide variety of pollinators including honeybees. Such a threat situation requires the development of effective management and prevention plans, which can greatly benefit from knowing both the origin and the genetic structure of the invading populations. Here we conduct a genetic study to shed light on both the origin and the phylogenetic relationships of V. velutina populations from Mallorca and mainland Spain using nuclear (STRs) and mitochondrial (cytochrome oxidase c subunit 1) gene markers. Our results show that Mallorca populations originated from invasive European specimens. Moreover, FST values, DAPC and genetic structure analysis suggest two independent incursions in the island with bottleneck and founder effect signatures. Finally, we contribute additional genetic evidence of the polyandrous behavior of this invasive species based on the inference of a mean number of mattings per nest of 3.94 (range 2–6.5). This study supports the human-mediated pathways of this species and highlights the importance of implementing effective biosecurity measures to prevent the spread of invasive alien species in island habitats. [ABSTRACT FROM AUTHOR]

Purpose: Strongyloides stercoralis is a parasite with special characteristics presenting it as a unique nematode. Iran is an endemic area for S.stercoralis. In this study, nested-qPCR-high resolution melting (HRM) technology was applied on some human isolates of S.stercoralis from this country by focusing on evolutionary genetics analysis. Methods: Twelve human isolates of S. stercoralis were collected from four endemic provinces of Iran. Genomic DNA was extracted from a single filariform larva for every isolate. Using specific primers targeting partial regions in cox1 gene, nested-qPCR-HRM was performed and melting-curve profiles were analyzed alongside the evaluation of genetic proximity and phylogenetic analysis using MEGA7 and DnaSP5 software. Results: The melting temperature (Tm) values of the isolates were 77.9 °C−78.3 °C. All isolates from Guilan, Mazandaran, and Khouzestan Provinces shared Tm values of 78.2 °C to 78.3 °C, while the isolates from Hormozgan Province showed Tm values of 77.9 °C, 78.0 °C, and 78.1 °C. The phylogenetic tree illustrated that the sequences of the current study included nine haplotypes. Tajima's D index analyses showed that cox1 gene in S. stercoralis isolates was negative (Tajima's D = − 0.27). Conclusion: The isolates were divided into five temperature groups. Although HRM assay compared to PCR sequencing identified more limited genetic changes, it revealed that the mean of Tm of the isolates from Hormozgan Province was lower than those of other provinces and represented specific haplotypes for this geographical region on the phylogenetic tree. [ABSTRACT FROM AUTHOR]

Purpose: Using molecular techniques, we have previously shown that carnivorous mammals of the family Mustelidae might be common definitive hosts for various protozoan Sarcocystis species. In the present study we aimed to unravel whether Sarcocystis species using ungulates as intermediate hosts and canids or felids as definitive hosts can be found in intestine of mustelids. Methods: Small intestine samples of 93 individual mustelids of five different species from Lithuania were examined. Sarcocystis species were identified based on species-specific PCR and subsequent cox1 sequencing. Results: Six Sarcocystis species (S. arieticanis, S. bertrami, S. capracanis, S. capreolicanis, S. linearis and S. morae) defined by ungulate-canid life cycle were detected for the first time in small intestines of mustelids. By contrast, the prevalence of Sarcocystis characterised by ungulate-felid life cycle was low (3.2%). Overall, 76% of the examined animals were positive for at least one of the studied Sarcocystis species. Four species, S. arieticanis, S. bertrami, S. capracanis and S. morae were most commonly found, with the detection rate of about 40%. Conclusions: The current finding, in addition to our previous studies, suggests that mustelids play an important role in the spread of various Sarcocystis species. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study is to conduct a molecular characterization of Spirometra tapeworm from jungle cat (Felis chaus) in Guilan Province, north of Iran using DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (Cox1) and 12S rDNA sequences. Methods: Morphological features of the adult tapeworm of Spirometra were evaluated using specific staining and light microscopy. The molecular characterization was performed using partial Cox1 and 12S rDNA regions. Genetic diversity was calculated and phylogenetic trees of the obtained sequences were constructed. Results: Morphological features were compatible with previous description of adult Spirometra erinaceieuropaei. The Cox1 sequence of the specimen showed 100% similarity with S. erinaceieuropaei sequences in GenBank from Korea, China and Iran. Also, the 12S rDNA sequence revealed 99.7% similarity with S. erinaceieuropaei isolates from China and Japan. Intra-species variation within isolates of S. erinaceieuropaei was 0–1.4% and 0–4.6% for Cox1 and 12S rDNA genes, respectively. Conclusion: This is the first report of molecular characterization of S. erinaceieuropaei in jungle cat, F. chaus in Iran. Jungle cat probably plays a major role as reservoir host in maintaining of this parasite in this area with favorable climate condition. Needs for further assessment on the role of appropriate hosts, especially intermediate/paratenic hosts as well as the potential risk of human infectivity with sparganosis is emphasized. [ABSTRACT FROM AUTHOR]

Purpose: Derogenes ruber Lühe, 1900, the type-species of the genus Derogenes Lühe, 1900, is a poorly known derogenid digenean. The original description of this species was not illustrated and aspects of the morphology of the parasite from the type-host remain scarce. Available records of this species were brief and/or lacked illustrations and were based on morphology alone. Additionally, molecular data for Derogenes spp. are warranted to untangle species complexes as they provide a better assessment of interspecific genetic divergence. Methods: Derogenes ruber is redescribed based on newly collected specimens from the gall bladder of its type-host Chelidonichthys lastoviza (Bonnaterre, 1788) collected in the Western Mediterranean off the Algerian coast during 2017–2019 and molecular data are provided using a partial fragment of the nuclear 28S ribosomal RNA gene (28S rRNA), the internal transcribed spacer 2 (ITS2) and a fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Results: We herein provide a detailed illustrated redescription and morphometric data of D. ruber from its type-host C. lastoviza. We report a new geographical record (off Algeria) for it. Derogenes ruber is also genetically characterised for the first time. Species/lineages of Derogenes were recovered in five strongly supported reciprocally monophyletic clades: (i) D. ruber from C. lastoviza off Algeria; (ii) D. lacustris from Galaxias maculatus (Jenyns) off Argentina; (iii) Lineage "D. varicus DV1" (D. varicus sensu stricto) from fish hosts in the White and Barents seas and the North Sea; (iv) Lineage "D. varicus DV2" from mollusc hosts in the White Sea; and (v) Lineage "D. varicus DV3" from Eumicrotremus fedorovi Mandrytsa. in the Pacific Ocean. Hence, comparison of the newly generated sequences with other available data for Derogenes species supports the distinction of D. ruber confirming its taxonomic status and helping assess interspecific variation. Comparison of D. ruber with the closely related species Derogenes latus revealed overlaps in morphometric data and the validity of the latter species is questioned. Conclusion: The combination of morphological and molecular data provided for D. ruber provides a firm foundation for further investigations of Derogenes spp. Although we do describe herein material of D. ruber from the type-host, given that the occurrence of a single Derogenes species in various hosts has been challenged by molecular data, and both D. lacustris and D. varicus sensu stricto had been genetically proven to occur in various hosts, D. ruber and D. latus may be indeed synonymous. Additional sequencing effort on Derogenes spp. will strengthen systematic comparative studies and evolutionary relationships within the Derogenidae in general. [ABSTRACT FROM AUTHOR]

Research on complete mitochondrial genomes can help in understanding the molecular evolution and phylogenetic relationships of various species. In this study, the complete mitogenome of Hua aristarchorum was characterized to supplement the limited mitogenomic information on the genus Hua. Three distinct assembly methods, GetOrganelle, NovoPlasty and SPAdes, were used to ensure reliable assembly. The 15,691 bp mitogenome contains 37 genes and an AT-rich region. Notably, the cytochrome c oxidase subunit I (COX1) gene, commonly used for species identification, appears to be slow-evolving and less variable, which may suggest the inclusion of rapidly evolving genes (NADH dehydrogenase subunit 6 [ND6] or NADH dehydrogenase subunit 2 [ND2]) as markers in diagnostic, detection, and population genetic studies of Cerithioidea. Moreover, we identified the unreliability of annotations (e.g., the absence of annotations for NADH dehydrogenase subunit 4L [ND4L] in NC_037771) and potential misidentifications (NC_023364) in public databases, which indicate that data from public databases should be manually curated in future research. Phylogenetic analyses of Cerithioidea based on different datasets generated identical trees using maximum likelihood and Bayesian inference methods. The results confirm that Semisulcospiridae is closely related to Pleuroceridae. The sequences of Semisulcospiridae clustered into three clades, of which H. aristarchorum is one; H. aristarchorum is sister to the other two clades. The findings of this study will contribute to a better understanding of the characteristics of the H. aristarchorum mitogenome and the phylogenetic relationships of Semisulcospiridae. The inclusion of further mitochondrial genome sequences will improve knowledge of the phylogeny and origin of Cerithioidea. [ABSTRACT FROM AUTHOR]

Cryptic diversity is common among marine macroalgae, with molecular tools leading to the discovery of many new species. To assign names to these morphologically similar species, the type and synonyms have to be examined, and if appropriate, new species must be described. The turf‐forming red alga Polysiphonia scopulorum was originally described from Rottnest Island, Australia, and subsequently widely reported in tropical and temperate coasts based on morphological identifications. A recent study of molecular species delineation revealed a complex of 12 species in Australia, South Africa, and Europe. These species are placed in a taxonomically unresolved lineage of the tribe Polysiphonieae. The aim of this study was to resolve the genus‐ and species‐level taxonomy of this complex and related species using molecular and morphological information. Three morphologically indistinguishable species of the complex were found at the type locality of P. scopulorum, preventing a straightforward assignment of the name to any of the molecular lineages. Therefore, we propose a molecularly characterized epitype. Polysiphonia caespitosa is reinstated for the only species found in its type locality in South Africa. We describe seven new species. Only one species of the complex can be morphologically recognized, with the other eight species indistinguishable based on morphometric analysis. The studied complex, together with another seven species currently placed in Polysiphonia and two Bryocladia species, formed a clade distinct from Polysiphonia sensu stricto. Based on observations of Bryocladia cervicornis (the generitype), we describe our seven new species in the genus Bryocladia and transfer another nine species from Polysiphonia to Bryocladia. [ABSTRACT FROM AUTHOR]

To determine the genotypes of the epidemic strains of Echinococcus granulosus in livestock in Tibet, samples of E. granulosus cysts were collected from 11 yaks and 62 sheep. Genomic DNA was extracted from these samples, and gene fragments of mitochondrial cytochrome c oxidase subunit I (cox1) and NADH dehydrogenase subunit I (nad1) were amplified by PCR and sequenced. DNASTAR and MAGA7.0 were employed for homology analysis and phylogenetic tree construction. Echinococcus granulosus cysts were detected in 56.2% (41/73) of the samples screened. Of these, 63.4% (26/41) were identified as E. granulosus G1 genotype (common sheep strain), 24.4% (10 /41) as G3 genotype (buffalo strain), and 12.2% (5/41) were G6 genotype (camel strain). The study concludes that yaks and sheep in Langkazi county, Tibet, carry three E. granulosus genotypes (G1, G3, and G6), with the G1 genotype the predominant genotype in the region. This study clarifies the distribution of E. granulosus genotypes, providing genetic data and insight for the surveillance and prevention of echinococcosis. [ABSTRACT FROM AUTHOR]

Gall mites (Eriophyoidea) are cosmopolitan microscopic phytoparasites that often transmit viruses and induce gallogenesis. The tribe Calacarini is diagnosed by a set of plesiomorphic and homoplastic traits, including elimination of setae sc shared with other lineages of Eriophyoidea. We reviewed data on the generic diversity of calacarines, revised the concept of the type genus Calacarus Keifer 1940, and proposed three zones (MZ, SMZ, LZ) in the prodorsal shields of calacarines to simplify descriptions of their shield patterns. We describe three new calacarine species (Calacarus baviensisn. sp., C. burchelliaen. sp., and Viginticus searsiaen. sp.) from indigenous dicotyledonous trees from South Africa and Vietnam and report on new findings of Paracalacarus podocarpi Keifer in Brazil, Jiangsuacarus sp. in the USA, and Calacarus pusillus Pye in Latvia and Russia. The latter represents the new most northern locality of Calacarini. Reinvestigating the type species of Jaranasia Chandrapatya & Boczek 2000 revealed that absence of setae l" II is the only character separating it from Jiangsuacarus Xue 2009. We proposed two new combinations: Jiangsuacarus sesleriae (Skoracka 2004) n. comb. (transferred from Jaranasia) and Procalacarus mussaendae (Keifer 1977) n. comb. (transferred from Calacarus). Partial sequences of Cox1 and 28S genes were obtained for six calacarines, some of them originating from old ethanol material kept at room temperature. Molecular phylogenetics revealed a stable cluster of "true" calacarine sequences comprising Calacarus, Jaranasia, Latitudo, and Viginticus and a polyphyletic group of erroneous sequences assigned to Calacarini in GenBank. All investigated females of calacarines have a pair of genital tubules associated with the vestibulum and hypothesized to participate in fertilization. This finding may contribute to resolving the question on how the fusion of gametes happens in gall mites. [ABSTRACT FROM AUTHOR]

Alectorobius species are soft ticks primarily infesting birds, such as swallows, while Dermacentor species are hard ticks mainly infesting mammals, such as small ruminants. This study for the first time reported on the morphological and molecular bases of two tick species, namely A. coniceps and a Dermacentor sp. in Pakistan. The former species was examined in swallows' nests in Khyber Pakhtunkhwa province, while the latter species was examined in small ruminants in Balochistan province. In total, 25 ticks were collected, with 14 ticks morphologically identified as A. coniceps (males = 9 and females = 5) and 11 ticks identified as Dermacentor sp. (males = 7 and females = 4). Following morphological identification, molecular identification was gained by obtaining 16S rDNA and cox1 sequences for these ticks. The BLAST results for the 16S rDNA and cox1 sequences from A. coniceps shared a maximum identity of 97.46% and 96.49% with the same species from Malta. The BLAST analysis of the 16S rDNA and cox1 sequences from Dermacentor sp. showed maximum identities of 98.42% and 97.45% with Dermacentor pavlovskyi from China. The phylogenetic analysis based on 16S rDNA and cox1 of A. coniceps showed a close evolutionary relationship with the same species. The case of Dermacentor sp., based on 16S DNA and cox1, indicated a close evolutionary relationship with Dermacentor pavlovskyi from China. [ABSTRACT FROM AUTHOR]

• We characterize the diplostomid parasite communities in two fish species. • We applied a quadrupled-indexing cox1 region-specific DNA metabarcoding method. • Diplostomid communities in temperate lakes show spatial and temporal heterogeneity. • We revealed novel associations between co-infecting diplostomid species. • This method proved to be effective to describe communities with cryptic species. Eye flukes (Diplostomidae) are diverse and abundant trematode parasites that form multi-species communities in fish with negative effects on host fitness and survival. However, the environmental factors and host-related characteristics that determine species diversity, composition, and coexistence in such communities remain poorly understood. Here, we developed a cost-effective cox1 region-specific DNA metabarcoding approach to characterize parasitic diplostomid communities in two common fish species (Eurasian perch and common roach) collected from seven temperate lakes in Estonia. We found considerable inter- and intra-lake, as well as inter-host species, variation in diplostomid communities. Sympatric host species characterization revealed that parasite communities were typically more diverse in roach than perch. Additionally, we detected five positive and two negative diplostomid species associations in roach, whereas only a single negative association was observed in perch. These results indicate that diplostomid communities in temperate lakes are complex and dynamic systems exhibiting both spatial and temporal heterogeneity. They are influenced by various environmental factors and by host-parasite and inter-parasite interactions. We expect that the described methodology facilitates ecological and biodiversity research of diplostomid parasites. It is also adaptable to other parasite groups where it could serve to improve current understanding of diversity, distribution, and interspecies interactions of other understudied taxa. [ABSTRACT FROM AUTHOR]