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Hepatic lipidosis (ie, fatty liver) occurs primarily during the first weeks of lactation in dairy cows because of excessive lipolysis overwhelming the concomitant capacity for beta-oxidation and hepatic export of triglycerides. Besides economic losses due to reduced lactational and reproductive performance, close associations with concomitantly occurring infectious and metabolic health disorders, in particular ketosis, exist. Hepatic lipidosis is not only a consequence from the postpartal negative energy balance but also acts as a disease component for further health disorders.

This study aimed to investigate the effect of the different dry-off strategies based on reducing feeding level (normal vs. reduced energy density), reducing milking frequency (twice vs. once daily), and administration of a dopamine agonist after last milking (i.e. saline vs. cabergoline injection) on blood metabolites, hormones, and minerals around dry-off. In this experiment, 119 Holstein dairy cows were used in a 2 × 2 × 2 factorial arrangement. In the last week before dry-off, cows were allocated to 1 of the 4 possible dry-off strategies based on feeding level and milking frequency. Within 3 h after last milking, cows were injected with either saline or a D2 dopamine agonist (cabergoline; Velactis, Ceva Santé Animale, Libourne, France; labeled for use only with abrupt dry-off, e.g., no preceding reduction in feeding level or milking frequency before last milking). After dry-off, all cows were fed the same dry cow diet and data collection continued for a week. Blood samples were collected from the coccygeal vein on d -9, -6, -5, -2, 1, 2, 5, and 7 relative to dry-off. Additionally, blood was sampled at 0, 3, and 6 h relative to injection of either cabergoline or saline, equivalent to d 0.125, 0.250, and 0.375 relative to last milking (dry-off). The reduced feeding level before dry-off caused reduced glucose and insulin concentrations as well as increased free fatty acid concentrations, particularly when reduced feeding level was combined with milking the cows 2× daily. The intramuscular injection of cabergoline caused the expected reduction in circulating prolactin concentrations. In addition, dopamine-agonist cabergoline induced an atypical simultaneous pattern of plasma metabolites (i.e., increased glucose and free fatty acid concentrations), hormones (i.e., reduced insulin and increased cortisol concentrations), and minerals (i.e., reduced calcium concentration), indicating that normal metabolic and mineral homeostatic regulations were hindered after the injection of ergot alkaloid cabergoline. In conclusion, reducing milking frequency seems the best management strategy to reduce milk production at dry-off among those tested in this study.

Meningioma is the most frequent intracranial neoplasm in cats. Here we describe the first case of chordoid meningioma (CM), a rare grade II meningioma subtype, in a 5.5-year-old European wildcat (Felis silvestris) from a Swiss zoo. The wildcat was found dead after a clinical history of neurological signs and clinical suspicion of a carcinoma in the right external ear canal with concurrent chronic otitis. Post-mortem examination revealed a large intracranial, extra-axial and intradural neoplasm that invaded into the right ear canal and had histological features compatible with CM, which has been only reported in humans and dogs. Neoplastic cells expressed vimentin but were negative for glial fibrillary acidic protein, S100 and pancytokeratin. Immunohistochemistry revealed epithelial membrane antigen (EMA) expression in neoplastic cells. To the best of our knowledge, we provide the first evidence of EMA expression in feline meningioma.

Increasing the productivity of Canadian dairy goats is critical to the competitiveness of the sector; however, little is known about the underlying genetic architecture of economically important traits in these populations. Consequently, the objectives of this study were as follows: (1) to perform a single-step GWAS for milk production traits (milk, protein, and fat yields, and protein and fat percentages in first and later lactations) and conformation traits (body capacity, dairy character, feet and legs, fore udder, general appearance, rear udder, suspensory ligament, and teats) in the Canadian Alpine and Saanen breeds; and (2) to identify positional and functional candidate genes related to these traits. The data available for analysis included 305-d milk production records for 6,409 Alpine and 3,434 Saanen does in first lactation and 5,827 Alpine and 2,632 Saanen does in later lactations; as well as linear type conformation records for 5,158 Alpine and 2,342 Saanen does. Genotypes were available for 833 Alpine and 874 Saanen animals. Both single-breed and multiple-breed GWAS were performed using single-trait animal models. Positional and functional candidate genes were then identified in downstream analyses. The GWAS identified 189 unique SNP that were significant at the chromosomal level, corresponding to 271 unique positional candidate genes within 50 kb up- and downstream, across breeds and traits. This study provides evidence for the economic importance of several candidate genes (e.g., CSN1S1, CSN2, CSN1S2, CSN3, DGAT1, and ZNF16) in the Canadian Alpine and Saanen populations that have been previously reported in other dairy goat populations. Moreover, several novel positional and functional candidate genes (e.g., RPL8, DCK, and MOB1B) were also identified. Overall, the results of this study have provided greater insight into the genetic architecture of milk production and conformation traits in the Canadian Alpine and Saanen populations. Greater understanding of these traits will help to improve dairy goat breeding programs.

50 Pág., In March 2022, following the annual International Committee on Taxonomy of Viruses (ICTV) ratifcation vote on newly proposed taxa, the phylum Negarnaviricota was amended and emended. The phylum was expanded by two new families (bunyaviral Discoviridae and Tulasviridae), 41 new genera, and 98 new species. Three hundred forty-nine species were renamed and/or moved. The accidentally misspelled names of seven species were corrected. This article presents the updated taxonomy of Negarnaviricota as now accepted by the ICTV, This work was supported in part through Laulima Govern ment Solutions, LLC prime contract with the US National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272201800013C. J.H.K. performed this work as an employee of Tunnell Government Services (TGS), a subcontractor of Laulima Gov ernment Solutions, LLC under Contract No. HHSN272201800013C. This work was also funded in part by Contract No. HSHQDC 15-C-00064 awarded by DHS S&T for the management and operation of The National Biodefense Analysis and Countermeasures Center, a federally funded research and development center operated by the Battelle National Biodefense Institute (V.W.); and NIH contract HHSN272201000040I/HHSN27200004/D04 and grant R24AI120942 (N.V., R.B.T.). S.S. acknowledges support from the Mississippi Agri cultural and Forestry Experiment Station (MAFES), USDA-ARS pro ject 58-6066-9-033 and the National Institute of Food and Agriculture, US Department of Agriculture, Hatch Project, under Accession Num ber 1021494.

Weber et al. provide evidence in support of a stress-diathesis model of functional neurological disorders. They identify trauma history in the form of emotional neglect as a psychological risk factor, and reduced hippocampus and amygdala volume as a predisposing biological trait marker., Stress is a well-known risk factor to develop a functional neurological disorder, a frequent neuropsychiatric medical condition in which patients experience a variety of disabling neurological symptoms. Only little is known about biological stress regulation, and how it interacts with predisposing biological and psychosocial risk factors. Dysregulation of the hypothalamic-pituitary-adrenal axis in patients with functional neurological disorders has been postulated, but its relationship to preceding psychological trauma and brain anatomical changes remains to be elucidated. We set out to study the hypothalamic-pituitary-adrenal axis analysing the cortisol awakening response and diurnal baseline cortisol in 86 patients with mixed functional neurological symptoms compared to 76 healthy controls. We then examined the association between cortisol regulation and the severity and duration of traumatic life events. Finally, we analysed volumetric brain alterations in brain regions particularly sensitive to psychosocial stress, acting on the assumption of the neurotoxic effect of prolonged cortisol exposure. Overall, patients had a significantly flatter cortisol awakening response (P < 0.001) and reported longer (P = 0.01) and more severe (P < 0.001) emotional neglect as compared to healthy controls. Moreover, volumes of the bilateral amygdala and hippocampus were found to be reduced in patients. Using a partial least squares correlation, we found that in patients, emotional neglect plays a role in the multivariate pattern between trauma history and hypothalamic-pituitary-adrenal axis dysfunction, while cortisol did not relate to reduced brain volumes. This suggests that psychological stress acts as a precipitating psychosocial risk factor, whereas a reduced brain volume rather represents a biological predisposing trait marker for the disorder. Contrarily, an inverse relationship between brain volume and cortisol was found in healthy controls, representing a potential neurotoxic effect of cortisol. These findings support the theory of reduced subcortical volumes representing a predisposing trait factor in functional neurological disorders, rather than a state effect of the illness. In summary, this study supports a stress-diathesis model for functional neurological disorders and showed an association between different attributes of trauma history and abnormalities in hypothalamus-pituitary-adrenal axis function. Moreover, we suggest that reduced hippocampal and amygdalar volumes represent a biological 'trait marker' for functional neurological disorder patients, which might contribute to a reduced resilience to stress.

Dry matter intake (DMI) is a fundamental component of the animal's feed efficiency, but measuring DMI of individual cows is expensive. Mid-infrared reflectance spectroscopy (MIRS) on milk samples could be an inexpensive alternative to predict DMI. The objectives of this study were (1) to assess if milk MIRS data could improve DMI predictions of Canadian Holstein cows using artificial neural networks (ANN); (2) to investigate the ability of different ANN architectures to predict unobserved DMI; and (3) to validate the robustness of developed prediction models. A total of 7,398 milk samples from 509 dairy cows distributed over Canada, Denmark, and the United States were analyzed. Data from Denmark and the United States were used to increase the training data size and variability to improve the generalization of the prediction models over the lactation. For each milk spectra record, the corresponding weekly average DMI (kg/d), test-day milk yield (MY, kg/d), fat yield (FY, g/d), and protein yield (PY, g/d), metabolic body weight (MBW), age at calving, year of calving, season of calving, days in milk, lactation number, country, and herd were available. The weekly average DMI was predicted with various ANN architectures using 7 predictor sets, which were created by different combinations MY, FY, PY, MBW, and MIRS data. All predictor sets also included age of calving and days in milk. In addition, the classification effects of season of calving, country, and lactation number were included in all models. The explored ANN architectures consisted of 3 training algorithms (Bayesian regularization, Levenberg-Marquardt, and scaled conjugate gradient), 2 types of activation functions (hyperbolic tangent and linear), and from 1 to 10 neurons in hidden layers). In addition, partial least squares regression was also applied to predict the DMI. Models were compared using cross-validation based on leaving out 10% of records (validation A) and leaving out 10% of cows (validation B). Superior fitting statistics of models comprising MIRS information compared with the models fitting milk, fat and protein yields suggest that other unknown milk components may help explain variation in weekly average DMI. For instance, using MY, FY, PY, and MBW as predictor variables produced a predictive accuracy (r) ranging from 0.510 to 0.652 across ANN models and validation sets. Using MIRS together with MY, FY, PY, and MBW as predictors resulted in improved fitting (r = 0.679-0.777). Including MIRS data improved the weekly average DMI prediction of Canadian Holstein cows, but it seems that MIRS predicts DMI mostly through its association with milk production traits and its utility to estimate a measure of feed efficiency that accounts for the level of production, such as residual feed intake, might be limited and needs further investigation. The better predictive ability of nonlinear ANN compared with linear ANN and partial least squares regression indicated possible nonlinear relationships between weekly average DMI and the predictor variables. In general, ANN using Bayesian regularization and scaled conjugate gradient training algorithms yielded slightly better weekly average DMI predictions compared with ANN using the Levenberg-Marquardt training algorithm.

We used genome-wide SNP data from 18 local cattle breeds from six countries of the Alpine region to characterize population structure and identify genomic regions underlying positive selection. The geographically close breeds Evolèner, Eringer, Valdostana Pezzata Nera, and Valdostana Castana were found to differ from all other Alpine breeds. In addition, three breeds, Simmental, and Original Braunvieh from Switzerland and Pinzgauer from Austria built three separate clusters. Of the 18 breeds studied, the intra-alpine Swiss breed Evolèner had the highest average inbreeding based on runs of homozygosity (FROH ) and the highest average genomic relationship within the breed. In contrast, Slovenian Cika cattle had the lowest average genomic inbreeding and the lowest average genomic relationship within the breed. We found selection signatures on chromosome 6 near known genes such as KIT and LCORL explaining variation in coat color and body size in cattle. The most prominent selection signatures were similar regardless of marker density and the breeds in the data set. In addition, using available high-density SNP data from 14 of the breeds we identified 47 genome regions as ROH islands. The proportion of homozygous animals was higher in all studied animals of local breeds than in Holstein and Brown Swiss cattle, the two most important commercial breeds in the Alpine region. We report ROH islands near genes related to thermoregulation, coat color, production, and stature. The results of this study serve as a basis for the search for causal variants underlying adaptation to the alpine environment and other specific characteristics selected during the evolution of local Alpine cattle breeds.

Univentricular heart malformations are rarely described in the veterinary literature. This case report describes a single-inlet univentricular heart with persistent truncus arteriosus as a cause of death of a captive-bred, 2-day-old, female Sumatran orangutan (Pongo abelii), a critically endangered species.

Equine pastern dermatitis has a high prevalence in the equine population, especially in draft breeds. This skin condition is difficult to treat, and it is suspected that owners often decide on a treatment without consulting a veterinarian. The objectives of this study were to describe owner-reported clinical signs, severity, and reasons to consult a veterinarian. Moreover, we inquired about preventive measures and treatments, both instituted by owners without previous consultation or prescribed by their veterinarians. A total of 123 horses (owners recruited over social media) were included in the study. All horses suffered from equine pastern dermatitis at least once in the two years preceding the study. Standardized questionnaires collecting information on management, housing conditions, clinical signs as well as preventive measures and treatments were filled out by participants. The data was recorded, and descriptive statistics were performed. Most horses (71 out of 115 available answers, 62 %) had shown clinical signs of equine pastern dermatitis at least four times in their lives. A total of 113 horses (92 % of all included horses) were affected by equine pastern dermatitis at the time of the interview. For 37 horses (32 %) out of 114 available answers the owners consulted their veterinarian only after the horse showed signs of pain or lameness. Usually, treatments that did not require a medical prescription were applied without consulting their veterinarian (e. g. only 9 % (14 out of 150 prescriptions) of topical creams promoting wound healing were prescribed by a veterinarian). A total of 31 treatment decisions (55 %) with creams containing anti-bacterial, anti-mycotic and/or anti-inflammatory agents and 100 % of systemic medications containing anti-bacterial, anti-parasitic or anti-inflammatory agents were prescribed by veterinarians. Overall, 69 % of treatment decisions were made without consulting a veterinarian, making it then more difficult to determine underlying causes for the pastern dermatitis and rendering the treatment often longer and more onerous. To raise owner awareness of possible consequences if a veterinarian is consulted too late in the disease process, specific information campaigns to improve animal welfare should be considered.Mauke hat eine hohe Prävalenz in der Pferdehaltung, insbesondere bei Kaltblutpferden. Die Therapie dieser Hauterkrankung gestaltet sich schwierig, und vermutlich entscheiden sich Besitzer oft für eine Behandlung ohne einen Tierarzt zu konsultieren. Die Ziele der vorliegenden Studie waren die klinische Beschreibung der Hautveränderungen durch die Besitzer, den Schweregrad und die Entscheidungsgrundlage einen Tierarzt zu konsultieren abzuklären. Zusätzlich wurden die Prophylaxe und Behandlungen durch Besitzer mit oder ohne tierärztliche Konsultation abgefragt. Insgesamt wurden die Daten von 123 Pferden (Besitzer über soziale Medien rekrutiert) analysiert. Alle Pferde erkrankten mindestens einmal in den zwei Studienjahren an Mauke. Die Resultate wurden mit standardisierten Fragebögen zu Management, Haltungsbedingungen, klinischen Symptomen sowie vorbeugenden Massnahmen und Behandlungen gesammelt. Die Daten wurden mittels deskriptiver Statistik dargestellt. Die meisten Pferde (71 von 115 verfügbaren Antworten, 62 %) hatten mindestens viermal in ihrem Leben klinische Anzeichen einer Mauke. Insgesamt 113 Pferde (113/123, 92 %) waren zum Zeitpunkt der Befragung an Mauke erkrankt. Bei 37 Pferden (37/114, 32 %) konsultierten die Besitzer ihren Tierarzt erst, nachdem das Pferd Anzeichen von Schmerzen oder Lahmheit zeigte. Behandlungen, die keiner tierärztlichen Verordnung bedurften, wurden in der Regel ohne Rücksprache mit dem Tierarzt durchgeführt (z. B. nur 9 % (14 /150) der topischen wundheilungsfördernden Cremes wurden von einem Tierarzt verschrieben). Insgesamt 31 Behandlungsentscheidungen (55 %) mit Cremes mit antibakteriellen, antimykotischen und/oder entzündungshemmenden Wirkstoffen und 100 % systemische Medikamente mit antibakteriellen, antiparasitären oder entzündungshemmenden Wirkstoffen wurden von Tierärzten verschrieben. 69 % der Behandlungen wurden ohne Rücksprache mit einem Tierarzt durchgeführt, welche die Bestimmung der Ursache oft schwieriger und die Behandlung der Mauke länger und belastender macht. Informationskampagnen sind für das Tierwohl notwendig, damit Besitzer die Vorteile einer frühen Konsultation eines Tierarztes erkennen.La dermatite du paturon a une prévalence élevée dans la population équine, en particulier chez les races de trait. Cette affection cutanée est difficile à traiter et on soupçonne que les propriétaires décident souvent d’un traitement sans consulter un vétérinaire. Les objectifs de cette étude étaient de décrire les signes cliniques rapportés par les propriétaires, la sévérité et les raisons de consulter un vétérinaire. De plus, nous nous sommes renseignés sur les mesures préventives et les traitements, qu’ils soient mis en place par les propriétaires sans consultation préalable ou prescrits par leurs vétérinaires. Un total de 123 chevaux (propriétaires recrutés via les médias sociaux) ont été inclus dans l’étude. Tous les chevaux ont souffert de dermatite du paturon au moins une fois au cours des deux années précédant l’étude. Des questionnaires standardisés, recueillant des informations sur la gestion, les conditions de logement, les signes cliniques ainsi que les mesures préventives et les traitements, ont été remplis par les participants. Les données ont été enregistrées et des statistiques descriptives ont été réalisées. La plupart des chevaux (71 sur 115 réponses disponibles, 62 %) avaient présenté des signes cliniques de dermatite du paturon au moins quatre fois dans leur vie. Au total, 113 chevaux (92 % de tous les chevaux inclus) étaient atteints de dermatite du paturon équin au moment de l’entretien. Pour 37 chevaux (32%) sur les 114 réponses disponibles, les propriétaires ont consulté leur vétérinaire uniquement après que le cheval a eu montré des signes de douleur ou de boiterie. En général, les traitements qui ne nécessitaient pas de prescription médicale étaient appliqués sans consulter leur vétérinaire (par exemple, seuls 9 % [14 sur 150 prescriptions] des crèmes topiques favorisant la cicatrisation des plaies étaient prescrites par un vétérinaire). Au total, 31 décisions de traitement (55%) avec des crèmes contenant des agents antibactériens, antimycosiques et/ou anti-inflammatoires et 100% des médicaments systémiques contenant des agents antibactériens, antiparasitaires ou anti-inflammatoires ont été prescrits par des vétérinaires. Dans l’ensemble, 69 % des décisions de traitement ont été prises sans consulter un vétérinaire, ce qui rend plus difficile la détermination des causes sous-jacentes de la dermatite du paturon et rend le traitement souvent plus long et plus onéreux. Pour sensibiliser les propriétaires aux conséquences possibles d’une consultation trop tardive d’un vétérinaire dans le processus de la maladie, des campagnes d’information spécifiques visant à améliorer le bien-être des animaux devraient être envisagées.La dermatite al pastorale (Equine pastern dermatitis) ha una grande prevalenza nella popolazione equina, specialmente in quella a sangue freddo. Questa condizione della pelle è difficile da trattare, e si sospetta che i proprietari spesso decidano di trattarla senza consultare un veterinario. Gli obiettivi di questo studio erano di descrivere i segni clinici riportati dai proprietari, la gravità e le ragioni della consultazione dal veterinario. Inoltre, abbiamo raccolto informazioni sulle misure preventive e i trattamenti, sia intrapresi dai proprietari senza consultazione precedente che prescritti dai veterinari. Un totale di 123 cavalli (i proprietari sono stati reclutati sui social media) sono stati inclusi nello studio. Tutti i cavalli hanno sofferto di dermatite al pastorale almeno una volta nei due anni precedenti lo studio. Ai partecipanti è stato richiesto di completare dei questionari standardizzati che raccoglievano informazioni sulla gestione, le condizioni di stabulazione, i segni clinici, le misure preventive e i trattamenti. I dati sono stati registrati e sono state eseguite statistiche descrittive. La maggior parte dei cavalli (71 su 115 risposte disponibili, 62%) aveva mostrato segni clinici di dermatite al pastorale almeno quattro volte nella sua vita. Un totale di 113 cavalli (92% di tutti i cavalli inclusi) erano affetti dalla dermatite al pastorale al momento dell’intervista. Per 37 cavalli (32%) su 114 risposte disponibili, i proprietari hanno consultato il loro veterinario solo dopo che il cavallo ha mostrato segni di dolore o di zoppia. Di solito, i trattamenti che non richiedevano una prescrizione medica sono stati applicati senza consultare il veterinario (ad esempio, solo il 9% (14 su 150 prescrizioni) delle creme topiche che promuovono la guarigione delle ferite sono state prescritte da un veterinario). Un totale di 31 decisioni di trattamento (55%) con creme contenenti agenti antibatterici, antimicotici e/o antinfiammatori e il 100% dei farmaci sistemici contenenti agenti antibatterici, antiparassitari o antinfiammatori sono stati prescritti da veterinari. Nel complesso, il 69% delle decisioni di trattamento sono state prese senza consultare un veterinario, rendendo così più difficile determinare le cause alla base della dermatite al pastorale e rendendo il trattamento spesso più lungo e oneroso. Alfine di sensibilizzare i proprietari sulle possibili conseguenze di una consultazione veterinaria tardiva nel processo della malattia, dovrebbero essere prese in considerazione campagne di informazione specifiche per migliorare il benessere degli animali.

Rhinoviruses and allergens, such as house dust mite are major agents responsible for asthma exacerbations. The influence of pre-existing airway inflammation on the infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is largely unknown. We analyse mechanisms of response to viral infection in experimental in vivo rhinovirus infection in healthy controls and patients with asthma, and in in vitro experiments with house dust mite, rhinovirus and SARS-CoV-2 in human primary airway epithelium. Here, we show that rhinovirus infection in patients with asthma leads to an excessive RIG-I inflammasome activation, which diminishes its accessibility for type I/III interferon responses, leading to their early functional impairment, delayed resolution, prolonged viral clearance and unresolved inflammation in vitro and in vivo. Pre-exposure to house dust mite augments this phenomenon by inflammasome priming and auxiliary inhibition of early type I/III interferon responses. Prior infection with rhinovirus followed by SARS-CoV-2 infection augments RIG-I inflammasome activation and epithelial inflammation. Timely inhibition of the epithelial RIG-I inflammasome may lead to more efficient viral clearance and lower the burden of rhinovirus and SARS-CoV-2 infections.

An 8-year-old female spayed Labrador retriever was presented for the evaluation of severe weight loss 10 weeks after starting an immunomodulatory treatment, including prednisolone and cyclosporine, for meningoencephalitis of unknown origin. Plasma biochemistry analysis showed mild to moderate increases in liver enzyme activities and a moderate decrease in urea concentration. Abdominal ultrasound revealed mild hepatomegaly and a large gall bladder with unremarkable wall and content. Cholecystocentesis was performed and bile was examined both cytologically and by molecular methods, which revealed the presence of Enterocytozoon bieneusi. Treatment was initiated with albendazole but was discontinued due to the development of severe neutropenia. The medical management was subsequently changed to fenbendazole and the dog made a complete recovery. This report describes the first case of clinical manifestation and successful treatment of biliary E. bieneusi infection in a dog.

The sesquiterpene lactone artemisinin and its amino-artemisinin derivatives artemiside (GC008) and artemisone (GC003) are potent antimalarials. The mode of action of artemisinins against Plasmodium sp is popularly ascribed to 'activation' of the peroxide group by heme-Fe(II) or labile Fe(II) to generate C-radicals that alkylate parasite proteins. An alternative postulate is that artemisinins elicit formation of reactive oxygen species by interfering with flavin disulfide reductases resposible for maintaining intraparasitic redox homeostasis. However, in contradistinction to the heme-activation mechanism, the amino-artemisinins are effective in vitro against non-heme-degrading apicomplexan parasites including T. gondii, with IC 50 values of 50-70 nM, and induce distinct ultrastructural alterations. However, T. gondii strains readily adapted to increased concentrations (2.5 μM) of these two compounds within few days. Thus, T. gondii strains that were resistant against artemisone and artemiside were generated by treating the T. gondii reference strain ME49 with stepwise increasing amounts of these compounds, yielding the artemisone resistant strain GC003R and the artemiside resistant strain GC008R. Differential analyses of the proteomes of these resistant strains compared to the wildtype ME49 revealed that 215 proteins were significantly downregulated in artemisone resistant tachyzoites and only 8 proteins in artemiside resistant tachyzoites as compared to their wildtype. Two proteins, namely a hypothetical protein encoded by ORF TGME49_236950, and the rhoptry neck protein RON2 encoded by ORF TGME49_300100 were downregulated in both resistant strains. Interestingly, eight proteins involved in ROS scavenging including catalase and superoxide dismutase were amongst the differentially downregulated proteins in the artemisone-resistant strain. In parallel, ROS formation was significantly enhanced in isolated tachyzoites from the artemisone resistant strain and - to a lesser extent - in tachyzoites from the artemiside resistant strain as compared to wildtype tachyzoites. These findings suggest that amino-artemisinin derivatives display a mechanism of action in T. gondii distinct from Plasmodium.

Ichthyoses represent a heterogeneous group of cornification disorders that are associated with skin barrier defects. We investigated a 9-month-old Chihuahua showing excessive scale formation. Clinical and histopathological examinations revealed non-epidermolytic ichthyosis and a genetic defect was suspected. We therefore sequenced the genome of the affected dog and compared the data with 564 genetically diverse control genomes. Filtering for private variants identified a homozygous missense variant in SDR9C7, c.454C>T or p.(Arg152Trp). SDR9C7 is a known candidate gene for ichthyosis in humans and encodes the short-chain dehydrogenase/reductase family 9C member 7. The enzyme is involved in the production of a functional corneocyte lipid envelope (CLE), a crucial component of the epidermal barrier. Pathogenic variants in SDR9C7 have been described in human patients with autosomal recessive ichthyosis. We assume that the identified missense variant in the affected Chihuahua of this study impairs the normal enzymatic activity of SDR9C7 and thus prevents the formation of a functioning CLE, resulting in a defective skin barrier. To the best of our knowledge, this is the first report of a spontaneous SDR9C7 variant in domestic animals.

The complete genome sequence of Lactobacillus melliventris strain IBH004, isolated from the gut of a honeybee worker ( Apis mellifera ) and containing two plasmids and a temperate phage, was determined using hybrid assembly of Oxford Nanopore and Illumina reads. Phage-sequence relationships were identified from the coding sequences, and a proteomic tree was constructed.

The effects of decoquinate (DCQ) and three O-quinoline-carbamate-derivatives were investigated using human foreskin fibroblasts (HFF) infected with Neospora caninum tachyzoites. These compounds exhibited half-maximal proliferation inhibition (IC50s) from 1.7 (RMB060) to 60 nM (RMB055). Conversely, when applied at 5 (DCQ, RMB054) or 10µM (RMB055, RMB060), HFF viability was not affected. Treatments of infected cell cultures at 0.5µM altered the ultrastructure of the parasite mitochondrion and cytoplasm within 24 h, most pronounced for RMB060, and DCQ, RMB054 and RMB060 did not impair the viability of splenocytes from naïve mice. Long-term treatments of N. caninum-infected HFF monolayers with 0.5µM of each compound showed that only exposure to RMB060 over a period of six consecutive days had a parasiticidal effect, while the other compounds were not able to kill all tachyzoites in vitro. Thus, DCQ and RMB060 were comparatively assessed in the pregnant neosporosis mouse model. The oral application of these compounds suspended in corn oil at 10 mg/kg/day for 5 d resulted in a decreased fertility rate and litter size in the DCQ group, whereas reproductive parameters were not altered by RMB060 treatment. However, both compounds failed to protect mice from cerebral infection and did not prevent vertical transmission/pup mortality. Thus, despite the promising in vitro efficacy and safety characteristics of DCQ and DCQ-derivatives, proof of concept for activity against neosporosis could not be demonstrated in the murine model.

Darier disease is caused by heterozygous loss of function variants in the ATP2A2 gene encoding the endoplasmic/sarcoplasmic reticulum Ca2+ pump ATP2A2. Defective intracellular calcium signaling in the epidermis results in a loss of desmosomal adhesion and the development of characteristic skin lesions. In this study, we investigated a Shih Tzu that developed erythematous papules on the ventrum and, over time, the dorsal neck and a nodule in the right ear canal with secondary ear infection. Histopathologic examination demonstrated discrete foci of acantholysis affecting suprabasal layers of the epidermis. Whole genome sequencing of the affected dog identified a heterozygous missense variant, p.N809H, affecting an evolutionarily conserved amino acid residue of the ATP2A2 protein. The highly characteristic clinical and histopathologic findings together with a plausible variant in the only known functional candidate gene establish the diagnosis of canine Darier disease in the studied dog and highlight the potential of genetic analyses as complementary diagnostic approach in veterinary medicine.

OBJECTIVE To describe the clinical signs, electroencephalographic (EEG) findings, treatment, and outcome in a dog after successful resuscitation from out-of-hospital cardiopulmonary arrest (OHCA) induced by pentobarbital intoxication. CASE SUMMARY A 10-year-old, male intact Jack Russell Terrier was referred for management of refractory status epilepticus and presented dead on arrival. After 7 minutes of cardiopulmonary resuscitation, return of spontaneous circulation was achieved, but the dog remained comatose, apneic, and lacked brainstem reflexes on neurological examination 6 hours following resuscitation. Magnetic resonance imaging showed polioencephalomalacia consistent with prolonged epileptiform activity, and EEG was initially concerning for electrocerebral inactivity. Following supportive care that included short-term mechanical ventilation, the dog made a full recovery and was discharged from the hospital alive 7 days postresuscitation. It was later revealed that the dog had been administered an unknown amount of pentobarbital during transportation, which likely contributed to the OHCA, clinical, and EEG findings. NEW INFORMATION PROVIDED This is the first report to describe the full recovery and hospital discharge of a dog suffering OHCA and the first description of EEG findings in a clinical veterinary patient following cardiopulmonary arrest and successful resuscitation. Factors likely contributing to successful patient outcome and potential benefits and limitations of EEG in monitoring postcardiac arrest patients are discussed.

Background Microbial communities in recirculating aquaculture systems (RAS) play a role in system success, nutrient cycling, and water quality. Considering the increasing socio-economic role of fish farming, e.g., regarding food security, an in-depth understanding of aquaculture microbial communities is also relevant from a management perspective, especially regarding the growth, development, and welfare of the farmed animal. However, the current data on the composition of microbial communities within RAS is patchy, which is partly attributable to diverging method choices that render comparative analyses challenging. Therefore, there is a need for accurate, standardized, and user-friendly methods to study microbial communities in aquaculture systems. Results We compared sequencing approach performances (3 types of 16S short amplicon sequencing, PacBio long-read amplicon sequencing, and amplification-free shotgun metagenomics) in the characterization of microbial communities in two commercial RAS fish farms. Results showed that 16S primer choice and amplicon length affect some values (e.g., diversity measures, number of assigned taxa or distinguishing ASVs) but have no impact on spatio-temporal patterns between sample types, farms and time points. This implies that 16S rRNA approaches are adequate for community studies. The long-read amplicons underperformed regarding the quantitative resolution of spatio-temporal patterns but were suited to identify functional services, e.g., nitrification cycling and the detection of pathogens. Finally, shotgun metagenomics extended the picture to fungi, viruses, and bacteriophages, opening avenues for exploring inter-domain interactions. All sequencing datasets agreed on major prokaryotic players, such as Actinobacteriota, Bacteroidota, Nitrospirota, and Proteobacteria. Conclusion The different sequencing approaches yielded overlapping and highly complementary results, with each contributing unique data not obtainable with the other approaches. We conclude that a tiered approach constitutes a strategy for obtaining the maximum amount of information on aquaculture microbial communities and can inform basic research on community evolution dynamics. For specific and/or applied questions, single-method approaches are more practical and cost-effective and could lead to better farm management practices.

Canine distemper virus (CDV) is an enveloped RNA morbillivirus that triggers respiratory, enteric, and high incidence of severe neurological disorders. CDV induces devastating outbreaks in wild and endangered animals as well as in domestic dogs in countries associated with suboptimal vaccination programs. The receptor-binding tetrameric attachment (H)-protein is part of the morbilliviral cell entry machinery. Here, we present the cryo-electron microscopy (cryo-EM) structure and supramolecular organization of the tetrameric CDV H-protein ectodomain. The structure reveals that the morbilliviral H-protein is composed of three main domains: stalk, neck, and heads. The most unexpected feature was the inherent asymmetric architecture of the CDV H-tetramer being shaped by the neck, which folds into an almost 90° bent conformation with respect to the stalk. Consequently, two non-contacting receptor-binding H-head dimers, which are also tilted toward each other, are located on one side of an intertwined four helical bundle stalk domain. Positioning of the four protomer polypeptide chains within the neck domain is guided by a glycine residue (G158), which forms a hinge point exclusively in two protomer polypeptide chains. Molecular dynamics simulations validated the stability of the asymmetric structure under near physiological conditions and molecular docking showed that two receptor-binding sites are fully accessible. Thus, this spatial organization of the CDV H-tetramer would allow for concomitant protein interactions with the stalk and head domains without steric clashes. In summary, the structure of the CDV H-protein ectodomain provides new insights into the morbilliviral cell entry system and offers a blueprint for next-generation structure-based antiviral drug discovery.

Background Intervertebral disc disease (IVDD) is a common diagnosis and well-investigated pathological condition in French Bulldogs with neurological deficiencies. However there is currently only one recently published retrospective descriptive study looking for recurrence rates of IVDD in French Bulldogs. Medical reports of French Bulldogs with a first episode of IVDD and surgical treatment were evaluated and reviewed for clinical signs of recurrence. Risk factors for Total-Recurrence, Cervical- and Thoracolumbar-Recurrence were evaluated by means of logistic regression models. The aim of this study was to assess frequency and risk factors associated with the recurrence of IVDD in French Bulldogs. Results One hundred twenty-seven French Bulldogs with a first episode of IVDD and surgical treatment were evaluated. 52.7% (67/127) of these patients showed signs of recurrence. The recurrence rate in the cervical spine was slightly lower (47%) compared to the thoracolumbar spine (56.6%). A significant association with recurrence could be found for the factor age: French Bulldogs with a first episode of IVDD ≤ 3 years seem to be prone for Total-Recurrence (P = 0.002) and Cervical-Recurrence (with ORs ranging from 0.02 to 0.03 for patients older than 3 years). 50% of the recurrences (median) occurred within the first 12 month after the first episode of IVDD. Conclusions Recurrence of IVDD can be expected in more than half of French Bulldogs affected by IVDD. Especially young French Bulldogs are prone for recurrence in cervical spine. Almost every fourth patient with IVDD suffers from a recurrence within 12 months. Future dog owners should be informed about the risk of IVDD and the early onset of recurrences in French Bulldogs.

Reduction of risk factors for bovine digital dermatitis (BDD) is crucial in current disease control. However, risk factors that might arise especially in mountainous regions are unknown until now, and an adapted BDD control program is consequently missing. The objective of this observational case-control study was to identify farm-level risk factors for BDD in dairy herds in mountainous regions. To investigate predictors for the occurrence of BDD, 100 farms were visited and information about herd characteristics and management practices, potentially relevant explanatory variables for either introduction or establishment of BDD, were gathered by completing a questionnaire with the farmer or herd manager. Within-herd prevalences of BDD assessed during 3 routine claw trimmings with an interval of 6 mo before the survey were used to define cases (BDD within-herd prevalence of ≥26% during each claw trimming) and controls (no BDD cases in each of the 3 routine claw trimmings before the survey). Data were analyzed using 2 separate binomial generalized linear models according to either establishment or introduction of BDD. After prescreening, 15 of 23 explanatory variables were included in the final analysis, which showed 3 variables related to introduction and establishment, each being significantly associated with the occurrence of BDD within a farm. Results of model 1 (i.e., aspects related to BDD introduction) revealed that access to mountain pastures during the summer season (odds ratio, 95% confidence interval: 0.12, 0.04-0.35), participation in dairy shows (0.32, 0.11-0.94), and the number of new animals introduced into the farm during the last 2 yr (1.28, 1.12-1.52) were significantly associated with the occurrence of BDD. Model 2 (i.e., aspects related to BDD establishment) showed that cows kept in freestalls were at higher risk for BDD compared with those kept in tiestalls (20.65, 1.59-649.37). Furthermore, number of days between diagnosis and treatment of a BDD lesion (10.31, 3.55-81.21) and the amount of concentrate feeding (median 5 kg) per cow and day (7.72, 2.46-6.47) were positively associated with BDD occurrence. In conclusion, the findings of this study provide a set of risk factors that are associated with BDD status within herds in mountainous regions. These results may help in development of adapted control programs for BDD in dairy cows.

A 3.5-year-old female cheetah (Acinonyx jubatus) died after a 10-day history of anorexia, regurgitation and diarrhoea despite symptomatic therapy. At gross post-mortem examination, the stomach was blood-filled with mucosal thickening and multifocal ulcerations. The intestinal mucosa was thickened and reddened, and the intestinal lumen was filled with dark red to black pasty content. Gastric histological lesions were compatible with gastritis due to Helicobacter infection, which was confirmed by polymerase chain reaction. Histology of the intestines revealed a severe necrotizing neutrophilic enterocolitis with abundant intralesional curved to spiral bacteria, corresponding to Campylobacter jejuni, which were subsequently isolated from both small and large intestinal contents. No other intestinal pathogens were detected despite thorough investigations. These findings suggest that C. jejuni may have played an aetiological role in the enterocolitis. Such an association has not been previously reported in non-domestic felids.

Although selection for increased milk production traits has led to a genetic increase in body weight (BW), the genetic gain in milk production has exceeded the gain in BW, so gross feed efficiency has improved. Nonetheless, greater gains may be possible by directly selecting for a measure of feed efficiency. Australia first introduced Feed Saved (FS) estimated breeding value (EBV) in 2015. Feed Saved combines residual feed intake (RFI) genomic EBV and maintenance requirements calculated from mature BW EBV. The FS EBV was designed to enable the selection of cows for reduced energy requirements with similar milk production. In this study, we used a reference population of 3,711 animals in a multivariate analysis including Australian heifers (AUSh), Australian cows (AUSc), and overseas cows (OVEc) to update the Australian EBV for lifetime RFI (i.e., a breeding value that incorporated RFI in growing and lactating cows) and to recalculate the FS EBV in Australian Holstein bulls (AUSb). The estimates of genomic heritabilities using univariate (only AUSc or AUSh) to trivariate (including the OVEc) analyses were similar. Genomic heritabilities for RFI were estimated as 0.18 for AUSc, 0.27 for OVEc, and 0.36 for AUSh. The genomic correlation for RFI between AUSc and AUSh was 0.47 and that between AUSc and OVEc was 0.94, but these estimates were associated with large standard errors (range: 0.18-0.28). The reliability of lifetime RFI (a component of FS) in the trivariate analysis (i.e., including OVEc) increased from 11% to 20% compared with the 2015 model and was greater, by 12%, than in a bivariate analysis in which the reference population included only AUSc and AUSh. By applying the prediction equation of the 2020 model, the average reliability of the FS EBV in 20,816 AUSb that were born between 2010 and 2020 improved from 33% to 43%. Previous selection strategies-that is, using the predecessor of the Balanced Performance Index (Australian Profit Ranking index) that did not include FS-have resulted in an unfavorable genetic trend in FS. However, this unfavorable trend has stabilized since 2015, when FS was included in the Balanced Performance Index, and is expected to move in a favorable direction with selection on Balanced Performance Index or the Health Weighted Index. Doubling the reference population, particularly by incorporating international data for feed efficiency, has improved the reliability of the FS EBV. This could lead to increased genetic gain for feed efficiency in the Australian industry.

Background Meningoencephalitis of unknown origin (MUO) is an inflammatory disease of the canine central nervous system (CNS) that shares several features with multiple sclerosis (MS) in humans. In approximately 95% of MS patients, ≥ two immunoglobulin G (IgG) oligoclonal bands (OCBs) are detectable exclusively in the cerebrospinal fluid (CSF). Hypothesis/objectives To investigate OCBs in CSF and serum in dogs affected by MUO, intervertebral disc disease (IVDD), idiopathic epilepsy (IE), intracranial neoplasia (IN), steroid-responsive meningitis-arteritis (SRMA), and diseases outside the CNS. We hypothesize that the highest prevalence of CSF-specific OCBs (≥ two OCBs uniquely in the CSF) would be found in dogs affected by MUO. Animals Client-owned dogs (n = 121) presented to the neurology service due to neurological deficits. Methods Prospective study. Measurement of IgG concentration in CSF and serum via a canine IgG ELISA kit. OCB detection via isoelectric focusing (IEF) and immunoblot. Results Presence of CSF-specific OCBs was significantly higher in dogs with MUO (57%) compared to 22% in IN, 6% in IE, 15% in SRMA, 13% in IVDD, and 0% in the non-CNS group (p < .001). Dogs with MUO were 9.9 times more likely to show CSF-specific OCBs than all other diseases together (95% confidence interval, 3.7–26.4; p < .001). Conclusions and clinical importance MUO showed the highest prevalence of CSF-specific OCBs, indicating an inflammatory B cell response. Future studies are needed to evaluate the prevalence in the specific MUO subtypes and a possible similarity with human MS.

While pathogens of the eye have been studied for a very long time, the existence of resident microbes on the surface of healthy eyes has gained interest only recently. It appears that commensal microbes are a normal feature of the healthy eye, whose role and properties are currently the subject of extensive research. This review provides an overview of studies that have used 16s rRNA gene sequencing and whole metagenome shotgun sequencing to characterize microbial communities associated with the healthy ocular surface from kingdom to genus level. Bacteria are the primary colonizers of the healthy ocular surface, with three predominant phyla: Proteobacteria, Actinobacteria, and Firmicutes, regardless of the host, environment, and method used. Refining the microbial classification to the genus level reveals a highly variable distribution from one individual and study to another. Factors accounting for this variability are intriguing - it is currently unknown to what extent this is attributable to the individuals and their environment and how much is artifactual. Clearly, it is technically challenging to accurately describe the microorganisms of the ocular surface because their abundance is relatively low, thus, permitting substantial contaminations. More research is needed, including better experimental standards to prevent biases, and the exploration of the ocular surface microbiome's role in a spectrum of healthy to pathological states. Outcomes from such research include the opportunity for therapeutic interventions targeting the microbiome.

Eight novel carbohydrate-tethered trithiolato dinuclear ruthenium(II)-arene complexes were synthesized using CuAAC ‘click’ (Cu(I)-catalyzed azide-alkyne cycloaddition) reactions, and there in vitro activity against transgenic T. gondii tachyzoites constitutively expressing β-galactosidase (T. gondii β-gal) and in non-infected human foreskin fibroblasts, HFF, was determined at 0.1 and 1 µM. When evaluated at 1 µM, seven diruthenium-carbohydrate conjugates strongly impaired parasite proliferation by >90%, while HFF viability was retained at 50% or more, and they were further subjected to the half-maximal inhibitory concentration (IC50) measurement on T. gondii β-gal. Results revealed that the biological activity of the hybrids was influenced both by the nature of the carbohydrate (glucose vs. galactose) appended on ruthenium complex and the type/length of the linker between the two units. 23 and 26, two galactose-based diruthenium conjugates, exhibited low IC50 values and reduced effect on HFF viability when applied at 2.5 µM (23: IC50 = 0.032 µM/HFF viability 92% and 26: IC50 = 0.153 µM/HFF viability 97%). Remarkably, compounds 23 and 26 performed significantly better than the corresponding carbohydrate non-modified diruthenium complexes, showing that this type of conjugates are a promising approach for obtaining new antiparasitic compounds with reduced toxicity.

Interdigital hyperplasia (IH) is a fold of fibrous tissue protruding into the interdigital space that rarely occurs in sheep. Interdigital hyperplasia secondary infected with bovine digital dermatitis (BDD) treponemes has been reported in cattle in the course of the increasing spread of classical BDD lesions. In this report, we describe proliferative/ulcerative interdigital lesions associated with contagious ovine digital dermatitis (CODD) treponemes and clinically scored as (IH+CODD), occurring in both hind limbs of a ram and the left hindlimb of a ewe. Both cases exhibited epidermal hyperplasia, parakeratosis and focal-extensive areas of epidermal necrosis with numerous infiltrating neutrophils. Treponema PCR and fluorescence in situ hybridization (FISH) were positive for Treponema phylotype 1 (PT1). In addition, Dichelobacter (D.) nodosus and Porphyromonas (P.) levii were detected in the biopsy by PCR. In three slaughter sheep, without claw lesions, which were kept together with both affected sheep, Treponema spp. were detected neither with PCR nor FISH; the PCRs for D. nodosus and P. levii were also negative. Complete clinical healing occurred in the ewe within 6 weeks after three local applications of a chlortetracycline spray in 2 weeks intervals. This report is the first description of IH+CODD in sheep as demonstrated by a combination of histopathological and molecular analyses.

Desmosome diseases are caused by dysfunction of desmosomes, which anchor intermediate filaments (IFs) at sites of cell–cell adhesion. For many decades, the focus of attention has been on the role of actin filament-associated adherens junctions in development and disease, especially cancer. However, interference with the function of desmosomes, their molecular constituents or their attachments to IFs has now emerged as a major contributor to a variety of diseases affecting different tissues and organs including skin, heart and the digestive tract. The first Alpine desmosome disease meeting (ADDM) held in Grainau, Germany, in October 2022 brought together international researchers from the basic sciences with clinical experts from diverse fields to share and discuss their ideas and concepts on desmosome function and dysfunction in the different cell types involved in desmosome diseases. Besides the prototypic desmosomal diseases pemphigus and arrhythmogenic cardiomyopathy, the role of desmosome dysfunction in inflammatory bowel diseases and eosinophilic esophagitis was discussed.

BACKGROUND Hyperlipasemia has been reported in dogs with acute kidney injury (AKI) but associations with AKI severity, hemodialysis (HD) treatment, and outcome have not been extensively evaluated. OBJECTIVES Investigate the prevalence and clinical relevance of hyperlipasemia in dogs with AKI, treated with and without HD. ANIMALS Client-owned dogs (n = 125) with AKI. METHODS Retrospective data extraction from medical records, including signalment, cause of AKI, duration of hospitalization, survival, plasma creatinine concentration, and 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6'-methyresorufin) ester (DGGR) lipase activity at admission and throughout hospitalization. RESULTS A DGGR-lipase activity >3× the upper reference limit (URL) was found in 28.8% and 55.4% of dogs at admission and during hospitalization, respectively, but only 8.8% and 14.9% of dogs, respectively, were diagnosed with acute pancreatitis. Hyperlipasemia >10 × URL was observed in 32.7% of dogs during hospitalization. The DGGR-lipase activity was higher in dogs with International Renal Interest Society (IRIS) Grades 4-5 than Grades 1-3, but correlation between DGGR-lipase activity and creatinine concentration was poor (rs = .22; 95% confidence intervals [CI], 0.04-0.38). Treatment with HD was not associated with DGGR-lipase activity independent of IRIS grade. Survival to discharge and 30 days after admission was 65.6% and 59.6%, respectively. High IRIS grades (P = .03) and high DGGR-lipase activity at admission (P = .02) and during hospitalization (P = .003) were associated with nonsurvival. CONCLUSIONS AND CLINICAL IMPORTANCE Hyperlipasemia is frequent and often marked in dogs with AKI despite only a minority being diagnosed with pancreatitis. Hyperlipasemia is associated with AKI severity but not independently with HD treatment. High IRIS grade and hyperlipasemia were associated with nonsurvival.

Epilepsy is a challenging multifactorial disorder with a complex genetic background. Our current understanding of the pathophysiology and treatment of epilepsy has substantially increased due to animal model studies, including canine studies, but additional basic and clinical research is required. Drug-resistant epilepsy is an important problem in both dogs and humans, since seizure freedom is not achieved with the available antiseizure medications. The evaluation and exploration of pharmacological and particularly non-pharmacological therapeutic options need to remain a priority in epilepsy research. Combined efforts and sharing knowledge and expertise between human medical and veterinary neurologists are important for improving the treatment outcomes or even curing epilepsy in dogs. Such interactions could offer an exciting approach to translate the knowledge gained from people and rodents to dogs and vice versa. In this article, a panel of experts discusses the similarities and knowledge gaps in human and animal epileptology, with the aim of establishing a common framework and the basis for future translational epilepsy research.

Avian haemosporidian parasites are widespread and infect birds from a broad variety of avian families with diverse consequences ranging from subclinical infections to severe and fatal disease. This study aimed to determine the occurrence and diversity of avian haemosporidia including associated clinical signs and pathomorphological lesions in captive and free-ranging, wild birds from two zoos and the near environment in Switzerland. Blood samples from 475 birds, including 230 captive and 245 free-ranging, wild individuals belonging to 42 different avian species from 15 orders were examined for the presence of avian haemosporidian DNA by a one-step multiplex PCR designed to simultaneously detect and discriminate the genera Plasmodium, Haemoproteus and Leucocytozoon by targeting mitochondrial genome sequences. Positive samples were additionally tested using a nested PCR targeting the cytochrome b gene of Plasmodium and Haemoproteus. The obtained amplicons were bidirectionally sequenced. This study revealed haemosporidian DNA in 42 samples, belonging to ten host species. The most commonly detected lineage was Plasmodium relictum SGS1, which was identified in 29 birds (Phoenicopterus roseus: n = 24, Alectoris graeca: n = 1, Lamprotornis superbus: n = 1, Somateria mollissima: n = 1, Spheniscus demersus: n = 1, Tetrao urogallus crassirostris: n = 1), followed by Haemoproteus sp. STRURA03 in six avian hosts (Bubo bubo: n = 5, Bubo scandiacus = 1), Plasmodium relictum GRW11 in four individuals (Phoenicopterus roseus: n = 3, Spheniscus demersus: n = 1) and Plasmodium elongatum GRW06 in one Alectura lathami lathami. A Phalacrocorax carbo was infected with Plasmodium relictum, but the exact lineage could not be determined. One mixed infection with P. relictum and Haemoproteus sp. was detected in a Bubo scandiacus. Only five individuals (Spheniscus demersus: n = 2, Somateria mollissima: n = 1, Bubo scandiacus: n = 1, Alectoris graeca: n = 1) showed clinical and pathomorphological evidence of a haemosporidian infection.

There is scant literature on primary nonhematopoietic malignant liver tumors (PMLT) in cats. In this retrospective study, medical data of 40 cats diagnosed with PMLT were reviewed over a period of 22 years (2000-2021). The most frequent epithelial tumors were hepatocellular (42.5%) and bile duct carcinomas (32.5%), only six (15%) cats had mesenchymal tumors. The median age was 13 years and clinical signs commonly included ano-/hyporexia (62.5%), apathy/lethargy (52.5%), weight loss (42.5%) and vomiting (35%). At initial diagnosis, metastases were confirmed in 1 (2.5%) and suspected in three (7.5%) cats. Massive was the most frequent morphology (75%). Most intrahepatic tumors were left-sided (54.2%) with the left medial lobe being primarily affected (25%). Extrahepatic tumors were rare (5%). In 34 (85%) cats, liver lobectomy was performed (surgery group), four (10%) were treated palliatively (non-surgery group), and two (5%) received no treatment. Intraoperative complications occurred in 11.8% with four (15.4%) postoperative deaths. Recurrence was detected in 28.6% at a median of 151 days (range, 79-684 days), while postoperative metastases were suspected in 21.4% at a median of 186 days (range, 79-479 days). The median survival time (MST) was significantly longer in cats of the surgery group (375 days) than in the non-surgery group (16 days) (P = 0.002). MST was 868 days for hepatocellular compared to 270 days for bile duct carcinomas (P = 0.06). In summary, liver lobectomy is associated with prolonged survival times and good prognosis in cats with hepatocellular, and an acceptable prognosis in cats with bile duct carcinoma.

Early and strong interferon type I (IFN-I) responses are usually associated with mild COVID-19 disease, whereas persistent or unregulated proinflammatory cytokine responses are associated with severe disease outcomes. Previous work suggested that monocyte-derived macrophages (MDMs) are resistant and unresponsive to SARS-CoV-2 infection. Here, we demonstrate that upon phagocytosis of SARS-CoV-2-infected cells, MDMs are activated and secrete IL-6 and TNF. Importantly, activated MDMs in turn mediate strong activation of plasmacytoid dendritic cells (pDCs), leading to the secretion of high levels of IFN-α and TNF. Furthermore, pDC activation promoted IL-6 production by MDMs. This kind of pDC activation was dependent on direct integrin-mediated cell‒cell contacts and involved stimulation of the TLR7 and STING signaling pathways. Overall, the present study describes a novel and potent pathway of pDC activation that is linked to the macrophage-mediated clearance of infected cells. These findings suggest that a high infection rate by SARS-CoV-2 may lead to exaggerated cytokine responses, which may contribute to tissue damage and severe disease.

Endometritis is a leading cause of sub- and infertility in domestic animal species. The healthy uterus is colonized by commensal bacteria, viruses, and yeast/fungi that represent the non-pathogenic microbiota. A shift in the number or type of organisms accompanied by immune dysfunction, however, may trigger uterine infection and inflammation. Metritis is associated with inflammation of all uterine layers (endometrium, myometrium, and perimetrium) whereas endometritis is a more superficial inflammation involving solely the endometrium. Endometritis generally occurs at two time points in domestic animal species, post-partum and post-mating. Post-partum endometritis may chronically persist, either as a low-grade disease that often manifests as a vaginal discharge but not a systemic illness (in some species termed clinical endometritis) or sometimes subclinical where features are only detected by endometrial sampling. Contamination of the uterus at the time of mating occurs by direct deposition of semen (ejaculated or artificially inseminated) into the uterus. Improper drainage of the ejaculatory fluid or an inadequate immune response may result in persistent mating-induced endometritis. Both post-partum and post-mating endometritis interferes with fertility by creating a suboptimal environment for embryo development and placentation and chronic endometritis may have an impact on sperm survival and fertilisation ability. In the post-partum animal, there may also be changes in milk production and maternal behaviour which can affect offspring health and survival. Preventive strategies for endometritis largely depend on monitoring their known risk factors, which are sometimes specific with regard to the species. Effective, non-antibiotic therapy for endometritis is not available to date. Overall, extensive research has been performed in cattle and horses to unravel key aspects of endometritis, but in sows and bitches, the available literature is scant. Thus, the need and opportunity to investigate the condition vary considerably among domestic species and necessitate their comparative assessment. This article reviews general and comparative aspects of the diagnosis and classification, pathogenesis, preventive strategies, and therapeutics of endometritis in domestic species with a specific focus on cows, mares, sows, and bitches.

Energy and nutrient deficiency in dairy cows in early lactation is considered to contribute to their increased susceptibility to mastitis. We have tested the hypothesis that feeding diets with high contents of either nitrogenic, glucogenic, or lipogenic components in early lactation affects both the endocrine and metabolic status, as well as the mammary immune competence. After calving, cows were fed increasing amounts of concentrate up to 10 kg/d rich in crude protein (nitrogenic, n = 10), glucogenic precursors (glucogenic, n = 11), or lipids (lipogenic, n = 11). In wk 3, one udder quarter was challenged with lipopolysaccharide (LPS) from Escherichia coli. Blood and milk were sampled on the day before LPS challenge (d -1), and on d 0, 1, 2, 3, and 9 after LPS challenge. On the day of LPS challenge additional samples were taken hourly for quarter milk and every 3 h for blood. Urea concentrations were higher in plasma and milk of cows fed the nitrogenic diet. However, plasma concentrations of glucose, cholesterol, triglycerides, β-hydroxybutyrate, nonesterified fatty acids, as well as insulin, glucagon, and insulin-like growth factor-1 were not affected by the different diets. The mammary immune challenge induced a substantial increase of somatic cell count (SCC) in the treated quarter, and a transient decrease of total milk yield and white blood cells similar in all diet groups for one day. The absolute phagocytosis of blood leukocytes was decreased; however, the phagocytosis per cell was increased in glucogenic-fed cows at 6 h after LPS challenge. During mammary inflammation an insulin resistance, shown by increased plasma glucose, insulin, and glucagon, developed similarly in all diet groups. β-hydroxybutyrate and nonesterified fatty acids were decreased at 1 d after LPS challenge in glucogenic-fed cows only. Cholesterol did not change, and triglycerides only decreased significantly in lipogenic-fed cows 6 h after challenge. On d 9 after LPS challenge, SCC and milk yield and metabolic factors were recovered in all groups. In conclusion, the endocrine and metabolic situation, and the immune response to intramammary LPS of dairy cows during early lactation was not substantially influenced by the elevated supply of nitrogenic, glucogenic, or lipogenic components due to the provided feed in this study.

International audience; During the course of the infectious disease alveolar echinococcosis (AE), the larval stage of Echinococcus multilocularis develops in the liver, where an initial Th1/Th17 immune response may allow its elimination in resistant individuals. In patients susceptible to infection and disease, the Th2 response initiates later, inducing tolerance to the parasite. The role of interleukin 33 (IL-33), an alarmin released during necrosis and known to drive a Th2 immune response, has not yet been described during AE. Wild-type (WT) and IL-33(-/-) C57BL/6J mice were infected by peritoneal inoculation with E. multilocularis metacestodes and euthanized 4 months later, and their immune response were analyzed. Immunofluorescence staining and IL-33 enzyme-linked immunosorbent assay (ELISA) were also performed on liver samples from human patients with AE. Overall, metacestode lesions were smaller in IL-33(-/-) mice than in WT mice. IL-33 was detected in periparasitic tissues, but not in mouse or human serum. In infected mice, endogenous IL-33 modified peritoneal macrophage polarization and cytokine profiles. Th2 cytokine concentrations were positively correlated with parasite mass in WT mice, but not in IL-33(-/-) mice. In human AE patients, IL-33 concentrations were higher in parasitic tissues than in distant liver parenchyma. The main sources of IL-33 were CD31(+) endothelial cells of the neovasculature, present within lymphoid periparasitic infiltrates together with FOXP3(+) T(reg)s. In the murine model, periparasitic IL-33 correlated with accelerated parasite growth putatively through the polarization of M2-like macrophages and release of immunosuppressive cytokines IL-10 and transforming growth factor β1 (TGF-β1). We concluded that IL-33 is a key alarmin in AE that contributes to the tolerogenic effect of systemic Th2 cytokines. IMPORTANCE Infection with the metacestode stage of Echinococcus multilocularis, known as alveolar echinococcosis, is the most severe cestodosis worldwide. However, less than 1% of exposed individuals, in which the immune system is unable to control the parasite, develop the disease. The factors responsible for this interindividual variability are not fully understood. In this in vivo study comparing wild-type and IL-33(-/-) infected mice, together with data from human clinical samples, we determined that IL-33, an alarmin released following tissue injury and involved in the pathogenesis of cancer and asthma, accelerates the progression of the disease by modulating the periparasitic microenvironment. This suggests that targeting IL-33 could be of interest for the management of patients with AE, and that IL-33 polymorphisms could be responsible for increased susceptibility to AE.

BackgroundHernia formation is a well-known abdominal wall disorder in calves and most often occurs in the umbilical region. In addition, it occurs in the abdominal wall outside the umbilical region. It has been reported to involve acquired factors, such as external force to the lower or lateral abdominal wall, trauma, muscle weakness, and, although rare, congenital or hereditary factors. Although there have been reports on the repair of abdominal wall hernias caused by abnormal muscle formation, there have been no reports on the treatment of abdominal wall hernias caused by abnormal vertebral and rib formation in cattle or other ruminants. In this study, for the first time, we describe a case of successful closure of a hernia in the left flank caused by malformation of the vertebrae and ribs. The repair was performed by narrowing the ribs using a surgical wire and covering the defect with a free omental graft.Case presentationA male Japanese Black calf showed a mild bulge of the left abdominal wall and abnormal morphology of the left ribs immediately after birth. At 3 months of age, computed tomography revealed fusion of the 9th, 10th, and 11th thoracic vertebrae and missing formation of the 10th and 11th left ribs at the thoracic vertebral fusion site. Additionally, a 15.2 × 24.4 cm abdominal hernia had formed in the abdominal wall. During surgery, the ribs forming the hernia were narrowed with a surgical wire, and the hernia was covered with a large autologous free omental graft to prevent intra-abdominal organ prolapse. At the monthly follow-up (11 months after surgery), the calf had developed similarly to other calves of the same age on the farm, and no complications were noted.ConclusionsThe current case shows that a combination of rib correction using a surgical wire combined with transplantation of a free autologous greater omentum graft was shown to be a possible treatment option for surgical repair of large abdominal wall hernias caused by rib malformation.

Brucellosis, Rift Valley fever (RVF) and Q fever are zoonoses prevalent in many developing countries, causing a high burden on human and animal health. Only a few studies are available on these among agro-pastoralist communities and their livestock in Chad. The objective of our study was to estimate brucellosis, RVF and Q fever seroprevalence among Chadian agro-pastoralist communities and their livestock, and to investigate risk factors for seropositivity. We conducted a multi-stage cross-sectional serological survey in two rural health districts, Yao and Danamadji (966 human and 1041 livestock (cattle, sheep, goat and equine) samples)). The true seroprevalence were calculated applying a Bayesian framework to adjust for imperfect diagnostic test characteristics and accounting for clustering in the study design. Risk factors for each of the zoonotic diseases were estimated using mixed effects logistic regression models. The overall prevalence for brucellosis, Q fever and RVF combined for both regions was estimated at 0.2% [95% credibility Interval: 0-1.1], 49.1% [%CI: 38.9-58.8] and 28.1% [%CI: 23.4-33.3] in humans, and 0.3% [%CI: 0-1.5], 12.8% [%CI: 9.7-16.4] and 10.2% [%CI: 7.6-13.4] in animals. Risk factors correlating significantly with the respective disease seropositivity were sex for human brucellosis, sex and Q fever co-infection for animal brucellosis, age for human Q fever, species and brucellosis co-infection for animal Q fever, age and herd-level animal RVF seroprevalence within the same cluster for human RVF, and cluster-level human RVF seroprevalence within the same cluster for animal RVF. In Danamadji and Yao, Q fever and RVF are notably seroprevalent among agro-pastoralist human and animal communities, while brucellosis appears to have a low prevalence. Correlation between the seroprevalence between humans and animals living in the same communities was detected for RVF, highlighting the interlinkage of human and animal transmissible diseases and of their health, highlighting the importance of a One Health approach.

BackgroundGastric dilatation volvulus (GDV) can lead to organ failure including acute kidney injury (AKI). Due to its cytoprotective, antioxidant and anti-inflammatory effects, lidocaine has a potential to prevent AKI in dogs with GDV.Design and settingProspective, observational cohort study in client-owned dogs with GDV.ObjectiveTo determine concentrations of renal biomarkers for AKI in dogs with GDV with and without intravenous (IV) lidocaine therapy.MethodsThirty-two dogs were randomized to receive either IV lidocaine (2 mg/kg, followed by a lidocaine constant rate infusion at a dose of 50 μg/kg/min over 24 h; n = 17) or no lidocaine (n = 15). Blood and urine samples were taken at admission (T0) (only blood), during or immediately after surgery (T1), and 24 (T24) and 48 (T48) h after surgery. Plasma creatinine (pCr), plasma neutrophil gelatinase-associated lipocalin (pNGAL), urinary NGAL (uNGAL), uNGAL to creatinine ratio (UNCR), and urinary gamma-glutamyl transferase to creatinine ratio (uGGT/uCr) were evaluated. Biomarker concentrations were compared between dogs with and without IV lidocaine and the course of each marker was determined in comparison to its admission value.ResultsIn the entire population, a significantly higher pCr at T0 (median, 95 μmol/L, interquartile range, 82–105) compared with T1 (69 μmol/L, 60–78), T24 (63 μmol/L, 52–78), and T48 (78 μmol/L, 65–87) (P < 0.001) was found. Plasma NGAL increased significantly between T0 (5.66 ng/mL, 3.58–7.43) and T24 (7.50 ng/mL, 4.01–11.89) (P = 0.006) and T48 (9.86 ng/mL, 5.52–13.92) (P < 0.001), respectively. Urinary NGAL increased significantly between T1 (0.61 ng/mL, 0.30–2.59) and T24 (2.62 ng/mL, 1.86–10.92) (P = 0.001) and T48 (4.79 ng/mL, 1.96–34.97 (P < 0.001), respectively. UNCR increased significantly between T1 (0.15 μg/mmol, 0.09–0.54) and T24 (1.14 μg/mmol, 0.41–3.58) (P = 0.0015) and T48 (1.34 μg/mmol, 0.30–7.42) (P < 0.001), respectively. Concentrations of uGGT/uCr increased significantly from T0 highest at T24 (6.20 U/mmol, 3.90–9.90) and significantly decreased at T48 (3.76 U/mmol, 2.84–6.22) (P < 0.001). No significant differences in any renal biomarker concentration were found between dogs with and without IV lidocaine therapy.Conclusion and clinical relevancePlasma NGAL, uNGAL and UNCR remained increased up to 48 h post-surgery. No evidence of lidocaine-associated renoprotection was found.

Swine dysentery and porcine intestinal spirochaetosis caused by Brachyspira (B.) hyodysenteriae and B. pilosicoli, respectively, are important diseases in swine production worldwide. The aim of this study was to assess the prevalence of both pathogens in farms with a history of diarrhoea within the last 12 months in Denmark, France, Germany, the Netherlands, Spain, and United Kingdom. In addition, risk factors for their prevalence and correlations between presence of different Brachyspira spp. and Lawsonia intracellularis were investigated. Therefore, faecal samples of 6355 nursery to finishing pigs out of 144 herds were sampled in 2017/2018 during a prevalence study on Lawsonia intracellularis, followed by polymerase chain reaction analysis for Brachyspira spp. detection. Herd prevalence differed significantly between countries, from 4.2% to 45.8% for B. hyodysenteriae and 8.3-87.5% for B. pilosicoli, respectively (p

Understanding the links between environmental and wildlife elemental concentrations is key to help assess ecosystem functions and the potential effects of legacy pollutants. In this study, livers from 448 European badgers (Meles meles) collected across the English Midlands were used to investigate the relationship between elemental concentrations in topsoils and wildlife. Mean soil sample concentrations within 2 km of each badger, determined using data from the British Geological Survey's 'Geochemical Baseline Survey of the Environment', were compared to badger liver elemental concentrations, focusing primarily on Ag, As, Cd, Cr, Cu, K, Mn, Pb, Se, Zn. Generally, the badgers appeared to have elemental concentrations comparable with those published for other related animals, though Cu concentrations tended to be lower than expected. While there was no relationship between soil and badger liver concentrations for most biologically essential elements, biologically non-essential elements, specifically Pb, Cd, As, and Ag, were positively correlated between soil and badger livers. Lead and Cd, the elements with the strongest relationships between soils and badger livers, were primarily elevated in badgers collected in Derbyshire, a county with a millennia-long history of Pb mining and significant Pb and Cd soil pollution. Cadmium concentrations in badgers were also, on average, almost nine times higher than the local soil concentrations, likely due to Cd biomagnification in earthworms, a dietary staple of badgers. While badgers are good models for studying associations between soil and wildlife elemental concentrations, due to their diet, burrowing behaviours, and site fidelity, all flora and fauna local to human-modified environments could be exposed to and impacted by legacy pollutants.

Here, we present a protocol to analyze the T cell profiles of the neonatal ovine lung during respiratory syncytial virus (RSV) infection. The protocol delivers standardized multiparameter flow cytometry (FCM) analysis of CD4+, CD8+, regulatory, and γδ T cells isolated from lung, lymph nodes, and bronchoalveolar lavages (BALs). We detail the preparation of RSV and transtracheal inoculation of newborn lambs. We then describe tissue isolation and preparation of cell suspensions, followed by FCM acquisition to identify different T cell subsets. For complete details on the use and execution of this protocol, please refer to Démoulins et al. (2021).

The oviduct provides optimum physiological and biochemical milieu essential for successful fertilization, early embryo development and facilitates functional maturation of spermatozoa. A study has revealed that spermatozoa alters the gene expression in bovine oviductal epithelial cells (BOECs) remotely via bio-active particles, thus acting as a cue to the oviduct prior to their arrival. However, very little attention has been paid to the question of whether spermatozoa could alter the cargo of extracellular vesicles (EVs) derived from BOECs. Therefore, the aim of this study was to investigate the alterations in small non-coding RNAs in EVs cargo derived from BOECs when incubated with spermatozoa in contact and non-contact co-culture models. After 4 h of incubation the EVs were isolated from the conditioned media, followed by small non-coding sequencing of the BOEC derived EVs. Our results revealed that EVs from both co-culture models contained distinct cargo in form of miRNA, fragmented mRNA versus control. The pathway enrichment analysis revealed that EV miRNA from direct co-culture were involved in the biological processes associated with phagocytosis, macroautophagy, placenta development, cellular responses to TNF and FGF. The mRNA fragments also varied within the different groups and mapped to the exonic regions of the transcriptome providing vital insights regarding the changes in cellular transcriptome on the arrival of spermatozoa. The findings of this study suggest that spermatozoa, in contact as well as remotely, alter the EV cargo of female reproductive tract epithelial cells which might be playing an essential role in pre and post-fertilization events. Graphical abstract

A farm belonging to a Swiss sow pool system reported increased cases of necrosis on the base of the tail or ears in their piglets. Therefore, herd examination was performed in February 2021, and it was found that about half of all examined litters included piglets with necrosis of different locations, and that the sows of these piglets were rather thin. Upon instruction, the farmer then documented the body condition score (BCS) and weight before farrowing and after weaning, and the number of liveborn piglets affected by necrosis of the tail or ear of the next four farrowing batches. In total, data of 97 sows with 1214 liveborn piglets were evaluated. Sows were retrospectively allocated into two groups: Those with piglets with ear and/or tail necrosis (NE), and those without (WN). Of the 97 litters, 40 included piglets with necrosis, with 28 of them having piglets only with tail necrosis, 8 only with ear necrosis, and 4 litters included piglets with both types of necrosis. The group NE lost significantly more weight and BCS points over the suckling period than the group WN, with a tendency of having a lower BCS after weaning (2,0 vs. 2,25/5,0). Blood samples of five sows were analyzed and tested positive for the Fusarium mycotoxin deoxynivalenol (DON). It could be possible that the sows previously consumed DON contaminated feed, which was then stored in their fat tissue, and released again into the blood stream during increased weight loss. Since DON can be transferred from the sow to her piglets during gestation or lactation, this release might have affected the piglets, leading to tail or ear necrosis. However, causative studies are needed to confirm this hypothesis.

Objectives The aim of this study was to evaluate the association between meningeal enhancement (MgE) and cerebrospinal fluid (CSF) analysis results, their individual association with bacteriology results from affected ear samples and whether these test results influenced clinicians’ therapeutic choice in cats with otitis media and interna (OMI). Methods This was a multicentre retrospective study carried out over an 8-year period. Cats diagnosed with OMI, with or without a nasopharyngeal polyp, leading to peripheral vestibular signs were included. Only cats for which MRI with postcontrast T1-weighted sequences and CSF analyses available were included. Cats with intra-axial MRI lesions or empyema were excluded. Results Fifty-eight cats met the inclusion criteria. MgE was reported in 26/58 cases, of which nine had an abnormal CSF result (increased total nucleated cell count [TNCC] or total protein); 32/58 cases had no MgE, of which 10 showed abnormal CSF results. There was no association between bacteriology results (external ear canal or bulla) and MgE or abnormal CSF results. CSF abnormalities were statistically significantly more common in acute cases (n = 16/37) than in chronic cases (n = 3/21; Fischer’s test P = 0.04). Prednisolone was prescribed in 10/16 cases with increased TNCC. Among the 42 cases with normal TNCC, 15 received prednisolone and 13 received non-steroidal anti-inflammatory drugs. Various antimicrobial drugs were prescribed in 53/58 cats. Duration of antimicrobial treatment was similar, regardless of positive bacterial culture (5.58 vs 4.22 weeks), abnormal CSF (5.83 vs 4.76 weeks) or MgE (5.33 vs 4.90 weeks). Conclusions and relevance No association was found between the CSF and MgE results. Furthermore, no association was found between MgE, CSF or bacteriology findings. In addition, abnormal CSF results might lead the clinician to treat with corticosteroids, but they did not have any impact on duration of antimicrobial treatment. CSF abnormalities were seen significantly less frequently in chronic cases. The outcome tended to be poorer when MgE was detected on MRI.

Background Urothelial carcinoma (UC) is the most common neoplasm of the canine lower urinary tract, affecting approximately 2% of dogs. Elderly female patients of certain breeds are predisposed, and clinical signs of UC can easily be confused with urinary tract infection or urolithiasis. Diagnosis and treatment are challenging given the lack of disease-specific markers and treatments. The S100A8/A9 complex and S100A12 protein are Ca2+-binding proteins expressed by cells of the innate immune system and have shown promise as urinary screening markers for UC. The neutrophil-to-lymphocyte ratio (NLR) can also aid in distinguishing certain neoplastic from inflammatory conditions. Our study aimed to evaluate the tissue expression of S100/calgranulins and the blood NLR in dogs with UC. Urinary bladder and/or urethral tissue samples from dogs with UC (n = 10), non-neoplastic inflammatory lesions (NNUTD; n = 6), and no histologic changes (n = 11) were evaluated using immunohistochemistry. Blood NLRs were analyzed in dogs with UC (n = 22) or NNUTD (n = 26). Results Tissue S100A12-positive cell counts were significantly higher in dogs with lower urinary tract disease than healthy controls (P = 0.0267 for UC, P = 0.0049 for NNUTD), with no significant difference between UC and NNUTD patients. Tissue S100A8/A9-positivity appeared to be higher with NNUTD than UC, but this difference did not reach statistical significance. The S100A8/A9+-to-S100A12+ ratio was significantly decreased in neoplastic and inflamed lower urinary tract tissue compared to histologically normal specimens (P = 0.0062 for UC, P = 0.0030 for NNUTD). NLRs were significantly higher in dogs with UC than in dogs with NNUTD, and a cut-off NLR of ≤ 2.83 distinguished UC from NNUTD with 41% sensitivity and 100% specificity. Higher NLRs were also associated with a poor overall survival time (P = 0.0417). Conclusions These results confirm that the S100/calgranulins play a role in the immune response to inflammatory and neoplastic lower urinary tract diseases in dogs, but the tissue expression of these proteins appears to differ from their concentrations reported in urine samples. Further investigations of the S100/calgranulin pathways in UC and their potential as diagnostic or prognostic tools and potential therapeutic targets are warranted. The NLR as a routinely available marker might be a useful surrogate to distinguish UC from inflammatory conditions.

Testudo hermanni is included as near‑threatened in the Red List of the International Union for Conservation of Nature, while T. hermanni hermanni is considered endangered in the Italian Red List. Appropriate management of smuggled or seized wild individuals is recommended before their reintroduction into the wild. Accordingly, a health monitoring study was carried out. During 2014‑2016, 133 oral swabs and 121 cloacal swabs were collected from a total of approximately 180 free‑ranging and rescued T. hermanni hermanni from eight different Italian regions to investigate the presence of DNA of Testudinid alphaherpesvirus (TeAHV), Chlamydia spp. and Mycoplasma spp. in the oral cavity, and Salmonella spp. isolates in the cloaca. Mycoplasma spp. was detected in 52 out of 87 (59.77%) of rescued and in 1 out of 46 free‑ranging (2.17%) individuals; 33 out of 53 (62.26%) Mycoplasma spp. positive samples were typed as M. agassizii by PCR. Salmonella spp. was isolated from 45 out of 121 (37.19%) cloacal swabs, typed into 14 serovars, and characterized for complete antimicrobial susceptibility. A significantly different distribution of Salmonella spp. isolates was found in 2016 in comparison with 2014 and 2015, without any difference between free‑ranging and rescued tortoises. All the tested tortoises were negative for TeAHV and Chlamydia spp. These results are considered a baseline information critical to monitor the dynamics of these microorganisms in free‑ranging and rescued populations of T. h. hermanni, and to correctly approach the management of rescued animals and possible relocation programs.