The genome of the black-footed cat: Revealing a rich natural history and urgent conservation priorities for small felids.

Habitat degradation and loss of genetic diversity are common threats faced by almost all of today's wild cats. Big cats, such as tigers and lions, are of great concern and have received considerable conservation attention through policies and international actions. However, knowledge of and conservation actions for small wild cats are lagging considerably behind. The black-footed cat, Felis nigripes, one of the smallest felid species, is experiencing increasing threats with a rapid reduction in population size. However, there is a lack of genetic information to assist in developing effective conservation actions. A de novo assembly of a high-quality chromosome-level reference genome of the black-footed cat was made, and comparative genomics and population genomics analyses were carried out. These analyses revealed that the most significant genetic changes in the evolution of the black-footed cat are the rapid evolution of sensory and metabolic-related genes, reflecting genetic adaptations to its characteristic nocturnal hunting and a high metabolic rate. Genomes of the black-footed cat exhibit a high level of inbreeding, especially for signals of recent inbreeding events, which suggest that they may have experienced severe genetic isolation caused by habitat fragmentation. More importantly, inbreeding associated with two deleterious mutated genes may exacerbate the risk of amyloidosis, the dominant disease that causes mortality of about 70% of captive individuals. Our research provides comprehensive documentation of the evolutionary history of the black-footed cat and suggests that there is an urgent need to investigate genomic variations of small felids worldwide to support effective conservation actions.

Severe bronchiectasis resulting from chronic bacterial bronchitis and bronchopneumonia in a jungle cat.

Bronchiectasis is irreversible bronchial dilation that can be congenital or acquired secondary to chronic airway obstruction. Feline bronchiectasis is rare and, to our knowledge, has not been reported previously in a non-domestic felid. An ~10-y-old female jungle cat (Felis chaus) was presented for evaluation of an abdominal mass and suspected pulmonary metastasis. The animal died during exploratory laparotomy and was submitted for postmortem examination. Gross examination revealed consolidation of the left caudal lung lobe and hila of the cranial lung lobes. Elsewhere in the lungs were several pale-yellow pleural foci of endogenous lipid pneumonia. On cut section, there was severe distension of bronchi with abundant white mucoid fluid. The remaining lung lobes were multifocally expanded by marginal emphysema. Histologically, ectatic bronchi, bronchioles, and fewer alveoli contained degenerate neutrophils, fibrin, and mucin (suppurative bronchopneumonia) with rare gram-negative bacteria. Aerobic culture yielded low growth of Proteus mirabilis and Escherichia coli. There was chronic bronchitis, marked by moderate bronchial gland hyperplasia, lymphoplasmacytic inflammation, and lymphoid hyperplasia. The palpated abdominal mass was a uterine endometrial polyp, which was considered an incidental, but novel, finding. Chronic bronchitis and bronchopneumonia should be considered as a cause of bronchiectasis and a differential diagnosis for respiratory disease in non-domestic felids.

The detection and phylogenetic characterization of Cryptosporidium, Cystoisospora, and Giardia duodenalis of cats in South Korea.

Introduction: Cryptosporidium, Cystoisospora, and Giardia duodenalis are gastrointestinal protozoa parasites that cause diarrhea in various animals. However, information regarding the detection and phylogenetic characterization of gastrointestinal protozoa parasites in cats is limited throughout South Korea. Therefore, this study aimed to determine the detection and identify subspecies of gastrointestinal protozoa parasites in cats from South Korea. Methods: A total of 290 fecal samples were collected from stray, companion, and shelter cats in six provinces. Cryptosporidium, Cystoisospora, and G. duodenalis were identified by PCR. All positive samples were subtyped by PCR and sequencing of gp60, ITS-1, tpi, bg, and gdh. Results: The overall detection of gastrointestinal protozoan parasitic infection was 17.93%. G. duodenalis was the most prevalent, with 7.93%, followed by Cystoisospora spp. (7.24%) and Cryptosporidium spp. (4.48%). In addition, C. felis (n=10), C. parvum (n=2), C. ryanae (n=1), Cystoisospora felis (n=14), Cystoisospora suis (n=5), Cystoisospora ohioensis (n=1), Cystoisospora spp. were identified in subspecies analysis of positive samples. C. felis showed a significant association with diarrhea (7.81%) and living condition (6.04%), and Cystoisospora felis in diarreha (9.38%) according to detection. Through phylogenetic analysis of the tpi, bg, and gdh genes from 23 G. duodenalispositive samples, it was confirmed that the samples of present study belonged to assemblage A, B, C, and D. Discussion: South Korean cats have a high rate of gastrointestinal protozoan parasites infection with cat-specific Cryptosporidium and Cystoisospora, which are associated with living conditions and diarrhea symptoms. Moreover, zoonotic and other animal-specific subtype of protozoan parasites have been detected in cat feces.

Schlafen4+-MDSC in Helicobacter-induced gastric metaplasia reveals role for GTPases.

Introduction: MDSCs express SCHLAFEN 4 (SLFN4) in Helicobacter-infected stomachs coincident with spasmolytic polypeptide-expressing metaplasia (SPEM), a precursor of gastric cancer. We aimed to characterize SLFN4+ cell identity and the role of Slfn4 in these cells. Methods: Single-cell RNA sequencing was performed on immune cells sorted from PBMCs and stomachs prepared from uninfected and 6-month H. felis-infected mice. Knockdown of Slfn4 by siRNA or PDE5/6 inhibition by sildenafil were performed in vitro. Intracellular ATP/GTP levels and GTPase activity of immunoprecipitated Slfn4 complexes were measured using the GTPase-Glo assay kit. The intracellular level of ROS was quantified by the DCF-DA fluorescent staining, and apoptosis was determined by cleaved Caspase-3 and Annexin V expression. Gli1CreERT2 x Slfn4 fl/fl mice were generated and infected with H. felis. Sildenafil was administered twice over 2 weeks by gavaging H. felis infected mice ~4 months after inoculation once SPEM had developed. Results: Slfn4 was highly induced in both monocytic and granulocytic MDSCs from infected stomachs. Both Slfn4 +-MDSC populations exhibited strong transcriptional signatures for type-I interferon responsive GTPases and exhibited T cell suppressor function. SLFN4-containing protein complexes immunoprecipitated from myeloid cell cultures treated with IFNa exhibited GTPase activity. Knocking down Slfn4 or PDE5/6 inhibition with sildenafil blocked IFNa induction of GTP, SLFN4 and NOS2. Moreover, IFNa induction of Slfn +-MDSC function was inhibited by inducing their reactive oxygen species (ROS) production and apoptosis through protein kinase G activation. Accordingly, in vivo disruption of Slfn4 in Gli1CreERT2 x Slfn4 fl/fl mice or pharmacologic inhibition by sildenafil after Helicobacter infection also suppressed SLFN4 and NOS2, reversed T cell suppression and mitigated SPEM development. Conclusion: Taken together, SLFN4 regulates the activity of the GTPase pathway in MDSCs and precludes these cells from succumbing to the massive ROS generation when they acquire MDSC function.

The cytosolic DNA sensor AIM2 promotes Helicobacter-induced gastric pathology via the inflammasome.

Helicobacter pylori (H. pylori) infection can trigger chronic gastric inflammation perpetuated by overactivation of the innate immune system, leading to a cascade of precancerous lesions culminating in gastric cancer. However, key regulators of innate immunity that promote H. pylori-induced gastric pathology remain ill-defined. The innate immune cytosolic DNA sensor absent in melanoma 2 (AIM2) contributes to the pathogenesis of numerous autoimmune and chronic inflammatory diseases, as well as cancers including gastric cancer. We therefore investigated whether AIM2 contributed to the pathogenesis of Helicobacter-induced gastric disease. Here, we reveal that AIM2 messenger RNA and protein expression levels are elevated in H. pylori-positive versus H. pylori-negative human gastric biopsies. Similarly, chronic Helicobacter felis infection in wild-type mice augmented Aim2 gene expression levels compared with uninfected controls. Notably, gastric inflammation and hyperplasia were less severe in H. felis-infected Aim2-/- versus wild-type mice, evidenced by reductions in gastric immune cell infiltrates, mucosal thickness and proinflammatory cytokine and chemokine release. In addition, H. felis-driven proliferation and apoptosis in both gastric epithelial and immune cells were largely attenuated in Aim2-/- stomachs. These observations in Aim2-/- mouse stomachs correlated with decreased levels of inflammasome activity (caspase-1 cleavage) and the mature inflammasome effector cytokine, interleukin-1ß. Taken together, this work uncovers a pathogenic role for the AIM2 inflammasome in Helicobacter-induced gastric disease, and furthers our understanding of the host immune response to a common pathogen and the complex and varying roles of AIM2 at different stages of cancerous and precancerous gastric disease.

DISEASE PROCESSES IDENTIFIED IN CAPTIVE ARABIAN SAND CATS (FELIS MARGARITA HARRISONI).

The objective of this retrospective study is to identify common and significant causes of mortality and disease processes in the Arabian sand cat (Felis margarita harrisoni) captive population at Al Ain Zoo (Abu Dhabi, United Arab Emirates). Complete postmortem records of 25 Arabian sand cats, dead between 2009 and 2022, were reviewed retrospectively. A complete postmortem examination was done in all cases, and information was recorded in the Al Ain Zoo database and files. Out of 25 animals dead, 11 were adults (4-12 yr) and 12 were classified as geriatric animals (>12 yr), with only two neonatal (0-4 mon) deaths and no recorded deaths in juveniles (4 mon to 4 yr). Interestingly, but also expected because of the age range, 24% of the cases had concurrent pathologies at the time of death. As expected in adult and geriatric felines, more than half of the cases (60%) developed nephropathies that were either one of the most important contributors or the main cause of death of the animal. Different neoplastic lesions were described in four cases and reported for the first time in this subspecies: benign peripheral nerve sheath tumor, hepatobiliary carcinoma, and two different thyroid neoplasia. A vasculoproliferative disorder of the liver, peliosis hepatis, was described in one of the cases. Additionally, in at least four cases, hyperthyroidism was strongly suspected in connection with thyroid neoplasia and hyperplasia, clinical signs, and other observed postmortem lesions. Traumatic causes of death also were reported in six cases, including the only two neonates recorded dead. This information will contribute to Arabian sand cat improved veterinary care by identifying common pathologies in this species, potentially allowing earlier diagnosis and, ultimately, improving their management and husbandry in the captive breeding populations.

Epithelial membrane antigen-reactive feline chordoid meningioma in a European wildcat (Felis silvestris).

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.

Experimental infection of cats with Cryptosporidium felis.

OBJECTIVES: The aims of this study were to experimentally inoculate cats with Cryptosporidium felis oocysts and compare fecal detection by fluorescent antibody assay (FA) and quantitative PCR (qPCR), and document clinical signs associated with infection. METHODS: Cryptosporidium felis oocysts were concentrated from the feces of a naturally infected cat and orally inoculated into six cats that tested negative for C felis by an FA and fecal flotation (FF). Cats were observed daily for the presence of clinical signs consistent with infection. Fecal samples from all cats on days 0 and 9, and one sample per cat (days 18-21), were evaluated by all assays. On day 31, two cats negative for C felis by FF and FA were administered methylprednisolone acetate and all assays were repeated on days 34, 36 and 38. Samples from all cats were tested by FF and FA on days 41, 43, 45 and 48. RESULTS: A total of 41 samples were tested, 25 of which were compared by FA and qPCR. Cryptosporidium felis was detected in 2/25 (8%) and in 19/25 (76%) samples by FA and by qPCR, respectively; the other 16 samples were tested by FF and FA. None of the cats was positive for C felis by FF or FA in samples collected on days 0, 9 or 18-21. One, five and six samples tested positive by qPCR on days 0, 9 and 18-21, respectively. The cats administered methylprednisolone acetate tested positive for C felis by FA on day 36 and by qPCR on days 31, 34, 36 and 38. None of the cats showed clinical signs of disease. CONCLUSIONS AND RELEVANCE: Clinical signs were not recognized in any of the cats for the duration of the study. FA was insensitive compared with qPCR for detecting cats with subclinical C felis infection.

Molecular detection of Hepatozoon species infections in domestic cats living in Germany.

OBJECTIVES: Three species of protozoal Hepatozoon species (H felis, H canis and H silvestris) are known to infect cats in Europe. The objective of this study was to determine the prevalence of Hepatozoon species in samples from cats living in Germany that were submitted to a veterinary laboratory. METHODS: The study included cats tested for Hepatozoon species by PCR between 2007 and 2020 by the Laboklin laboratory. Travel history and haematological results were documented for cats with positive test results. From 2018 onwards, a partial 18S rRNA Hepatozoon gene fragment was sequenced from cats with positive PCR results. RESULTS: Sixty-four of 931 cats (7%) tested positive for Hepatozoon species. Sex and age did not have a statistically significant impact. Sequencing was carried out for 16 samples and revealed H felis in all cases. All cats with positive test results and a relevant travel history had been imported from the Mediterranean or south-eastern Europe. There were no autochthonous infections with Hepatozoon species. Leukocytosis, haemoconcentration and anaemia were the most common haematological abnormalities. CONCLUSIONS AND RELEVANCE: Although infections with Hepatozoon species in cats are usually subclinical, it may be useful to screen cats imported from the Mediterranean and south-eastern Europe for these pathogens to prevent local transmission cycles. There was no evidence of autochthonous infections in Germany; however, further investigations regarding a possible transmission of Hepatozoon species from infected cats to blood-feeding arthropods in Germany may be of interest. To avoid potential spread of the pathogens, ectoparasite prophylaxis is advisable.

Comparación del neurocráneo entre el gato doméstico (Felis catus Linneas 1758) y el montés (Felis silvestris Schreber, 1777) mediante técnicas de morfometría geométrica / Comparison of neurocranium between domestic cat (fFlis catus Linneas 1758) and wild cat (Felis silvestris Schreber, 1777) by means of geometric morphometric techniques

RESUMEN: El objetivo de este estudio fue comparar objetivamente las diferencias en la forma total (tamaño y forma pura) del neurocráneo entre un grupo de cráneos de gatos domésticos (Felis catus, n=81), y un grupo de gato montés (Felis silvestris, n=12) mediante técnicas de morfometría geométrica. Para ello se obtuvieron fotografías digitales del lado izquierdo de los cráneos de un modo estandarizado, y en las que se ubicaron 8 landmarks y 9 semilandmarks. Después de una superposición generalizada de Procrustes se procedió a un análisis de componentes principales y a otro de canónico a fin de comparar y detectar las posibles diferencias. Los tamaños neurocraneales no mostraron diferencias entre ambos grupos, pero sí que aparecieron diferencias significativas en la forma. Los cambios de forma afectarían principalmente la sutura escamoso-parietal-alisfenoidal y, en menor medida, puntos caudales del cráneo, lo que sugeriría cambios a nivel de músculos masticatorios. A conocimiento del autor, constituye esta la primera investigación que caracteriza y compara el cráneo de los gatos mediante métodos de morfometría geométrica, recomendándose un mayor uso de esta técnica dada la escasez de publicaciones al respecto en mamíferos domésticos.

SUMMARY: The objective of this study was to objectively compare the differences in the total shape (size and pure shape) of the neurocranium between a group of domestic cat skulls (Felis catus, n=81), and wildcats (Felis silvestris, n=12) by means of geometric morphometric techniques. To achieve it, we obtained digital photographs of the left side of the skulls in a standardized form, in which 8 landmarks and 9 semilandmarks were located. After a Generalized Procrustes Superimposition, a Principal Component Analysis and a Canonical Analysis one were performed to compare and detect possible differences. Neurocranial sizes showed no differences between the two groups, however, significant differences in shape did appear. Shape changes would mainly affect the squamosal-coronal suture and, to a lesser extent, caudal skull points, therefore suggesting changes in masticatory muscles. As far as this author´s knowledge, this is the first research that characterizes and compares the skull of cats using geometric morphology methods, recommending greater use of this technique given the scarcity of publications on this subject in domestic mammals.

Microstructural changes of the wall of uterine horn in the domestic cat (Felis silvestris catus) during postnatal development.

The study describes the details of histological changes occurring during the postnatal development of the wall of the uterine horn in the European shorthair domestic cat. Light microscope observations and morphometric analysis were carried out to investigate the rate of development of the wall of the uterine horns in order to estimate the maturing time. Uterine horns of newborn and 1week-old cats are fetal-like and showing smooth, nonfolded endometrium lined with pseudostratified epithelium, deprived of uterine glands, and poorly developed two-layered myometrium. The lamina propria of the mucosa becomes folded in the first month of postnatal life. At the same time, the primordia of the uterine glands appear, elongating, doubling in number and reaching the basal layer by sixth month of age. In six-month-old cats, the lamina propria of the mucosa is formed into the functional and basal layer and its thickness increases rapidly. Until the sixth month of age, the inner circular muscle layer is the widest part of the myometrium, the outer longitudinal layer is the thinnest, while the width of the vascular layer is an intermediate value between the width of the inner and outer layer. From the seventh month of age, the proportion of the thickness of the inner and outer layer is reversed and the vascular layer becomes the thinnest part of the myometrium. Between seventh and twelfth month of age, the structure of the wall of the uterine horn does not change significantly, as the longitudinal mucosal folds limiting the narrow lumen of the uterine horns, and the arrangement of the layers in the wall is assigned to a pattern according to which the endometrium represents about 40% of the total width of the wall, while the myometrium is about 60%. In the studied time span of postnatal life, the average thickness of the wall of the uterine horn increases 9.4 -fold, the thickness of the lamina propria and mucosal folds increases 6.9-fold and 3.8-fold, respectively, while the thickness of the myometrium increases 17.3-fold. Summing up, the process of maturation of the wall of the uterine horns in the domestic cat lasts up to sixth month of postnatal life and includes the formation of the endometrial longitudinal folds, appearance and development of the uterine glands, division of the lamina propria of the mucosa into the functional and basal layers, and continued development of the circular, vascular and longitudinal muscle layers.

Experimental infection of cats with Cystoisospora felis.

BACKGROUND: Cystoisospora felis is a common parasite of cats and is diagnosed by fecal flotation, but false-negative results can be common. HYPOTHESIS/OBJECTIVES: To experimentally inoculate cats with C. felis oocysts, to compare fecal flotation and polymerase chain reaction (PCR) results, and to describe any clinical signs consistent with infection. ANIMALS: Six cats. METHODS: Cystoisospora felis oocysts were identified morphologically from feces of a naturally infected kitten with diarrhea, sporulated oocysts (5000) were inoculated to 6 cats that were negative for fecal parasites by fecal flotation and by a fluorescent antibody assay (FA) for Giardia spp. and Cryptosporidium spp. Cats were observed daily for the presence of clinical signs consistent with infection. Fecal samples were evaluated by fecal flotation and FA up to 3 times per week post inoculation (PI) to Day 27. Thirty-six samples collected before inoculation and from Days 8, 10, 13, 15, and 20 PI were assayed using an internal transcribed spacer 1 (ITS1) PCR that amplifies DNA of C. felis. RESULTS: All cats were negative for C. felis by both assays before inoculation. All cats shed C. felis oocysts by Day 10 PI, oocysts were not detected by fecal flotation after Day 15 PI. Cystoisospora felis DNA was amplified from 24/36 (66.6%) fecal samples from 6/6 (100%) of the cats. Oocysts were not detected by fecal flotation in 4 of the samples that were positive for C. felis DNA by PCR. Clinical signs were not recognized in any of the study cats. CONCLUSIONS AND CLINICAL IMPORTANCE: Fecal flotation is a convenient assay for detection of C. felis but could occasionally give false-negative results when compared to this ITS1 PCR.

Reproductive hormones monthly variation in free-ranging European wildcats: Lack of association with faecal marking.

Integrating ecophysiological and behavioural discoveries in conservation and management plans is essential to preserve scarce and elusive species such as the European wildcat (Felis silvestris). The purpose of this study was to characterize the monthly variation in the steroid reproductive hormone metabolite levels (oestradiol, progesterone and testosterone) in this species and to test its possible association with a monthly pattern of faecal marking. By collecting fresh faecal samples in Montes do Invernadeiro Natural Park (Galicia, Northwest Spain) each month, we obtained a total of 110 samples belonging to 25 different individuals. We conducted enzyme immunoassays which allowed us to track the annual variation in reproductive hormone excretion patterns in wildcat scats. Furthermore, we also evaluated the possible relation between the faeces used as marks and the reproductive hormone levels. We found that oestradiol and progesterone metabolite levels exhibited a distinct pattern, both increasing during the breeding months. Oestradiol metabolite larger peaks were found during March and April, whereas the highest concentration of progesterone metabolites appeared in July. On the contrary, testosterone metabolite levels did not significantly change depending on the month. Moreover, we did not find any evidence that the faecal marking behaviour pattern was associated with reproductive hormone metabolite levels. It seems that other factors related to habitat and food resources could be more important in the performance of this behaviour.

Clinical investigations and treatment outcome in a European wildcat (Felis silvestris silvestris) infected by cardio-pulmonary nematodes.

Recently, feline cardio-pulmonary nematodes have attracted high scientific interest, as they are increasingly reported from various areas. Most of these parasites have similar transmission patterns and/or host reservoirs, thus they may affect domestic and wild felids living in sympatry. In the present study, a case of multiple cardio-pulmonary parasitism in co-infection with other parasites in a European wildcat is presented. The animal, found exhausted, was hospitalised for recovery and parasitological, haematological, clinical and imaging examinations were performed. The parasitological examinations revealed 4 cardio-pulmonary nematodes, i.e. Aelurostrongylus abstrusus, Troglostrongylus brevior, Eucoleus aerophilus, Angiostrongylus chabaudi, 3 intestinal parasites, i.e. Toxocara cati, ancylostomatids, Cystoisospora felis, 2 haemoparasites, i.e. Hepatozoon felis and elements morphologically compatible with small Babesia/Cytauxzoon spp., and Ixodes ricinus and Haemaphysalis erinacei ticks. Treatment with a spot-on formulation containing imidacloprid 10% and moxidectin 1% (Advocate® spot-on solution for cats, Bayer) was decided and follow-up faecal examinations were performed until the release of the animal. By the end of the hospitalisation, all metazoan endoparasites were no longer detectable in faecal examinations, with the exception of a low number of A. abstrusus larvae. Thus, the animal was released after a second treatment with the same product. This is the first description of an apparently successful treatment of multiple cardio-respiratory parasitosis in a naturally infected wildcat showing compatible clinical signs. The evidence that Advocate® may be effective against A. chabaudi could be useful for treating infected, hospitalised, wildcats and it is promising in the case A. chabaudi infection will spread to domestic cats in a near future.

Severe Verminous Pneumonia Caused by Natural Mixed Infection with Aelurostrongylus abstrusus and Angiostrongylus chabaudi in a European Wildcat from Western Balkan Area.

AIMS: Aelurostrongylus abstrusus (Railliet, 1898) and Angiostrongylus chabaudi (Biocca, 1957) are important cardiopulmonary metastrongyloids in felids. This case report describes, for the first time, a natural and patent mixed infection caused by A. abstrusus and A. chabaudi in a European wildcat (Felis silvestris silvestris) from Bosnia and Herzegovina. Most important, this is a rare report of fatal parasitism in wildlife, involving a severe verminous bronchopneumonia and gastrointestinal nematode and cestode infection with Toxocara cati, Taenia taeniaeformis, Aonchotheca putorii, and Ancylostoma spp. RESULTS: Emphasis is set to detailed description of granulomatous, interstitial verminous bronchopneumonia, morphological description of A. abstrusus and A. chabaudi, and molecular confirmation of diagnosis by triplex PCR. CONCLUSION: The data provided in this study contribute to the knowledge on the epizootiology and pathological effect of these neglected metastrongyloids in European wildcat.

Concurrent Transitional Meningioma and Ceruminous Gland Adenocarcinoma in a Scottish Wildcat Hybrid (Felis silvestris).

The Scottish wildcat (Felis silvestris) is an iconic and endangered subpopulation of the European wildcat (F. silvestris silvestris). There is much research devoted to the ecology, genetics and conservation of this animal, but little published information on pathology and disease. The investigation and reporting of such information is vital to furthering understanding of the effects of hybridization, a factor that is crucial if we are to secure a future for the Scottish wildcat. This report describes the clinical presentation, gross post-mortem and histological findings in an elderly Scottish wildcat hybrid with concurrent transitional meningioma and ceruminous gland adenocarcinoma.

First evidence of the European wildcat (Felis silvestris silvestris) as definitive host of Angiostrongylus chabaudi.

Angiostrongylus chabaudi (Strongylida, Angiostrongylidae) is a parasitic nematode described for the first time last century from the pulmonary arteries of six European wildcats (Felis silvestris silvestris) in central Italy. Since then, this parasite remained practically unknown until recently, when immature A. chabaudi have been reported from one wildcat in Germany and two domestic cats (Felis silvestris catus) in Italy. The present report describes the first record of A. chabaudi in Greece and, most importantly, the first known case of patent infection by A. chabaudi. The necropsy of a road-killed F. s. silvestris found near the lake Kerkini, in the municipality of Serres (Macedonia, Greece), revealed the presence of nematodes of both sexes in the right ventricle and the pulmonary artery of the heart. All parasites were mature adults and numerous eggs were present in the uteruses of females. The morphological characteristics of the parasites were consistent with those of A. chabaudi. Moreover, Angiostrongylus-like first stage larvae (L1) were present in the faeces of the animal that was negative for any other cardio-pulmonary parasite. Genetic examination of adult parasites and L1 confirmed the morphological identification as A. chabaudi. Histopathological examination of the lungs showed severe, multifocal to coalescing, chronic, interstitial granulomatous pneumonia due to the presence of adult parasites, larvae and eggs. These findings demonstrate for the first unequivocal time that this nematode reproduces in the European wildcat which should be ultimately considered a definitive host of A. chabaudi. Finally, the L1 of A. chabaudi are described here for the first time, opening new prospects for further studies on this neglected parasite.

A RARE CASE OF HEPATOCELLULAR CARCINOMA IN THE ARABIAN SAND CAT (FELIS MARGARITA HARRISONI).

Primary liver cancer is uncommon and is generally classified as hepatocellular carcinoma (HCC), bile duct carcinoma, neuroendocrine (or carcinoid) tumor, and mesenchymal tumor (sarcoma). Here we describe the gross and histopathology characteristics of a rare HCC in a captive Arabian sand cat (Felis margarita harrisoni) held at Al Ain Zoo, United Arab Emirates. The description of this case in the Arabian sand cat adds to the current knowledge of hepatocellular carcinoma in captive nondomestic felids.

Characterization and Multilineage Differentiation of Domestic and Black-Footed Cat Mesenchymal Stromal/Stem Cells from Abdominal and Subcutaneous Adipose Tissue.

Transplantation of mesenchymal stem cells (MSCs) isolated from bone marrow or adipose tissue is emerging as a promising tool for cell replacement therapy and regenerative medicine in domestic and endangered animal species. Defining the differentiation capability of adipose-derived mesenchymal stromal/stem cells (AMSCs) collected from different depot sites of adipose tissue will be essential for developing strategies for cell replacement therapy. In the present study, we compared the biological characteristics of domestic cat AMSCs isolated from visceral fat of the abdominal cavity (AB) with AMSCs from subcutaneous (SQ) tissue, and the functional capability of domestic and black-footed cat (Felis nigripes) AMSCs to differentiate into other cell types. Our results showed that both domestic and black-footed cat adipose-derived stromal vascular fractions contained AMSCs. Both domestic cat AB- and SQ-AMSCs showed important clonogenic ability and the minimal MSC immunophenotype as defined by the International Society for Cellular Therapy in humans. However, domestic cat AB-AMSCs had higher percentages of cells positive for MSCs-associated cluster of differentiation (CD) markers CD90(+) and CD105(+) (92% and 80%, respectively) than those of SQ-AMSCs (77% and 58%, respectively). Although these results may suggest that AB-AMSCs may be more multipotent than SQ-AMSCs, both types of cells showed similar expression of pluripotent genes Oct-4 and Klf4, except for higher expression of Nanog than in AB-AMSCs, and equivalent in vitro multilineage differentiation. Under appropriate stimuli, the black-footed cat and both domestic cat AB- and SQ-AMSCs differentiated not only toward mesoderm cell lineages but also toward ectoderm cell lineage, such as neuron cell-like cells. Black-footed cat AMSCs had more capability to differentiate toward chondrocytes. These results suggest that the defined AMSC population (regardless of site of collection) could potentially be employed as a therapeutic agent for both domestic and endangered diseased or injured felids.

Echinococcus multilocularis detection in the intestines and feces of free-ranging domestic cats (Felis s. catus) and European wildcats (Felis s. silvestris) from northeastern France.

Experimental studies have demonstrated that cats can be infected by Echinococcus multilocularis, although few data are available concerning their natural infection. This study was designed to compare experimental findings with information on the prevalence of natural E. multilocularis infections of cats in a rural high endemic area. Of 19 intestines of domestic cats (Felis s. catus) and five of European wildcats (Felis s. silvestris) analyzed by segmental sedimentation and counting technique (SSCT), infection by E. multilocularis was observed for one individual of each species, resulting in a prevalence estimated at 5%, (CI95%: 1-26) in domestic cats and at 20% (CI95%: 1-72) in wildcats. High worm burdens (680 and 7040) were noted, but comprised only immature worms. The same EmsB microsatellite profile obtained from the worms' DNA was observed in the two cats as in foxes from the same area and from other European countries. The presence of E. multilocularis DNA was diagnosed in 3.1% (10/321) of the domestic cat feces collected on the field in two villages. However, no E. multilocularis eggs were found after flotation with zinc chloride of the positive feces. The detection of DNA from E. multilocularis was thought to be due to the presence of cells from worms untied from the intestine and corresponding to prepatent infection or due to the digested metacestode. These results from E. multilocularis presence in wild and domestic cat populations agree with those previously obtained by experimental infections. These findings support that these cats play an insignificant role in E. multilocularis transmission, even in a "highly endemic" region. Nevertheless, since the presence of thick-shelled E. multilocularis eggs from cats has already been reported, the associated zoonotic risk cannot be totally ruled out, even if it is very low.