Molecular detection using hybridization capture and next-generation sequencing reveals cross-species transmission of feline coronavirus type-1 between a domestic cat and a captive wild felid.

Feline coronavirus (FCoV) infection normally causes mild or subclinical signs and is common in domestic cats. However, in some cats, FCoV infection can also lead to the development of feline infectious peritonitis (FIP)-a typically lethal disease. FCoV has two serotypes or genotypes, FCoV-1 and FCoV-2, both of which can cause FIP. The main difference between the genotypes is the viral spike (S) protein that determines tropism and pathogenicity, crucial mechanisms in the development of FIP. Subclinical infection and FIP have both been reported in wild felids, including in threatened species. Due to the high genetic variability of the S gene and the technical challenges to sequencing it, detection and characterization of FCoV in wild felids have mainly centered on other more conserved genes. Therefore, the genotype causing FIP in most wild felids remains unknown. Here, we report a retrospective molecular epidemiological investigation of FCoV in a zoological institution in the U.Ss. In 2008, a domestic cat (Felis catus) and a Pallas' cat (Otocolobus manul) sharing the same room succumbed to FIP. Using in situ hybridization, we detected FCoV RNA in different tissues of both felids. Using hybridization capture and next-generation sequencing, we detected, sequenced, and characterized the whole genome of the FCoV infecting both felids. Our data show for the first time that FCoV-1 can be transmitted between domestic and wild felids and extends the known host range of FCoV-1. Our findings highlight the importance of identifying the genotype causing FIP, to develop effective control measures. IMPORTANCE: Feline coronavirus (FCoV) is highly prevalent in domestic cats worldwide and has also been reported in wild felids, including endangered species, in which it has caused substantial population declines. Characterizing the genetic diversity of FCoV is crucial due to recent reports of novel pathogenic recombinant variants causing high mortality in feral cats in Cyprus. In this retrospective molecular epidemiology study, we used archived samples collected in a zoological institution in the U.S. in which a domestic and a wild felid succumbed to FCoV. Using hybridization capture (HC) and next-generation sequencing, we show for the first time that FCoV can be naturally transmitted between domestic and wild felids. We demonstrate the efficacy of HC for detecting and sequencing the whole genome of FCoV, which is essential to characterize its different genotypes.

Clinical Evaluation of Metal Dental Prostheses in Captive Wild Felids.

Captive felids are prone to dental fractures, and the recommended treatment involves endodontics (root canal treatment), prosthodontics (coronal restoration), or tooth extraction. A single-unit dental prosthesis can restore the crown of a tooth whose structure has been partially or completely lost. Nine animals out of 35 captive wild felids kept under human care in 7 different Brazilian institutions were selected for the study. The animals selected included the little spotted cat (Leopardus tigrinus), ocelot (L. pardalis), margay cat (L. wiedii), pampas cat (L. colocolo), jaguarundi cat (Puma yagouaroundi), puma (P. concolor), jaguar (Panthera onca), lion (P. leo), and tiger (P. tigris). Nineteen canine prosthodontic crowns (18 full and one 3/4) were cemented using dual-cure resin cement and their feasibility was evaluated for 30 months. Fourteen (73%) remained fixed to the tooth and 5 (27%) were lost after the evaluation period. This study aimed to assess the longevity of prosthodontic crowns in the canine teeth of wild felids. Furthermore, it aimed to determine the functionality and longevity of teeth which had received prosthodontic crowns.

Cheetah Acinonyx pardinensis (Felidae, Carnivora) from the Early Pleistocene of Crimea (Taurida Cave).

A mandible fragment of Acinonyx pardinensis (Croizet et Jobert, 1828) is described from the Early Pleistocene locality in the Taurida cave (Crimea, Late Villafranchian, about 1.8-1.5 Ma). This is the first discovery of the genus Acinonyx in the Pleistocene of Crimea. Along with other felids, such as Homotherium, Megantereon, Panthera, Lynx, and Puma, cheetahs were typical representatives of the Late Villafranchian faunas of Eurasia. In the Taurida locality, Acinonyx pardinensis is co-occurred with Homotherium crenatidens, Megantereon adroveri, and Lynx issiodorensis.

Semen Collection and Evaluation in Two Tigers (Panthera tigris) and Two Leopards (Panthera pardus).

Assisted reproduction techniques play a significant role in veterinary medicine, and although they are widely used in domestic animals, they are also becoming increasingly relevant in clinical practice for wild felids, especially in the conservation efforts for endangered species. In this study, the result of two semen collection techniques, urethral catheterization after pharmacological induction (Ur.Ca.P.I.) and electroejaculation, are described, aiming to provide new practical information about sperm collection using the Ur.Ca.P.I. technique and electroejaculation in tigers and leopards, describing the authors' experience and presenting new data and observations. The following descriptive study included two subjects of Panthera tigris species and two of Panthera pardus. These subjects, after general anesthesia, underwent sperm collection initially with Ur.Ca.P.I. and, subsequently, with electroejaculation. Sampling was made possible in both species thanks to the use of electroejaculation. Sperm volumes in leopards ranged from 0.3 to 0.5 mL and in tigers from 0.5 to 2.177 mL. Sperm concentration in leopards ranged from 136 × 106 to 280 × 106 sperm/mL, and in tigers, from 21.5 × 106 to 354 × 106 sperm/mL. Urethral catheterization gave positive results in leopards, with sperm volumes ranging from 25 up to 150 µL and a concentration ranging from 110 × 106 up to 1082 × 106 sperm/mL. In tigers, unlike in leopards, the use of the Ur.Ca.P.I. technique encountered difficulties that did not allow satisfactory results to be obtained. Therefore, it would be useful to test the feasibility of urethral catheterization on a larger group of individuals in order to have more meaningful feedback. Finally, because electroejaculation always allowed semen collection in tigers, with a higher sperm quality than samples collected by Ur.Ca.P.I., we currently consider it the technique of choice for the collection of semen material in this species.

Metabarcoding analysis provides insight into the link between prey and plant intake in a large alpine cat carnivore, the snow leopard.

Species of the family Felidae are thought to be obligate carnivores. However, detection of plants in their faeces raises questions about the role of plants in their diet. This is particularly true for the snow leopard (Panthera uncia). Our study aimed to comprehensively identify the prey and plants consumed by snow leopards. We applied DNA metabarcoding methods on 90 faecal samples of snow leopards collected in Kyrgyzstan, employing one vertebrate and four plant markers. We found that argali (Ovis ammon) was detected only from male snow leopards. Myricaraia sp. was the most consumed among 77 plant operational taxonomic units found in snow leopard samples. It frequently appeared in samples lacking any prey animal DNA, indicating that snow leopards might have consumed this plant especially when their digestive tracts were empty. We also observed differences in the patterns of plant consumption between male and female snow leopards. Our comprehensive overview of prey and plants detected in the faeces of snow leopards and other sympatric mammals will help in formulating hypotheses and guiding future research to understand the adaptive significance of plant-eating behaviour in felids. This knowledge supports the enhancement of their captive environments and the conservation planning of their natural habitats.

Protein quality of a small mammal prey and its body organs for felids.

This study evaluated the protein quality of small mammalian prey and its body organs by analyzing amino acid (AA) composition and digestibility of wild adult rats and their body organs (skin/fur, bone, muscle, intestine, liver, kidney, spleen, brain, heart, and lung) utilizing an in vitro digestion method. The average dry matter (DM) digestibility of whole rats was 89.9%. The digestibility of total AA (TAA), total indispensable AA (TIAA), and total dispensable AA (TDAA) in whole rats was 85.6, 87.0, and 87.6%, respectively. Differences in DM digestibility were observed among rat organs, ranging from 59.0% in bone to 99.8% in muscle (P < 0.001). Highly digestible organs generally exhibited AA digestibility exceeding 90%, except for cysteine (Cys) in the intestine and kidney (83.8% and 88.9%, respectively). The digestibility of AAs in skin/fur ranged from 19.7% for Cys to 81.0% for glycine (Gly). In bone, the digestibility spanned from 56.9% for Gly to 81.1% for tyrosine (Tyr). Additionally, examining the digestible indispensable AA score (DIAAS) gives us an idea of the protein quality of small mammalian prey and their body organs. Our results complement information on AA supply and digestion during prey ingestion by felids.

As obligate carnivores, free-ranging felids consume prey and rely on nutrients from animal organs. Studies in adult carnivores such as domestic cats have demonstrated the importance of the dietary amino acid profile. Therefore, this research used rats as a small prey model to analyze the amino acid composition and digestibility of whole prey and its body organs through in vitro digestion methods. Our results add information on amino acid supply and digestion during natural food intake in felids.

Identification and characterization of a novel circovirus in Iberian lynx in Spain.

Circoviruses cause severe disease in pigs and birds. Canine circovirus has thus far only been associated with respiratory and gastrointestinal disorders and systemic disease in dogs. The Iberian lynx (Lynx pardinus) is one of the most endangered carnivores in Europe and the most endangered felid worldwide. Exploring the virome of these animals may be important in terms of virus discovery and assessing the interspecies-circulation of viruses from related carnivores. In this study, 162 spleen samples from Iberian lynx were screened for CRESS DNA viruses. Overall, 11 (6.8%) of 162 samples tested positive using a consensus PCR. Partial rep sequences were tightly related to each other (96.6-100%). Specific molecular protocols were designed on the partial rep sequences of the novel virus, Iberian lynx-associated circovirus-1 (ILCV-1). By screening a subset of 45 spleen samples, the infection rate of ILCV-1 in Iberian lynxes was 57.8% (26/45). ILCV-1 strains formed a separate cluster intermingled with bat, rodent, mongoose, and felid circoviruses. The genome of the novel virus displayed the highest nucleotide identity (64.3-65.3%) to mongoose circoviruses, thus representing a novel candidate circovirus species. The detection of these viruses in the spleen tissues could suggest systemic infection in the animal host. Overall, these findings suggest that this novel circovirus is common in the Iberian lynx. Further studies are warranted to assess the possible health implications of ILCV-1 in this endangered species.

Renal candidiasis associated with papillary necrosis in a captive tiger (Panthera tigris).

A 14-year-old male tiger developed anorexia with elevated blood urea nitrogen and creatinine levels. The patient had a palpable abdominal mass and demonstrated neutrophilic leukocytosis and anaemia. Leukocytes, yeast and bacteria were present in the urine. The animal was non-responsive to therapy and was subsequently euthanised. Extensive acute renal papillary necrosis (RPN) with pyelonephritis, chronic nephritis and polycystic renal disease were evident during gross and microscopic pathology examinations. The histologic occurrence of fungal spores and pseudohyphae morphologically consistent with Candida species were observed within the necrotic papillary regions of the kidney and within multiple foci of mild parakeratotic hyperkeratosis present in the gingiva and tongue. Candida albicans along with a slight growth of Escherichia coli were recovered from kidney cultures. Possible contributory factors for the renal candidiasis and associated RPN include predisposing oral candidiasis, polycystic renal disease, ischaemic nephrosclerosis, age-associated or other forms of immunodeficiency and therapy with meloxicam, a non-steroidal anti-inflammatory drug. The absence of apparent lower urinary tract involvement coupled with the presence of intravascular renal 'Candida emboli' suggest that chronic oral candidiasis was the probable source of the kidney infection.

Nutraceutical supplementation increases mobility in aged captive non-domesticated felids.

BACKGROUND: Musculoskeletal diseases (MSDs) are an increasing issue as the lifespan of captive animals increases. Extracts of green-lipped mussels have been linked to alleviation of MSDs in domestic carnivores. Understanding their efficacy in non-domestic felids could provide another tool to improve the welfare of aged individuals in collections. METHODS: A within-subject study design quantified steps per minute in each of 18 cats of 13 species before and after the addition of a nutraceutical containing green-lipped mussel extract (Antinol) to their diets. The age structure of four commonly kept subspecies of non-domestic cats was quantified to provide a demographic context to the need for managing aged individuals. RESULTS: Each of the 18 cats exhibited a higher number of steps per minute after the addition of Antinol to their diet. At the group level, a paired t-test showed that the step rate was significantly increased after the addition of Antinol to the diet. LIMITATIONS: While our results showed a strong significant increase in step rate following Antinol supplementation, further studies that incorporate a placebo, more individuals and more detailed metrics of mobility would provide a more detailed evidence base for practitioners. CONCLUSION: Nutraceuticals may yield benefits to aged individual felids, including species kept widely in European collections. Their use warrants further, detailed research in collections.

Perspectives on SARS-CoV-2 Cases in Zoological Institutions.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections in a zoological institution were initially reported in March 2020. Since then, at least 94 peer-reviewed cases have been reported in zoos worldwide. Among the affected animals, nonhuman primates, carnivores, and artiodactyls appear to be most susceptible to infection, with the Felidae family accounting for the largest number of reported cases. Clinical symptoms tend to be mild across taxa; although, certain species exhibit increased susceptibility to disease. A variety of diagnostic tools are available, allowing for initial diagnostics and for the monitoring of infectious risk. Whilst supportive therapy proves sufficient in most cases, monoclonal antibody therapy has emerged as a promising additional treatment option. Effective transmission of SARS-CoV-2 in some species raises concerns over potential spillover and the formation of reservoirs. The occurrence of SARS-CoV-2 in a variety of animal species may contribute to the emergence of variants of concern due to altered viral evolutionary constraints. Consequently, this review emphasizes the need for effective biosecurity measures and surveillance strategies to prevent and control SARS-CoV-2 infections in zoological institutions.

Prolonged cold-preservation of domestic cat ovarian tissue is improved by extracellular solution but impaired by the fragmentation of ovary.

For domestic cats ovaries, recommended cold-storage limit is 24 h in Phosphate Buffered Saline (PBS) or Dulbecco`s PBS (DPBS). Here, we attempted to verify wheatear cat ovaries may benefit from more complex solutions during prolonged cold-storage (>24 h). First, the preservation capabilities of extracellular (SP+), intracellular (UW) solutions and DPBS supplemented with glutathione (DPBS+GSH) were compared using ovary fragments from the same ovary (n=10). Intact ovary stored in DPBS served as a control. Ovaries were kept at 4 °C for 48 h, and 72 h. In the second experiment, first ovary was stored in DPBS, second in SP+ or UW solution for 48 h (n = 12). Ovaries pairs stored in DPBS for 24 h served as a control (n=8). Tissue samples were evaluated directly after cold-storage and after following 24 h in vitro culture. Ovarian follicle morphology, apoptosis rates (cleaved caspase-3, TUNEL), and follicular growth activation (Ki-67) were assessed. Ovary fragmentation impaired follicular morphology preservation upon cold-storage comparing to intact ovary. However, ovarian fragments stored in UW for 48 h and in SP+ for 72 h presented better morphology than DPBS+GSH group. Comparison of intact ovaries cold-storage for 48 h showed that SP+ provided superior follicular morphology over DPBS, and it was comparable to the outcome of 24-hour storage. No follicular activation after in vitro culture was observed. Nevertheless, tissue culture increased considerably caspase-3 cleavage and TUNEL detection. The ovary fragmentation prior to cold-storage is not recommended in domestic cats. Replacement of DPBS with SP+ solution for whole ovary and UW solution for ovarian tissue fragments improves follicular structure preservation during 48-hour cold-storage.

A multidisciplinary approach to analyze the antimicrobial resistance in natural ecosystems.

Antimicrobial Resistance (AMR) poses a global threat to both human health and environmental well-being. Our study delved into Costa Rican wildlife reserves, uncovering a substantial human impact on these ecosystems and underscoring the imperative to pinpoint AMR hotspots. Embracing a One Health perspective, we advocated for a comprehensive landscape analysis that intricately intertwined geographic, climatic, forest, and human factors. This study illuminated the link between laboratory results and observed patterns of antimicrobial use, thereby paving the way for sustainable solutions. Our innovative methodology involved deploying open-ended questions to explore antimicrobial usage across livestock activities, contributing to establishing a comprehensive methodology. Non-invasive sampling in wildlife emerged as a critical aspect, shedding light on areas contaminated by AMR. Feline species, positioned at the apex of the food chain, acted as sentinels for environmental health due to heightened exposure to improperly disposed waste. Regarding laboratory findings, each sample revealed the presence of at least one antimicrobial resistance gene (ARG). Notably, genes encoding resistance to tetracyclines dominated (94.9%), followed by beta-lactams (75.6%), sulfonamides (53.8%), aminoglycosides (51.3%), quinolones (44.9%), phenicols (25.6%), and macrolides (20.5%). Genes encoding polymyxins were not detected. Moreover, 66% of samples carried a multi-resistant microbiome, with 15% exhibiting resistance to three antimicrobial families and 51% to four. The absence of a correlation between forest coverage and ARG presence underscored the profound human impact on wildlife reserves, surpassing previous estimations. This environmental pressure could potentially modify microbiomes and resistomes in unknown ways. As not all antimicrobial families encoding ARGs were utilized by farmers, our next step involved evaluating other human activities to identify the primary sources of contamination. This comprehensive study contributed crucial insights into the intricate dynamics of AMR in natural ecosystems, paving the way for targeted interventions and sustainable coexistence.

Detection of Trypanosoma evansi in jaguars (Panthera onca): insights from the Brazilian Pantanal wetland.

Trypanosoma evansi is a widespread and neglected zoonotic parasite that affects domestic and wild animals, causing a disease commonly known as "surra." The Brazilian Pantanal wetland is recognized as an enzootic area for this protozoan, yet recognizing the importance of reservoir hosts also in order to prevent zoonotic outbreaks. This study aimed to assess the occurrence of T. evansi in jaguars (Panthera onca) from the Brazilian Pantanal wetland and explore associated clinical and hematological manifestations. A total of 42 animals were screened by PCR and sequenced for species identification when positive. Trypanosoma evansi was detected in six free-ranging jaguars (six positive animals of 42 captures and 16 recaptures), representing the first molecular evidence of such infection in this animal species. Our findings suggest that jaguars may act as reservoir hosts of T. evansi in the Brazilian Pantanal wetland. The better understanding of the role of wildlife in the epidemiology of T. evansi is also of importance to future reintroduction and translocation programs toward wildlife conservation efforts.

Current practice for the chemical immobilisation of non-domestic feline species: An online survey study.

BACKGROUND: Safe chemical immobilisation of wild felids is essential for both conservational management and clinical purposes. However, little is known about drug protocols and current practice. METHODS: This study was designed as an online survey based on a questionnaire. Descriptive/correlation statistics and analysis of proportions were used for data analysis. RESULTS: The preferred immobilisation technique was the use of darts (37% of the respondents), while the most popular drug combination was a mixture of benzodiazepines, alpha-2 adrenoreceptor agonists and dissociative anaesthetics (27%). The inclusion of ketamine in the drug mixture was associated with a quicker anaesthetic onset, as estimated by the participants (p < 0.001). Common complications were prolonged recovery (46%), bradycardia (35%), hypoventilation (32%), hypothermia (26%) and arousal (26%). Commonly encountered problems were inappropriate equipment (39%), lack of suitable drugs (27%) and inadequate knowledge of species-specific pharmacology (29%) and physiology (24%). LIMITATIONS: Incomplete adherence to the Checklist for Reporting Results of Internet E-Surveys is acknowledged. CONCLUSIONS: Drug protocols including both alpha-2 adrenoreceptor agonists and dissociative anaesthetics are preferred in wild felids, and the inclusion of ketamine may be useful to achieve a quick onset. Equipment/drug availability and species-specific knowledge are potential areas of improvement to improve wild felid anaesthesia.

Trabecular bone morphology in big cats reflects the complex diversity of limb use but not home range size or daily travel distance.

A relationship exists between mechanical loading and bone morphology. Although studies show a relationship between trabecular bone morphology and locomotor strategy in mammals, none of them have studied trabecular bone morphology in felid species occupying disparate and overlapping habitats. We investigate trabecular bone volume fraction (BVF) in the femoral and humeral heads, and distal tibia of four felid species (mountain lions, jaguars, cheetahs, and leopards) to identify whether there is a relationship between BVF and locomotor behavior. This study's goals are to identify whether felid species with high daily travel distance or large home range size have greater BVF compared with those with small daily travel distance or home range size, and whether BVF is correlated among the three elements of the fore and hindlimb studied. We quantified BVF in micro- and peripheral computed tomography images and found no significant differences across species in the femoral and humeral head (p > 0.05). However, in the distal tibia, results showed that leopards, mountain lions, and cheetahs have significantly greater (p < 0.05) BVF than jaguars. Despite differences in home range size and daily travel distance, the proximal elements did not reflect differences in BVF; however, the distal-most element did, suggesting decreased loading among jaguars. These findings suggest that the observed pattern of trabecular bone morphology is potentially due to the diversity in locomotor strategy of the forelimb. Additionally, these results imply that neither home range size nor daily travel distance are clear indicators of activity levels. A cautious approach is warranted in studying how loading influences trabecular morphology.

Draft genomes of 16 Bartonella henselae strains from cats in Valdivia, Chile.

Bartonella henselae is a primary zoonotic agent, having cats as asymptomatic reservoirs. In humans, it causes cat scratch disease. Here, we report the whole genome sequences of 16 strains isolated from cats in Valdivia city, Southern Chile. Strains showed little variability in the multilocus sequence typing profiles.

Systemic histiocytic disorder in a jaguar (Panthera onca).

A 7-year-old captive female jaguar (Panthera onca) was presented with a 7-day history of dyspnoea and weight loss. Clinical examination revealed hepatomegaly and elevated serum alanine aminotransferase activity. Pulmonary ultrasonography revealed comet-tail images and an alveolar pattern was detected on thoracic radiography. Due to the poor prognosis, the jaguar was euthanized after 10 days. At necropsy, the main gross findings were hepatomegaly, splenomegaly and multifocal to coalescent, slightly elevated grey areas in the lungs. Histological examination revealed neoplastic proliferation of pleomorphic histiocytes arranged in cohesive sheets in the lungs, liver, spleen, kidneys and lymph nodes. Neoplastic cells had intense immunolabelling for vimentin and ionized calcium-binding adaptor molecule 1, and were immunonegative for pancytokeratin, E-cadherin, CD20, CD3 and CD79α. These findings were compatible with a systemic histiocytic disorder, distinct from any well-defined histiocytic proliferative disease in domestic animals.

Molecular survey of Cytauxzoon spp. and Hepatozoon spp. in felids using a novel real-time PCR approach.

Tick-transmitted apicomplexans of the genera Cytauxzoon and Hepatozoon affect a wide range of felids worldwide, but little is known about them. Recently, several studies addressed the species circulating in Europe, their distribution, and their hosts. Molecular assays are the method of choice for their detection. Unfortunately, conventional PCRs already described are time- and cost-consuming and specific for either Hepatozoon or Cytauxzoon detection. This study was developed to evaluate (i) the occurrence of Cytauxzoon and Hepatozoon in felids using a fast and cost-saving real-time PCR capable of detecting both protozoa simultaneously, (ii) the distribution of Cytauxzoon and Hepatozoon species in north-eastern Italy, and (iii) the involvement of other susceptible felid hosts in the same area. An SYBR® Green-based real-time PCR with primers targeting the 18S-rRNA was validated and applied to 237 felid samples, i.e., whole blood from 206 domestic cats and 12 captive exotic felids, and tissues from 19 wildcats. Positive results were obtained by melting temperature curve analysis due to the specific melting peak (i.e., 81°C Cytauxzoon spp.; 78-78.5°C Hepatozoon spp.). Positive samples were subjected to conventional PCR, followed by sequencing for species identification. Phylogenetic analyses were performed to assess relatedness among European isolates. Data on domestic cats (age class, sex, origin, management, and lifestyle) were recorded, and statistical analyses were performed to identify potential risk factors. A total of 31 (15%) domestic cats were positive for Hepatozoon spp. (i.e., 12 for H. felis, 19 for H. silvestris), while six (2.9%) for C. europaeus. The prevalence of Hepatozoon felis was significantly (p < 0.05) higher in domestic cats, while H. silvestris was higher in strays and animals from the Eastern region (i.e., Friuli-Venezia Giulia). Cytauxzoon europaeus was detected only in stray cats from Friuli-Venezia Giulia (province of Trieste). Among captive felids, one tiger was infected with H. felis and another with H. silvestris; eight out of 19 (42%) wildcats were positive for Hepatozoon spp. (i.e., six with H. felis, two with H. silvestris) and four out of 19 (21%) for Cytauxzoon europaeus. Outdoor lifestyle and origin (i.e., Friuli-Venezia Giulia region) were the most relevant risk factors for H. silvestris and C. europeus infections. Conversely, H. felis was most frequently isolated from domestic cats, suggesting different modes of transmission.

Bartonella spp. in households with cats: Risk factors for infection in cats and human exposure.

The aim of this study was to estimate the occurrence of Bartonella spp. per household in cats and the risk factors for Bartonella spp. positivity in cats and their owners from Valdivia, Chile. A total of 464 cats (distributed within 324 households) and 326 humans (control group [n = 112] and cat owner [n = 214]) distributed in 262 households were sampled. From the cat owners (n = 214), 128 humans were in households where the cat was also sampled, totaling 84 households with dual sampling. Real-time PCR (qPCR) was used for Bartonella spp. detection in blood from cats and humans, and immunofluorescent immunoassay (IFA) anti-Bartonella henselae was performed in human serum samples. Out of the total of 324 households, 20.43% presented at least one Bartonella positive cat. From the households with dual sampling, 29.7% (25/84) presented at least one qPCR-Bartonella spp. positive cat. However, Bartonella DNA was not amplified in humans, and in 7.3% (6/82) of the households was found at least one of the cat's owners exposed to B. henselae. Cats younger than one year (Odds Ratio (OR) = 5.3), non-neutered (OR 3.46), sampled at home (OR 5.82), and with improper application of tick/flea control products (OR 3.13) showed a higher risk for Bartonella spp. presence. Humans with occupational exposure involving animal contact, were more likely to exhibit B. henselae seropositivity (OR 7.5). Bartonella spp. was present in the cats a moderate number of households, but Bartonella DNA was not detected in owners' blood, inferring that there is a low risk of recent human infection in the studied population.

Do not disturb: Visitors' effect on the behavior and welfare of female and male big cats in zoos.

The effect of visitor presence on animal behavior needs to be evaluated to ensure the welfare and improve the husbandry of the individuals in zoos. This study aims to assess the effect of visitor presence on the behavior and welfare of pairs of Amur tiger, snow leopard, and Eurasian lynx at Parco Natura Viva, Italy. The study was made of two periods: the baseline, in which the zoo was closed, and the visitor presence period, in which the zoo was opened. Per period and per subject 12 30-min observations were done. Continuous focal animal sampling method was used to collect behavior duration of the big cats. The main results of the study highlighted that, when visitors were present, all felids except for the female lynx were significantly more inactive than in the baseline. Moreover, despite interindividual and inter-species variability in the significance level of findings, natural behaviors such as attentive behavior, exploration/marking, locomotion, and positive social interactions were performed more in the baseline than in the visitor presence period. Finally, when visitors were present, as the studied subjects experienced daily longer exposure to visitors, inactivity increased whereas individual species-typical behaviors (e.g., locomotion) and positive social interactions decreased. Thus, visitors seem to partially alter the behavioral time-budgets of the study big cats, increasing inactivity and decreasing the performance of species-specific behavior, at least in some individuals.