Genotyping of Enterocytozoon bieneusi isolates detected in stray cats of Izmir, Türkiye.

The phylum Microsporidia includes obligate intracellular parasites that can infect humans and various animals. To date, 17 different species within the phylum have been reported to infect humans. Among them, Enterocytozoon bieneusi (E. bieneusi) is one of the most frequently detected species in humans. Identification of E. bieneusi as well as its genotypes in humans and animals is important to reveal their role in transmission to each other. Cats are blamed as the source of E. bieneusi transmission to humans. In this study, we aimed to genotype 170 E. bieneusi positive samples isolated from stool of stray cats living in Izmir province of Türkiye. According to the results, 47 samples were amplified by nested PCR protocol targeting ITS region and successfully sequenced. The phylogenetic analysis showed the presence of zoonotic genotype D and type IV in stray cats, which are also frequently detected in humans. Among the E. bieneusi genotypes detected, the prevalence of type IV (93.6%; 44/47) was very high compared to genotype D. Overall, the identification of zoonotic genotypes of E. bieneusi supports that stray cats can play an important role in the transmission of E. bieneusi to humans in Izmir.

Molecular characterization of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene and frequency of blood types in stray cats of Izmir, Turkey.

BACKGROUND: Cytidine monophospho-n-acetylneuraminic acid hydroxylase (CMAH) gene associated with blood groups in cats encodes CMAH enzyme that converts Neu5Ac to Neu5Gc. Although variations in CMAH gene of pedigree cats have been revealed, the presence/lack of them in non-pedigree stray cats is unknown. Therefore, the present study aimed to investigate the variations in CMAH gene and the quantity of Neu5Ac and Neu5Gc on erythrocytes of non-pedigree stray cats (n:12) living in Izmir, Turkey. Also, the frequency of blood types was determined in 76 stray cats including 12 cats that were used for CMAH and Neu5A/Neu5Gc analysis. RESULTS: In total, 14 SNPs were detected in 5'UTR as well as in exon 2, 4, 9, 10, 11 and 12 of CMAH gene. Among these SNPs, -495 C > T in 5'UTR was detected for the first time as heterozygous in type A and AB cats, and homozygous and heterozygous in type B cats. The remaining 13 that have been detected in previous studies were also found as homozygous or heterozygous. Both Neu5Gc and Neu5Ac were detected in type A and AB cats. In type B cats, only Neu5Ac was detected. Among two type AB cats, the level of Neu5Ac was found higher in cat carrying heterozygous form (T/C) of 1392T > C. The prevalence of type B cats (67.1 %) was higher than others. CONCLUSIONS: The presence of a new SNP as well as previous SNPs indicates that more variations can be found in stray cats with a more comprehensive study in the future. Also, the high prevalence of type B cats demonstrates the possible risk of neonatal isoerythrolysis among stray cats living in Izmir, Turkey.

A survey of opinions towards dog and cat management policy issues in New Zealand.

Aims: To describe the opinions of respondents to an online survey on desexing, microchipping and pet registration, and the management of cats, and aggressive dogs in New Zealand. Methods: A cross-sectional survey was conducted online from 18-22 June 2015 using a permission-based panel of New Zealand residents aged ≥18 years. Questions included demographics of respondents, number of dogs and cats owned, and opinions on desexing, microchipping, pet registration, management of cats and aggressive dogs. Results: Of the 1,572 survey respondents, 216 (13.7%) owned ≥1 dog and ≥1 cat, 227 (14.4%) owned ≥1 dog and no cats, 480 (30.5%) owned ≥1 cat and no dogs, and 559 (35.6%) did not own any pets. The majority of dogs (456/613; 74.8%) and cats (974/1,045; 93.2%) were desexed. The most common reasons for not desexing pets were cost, feeling that it was not needed, or keeping the dog for breeding. Of the 613 dogs, 557 (90.9%) were registered with the local council, and 434 (71.0%) were microchipped, but only 290 (47.3%) were registered in the national database. Of the 1,045 cats, 326 (31.2%) were microchipped and 279/486 (57.4%) owners felt that it was unnecessary. Of the 1,572 respondents, 947 (60.2%) were unaware of stray cats in their local community, and 479 (30.5%) stated that local councils, or animal welfare organisations (546/1,572; 34.7%) should be responsible for managing strays. Among all 1,572 respondents, 787 (50.1%) thought stray cats should be assessed and subjected to euthanasia. Compared with non-pet owners, a lower percentage of cat owners agreed that cats should be confined (p < 0.001). When asked to choose the most appropriate course of action for dogs that had bitten people or other animals, 849 (54.0%) and 820 (52.5%) respondents, respectively, agreed that the dog should be assessed by an expert who would then determine the appropriate action. Compared with non-pet owners, a lower percentage of dog owners supported registration and education of dog owners, aggressive dogs being destroyed, and giving more power to local councils (p < 0.05). Conclusions: The results highlight opportunities to improve owner compliance with desexing, microchipping and registration of dogs and cats. Opinions towards management of stray cat and aggressive dogs varied between pet owners and non-owners. Further research is needed to better understand how to engage the public in important dog and cat management policy issues.

Seroprevalence of Leishmania infection and molecular detection of Leishmania tropica and Leishmania infantum in stray cats of Izmir, Turkey.

Leishmaniasis caused by more than 20 species of genus Leishmania is transmitted by the bite of infected phlebotomine sand flies. The studies on Leishmania infection in cats is very few in Turkey and therefore we aimed to screen stray cats living in city of Izmir located in western Turkey using nested PCR targeting kinetoplast DNA and serological techniques (ELISA and IFA). Leishmania DNA positive samples were also studied by ITS1 real time PCR. Whole blood and serum samples were obtained from stray cats (n: 1101) living in different counties of Izmir. In serological assays, a serum sample was considered positive in 1:40 dilution in IFA and for ELISA a serum sample was accepted positive when the absorbance value (AV) exceeded the mean AV + Standard Deviation (SD) of the negative control serum samples. According to the results, the seropositivity rates were 10.8% (119/1101) and 15.2% (167/1101) by in house ELISA and IFA, respectively. Among serology coherent samples, the seropositivity rate was 11.1% (116/1047) as detected by both assays after discordant samples (n: 54) were discarded. Of the 1101 stray cats, six (0.54%) were positive by nested PCR while only one of these six samples was positive by ITS1 real time PCR. During PCR, three controls designated as Leishmania infantum, Leishmania tropica, and Leishmania major were used for species identification. According to nested PCR results, L. tropica was identified in two cats (no.76 and 95). In another cat (no. 269), there were two bands in which one of them was well-matched with L. infantum and the other band had ∼850 bp size which does not match with any controls. Remaining three cats (no. 86, 514, and 622) also had the ∼850 bp atypical band size. ITS1 real time PCR detected L. tropica in only one cat (no. 622) which showed an atypical band size in nested PCR. These results indicated that three cats with only one atypical band (no. 86, 514, and 622) and the cat with mixed infection (no. 269) were infected with L. tropica. Altogether, L. tropica was detected in all six DNA positive cats and L. infantum was detected in one cat with mixed infection. In conclusion, although the reservoir role of cats in nature is still unclear the high seroprevalence rate against Leishmania parasites and detecting parasite DNA in stray cats in Izmir indicates that the stray cats are frequently bitten by infected sand flies. Further research activities are required to reveal the frequency of leishmaniasis in cats in different regions of Turkey where Leishmania species are endemic.

Molecular detection and genotyping of Toxoplasma gondii in stray cat feces from Khorramabad, West Iran.

Cats, being the definitive host of Toxoplasma gondii, have a significant impact on the spread and outbreaks of the parasite. An essential factor in comprehending the transmission pattern of this parasite is an analysis of the genetic diversity distribution in cats infected with T. gondii. This study was aimed at determining the prevalence rate and genotyping of T. gondii in stray cat feces from Khorramabad, West Iran. In the years 2016-2017, 200 cats were sampled to get fresh feces specimens. Parasitological methods were utilized for the identification of oocysts. The DNA was isolated from the feces using a commercially available Genomic Mini Kit. In order to identify the genetic composition of T. gondii, we employed PCR-RFLP, sequencing, and phylogenetic analysis of the GRA6 target gene. No one of the samples tested positive for parasitology techniques. A total of 6.5 % (13/200) samples were positive when using the GRA6-PCR method. Based on PCR-RFLP results, all 13 samples were of T. gondii type III genotype. The nucleotide sequences of two samples from this study were found to be 5 % different from those of 12 references of T. gondii and one strain of Hammondia hamondi that was used as an external control. Based on the findings, molecular tests are more sensitive than parasitological methods. The RFLP approach revealed that type III of T. gondii is the prevailing and important genotype in Khorramabad, West Iran.

First report of Bartonella henselae and Bartonella clarridgeiae carriage in stray cats from Ecuador and its link to a cat scratch disease outbreak in 2022.

INTRODUCTION: The genus Bartonella includes species and subspecies of fastidious, facultative intracellular Gram-negative bacilli that infect a wide variety of mammalian reservoirs including cats and humans. In 2022, the Ecuadorian Ministry of Health reported an outbreak of cat scratch disease caused by B. henselae in the city of Guayaquil. Therefore, we aimed to characterize the presence of Bartonella spp. in domestic and stray cats from the area of Guayaquil where the outbreak happened in 2022. METHODS: Whole blood samples of 100 domestic and stray cats were collected. Riboflavin synthase (ribC) and 16S rRNA genes detection was performed by PCR using Bartonella spp. specific primers, followed by Sanger sequencing and phylogenetic analysis. RESULTS: 14 cats were positive for Bartonella spp. carriage. Phylogenetic analysis confirmed the presence of 12 cats infected with B. henselae and 2 cats with B. clarridgeiae. CONCLUSIONS: There is a high prevalence of Bartonella spp. carriage in cats in the city of Guayaquil within the area where a recent cat scratch disease outbreak happened. Considering the high presence of cats and other domestic and stray animals in the city of Guayaquil, a One Health approach for surveillance and prevention of zoonotic diseases like cat scratch disease is needed.

Prospective Investigation of Feline Leukemia Virus Infection in Stray Cats Subjected to a Trap-Neuter-Return Program in Switzerland.

Feline leukemia virus (FeLV) remains a serious concern in some countries despite advances in diagnostics and vaccines. FeLV-infected cats often have reduced lifespans due to FeLV-associated diseases. The infection is transmitted through social interactions. While Northern European countries have reported a decrease in FeLV among pet cats, Switzerland's rates remain stagnant at 2.7% (2016/17: 95% CI 1.4-5.2%). Research on FeLV in Swiss stray cats has been lacking, even though these animals could serve as a virus reservoir. Sampling stray cats that do not receive regular veterinary care can be challenging. Collaboration with the Swiss Network for Animal Protection (NetAP) allowed for the prospective collection of saliva samples from 1711 stray cats during a trap-neuter-return program from 2019 to 2023. These samples were tested for FeLV RNA using RT-qPCR as a measure for antigenemia. Viral RNA was detected in 4.0% (95% CI 3.1-5.0%) of the samples, with 7.7% (95% CI 4.9-11.3%) in sick cats and 3.3% (95% CI 2.4-4.4%) in healthy ones. We identified three geographically independent hotspots with alarmingly high FeLV infection rates in stray cats (up to 70%). Overall, including the previous data of privately owned cats, FeLV-positive cats were scattered throughout Switzerland in 24/26 cantons. Our findings underscore welfare concerns for FeLV infections among stray cats lacking veterinary attention, highlighting the potential risk of infection to other free-roaming cats, including those privately owned. This emphasizes the critical significance of vaccinating all cats with outdoor access against FeLV and developing programs to protect cats from FeLV infections.

Seroprevalence of zoonotic pathogens in stray cats in an urban area of northeast Spain.

The feline population is extensive in urban areas worldwide, comprising stray and domestic cats. Cats, acting as reservoirs, can transmit various zoonotic organisms to humans, which can cause significant public health issues. We evaluated the seroprevalence of zoonotic pathogens in stray cats in an urban area of northeast Spain (the city of Zaragoza) to assess potential risks to human health. A total of 88 sampled cats (52 females and 36 males) underwent antibody evaluation using the indirect immunofluorescence technique. Seroprevalence rates were determined for IgG antibodies to Bartonella henselae (36.3%), Toxoplasma gondii (31.8%), Rickettsia felis (14.7%), Rickettsia typhi (9%), and Leishmania infantum (10.2%). Our results confirmed the presence in stray cats of antibodies against all those pathogens, indicating that they all circulate in the feline population in Zaragoza. Male cats exhibited a higher predisposition to T. gondii, whereas females showed an increased likelihood of contracting B. henselae. This difference may be attributed to distinct behaviors according to sex. Our findings underscore the importance of maintaining and intensifying surveillance coupled with preventive measures against zoonotic pathogens in cats. They highlight the need for comprehensive control strategies designed to mitigate public health risks associated with feline populations.

High prevalence of antibodies against feline calicivirus in Australian feral and stray cat (Felis catus) populations.

INTRODUCTION: Feline calicivirus (FCV) commonly causes upper respiratory tract, oral and ocular infections in species of the family Felidae, with high prevalence amongst domestic cat (Felis catus) populations worldwide. Detection of FCV-specific antibodies in serum provides evidence of previous infection with FCV and an indication of whether a cat may be protected against clinical FCV disease. This study describes the most extensive sampling for anti-FCV antibodies in feral and stray cat populations in Australia, and examines variation in prevalence associated with cat age, sex and location. METHODS: Blood samples were opportunistically collected from 669 feral, stray or Indigenous community cats from the Northern Territory, South Australia, Victoria, south-east Tasmania and south-west New South Wales. The sera were harvested and tested for antibodies capable of neutralising the FCV vaccine strain F9 by serum-virus neutralisation assay. RESULTS: Of the 669 cats tested, 69.7% had detectable FCV-F9-neutralising antibodies (titres ≥5). Maturity was significantly associated with higher seroprevalence and higher antibody titres, with adult cats being more than twice as likely to have detectable FCV-neutralising antibodies than subadults. Male cats had a higher seroprevalence and slightly higher antibody titres than females. Cats living in closer proximity to humans had significantly higher seroprevalences and higher FCV-neutralising antibody titres than feral cats from more remote regions of Australia. CONCLUSION: Australian feral and stray cats have a high risk of natural exposure to and infection with FCV, with the prevalence and levels of pre-existing immunity to FCV being highest amongst adult cats living in highly modified urban, peri-urban and agricultural environments.

Epidemiological Surveillance of Respiratory Diseases in Urban Stray Cats in Shanghai.

Urban stray cats are cats without owners that survive in the wild for extended periods of time. They are one of the most common stray animals in cities, and as such, monitoring the pathogens carried by urban stray cats is an important component of urban epidemiological surveillance. In order to understand the prevalence of respiratory diseases in urban stray cats in Shanghai and provide scientific evidence for the development of targeted prevention and control strategies for respiratory diseases in stray cats, we collected 374 ocular, nasal, and oropharyngeal swabs from urban stray cats in Shanghai from January 2022 to December 2022. After RNA extraction, we used real-time PCR to detect six respiratory pathogens, including influenza A virus, feline calicivirus, feline herpesvirus type 1, Mycoplasma, Chlamydia, and Bordetella bronchiseptica. The results showed that among the 374 samples, 146 tested positive, with a positivity rate of 39.04%. The highest positivity rate was observed for Mycoplasma felis at 18.72% (70/374), followed by Chlamydia felis at 11.76% (44/374), feline calicivirus at 3.74% (14/374), feline herpesvirus 1 at 3.48% (13/374), Bordetella bronchiseptica at 1.34% (5/374), and influenza A virus was not detected. The highest positivity rate for Mycoplasma felis was in Minhang District at 31.94% (23/72), while Chlamydia felis and Bordetella bronchiseptica had the highest positivity rates in Jiading District at 23.53% (8/34) and 5.88% (2/34), respectively. The highest positivity rates for feline calicivirus and feline herpesvirus 1 were both observed in Qingpu District, at 14.46% (12/83) and 9.64% (8/83), respectively. A total of 36 samples showed mixed infections with two or more pathogens, with Mycoplasma felis being involved in 32 of these mixed infections, with the highest number of mixed infections being with Chlamydia felis at 25 samples. Respiratory pathogen positivity was detected throughout the year, with peak detection rates in summer and winter. The positivity rates of cat respiratory pathogens in different seasons showed statistical differences (χ2 = 27.73, p < 0.01). There was no statistical difference in the positivity rates of respiratory pathogens between cats of different genders (χ2 = 0.92, p > 0.05). The positivity rates of respiratory pathogens in cats of different age groups showed statistical differences (χ2 = 44.41, p < 0.01). Mycoplasma felis and Chlamydia felis were the main pathogens causing respiratory infections in stray cats, with Mycoplasma felis showing a much higher positivity rate than other respiratory pathogens and often co-infecting with Chlamydia felis and feline calicivirus. The positivity rate of Mycoplasma felis was high in summer, autumn, and winter, with no statistical difference between seasons. These results indicate a serious overall prevalence of respiratory pathogens in urban stray cats in the Shanghai area, showing seasonal trends and mixed infections with other pathogens. These findings suggest the need for comprehensive prevention and control measures to address respiratory pathogen infections in urban stray cats in the Shanghai area.

Occurrence of Toxoplasma gondii and other gastrointestinal parasites in free-roaming cats from the Rio de Janeiro zoo.

This communication aimed to determine the frequency of infection by T. gondii and other gastrointestinal parasites in free-living cats captured in BioParque do Rio (Fundação RioZoo), Brazil. To this end, 58 blood and 51 fecal samples were collected from 68 cats from July 2019 to September 2020. The serum obtained was analyzed by indirect fluorescent antibody test for anti-T. gondii IgG. Fecal samples were examined by microscopic parasitological techniques. Of the total, 10.3% of the animals presented anti-T. gondii IgG. Parasitic structures were observed in 39.2% of the fecal samples. Hookworms were the most detected parasites (19.6%) followed by Cystoisospora sp. (11.7%), Dipylidium caninum (7.8%) and Toxocara cati (3.9%). These results indicated the exposure of this population of stray cats to potentially zoonotic parasites, which in addition to causing possible damage to domestic and wild animal health, also pose risks to public health.

Esta comunicação teve como objetivo determinar a frequência de infecção por T. gondii e outros parasitas gastrointestinais em gatos de vida livre capturados no BioParque do Rio (Fundação RioZoo), Brasil. Para isso, foram coletadas 58 amostras de sangue e 51 amostras fecais de 68 gatos capturados entre julho de 2019 e setembro de 2020. O soro obtido foi submetido à reação de imunofluorescência indireta para IgG anti-T. gondii. As amostras fecais foram submetidas a técnicas parasitológicas microscópicas. Do total, 10,3% dos animais apresentaram IgG anti-T. gondii. Estruturas parasitárias foram observadas em 39,2% das amostras fecais. Os ancilostomídeos foram os parasitas mais detectados (19,6%), seguidos por Cystoisospora sp. (11,7%), Dipylidium caninum (7,8%) e Toxocara cati (3,9%). Esses resultados indicaram a exposição dessa população de gatos de rua a parasitas potencialmente zoonóticos, que, além de causar possíveis danos à saúde dos animais domésticos e selvagens, também representam riscos à saúde pública.

Antibiotic Resistance Genes Carried by Commensal Escherichia coli from Shelter Cats in Italy.

Antimicrobial resistance is a widespread global health problem. The presence of resistant bacteria and antibiotic resistance genes has been demonstrated not only in humans but also in animals, including pets. Stray cats share the urban environment with people and pets. This may facilitate transmission of resistant bacteria and resistance genes between stray animals, people and domestic animals. Several studies have investigated the role of stray cats as a fecal carrier of ESBL-producing bacteria. However, there are many genes and resistance mechanisms that can be detected in commensal E. coli, which, because of its genetic plasticity, is considered an indicator for monitoring antibiotic resistance. In this study, rectal swabs were collected from stray cats from colonies and shelters in the city of Monza (Monza Brianza, Italy) to isolate commensal E. coli. Phenotypic tests, such as the minimum inhibitory concentration (MIC) and the double disc test (DDST), and molecular analyses to detect antimicrobial resistance genes (ARGs) were used to study the resistance of these isolates. The results obtained confirm that stray cats can carry ESBL-producing E. coli (6.7%) and genes conferring resistance to other important antibiotic classes such as tetracyclines and sulfonamides.

Molecular prevalence and genetic diversity of Hepatozoon spp. in stray cats of Izmir, Türkiye.

Hepatozoon spp. are an apicomplexan protozoan parasites that infect vertebrates including mammals, marsupials, birds, reptiles, and amphibians. Among Hepatozoon species, H. canis and H. felis are causative agents of hepatozoonosis in dogs and cats, respectively and have veterinary importance. This study aimed to determine the prevalence of Hepatozoon spp. in stray cats living in Izmir and investigate genetic diversity among positive samples. To achieve this aim, the prevalence of Hepatozoon spp. 18S rRNA gene was screened by PCR in DNA samples extracted from blood samples of stray cats (n = 1012). Then, Hepatozoon-positive samples were sequenced and the generated data were used for species identification, phylogenetic and haplotype analyses. According to the results, among the samples screened, 2.37 % (24/1012) of them were found to be Hepatozoon-positive, and of these positive samples, 18 (18/24; 75 %) were successfully sequenced. BLAST and phylogenetic analyses revealed that all of these samples were H. felis. Also, phylogenetic analysis showed that H. felis samples were genotype I. Within H. felis samples isolated from cats living in different countries/regions, 9 haplotypes were detected and among these haplotypes, H-1 was found to be prevalent (n = 20 H. felis isolates). In conclusion, this study showed that the prevalence of Hepatozoon spp. was low in stray cats analyzed. Also, H. felis genotype I was predominant in comparison to other Hepatozoon species.

Detection of Leishmania major and Leishmania infantum in cats during an outbreak of cutaneous leishmaniosis in Southern Israel.

Prevalence of Leishmania spp. infection was studied in stray cats in two military bases in Southern Israel during a cutaneous leishmaniosis (CL) human outbreak caused by Leishmania major. Human CL cases increased from 0/100 in 2008 to 1.28/100 in 2022 in camp #1, and from 0.17/100 in 2008 to 6.4/100 in 2022, in camp #2. Eight out of 29 cats sampled were Leishmania-seropositive (28 %) and 7/29 (24 %) were internal transcribed spacer 1 (ITS1) PCR-positive, out of which four (14 %) were positive for L. major and three (10 %) for L. infantum. Five positive-cats had skin lesions including ulcers, alopecia and scabs, and five had eye lesions. This is the first report of L. major infection in cats in Israel and one of the first descriptions in felines worldwide. A larger cohort of cats and vector studies are necessary to determine if felids may act as reservoirs or sentinels of human L. major infection.

Molecular prevalence of Enterocytozoon bieneusi in stray cats of Izmir, Türkiye.

The phylum Microsporidia contains obligate single celled parasites that can infect many vertebrate hosts including humans. Enterocytozoon bieneusi is considered as the most diagnosed species in humans. E. bieneusi has also been detected in many animals such as cats, dogs and cattle. Among these animals, cats are carriers of type D and IV which are the most common human pathogenic genotypes of E. bieneusi. In Türkiye, the prevalence of E. bieneusi in stray cats is not well known. Therefore, in this study, the molecular prevalence of E. bieneusi in stray cats (n = 339) was determined by Real-Time PCR targeting ribosomal DNA ITS (internal transcribed spacer) region of E. bieneusi. Initially, the analytical sensitivity of Real-Time PCR was determined by a plasmid control and then E. bieneusi DNA was investigated in fecal samples of stray cats. The results showed that the analytical sensitivity of Real-Time PCR targeting ITS region of E. bieneusi was ≤1 copy plasmid/reaction. Analysis of fecal samples revealed that the molecular prevalence of E. bieneusi was 50.15% (170/339). Overall, these results showed that the Real-Time PCR successfully detected E. bieneusi in cat's fecal samples and stray cats can be an important source for transmission of E. bieneusi to humans and other animals.

A cross-sectional serosurvey of SARS-CoV-2 and co-infections in stray cats from the second wave to the sixth wave of COVID-19 outbreaks in Spain.

Severe Acute Respiratory Syndrome Coronavirus 2 is the causative agent of Coronavirus Disease 2019 in humans. Among domestic animals, cats are more susceptible to SARS-CoV-2 than dogs. The detection of anti-SARS-CoV-2 antibodies in seemingly healthy cats and/or infected cats which are in close contact with infected humans has been described. The presence of animals that tested positive by serology or molecular techniques could represent a potential transmission pathway of SARS-CoV-2 that can spill over into urban wildlife. This study analyses the seroprevalence variation of SARS-CoV-2 in stray cats from different waves of outbreaks in a geographical area where previous seroepidemiological information of SARS-CoV-2 was available and investigate if SARS-CoV-2-seropositive cats were exposed to other co-infections causing an immunosuppressive status and/or a chronic disease that could lead to a SARS-CoV-2 susceptibility. For this purpose, a total of 254 stray cats from Zaragoza (Spain) were included. This analysis was carried out by the enzyme-linked immunosorbent assay using the receptor binding domain of Spike antigen and confirmed by serum virus neutralization assay. The presence of co-infections including Toxoplasma gondii, Leishmania infantum, Dirofilaria immitis, feline calicivirus, feline herpesvirus type 1, feline leukemia virus and feline immunodeficiency virus, was evaluated using different serological methods. A seropositivity of 1.57% was observed for SARS-CoV-2 including the presence of neutralizing antibodies in three cats. None of the seropositive to SARS-CoV-2 cats were positive to feline coronavirus, however, four SARS-CoV-2-seropositive cats were also seropositive to other pathogens such as L. infantum, D. immitis and FIV (n = 1), L. infantum and D. immitis (n = 1) and L. infantum alone (n = 1).Considering other pathogens, a seroprevalence of 16.54% was detected for L. infantum, 30.31% for D. immitis, 13.78%, for T. gondii, 83.86% for feline calicivirus, 42.52% for feline herpesvirus type 1, 3.15% for FeLV and 7.87% for FIV.Our findings suggest that the epidemiological role of stray cats in SARS-CoV-2 transmission is scarce, and there is no increase in seropositivity during the different waves of COVID-19 outbreaks in this group of animals. Further epidemiological surveillances are necessary to determine the risk that other animals might possess even though stray cats do not seem to play a role in transmission.

Public Opinions on Stray Cats in China, Evidence from Social Media Data.

The management of stray cats is often contentious because public perceptions about these animals are different. Using user-generated content from Weibo, this study investigated Chinese citizens' opinions on stray cats on a large scale. Through the techniques of natural language processing, we obtained each Weibo post's topics and sentiment propensity. The results showed that: (1) there were some irresponsible feeding behaviors among citizens; (2) public perceptions of the ecological impacts caused by stray cats were unlike; (3) the trap-neuter-return (TNR) method served high support in public discussion; (4) knowledge about stray cats' ecological impacts was positively correlated with support for the lethal control methods in management. Based on these findings, we suggested that management policies should be dedicated to (1) communicating to the (potential) cat feeders about the negative aspects of irresponsible feeding behaviors; (2) raising "ecological awareness" campaigns for the public as well as highlighting the environmental impacts caused by stray cats; (3) understanding citizens' perceptions toward different management scenarios and making decisions accordingly. In addition, this study also suggested that social media data can provide useful information about people's opinions on wild animals and their management. Policies would benefit by taking this source of information into the decision-making process.

Occurrence of Chlamydia spp. in Conjunctival Samples of Stray Cats in Timișoara Municipality, Western Romania.

Despite the widespread public health concern about stray cats serving as reservoirs for zoonotic agents, little is known about the effect of urban and peri-urban landscapes on exposure risk. We conducted this study to monitor the presence of Chlamydia spp. in stray cats, with or without conjunctivitis, living in Timișoara Municipality, Western Romania, using staining and PCR methods. A total of 95 cats were enrolled, and conjunctival samples were harvested from 68 clinically healthy cats and another 27 cats presenting with clinical signs of conjunctivitis. Overall, we found that 65.3% (62/95) of the cats tested positive for Chlamydia spp. by PCR. Chlamydia spp. were detected in 45/95 conjunctival samples using a standard Giemsa stain, compared with 62/95 using PCR (Cohen's kappa index = 0.308; p = 0.0640). Of the cats that tested positive by PCR, 72.6% (45/62) were asymptomatic, and another 27.4% (17/62) expressed clinical signs of conjunctivitis. We found no significant difference between (p > 0.05) the distribution of infection and the recorded epidemiological data (sex, breed, age, territorial distribution, or sampling season). However, the Chlamydia spp. detection frequency was significantly higher in asymptomatic than in symptomatic cats (p = 0.0383). The obtained results increase the level of concern and awareness about the possible zoonotic potential of this pathogen and highlight that urban stray cats can be essential sources of feline chlamydiosis.

Absence of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in stray cats.

SARS-CoV-2 positive or seropositive owned cats have been reported worldwide. The detection of seropositive stray cats in the proximity of farms of infected minks, coupled with the demonstration of cat-to-cat transmission in experimental settings, raise the question whether stray cats may have an epidemiological role in the COVID-19 pandemic and may act as sentinel for the circulation of SARS-CoV-2. The aim of this study was to evaluate the presence of SARS-CoV-2 RNA and anti-SARS-CoV-2 antibodies in free roaming cats belonging to colonies located in an area highly affected by the COVID-19 pandemic and to correlate the results with the positivity rate in people sharing the same area. Interdigital, cutaneous, oropharyngeal, nasal and rectal swabs, as well as blood samples, were collected from 99 cats living in colonies and admitted to our hospital for neutering. This caseload corresponds to the 24.2% of the feline population living in the 25 sampled colonies and to the 5.6% of all the free-roaming registered cats. The presence of SARS-CoV-2 RNA in swabs was assessed using real time RT-PCR. Anti-SARS-CoV-2 serum antibodies were assessed using commercially available ELISA kits and confirmed by serum virus neutralization. In people, the SARS-CoV-2 positivity rate ranged from 3.0% to 5.1% (mean rate: 4.1%) and the seropositive rate from 12.1% to 16.3% (mean rate: 14.2%). Most of the colonies were in urban areas and resident cats had frequent contacts with external cats or people. A COVID-19 positive caretaker was found, whereas all the cats were negative for SARS-CoV-2 RNA and seronegative. Although the negative results cannot exclude previous infections followed by decrease of antibodies, this study suggests that colony cats do not have an important epidemiological role in SARS-CoV-2 transmission dynamics. Further studies on larger caseloads are warranted, also in the light of the emerging new viral variants, on a One Health perspective.

One-Year Surveillance of SARS-CoV-2 Exposure in Stray Cats and Kennel Dogs from Northeastern Italy.

Dogs and cats are susceptible to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). During the pandemic, several studies have been performed on owned cats and dogs, whereas limited data are available on the exposure to stray animals. The objective of this study was to investigate the exposure to SARS-CoV-2 of feral cats and kennel dogs in northeastern Italy, through serological and molecular methods. From May 2021 to September 2022, public health veterinary services collected serum, oropharyngeal, and rectal swab samples from 257 free-roaming dogs newly introduced to shelters, and from 389 feral cats examined during the routinely trap-neutered-return programs. The swabs were analyzed for viral RNA through a real-time reverse transcriptase PCR (rRT-PCR), and sera were tested for the presence of the specific antibody against SARS-CoV-2 (enzyme-linked immunosorbent assay). Serology was positive in nine dogs (9/257) and three cats (3/389), while two asymptomatic cats tested positive to rRT-PCR. One cat turned out to be positive both for serology and molecular analysis. In addition, this study described the case of a possible human-to-animal SARS-CoV-2 transmission in a cat that travelled in close contact to a COVID-19-positive refugee from Ukraine. This study shows that SARS-CoV-2 can infect, in natural conditions, stray cats and kennel dogs in northeastern Italy, although with a low prevalence.