Feline Leukemia Research Articles

Animal shelter management of feline leukemia virus and feline immunodeficiency virus infections in cats.

Approximately 5% of cats in animal shelters in the United States test positive for either feline leukemia virus (FeLV) or feline immunodeficiency virus (FIV), which translates to more than 100,000 positive cats managed by shelters each year. Little is known about the current status of retroviral management in animal shelters, particularly in regions burdened by chronic pet overpopulation and high shelter admissions, such as the southern United States. The purpose of this study was to describe feline retroviral management in Florida shelters. Shelters were surveyed on practices including selection of cats for testing, diagnostic techniques, and outcome options for cats with positive test results. Responses were received from 139 of 153 animal shelters known to admit cats, including 55 municipal shelters (40%), 70 private shelters (50%), and 14 private shelters with municipal contracts (10%). A total of 115 shelters (83%) performed at least some testing, most using combination point-of-care devices for simultaneous FeLV antigen and FIV antibody screening. Of shelters that performed any testing, 56 (49%) tested all cats for FeLV and 52 (45%) tested all cats for both FeLV and FIV. The most common reason for testing was screening adoptable cats (108 shelters; 94%) and cats available for transfer to other organizations (78; 68%). Testing cats in trap-neuter-return/return-to-field programs was least common (21; 18%). Most common outcome options for positive cats included adoption (74; 64%), transfer (62; 54%), and euthanasia (49; 43%). Euthanasia following a positive test result was more common for cats with FeLV (49; 43%) than for cats with FIV (29; 25%) and was more common in municipal shelters, rural shelters, shelters taking in <500 cats a year, and shelters with overall live outcome rates for cats <70%. Although Florida shelter compliance with national guidelines for identification and management of FeLV and FIV positive cats was variable, most had live outcome options for at least some of their cats with positive test results. Increased access to training and practical programmatic tools may help more shelters implement cost-effective testing protocols, reduce risk for transmission to other cats, and support the best outcomes for this vulnerable population of cats.

Endothelial cells require functional FLVCR1a during developmental and adult angiogenesis

The Feline Leukemia Virus Subgroup C Receptor 1a (FLVCR1a) is a transmembrane heme exporter essential for embryonic vascular development. However, the exact role of FLVCR1a during blood vessel development remains largely undefined. Here, we show that FLVCR1a is highly expressed in angiogenic endothelial cells (ECs) compared to quiescent ECs. Consistently, ECs lacking FLVCR1a give rise to structurally and functionally abnormal vascular networks in multiple models of developmental and pathologic angiogenesis. Firstly, zebrafish embryos without FLVCR1a displayed defective intersegmental vessels formation. Furthermore, endothelial-specific Flvcr1a targeting in mice led to a reduced radial expansion of the retinal vasculature associated to decreased EC proliferation. Moreover, Flvcr1a null retinas showed defective vascular organization and loose attachment of pericytes. Finally, adult neo-angiogenesis is severely affected in murine models of tumor angiogenesis. Tumor blood vessels lacking Flvcr1a were disorganized and dysfunctional. Collectively, our results demonstrate the critical role of FLVCR1a as a regulator of developmental and pathological angiogenesis identifying FLVCR1a as a potential therapeutic target in human diseases characterized by aberrant neovascularization.

Feline Susceptibility to Leptospirosis and Presence of Immunosuppressive Co-Morbidities: First European Report of L. interrogans Serogroup Australis Sequence Type 24 in a Cat and Survey of Leptospira Exposure in Outdoor Cats.

Leptospirosis is one of the most widespread zoonotic diseases and can infect both humans and animals worldwide. The role of the cat as a susceptible host and potential environmental reservoir of Leptospira is still not well understood, due to the lack of obvious clinical signs associated with Leptospira spp. infection in this species. This study aims to describe the first European detection of Leptospira interrogans serogroup Australis ST 24 in a young outdoor cat with a severe comorbidity (feline panleukopenia virus). In addition, the results of a preliminary study conducted in 2014-2016 are presented (RC IZSVE 16/12), which reports an investigation of Leptospira exposure of outdoor cats in Northeast Italy by means of serological investigation and molecular evaluation of urine. The animals included in the survey are part of samples collected during active and passive surveillance (diagnostic samples). The study reported a seroprevalence of 10.5% among outdoor cats and the serogroups identified were Grippotyphosa, Icterohaemorrhagiae, Bratislava, Canicola and Ballum. Symptomatic cats reported high MAT titres (ranging from 1:800 to 1:1600) towards antigens belonging to the serovars Grippotyphosa (1:800), Bratislava (1:1600), Icterohaemorrhagiae (1:200) and Copenhageni (1:200-1:800). In one subject, urine tested positive for Leptospira PCR. Cats with high antibody titres for Leptospira and/or positivity on molecular test suffered from immunosuppressive comorbidities (feline immunodeficiency virus and feline leukaemia virus; feline herpesvirus and lymphoma; hyperthyroidism). The overall prevalence of serum antibodies against Leptospira found in free-ranging cats (10.53%, 95% CI: 4.35-16.70%) and the identification of L. interrogans ST 24 in a young cat with immunosuppressive disease (feline panleukopenia virus) suggest the possibility of natural resistance to clinical leptospirosis in healthy cats. In a One Health perspective, further studies are needed to better define the pathogenesis of leptospirosis in cats and their epidemiological role as environmental sentinels or possible carriers of pathogenic Leptospira.

Molecular detection of feline hemoplasmas and retroviruses in free-roaming and shelter cats within a university campus.

The aim of the present study was to assess the frequency of hemoplasma, feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) infections in cats living in an on-campus shelter and free-roaming cats within a university campus in Brazil. Blood samples were tested using quantitative PCR for hemoplasma, FIV and FeLV. Positive hemoplasma samples were sequenced. Associations between hemoplasma detection and living situation, sex, flea and/or tick parasitism, and coinfection with FIV and FeLV, were assessed using Fisher's exact test and the respective odds ratios were calculated. Overall, 6/45 (13.3%) cats tested positive: four (8.9%) were infected with 'Candidatus Mycoplasma haemominutum' and two (4.4%) with Mycoplasma haemofelis. All positive samples were from free-roaming cats (6/15; 40.0%) and had statistically significantly lower packed cell volumes (P = 0.037). Although 5/23 (21.7%) males and 1/22 (4.6%) females were positive, no statistically significant association between sex and hemoplasma infection was found (P = 0.19). Viral quantitative PCR (qPCR) was performed on 43/45 samples, among which 2/43 (4.7%) were positive for FIV and none for FeLV. Only one cat (2.3%) was coinfected with hemoplasma and FIV (P = 0.26). In addition, 4/6 (66.7%) cats that tested positive for hemoplasmas were infested by fleas (P = 0.0014) and/or ticks (P = 0.25). These results show that even if the free-roaming cat population is clinically healthy and has adequate access to food, it may present flea infestation and hemoplasma infection with lower packed cell volume values.

Feline leukemia as a still relevant problem in domestic and wild animals – the current state of knowledge

The aim of this review is to present information about leukemia caused by feline leukemia virus (FeLV) infection in felids. Despite widespread vaccinations, leukemia is still one of the most common diseases in cats. FeLV infection occurs by direct contact between animals, especially through contact with saliva. Infection with this highly contagious virus is characterized by immunosuppression and non-specific signs, such as cytopenia or tumorigenesis. The virus is transmitted horizontally through close contact between cats and vertically from infected queens to their kittens. Persistently infected cats are the reservoir of the virus. Rapid ELISA tests and immunochromatography play a major role in diagnosis, but RT-PCR is the most reliable diagnostic method. There is no specific therapy against FeLV, so treatment is mainly symptomatic. Research on the use of antiviral drugs, i.e. tenofovir, raltegravir, and fozyvudine, is underway. Proper prophylaxis, including regular vaccination of animals, is important to minimize the risk and consequences of infection

Prevalence of and exposure factors for infectious diseases in free-roaming cats from two Florida counties

Introduction: Ninety-nine deceased free-roaming cats (FRCs) from two nonadjacent counties (Volusia and Alachua) in Florida were used to determine the prevalence of and associated exposure to pathogens postmortem, including Feline Leukemia Virus (FeLV), Feline Immunodeficiency Virus (FIV), Dirofilaria immitis, Mycoplasma haemofelis, Mycoplasma haemominutum, and Cytauxzoon felis. Methods: Humanely euthanized FRCs or those FRCs found dead in the community were submitted for postmortem examinations. Blood samples from these FRCs were analyzed using a combination of antibody, antigen, and polymerase chain reaction (PCR) assays. Results: Male cats were at higher risk of infection for FeLV and FIV, intact cats were less likely to be infected with FeLV, and cats in the Volusia county were more likely to be infected with FIV. Mycoplasma haemominutum had the highest prevalence of all surveyed pathogens in this study, and infections were only identified in male cats. Conclusion: FRCs in this study had similar or higher prevalence rates of infections compared to studies assessing FRCs enrolled in trap-neuter-return programs from Florida.

Pyothorax caused by Nocardia sp. in a cat Case report

Nocardiosis is an infection caused by ubiquitous opportunistic bacteria and is rare in felines. In retrospective studies of the infection in the species, pyothorax was found in only about 5% of the cases described, with the most frequent forms being cutaneous and subcutaneous. This report describes Nocardia spp. as an etiologic agent of pyothorax in a feline leukemia virus (FeLV)-positive female cat. A 10-year-old female mixed-breed cat, positive for FeLV, with chronic kidney disease IRIS stage 2, was examined with a severe dyspneic onset, suspected of pleural effusion. It also had skin lesions in the interscapular region. The patient underwent thoracentesis, and a dense, creamy, fetid, and flocculated liquid was collected. Radiography was performed after draining the pleural fluid, which showed images suggestive of atelectasis of the left lung lobes and pneumonia. The collected material was sent for analysis, which found that it was a septic exudate. In addition, it was referred for bacterial culture in a special medium due to suspicion of nocardiosis. Long-term treatment with amoxicillin + clavulanic acid resolved the clinical case.

Characterization of ferret Pit1 as a receptor of feline leukemia virus subgroup B.

Feline leukemia virus (FeLV) is a retrovirus that causes immune suppression and immunodeficiency, leading to opportunistic infections and leukemia/lymphoma in cats. Today, a variety of domestic mammals are kept in houses, and it is important to evaluate the possibility of interspecies transmission of FeLV. In this study, we assessed the infectivity of FeLV-B in ferrets that belong to Mustelidae. By pseudotype virus infection assay, we revealed that a ferret cell line, Mpf cells, is resistant to FeLV-B infection. The mRNA expression level of the FeLV-B receptor, Pit-1, was approximately half that of cat FEA cells in ferret Mpf cells. There was no significant difference in receptor usage between ferret's and cat's Pit1. These data may indicate the presence of the post-transcriptional modification and/or the restriction factor(s) against the FeLV-B infection in ferrets.

Feline leishmaniosis: hematological and biochemical analysis.

One hundred and sixty-six cats from two animal shelters were subjected to enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence antibody test (IFAT), conventional polymerase chain reaction (cPCR), quantitative PCR (qPCR) and parasitological tests (PA) for the diagnosis of Leishmania spp. Among them, 15% (25/166), 53.6% (89/166), 3.6% (06/166) and 1.8% (03/166) were positive by ELISA, IFAT, both PCRs and PA, respectively. The sequencing of ITS-1 PCR amplicons revealed a 100% match with Leishmania infantum. After the Leishmania spp. survey, 12 cats were selected and divided into two groups for clinical, hematological, and biochemical analysis: six L. infantum positive cats (G1) and six Leishmania spp. negative cats (G2). All the cats were negative for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). A statistical analysis indicated significantly low platelet counts and significant hyperproteinemia associated with hypoalbuminemia in positive cats (p<0.05). Our results suggest that in endemic areas, cats with clinical signs of feline leishmaniosis (such as skin lesions, weight loss and/or enlarged lymph nodes) and that exhibit hematological and biochemical changes, such as low platelet counts and hyperproteinemia with hypoalbuminemia, should be tested for Leishmania spp. infection.

Mycoplasma haemofelis infection and its correlation with feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) in cats in Southern Brazil

A cross-sectional study was conducted on 274 cats in Southern Brazil to estimate the prevalence of Mycoplasma haemofelis PCR, associated factors, and its correlation with ELISA for FeLV and FIV. The apparent prevalence of M. haemofelis was 6.6% (18/274) (95% CI: 3.6–9.5%), of which 33.3% (6/18) had co-infection with FeLV, 5.6% (1/18) with FIV, and 5.6% (1/18) with both. Male cats were more likely to be positive for M. haemofelis [OR: 7.07 (1.97–25.34)]. Only three M. haemofelis-positive cats showed related clinical changes, such as mucosal pallor. A statistically significant difference was observed between M. haemofelis-positive cats and the negative control group for age, hemoglobin concentration, packed cell volume, and rod neutrophil counts. Mycoplasma haemofelis is prevalent in southern Brazil, with a higher risk in male cats. Most cats could be classified as asymptomatic carriers since they were healthy.

Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts

Respiratory tract and intestinal diseases are common threats to feline health. Coinfection with multiple pathogens is not rare among clinical infectious cases. Rapid diagnosis of these coinfections is of great significance for timely and effective clinical treatment. In this study, two novel multiplex polymerase chain reactions (mPCRs) were established for simultaneous detection of four pathogens associated with the feline intestinal tract (feline coronavirus (FCoV), feline astrovirus (FeAstV), feline panleukopenia virus (FPV) and feline kobuvirus (FeKoV)) and five pathogens associated with the respiratory tract (feline calicivirus (FCV), feline herpesvirus 1 (FHV-1), feline leukemia virus (FeLV), Chlamydia felis (C. felis) and influenza A virus (IAV)). The results of sensitivity analysis revealed that the detection limits for FeKoV, FPV, FeAstV, FCoV, IAV, C. felis, FeLV, FHV-1 and FCV were 103, 104, 103, 103, 103, 104, 104, 105 and 105 copies/µL, respectively. Moreover, the specificity of the two mPCRs was high. When the two mPCRs were applied to clinical samples, the assay worked well. In conclusion, we established two mPCR methods that provide an excellent tool for the diagnosis and monitoring of pathogens associated with the feline respiratory and intestinal tracts.

Detection and genetic characterization of feline retroviruses in domestic cats with different clinical signs and hematological alterations.

Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) are globally distributed retroviruses that infect domestic cats and cause various syndromes that can lead to death. The aim of this study was to detect and genotype feline retroviruses in Mexican domestic cats. We used PCR assays to identify proviral DNA and viral RNA in 50 domestic cats with different clinical signs and hematological alterations. Endogenous FeLV (enFeLV) was identified in the genomic DNA of all cats in the study, and we detected transcripts of the LTR region of enFeLV in 48 individuals. Exogenous FeLV (exFeLV) was found in 13 cats. Furthermore, we detected FIV proviral DNA in 10 cats. The enFeLV sequences were shown to be the most variable, while the exFeLV sequences were highly conserved and related to previously reported subgroup A sequences. Sequencing of the FIV gag gene revealed the presence of subtype B in the infected cats.

Occurrence and risk factors of feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) in cats of Lebanon

A cross-sectional study was carried out between April 2020 and August 2021. Blood samples were collected from 260 household cats recruited in different clinics in North and Mount Lebanon, with the aim of determining the seroprevalence of feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). The seroprevalence reported reached 18.84% (95% CI 0.14–0.24) and 13.84% (95% CI 0.09–0.18) for FIV and FeLV, respectively. FIV seropositivity was associated with a younger age, health and neuter status, lymphoma, lethargy, and vomiting. Furthermore, male sex, neuter status, lymphoma, anaemia, lethargy, and vomiting were significantly associated with FeLV seropositivity. This first data from Lebanon emphasizes the need for implementing preventive programmes to cope with FIV- and FeLV-associated morbidity and mortality among cats.

Feral Cat Populations and Feline Retrovirus Prevalence in San Mateo County, California in Three Time Periods between 2001 and 2016.

This study was initiated in 2004 because the prevalence of feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) infections in feral cats in San Mateo County (SMC) was not known. The cities attributed to the feral cat population presented to the Peninsula Humane Society & SPCA’s Spay/Neuter Clinic and to the Shelter itself were analyzed to examine potential geographic concentrations of feral cats with positive retroviral status. Trends in FIV and FeLV status were examined in three 3-year periods (2001−2003, 2005−2007, and 2014−2016). Population trends over the 15 years of this study for feral cats admitted to the Shelter were also examined. In each study period, more female feral cats were presented to the S/N Clinic (54.06%, 57.37%, 54.89%). FIV prevalence increased from 5.52% to 6.41% (p = 0.29) from the first to third period; FeLV prevalence decreased significantly from 1.73% to 0.29% (p = 0.01). Significantly more FIV-positive males than females were identified each year and for each period (p < 0.01). The four largest SMC cities were the major source of feral cats to the Shelter, S/N Clinic, and of FIV- and FeLV-positive cats in the first two periods; in the third period, 50% of feral cats to the Shelter and of FeLV-positive cats were from these cities. Despite a 61.63% reduction in feral cat admissions to the S/N Clinic, the FIV prevalence for males remained similar and increased for females. The retrovirus prevalence suggests the need for continued testing and surveillance of FIV among SMC free-living cats.

Clinicopathological Findings in Cats Tested for Feline Immunodeficiency Virus (FIV) and Feline Leukaemia Virus (FeLV)

Abstract This retrospective study aimed to evaluate the clinicopathological changes in a population of cats tested for feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV), in an Italian Veterinary University Hospital, in the period between January 2002 and May 2016. During the period of 14 years, 1834 cats were tested, and of these 241/1834 (13.1%) were positive for FIV antibodies and 92/1834 (5%) cats were positive for FeLV antigen. These data confirm the presence of a high prevalence of these viruses on Italian territory. To the authors’ knowledge, this study describes findings that have never been evaluated before, such as iron status in retrovirus-infected cats and urinalysis in FeLV-positive cats. In this study, FIV-positive cats were more likely to have higher serum protein concentration and lower albumin-globulin ratio than other groups of cats. Lower urine specific gravity and higher urine protein to creatinine ratio were also detected for FIV-positive cats when compared with negative and healthy cats. FeLV-positive cats were more likely to have cytopenia, decreased haemoglobin, haematocrit and RBC compared with other groups of cats. The data obtained underline the importance of considering retroviral infections in the presence of a broad spectrum of risk factors and laboratory anomalies.

A Retrospective Study of Viral Molecular Prevalences in Cats in Southern Italy (Campania Region).

From 2019 to 2021, a retrospective molecular study was conducted in the Campania region (southern Italy) to determine the prevalence of viral diseases in domestic cats. A total of 328 dead animals were analyzed by Real-Time PCR for the presence of feline panleukopenia virus (FPV), feline leukemia virus (FeLV), feline enteric coronavirus (FCoV), rotavirus (RVA), feline herpesvirus type 1 (FHV-1), and feline calicivirus (FCV). The possible presence of SARS-CoV-2 was also investigated by Real-Time PCR. The cats included in this study were specifically sourced and referred by local veterinarians and local authorities to the Zooprofilactic Experimental Institute of Southern Italy (IZSM) for pathological evaluation. The samples consisted of owners, catteries, and stray cats. Results revealed: 73.5% positive cats for FPV (189/257), 23.6% for FeLV (21/89), 21.5% for FCoV (56/266), 11.4% for RVA (16/140), 9.05% for FeHV-1 (21/232), and 7.04 for FCV (15/213). In contrast, SARS-CoV-2 was never detected. FPV was more prevalent in winter (p = 0.0027). FCoV FHV-1, FCV, and RVA predominated in autumn, whereas FeLV predominated in summer. As expected, viral infections were found more frequently in outdoor and shelter cats than in indoor ones, although no statistical association was found between animal lifestyle and viral presence. The study showed a high prevalence of FPV, FeLV, and FCoV and a moderate prevalence of RVA, FHV-1, and FCV. Moreover, the prevalence of these pathogens varied among the cat populations investigated.

Identification of crucial genes and metabolites regulating the eggshell brownness in chicken

BackgroundProtoporphyrin IX (Pp IX) is the primary pigment for brown eggshells. However, the regulatory mechanisms directing Pp IX synthesis, transport, and genetic regulation during eggshell calcification in chickens remain obscure. In this study, we investigated the mechanism of brown eggshell formation at different times following oviposition, using White Leghorn hens (WS group), Rhode Island Red light brown eggshell line hens (LBS group) and Rhode Island Red dark brown eggshell line hens (DBS group).ResultsAt 4, 16 and 22 h following oviposition, Pp IX concentrations in LBS and DBS groups were significantly higher in shell glands than in liver (P < 0.05). Pp IX concentrations in shell glands of LBS and DBS groups at 16 and 22 h following oviposition were significantly higher than WS group (P < 0.05). In comparative transcriptome analysis, δ-aminolevulinate synthase 1 (ALAS1), solute carrier family 25 member 38 (SLC25A38), ATP binding cassette subfamily G member 2 (ABCG2) and feline leukemia virus subgroup C cellular receptor 1 (FLVCR1), which were associated with Pp IX synthesis, were identified as differentially expressed genes (DEGs). RT-qPCR results showed that the expression level of ALAS1 in shell glands was significantly higher in DBS group than in WS group at 16 and 22 h following oviposition (P < 0.05). In addition, four single nucleotide polymorphisms (SNPs) in ALAS1 gene that were significantly associated with eggshell brownness were identified. By identifying the differential metabolites in LBS and DBS groups, we found 11-hydroxy-E4-neuroprostane in shell glands and 15-dehydro-prostaglandin E1(1-) and prostaglandin G2 2-glyceryl ester in uterine fluid were related to eggshell pigment secretion.ConclusionsIn this study, the regulatory mechanisms of eggshell brownness were studied comprehensively by different eggshell color and time following oviposition. Results show that Pp IX is synthesized de novo and stored in shell gland, and ALAS1 is a key gene regulating Pp IX synthesis in the shell gland. We found three transporters in Pp IX pathway and three metabolites in shell glands and uterine fluid that may influence eggshell browning.

Feline Leukemia Virus Frequently Spills Over from Domestic Cats to North American Pumas.

Feline leukemia virus (FeLV) is a gammaretrovirus with horizontally transmitted and endogenous forms. Domestic cats are the primary reservoir species, but FeLV outbreaks in endangered Florida panthers and Iberian lynxes have resulted in mortalities. To assess prevalence and interspecific/intraspecific transmission, we conducted an extensive survey and phylogenetic analysis of FeLV infection in free-ranging pumas (n = 641) and bobcats (n = 212) and shelter domestic cats (n = 304). Samples were collected from coincident habitats across the United States between 1985 and 2018. FeLV infection was detected in 3.12% of the puma samples, 0.47% of the bobcat samples, and 6.25% of the domestic cat samples analyzed. Puma prevalence varied by location, with Florida having the highest rate of infection. FeLV env sequences revealed variation among isolates, and we identified two distinct clades. Both progressive and regressive infections were identified in cats and pumas. Based on the time and location of sampling and phylogenetic analysis, we inferred 3 spillover events between domestic cats and pumas; 3 puma-to-puma transmissions in Florida were inferred. An additional 14 infections in pumas likely represented spillover events following contact with reservoir host domestic cat populations. Our data provide evidence that FeLV transmission from domestic cats to pumas occurs widely across the United States, and puma-to-puma transmission may occur in genetically and geographically constrained populations. IMPORTANCE Feline leukemia virus (FeLV) is a retrovirus that primarily affects domestic cats. Close interactions with domestic cats, including predation, can lead to the interspecific transmission of the virus to pumas, bobcats, or other feline species. Some infected individuals develop progressive infections, which are associated with clinical signs of disease and can result in mortality. Therefore, outbreaks of FeLV in wildlife, including the North American puma and the endangered Florida panther, are of high conservation concern. This work provides a greater understanding of the dynamics of the transmission of FeLV between domestic cats and wild felids and presents evidence of multiple spillover events and infections in all sampled populations. These findings highlight the concern for pathogen spillover from domestic animals to wildlife but also identify an opportunity to understand viral evolution following cross-species transmissions more broadly.

Evaluation of barcoded magnetic bead-based immunoassays for the simultaneous detection of feline leukemia virus antigen and antibody against feline immunodeficiency virus.

Testing platforms that leverage automation, require minimal sample volume, and enable various tests to be performed simultaneously on a single sample have the potential to improve workflow and efficiency in veterinary diagnostic laboratories. We evaluated a barcoded magnetic bead (BMB) technology using established immunoassays for detection of feline leukemia virus (FeLV) p27 antigen and antibody against feline immunodeficiency virus (FIV). Analytical sensitivity, limit of blank, and limit of detection were used to establish a functional sensitivity of 1.00 ng/mL of inactivated FeLV antigen and 35.7 ng/mL of anti-FIV monoclonal antibody. Common interferents, such as hemoglobin, lipid, and bilirubin, were not found to interfere with the performance of the assay. Intra- and inter-assay CVs were <13% for both assays using manufactured samples. Using a set of 116 feline samples, the diagnostic accuracy of our multiplex assay was 100% compared to reference assays. Performance in a convenience set of 1,000 feline samples submitted to a commercial diagnostic laboratory revealed a proportion of positive results of 1.3% for FeLV and 3.7% for FIV. BMB technology should enable rapid screening of samples for various markers in a single immunoassay well.

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.