Canine leishmaniosis, caused by Leishmania infantum, is a vector-borne disease in which early treatment monitoring remains challenging. This prospective study evaluated short-term changes in serological, inflammatory, and serum protein electrophoretic parameters during first-line therapy with meglumine antimoniate plus allopurinol. Eighteen dogs with confirmed clinical disease were enrolled at diagnosis, and fifteen dogs completing all follow-up visits (Days 0, 7, 14, 21, and 29) were included. Infection was confirmed by compatible clinical and/or laboratory abnormalities, high anti-Leishmania antibody levels, and successful parasite isolation. Anti-Leishmania antibodies were quantified using in-house single-dilution and two-fold serial dilution ELISA; circulating immune complexes, C-reactive protein, a complete blood count, a biochemical profile, and serum protein electrophoresis were also assessed. Both ELISA methods showed a significant decline in antibody levels during treatment, with serial dilution ELISA showing greater discrimination among highly positive samples. C-reactive protein concentrations decreased significantly from diagnosis to Day 29, indicating reduced systemic inflammation. Serum protein electrophoresis showed significant increases in albumin and the albumin:globulin ratio, along with a significant decrease in gamma globulins. Circulating immune complexes concentrations remained stable. These findings support quantitative serology, C-reactive protein, and serum protein electrophoresis as useful tools for early monitoring of therapeutic response in clinical canine leishmaniosis.

Ferrets are susceptible to infections with Leishmania infantum , Toxoplasma gondii , and Dirofilaria immitis ; however, epidemiological data on these infections are limited in Europe. A total of 448 serum samples from household ferrets were collected between December 2019 and December 2023 in Spain. In this study, we assessed the seroprevalence of L. infantum and D. immitis using an in-house enzyme-linked immunosorbent assay (ELISA) and T. gondii using an in-house immunofluorescence antibody test (IFAT). Among the ferrets tested, the seroprevalence was 10.49% (47/448) for L. infantum , 2.68% (12/448) for T. gondii and 10.27% (46/448) for D. immitis . There was no significant association between seropositivity and age, gender, neutering status, cohabitation, lifestyle, and collection date. Ferrets classified as sick animals related to the presence of compatible or non-compatible signs showed a higher seropositivity rate for L. infantum (15.90%) compared to subclinical animals (4.76%). Overall, 23.44% (105/448) of the samples were seropositive for at least one of the three parasitic agents, 3.12% (15/448) were positive for two agents, and 0.22% (1/448) tested positive for all three agents. Co-infections were also evaluated, revealing that 12.76% (6/47) of L. infantum seropositive ferrets were also positive for T. gondii ( p = 0.011) and 21.27% (10/47) for D. immitis ( p = 0.009). To the best of our knowledge, this is the first report on the seroprevalence of L. infantum , D. immitis , and T. gondii within the ferret population in Spain. Understanding the epidemiological status of these and other zoonotic pathogens is crucial for enhancing surveillance in both veterinary and public health sectors, as well as for strengthening prevention and control strategies.

Leishmania infantum infection has recently been identified in non-human primates in Spain, including clinically affected individuals from critically endangered species such as the Bornean orangutan (Pongo pygmaeus pygmaeus). Despite these emerging cases, clinicopathological data on leishmaniosis in captive wildlife remain limited. Here, we report the first confirmed case of clinical visceral leishmaniosis in a zoo-captive white-naped mangabey (Cercocebus lunulatus) in Spain. The affected animal presented with progressive weight loss, generalized lymphadenopathy, polyclonal gammopathy, and cytological evidence of pyogranulomatous lymphadenitis containing intralesional protozoal amastigotes consistent with Leishmania spp. The animal eventually died, and a detailed necropsy was performed. Moreover, to confirm L. infantum infection, histopathological analysis, serological assays, including immunofluorescent antibody test (IFAT), Western blot (WB), and enzyme-linked immunosorbent assay (ELISA), as well as polymerase chain reaction (PCR) and immunohistochemical analysis of affected tissues were conducted. Gross pathological examination revealed disseminated visceral involvement, characterized by multiple white nodules in the lungs, spleen, liver, and lymph nodes. Microscopically, these lesions corresponded to areas of coagulative necrosis surrounded by macrophage-rich inflammatory infiltrates, many of which contained protozoal amastigotes compatible with Leishmania spp. L. infantum infection was confirmed by all aforementioned serological tests, PCR, and immunohistochemistry. This case highlights the expanding host range of L. infantum and underscores the need to include leishmaniosis in the differential diagnosis of primates in zoological institutions, especially in endemic regions.

Leishmaniosis, caused by Leishmania infantum and transmitted by sand flies, is a significant zoonotic disease. Dogs are primary reservoirs, but other domestic animals, such as cats and ferrets, and wild species, including Eurasian otters and European mink, can be hosts. This study focused on European mink, a critically endangered species, investigating the seroprevalence of L.infantum antibodies and its correlation with serum protein profiles over 2021-2024. A total of 68 captive minks in Spain were monitored, yielding 105 serum samples. ELISA tests identified 44 seropositive samples (41.9 %), with seroprevalence varying annually. While no significant association was found between seropositivity and age, gender, or body condition, seropositivity correlated significantly with the year of collection. Some animals exhibited fluctuating antibody levels across sampling periods. Serum protein electrophoresis analyzed 96 samples, revealing that L.infantum seropositivity significantly affected total protein concentrations, β-globulins, γ-globulins, and albumin:globulin ratio. Variations were also observed in specific protein fractions among both seropositive and seronegative minks. These findings highlight European mink's potential epidemiological role in L.infantum transmission. Monitoring antibody levels and protein profiles in captive populations can inform conservation strategies and disease management efforts.

Leishmania infantum causes disease endemic in the Mediterranean Basin that affects humans and various mammals, including mustelids. Clinical leishmaniosis has been documented in domestic ferrets (Mustela putorius furo) and in a captive Eurasian otter (Lutra lutra). The European mink (Mustela lutreola), one of the world's most endangered mammals, is experiencing severe population declines linked to habitat degradation, climate change, the introduction of non-native species, and diseases. This report presents the first documented case of clinical leishmaniosis in a captive European mink, detailing its clinical presentation, anti-Leishmania therapy, and follow-up. The affected animal showed weight loss, pyogranulomatous blepharitis, and enlarged popliteal lymph nodes. Haematological and biochemical analyses revealed anaemia, hypergammaglobulinaemia, and elevated renal parameters. Diagnosis was supported by clinical signs, pathological findings, moderate anti-Leishmania antibody levels, and cytological detection of amastigotes in skin and lymph node samples, further confirmed by parasite culture. An initial oral regimen of miltefosine and allopurinol produced limited clinical improvement, likely due to inadequate drug intake. Subsequently, treatment with meglumine antimoniate and allopurinol resulted in a marked clinical response and resolution of the main signs. These findings underscore the need for screening for L.infantum and prompt therapeutic intervention to protect Mustela lutreola populations.

Panniculitis is a rare condition in ferrets (Mustela putorius furo), previously linked to nutritional deficiencies, infections, trauma, and injections. Pancreatic panniculitis, caused by the systemic release of pancreatic enzymes during pancreatic injury, has been documented in humans and other animal species, but not in ferrets. This report describes the first known case in a domestic ferret. A 2-year-old male ferret presented with ulcerated cutaneous nodules predominantly affecting the hind limbs and inguinal abdominal region. The ferret had been treated with rifampicin and clarithromycin for suspected mycobacterial infection. Hyperglycemia, hyperglobulinemia, and elevated alkaline phosphatase were noted. Cytology and culture of the lesions revealed neutrophilic inflammation and Pseudomonas aeruginosa, respectively. The animal developed acute abdominal discomfort and died shortly after. Post-mortem examination revealed severe pancreatic necrosis and peripancreatic panniculitis, with splenic pyogranulomatous inflammation. Immunohistochemistry detected systemic coronavirus antigen (clone FCV3-70) only in the spleen. In this patient, prolonged use of rifampicin and clarithromycin is considered a potential contributing factor to the development of acute pancreatic necrosis. This case highlights the clinical relevance of pancreatic panniculitis in ferrets and suggests a possible association between extended rifampicin/clarithromycin therapy and pancreatic necrosis. Ultrasonography and serum glucose, lipase, and amylase measurements are recommended for early diagnosis. Histopathological evaluation remains essential for confirmation.

Nocardia spp are ubiquitous, gram-positive, variably acid-fast, branching and beaded filamentous, facultative intracellular bacteria that are resistant to phagocytosis and can cause localized or systemic disease in a variety of mammals, including humans, as well as in birds, fish and reptiles. Seventeen pet domestic ferrets (Mustela putorius furo) were diagnosed with nocardiosis by several methods including cytological evaluation, histopathology, Ziehl-Neelsen staining and polymerase chain reaction (PCR). All except two ferrets were 2 years old or older at the time of clinical presentation. Clinical findings included anorexia, weight loss, lymphadenomegaly, splenomegaly, masses/nodules in internal organs, hindlimb weakness or paresis, vomiting, dyspnoea, central nervous system signs, diarrhoea, abdominal pain and coughing. Anaemia, leucocytosis, neutrophilia, hypoalbuminaemia and hyperglobulinaemia were frequent. All ferrets had granulomatous or pyogranulomatous inflammation involving most commonly the lymph nodes, spleen, lungs, liver and adipose tissue. Intralesional acid-fast, branching filamentous bacilli were detected in 15 of 17 ferrets. PCR for the 16S rRNA gene of Nocardia spp was positive in 13 of 15 ferrets, including the two that were negative by acid-fast staining; of these two ferrets, one had intralesional coronavirus antigen. BLASTn analysis of eight sequences revealed six different Nocardia spp, including N. globerula, N. seriolae and N. donostiensis, that have rarely been reported to cause disease in humans and terrestrial animals. Antibiotic treatment (most commonly trimethoprim-sulfamethoxazole alone or in combination with clarithromycin and marbofloxacin) was followed by a marked improvement in several patients, although relapses were frequent. Six ferrets had concurrent neoplasia as a potential predisposing factor. Nocardiosis should be included in the differential diagnosis of granulomatous/pyogranulomatous inflammation in ferrets, particularly in those over 2 years of age. (c) 2024 Elsevier Ltd. All rights are reserved, including those for text and data mining, AI training, and similar technologies.

Pseudomonas luteola (P.luteola), formerly called Chryseomonas luteola, is a strict aerobic gram-negative bacillus, 0.8 to 1.0 µm wide and 1.5 to 2.5 µm long, considered an opportunistic pathogen found ubiquitously in humid environments, both in soil and water. It sporadically causes disease in animals and immunosuppressed humans or those subjected to invasive procedures such us peritoneal dialysis or catheterization. In ferrets, this infection was first described in Spain in 2012 and since then, cases have appeared occasionally in Finland, Austria, Australia, France, the United States and also in Spain. This pathogen is considered an emerging zoonotic disease in ferrets, causing respiratory disease, panniculitis, and abscesses due to pyogranulomatous or suppurative inflammation predominantly of the pleura, lung, mediastinum, panniculus or salivary glands, frequently with lethal consequences. The clinical case of a ferret, infected by Pseudomona luteola, presenting with ulcerative suppurative pododermatitis and ipsilateral popliteal purulent lymphadenitis, is described. Together with a complete resolution of the clinical case by means of a non-invasive medical management likely due to the rapid detection, identification, and treatment of the infection.

Leishmaniosis in domestic ferrets is a vector-borne disease caused in Europe by the protozoan parasite Leishmania infantum. There is limited information on clinical signs and laboratory abnormalities in ferrets due to leishmaniosis. This clinical case report described a domestic ferret (Mustela putorius furo) with severe hyperbetaglobulinemia, anaemia, thrombocytopenia, and abnormal renal parameters. A good clinical response following an anti-Leishmania infantum treatment protocol was achieved. However, the presence of pain at the site of injection was the main side effect due to meglumine antimoniate administration. Xanthine crystalluria was not observed in urine sediment with no other urine alterations detected by urine analysis during the follow-up. Initially, clinical signs noted in this ferret could not initially be attributed to leishmaniosis. However, no causes were found that could have caused the hyperglobulinemia in this patient. A reduction of the levels of anti-L. infantum serum antibodies and the concentrations of beta-globulin fraction was detected in this patient after anti-Leishmania treatment administered as well as the disappearance of thrombocytopenia. To extent of the knowledge of leishmaniosis in ferrets, this is the fourth case report of leishmaniosis documented in this species.

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.

BACKGROUND Toxoplasmosis is a protozoan disease caused by Toxoplasma gondii. Different T. gondii confirmatory techniques, including serologic methods, are available to detect the presence of the parasite. Among serology techniques, immunochromatographic rapid testing could be a reliable alternative to serologic laboratory techniques. OBJECTIVE This study evaluated a commercial immunochromatographic test (FASTest TOXOPLASMA g) in seronegative and seropositive cats. METHODS Two indirect immunofluorescence antibody reference tests, an in-house technique, and a commercial test were used to classify 292 feline serum samples. The rapid test was evaluated in different groups of cats, including healthy seronegative cats (n = 121), seropositive cats with variable anti-Toxoplasma antibodies (n = 146), and cats with positive serologic results for other pathogens (n = 25). The sensitivity, specificity, accuracy, receiver operating characteristic curves, and kappa statistics were analyzed as performance measures. RESULTS Of the 292 samples, 146 were classified as T. gondii seropositive and 146 as T. gondii seronegative. Concordant results were obtained for all samples using immunofluorescence antibody tests. The diagnostic measures of this rapid test showed 98.63% sensitivity and 100% specificity, and 99.32% accuracy. The kappa statistics value was 0.986, and the area under the receiver operating characteristic curve was 0.993. CONCLUSIONS This rapid test showed diagnostic measurements similar to those of traditional quantitative serologic methods. In situations where laboratory techniques are not available, this test, under clinical conditions, could be a useful alternative to obtain accurate results rapidly.

SARS-CoV-2 is the causal agent of Coronavirus Disease 2019 (COVID-19) in humans that emerged in late 2019. This virus is able to infect humans and different animal species. Among pets, cats and ferrets are more susceptible to be infected by the SARS-CoV-2. Epidemiological studies are an important tool to provide information under natural conditions of exposure to SARS-CoV-2 virus. In comparison to cats, limited epidemiological studies have been performed in domestic ferrets ( Mustela putorius furo) reporting the presence of antibodies in this species. This study analysed the presence of anti-SARS-CoV-2 antibodies in 432 cliend-owned ferrets from different geographical areas of Spain during the different waves of COVID-19 outbreaks from December 2019 to May 2023 (42 months). For this purpose, anti-SARS-CoV-2 antibodies were detected by an enzyme-linked immunosorbent method (ELISA) using the receptor binding domain (RBD) of Spike antigen and confirmed by serum virus neutralization assay. Eighteen of the 432 ferrets included were seroreactive by the in-house ELISA (4.17%, 95% Confidence Interval (CI): 2.65–6.49). In this sense, the wave of COVID-19 with the higher number of seropositive ferrets occurred during the seventh wave when the different Omicron subvariants were the dominant virus variants. Our results suggest that the risk of SARS-CoV-2 transmission in domestic ferrets in natural conditions is low. Further research is need to evaluate the potential risk of transmission of SARS-CoV-2 from human to pets.

Leishmaniosis is a vector-borne disease caused by different Leishmania species and transmitted by phlebotomine sand flies under natural conditions in Europe. Scientific information related to Leishmania infantum in dogs is extensive, where less information is available in cats and other companion animals. Recently, first clinical cases of L.infantum infection in domestic ferrrets ( Mustela putorius furo ) have been described. However, clinical information on leishmaniosis in this species is limited A 15-month-old male neutered domestic ferret was presented with chronic weight loss and the presence of coalescent, erythematous and firm subcutaneous nodules in the ventral abdominal subcutis. A fine-needle aspiration of these nodules was performed and the cytological examination revealed a granulomatous inflammation with the presence of macrophages contained a number of oval organisms with an eccentric nucleus and pale cytoplasm, compatible with Leishmania spp. amastigotes compatible with Leishmania spp. amastigotes. The nodules were surgically excised and histological examination showed a severe multifocal pyogranulomatous panniculitis. Specific immunohistochemistry and qPCR for L. infantum from excised nodules were positive. Additionally, L. infantum was cultured and isolated from the nodules by a fine-needle aspiration. An in-house Western Blot test for L. infantum was performed in serum sample and a positive result was obtained. This is the first reported case of nodular pyogranulomatous panniculitis due to L. infantum infection in a domestic ferret. Further studies are necessary to determine the relevance of domestic ferrets in the transmission of leishmaniosis. The description of new clinical forms of the disease is important as it can assist veterinarians in identifying these new clinical presentations.

The information about the clinical features of Leishmania infantum infection in cats is scarce. In this study, we evaluated the serum protein electrophoresis of samples from 19 infected but apparently healthy cats. To detect L. infantum infection, two serological tests, i.e. western blot (WB) and enzyme-linked immunosorbent assay (ELISA) as well as quantitative polymerase chain reaction (qPCR) on the blood samples were performed. Eventual infection by several selected bacterial and viral pathogens was also tested. All but one of the cats were found positive with WB. The WB-negative cat was positive by ELISA only. From the 18 WB-positive cats, only three were positive also by ELISA and eight with qPCR, including the only animal which was positive in all the three tests. No concomitant infections were detected in any of the cats. The main alteration of the proteinogram was characterised by an increase of the α- 2 fraction. In the five cats with hypergammaglobulinaemia, the pattern detected was polyclonal. None of the cats were seropositive to any other pathogens tested. The presence of polyclonal gammopathy and elevation of the α -2 fraction could suggest the presence of active infection. In contrast, the only detection of an increase of the α -2 fraction alone with the presence of positive serological result could be associated by immune response activation against L. infantum.

The impact of the SARS-CoV-2 pandemic on wildlife is largely unevaluated, and extended surveillance of animal species is needed to reach a consensus on the role of animals in the emergence and maintenance of SARS-CoV-2. This infection has been detected in farmed and domestic animals and wild animals, mainly in captivity. The interactions or shared resources with wildlife could represent a potential transmission pathway for the SARS-CoV-2 spill over to other wild species and could lead to health consequences or the establishment of new reservoirs in susceptible hosts. This study evaluated the presence of SARS-CoV-2 in European mink (Mustela lutreola) and American mink (Neogale vison) in Spain by enzyme-linked immunosorbent assay (ELISA) using the receptor binding domain (RBD) of Spike antigen in serum samples and/or by RT-qPCR assays in oropharyngeal and rectal swabs. From January 2020 to February 2022, a total of 162 animals (127 European mink and 35 American mink) with no evidence of SARS-CoV-2 infection were included in the study. Antibodies against the SARS-CoV-2 were not found in the serum samples analysed (n = 126), nor was the virus amplified by RT-qPCR (n = 160 swabs). Our results suggest that the potential role of wild mink and the European mink bred in captivity and released to the wild as dispersers of SARS-CoV-2 is so far low. However, wildlife surveillance for early detection of human and animal risks should be continued. In this sense, epidemiological monitoring measures, including serology and molecular analysis, are necessary.

The European mink (Mustela lutreola) is listed as a critically endangered species because of ongoing population reduction from habitat degradation and the effects of introduced species, such as American mink (Neovison vison). This small, fragmented population becomes vulnerable to many other threats, including diseases. Leishmaniosis is a zoonotic disease caused by the protozoan parasite Leishmania infantum found in the Mediterranean area, which affects many mammals, including wild small mammals. Furthermore, clinical disease caused by L. infantum has recently been described in other mustelids. To assess the exposure to Leishmania sp. infection in mink species in northern Spain, blood samples from 139 feral American mink and 42 native European mink from north Spain were evaluated for Leishmania sp. infection using enzyme-linked immunosorbent assays against Leishmania spp. antibodies, with 52.4% of American mink and 45.3% of European mink being found seropositive. This finding raises questions regarding how the disease may affect these species and the potential repercussions for conservation efforts. Despite a high seroprevalence being observed in wild mink of both species in this study, association with clinical or pathologic signs of disease has yet to be elucidated.

Canine leishmaniosis is a vector-borne disease caused by Leishmania parasites. Serological methods are the most common tests used for the diagnosis. This study aimed to evaluate and compare different serological commercial immunochromatographic rapid tests available in Spain to detect anti-Leishmania canine antibodies. The immunochromatographic tests were evaluated in different groups of dogs (healthy seronegative dogs (n = 21), naturally-sick dogs with moderate anti-Leishmania antibodies (n = 39), naturally-sick dogs with high antiLeishmania antibodies (n = 37), dogs with the serological result of other pathogens infection (n = 20) and exposed dogs (n = 33)) admitted to the Veterinary Teaching Hospital of the University of Zaragoza (Spain) according to the clinical information sent with the sample to the laboratory for diagnostic purposes. The serology status was also routinely recorded through an in-house enzyme-linked immunosorbent assay (ELISA) and an inhouse indirect immunofluorescence test (IFAT). The qualitative commercial serological immunochromatographic tests used were: FASTest LEISH, Uranotest Leishmania, Uranotest Leishmania 2.0, Speed Leish K, Witness Leishmania, and DFV Test Leishmania. Performance measures analyzed for each test were: sensitivity, specificity, and area under the receiver-operating (ROC) curve. The maximum specificity (1.00) was attained for Uranotest Leishmania and DFT Test Leishmania, followed by FASTest LEISH (0.98), Uranotest Leishmania 2.0 (0.98), Speed Leish K (0.98), and Witness Leishmania (0.95). The maximum sensitivity was attained for FASTest LEISH (1.00), followed by Uranotest 2.0 (0.97), Speed Leish K (0.97), Uranotest (0.96), and the lowest results with Witness (0.84) and DFV Test (0.59). Regarding the ROC curve, the maximum value was attained with the FASTest LEISH (0.99), followed by Uranotest (0.98), Uranotest 2.0 (0.97), Speed Leish K (0.97), Witness (0.90), and the lowest result with DFV Test (0.79). Efforts in the field of diagnosis should focus on establishing a commercial immunochromatographic test with high sensitivity and specificity with a reasonable cost-benefit balance.

Abstract Background Leishmaniosis, a vector-borne disease caused by Leishmania infantum, is one of the most important parasitic zoonoses in Europe. The transmission cycle of leishmaniosis is maintained by both domestic and wild animals. However, few data are available on the role of wild mammals in transmitting the parasite in the European Mediterranean basin. As feline leishmaniosis, diagnosis of the infection in ferrets can be a challenge, the use of different serological and molecular methods combined is a recommended approach. Our aim was to investigate the prevalence of infection of L. infantum in apparently healthy domestic ferrets (Mustela putorius furo) in an endemic region of Spain (Community of Valencia), using serological and molecular methods and to evaluate the results comparing the different techniques. Methods The prevalence of Leishmania infection was studied in domestic ferrets. Blood was collected from each animal for serology and molecular analysis. Two serological methods, enzyme-linked immunosorbent assay (ELISA) and western blot (WB), were used for the detection of L. infantum antibodies, and real-time polymerase chain reaction (qPCR) was used for the detection of L. infantum DNA. Results Blood samples from 102 apparently healthy ferrets were analyzed. In the serological study, 25.5% of the animals tested positive by western blot, and 9.0% by enzyme-linked immunosorbent assays. The seroprevalence of L. infantum infection, based on a positive result in any serological test, was 28.4% (95% confidence interval [CI] 20.6–S37.9%). No kinetoplast DNA (kDNA) was detected by qPCR in peripheral blood samples from the ferrets tested. Conclusions The immunological response revealed by these tests indicates that the ferrets are exposed to repeated inoculations with the endemic parasite L. infantum. Although the low population of domestic ferrets means their reservoir potential is limited in the absence of a primary host, it would be of interest to carry out further studies using xenodiagnosis to determine whether they are accidental or reservoir host species capable of spreading infection. Graphical Abstract

Dirofilaria immitis is an endemic mosquito-borne pathogen widely spread throughout Europe as well as North and South America. Infection by D. immitis has been reported in domestic ferrets, although little is known about the occurrence and the epidemiological features of this nematode in this species. The aim of the present retrospective study was to assess the prevalence of D. immitis antibodies using an in-house enzyme-linked immunosorbent assay specifically developed for use in ferrets. One hundred and eighty-six serum samples were obtained from the Province of Valencia (Spain), an area endemic for dirofilariosis. Of the 186 serum samples included in the study, 27 (14.51%) were classified as D. immitis seropositive and 159 samples as D. immitis seronegative. The results provide valuable information on the seroprevalence of D. immitis infection in domestic ferrets in an area endemic for this vector-borne pathogen. The presence of seropositive ferrets should be taken into account and preventive measures should be implemented, including the possibility of serological screening for the early detection of Dirofilaria antibodies as a serological marker of exposure. This is the first study that demonstrates the presence of D. immitis exposure in ferrets in Spain. Veterinarians working in endemic areas should be aware of this infection in ferrets and their susceptibility.

A new coronavirus known as SARS-CoV-2 emerged in Wuhan in 2019 and spread rapidly to the rest of the world causing the pandemic disease named coronavirus disease of 2019 (COVID-19). Little information is known about the impact this virus can cause upon domestic and stray animals. The potential impact of SARS-CoV-2 has become of great interest in cats due to transmission among domestic cats and the severe phenotypes described recently in a domestic cat. In this context, there is a public health warning that needs to be investigated in relation with the epidemiological role of this virus in stray cats. Consequently, in order to know the impact of the possible transmission chain, blood samples were obtained from 114 stray cats in the city of Zaragoza (Spain) and tested for SARS-CoV-2 and other selected pathogens susceptible to immunosuppression including Toxoplasma gondii, Leishmania infantum, feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV) from January to October 2020. Four cats (3.51%), based on enzyme-linked immunosorbent assay (ELISA) using the receptor binding domain (RBD) of Spike antigen, were seroreactive to SARS-CoV-2. T. gondii, L. infantum, FeLV and FIV seroprevalence was 12.28%, 16.67%, 4.39% and 19.30%, respectively. Among seropositive cats to SARS-CoV-2, three cats were also seropositive to other pathogens including antibodies detected against T. gondii and FIV (n = 1); T. gondii (n = 1); and FIV and L. infantum (n = 1). The subjects giving positive for SARS-CoV-2 were captured in urban areas of the city in different months: January 2020 (2/4), February 2020 (1/4) and July 2020 (1/4). This study revealed, for the first time, the exposure of stray cats to SARS-CoV-2 in Spain and the existence of concomitant infections with other pathogens including T. gondii, L. infantum and FIV, suggesting that immunosuppressed animals might be especially susceptible to SARS-CoV-2 infection.