Anti-Leishmania spp. antibody detection in domestic cats from a visceral leishmaniasis transmission area.

Feline leishmanial infection is reported worldwide, but the epidemiological role of domestic cats in the leishmaniasis cycle remains unclear, and cats might act as cryptic reservoir hosts in endemic areas with no feline leishmaniosis cases. Considering that, a serological screening for anti-Leishmania spp. antibodies was performed by indirect immunofluorescence antibody test (IFAT) in 389 necropsied cats' serum samples from a new visceral leishmaniasis transmission area with no feline leishmanial infection reported to unveil if the cats are being exposed to the parasite. The overall seroprevalence for Leishmania spp. was 11.05% (43/389). No association was found between sex, neutering status, age group, breed, coat length, feline immunodeficiency virus (FIV) infection, and Leishmania spp. antibody detection. A positive association was found with coat color (cats within the orange spectrum with white [particolor]) (OR = 2.47, CI 95% 1 - 6.13, P = 0.044) and a negative association (OR = 0.38, CI 95% 0.18 - 0.79, P = 0.01) between feline leukemia virus (FeLV) infection and IFAT positivity for Leishmania spp. Therefore, it is concluded that the seroprevalence found was greater than 10%, indicating contact of the protozoan with cats in the region served.

Prevalence of Leishmania infection in 205 cats from a referral hospital population in Brazil (2021-2022).

Leishmaniases are a group of neglected diseases of significant public health concern, with Brazil being the primary focus of this disease in the Americas. The municipality of Sobral, in the state of Ceará, is a historical focus of visceral leishmaniasis in both humans and dogs, but data on Leishmania spp. infections in cats are limited. Between April 2021 and February 2022, 205 cats from a referral hospital population were sampled and tested for Leishmania spp. by real-time PCR. Eight cats (3.9%; 95% CI: 1.7-7.5%) tested positive. Among these, three (37.5%) displayed clinical signs compatible with feline leishmaniosis. Non-domiciled cats showed significantly higher positivity compared to domiciled ones (Fisher's exact test, P = 0.0124). Considering their potential role as reservoirs of L. infantum, it is crucial to conduct further studies to understand the Leishmania spp. circulating among cats in Sobral and to implement measures for reducing their exposure to phlebotomine sand fly vectors in this important focus of leishmaniases.

First Molecular Evidence of Leishmania Infantum in Domestic Cats and Associated Risk Factors from the Black Sea Region of Türkiye.

PURPOSE: The objectives of the present study are to determine the molecular prevalence of Leishmania spp. in the owned domestic cats in the Black Sea Region of Türkiye and analyze the associated risk factors in FeL. METHODS: Conjunctival swabs (CS), blood, demographic, and clinical data were collected from 150 owned cats brought to the Veterinary Teaching Hospital during 2020-2022. Leishmania kinetoplast DNA (kDNA) from CS was screened by TaqMan Real-Time PCR (qPCR) with the genus-specific primers and a probe. RESULTS: All qPCR positive products were also amplified and sequenced to identify Leishmania species by ITS1 primers. Molecular prevalence of L. infantum found as 12.6% (19/150) in the observed cats in the Black Sea Region of Türkiye. There was a significant difference (p < 0.05) between neutered and intact cats with regarding to L. infantum positivity. Intact cats found to be 0.368 times more prone to be L. infantum-positive (L+). Dermatological lesions were found the most common (26.3%) problems in the L + cats. The median leucocyte count was the only parameter that was found statistically (p < 0.05) lower in the L + group (6.60) than the negative group (L-) (8.96), when comparing the WBC, NEU/LYM, MONO/LYM, EOS/LYM and PLT/LYM values. CONCLUSION: This study presented the molecular occurrence of FeL in the Black Sea Region of Türkiye for the first time indicating that the carrier status of the cats makes them alternative reservoirs for possible zoonotic transmission of L. infantum in this zone.

Feline Leishmaniosis: An Emerging Public Health Problem.

Leishmaniosis is the third most important vector-borne disease in humans, preceded by malaria and lymphatic filariasis, and it is considered endemic in tropical and subtropical areas, where higher temperatures favor development of its vector, sandflies. This zoonotic disease is caused by infection of protozoa Leishmania spp. and the most serious mucocutaneous and visceral form is produced by Leishmania infantum, which predominates in the Mediterranean region. The usual hosts for this parasite are dogs and humans, but an increment in cases of L. infantum infection has been observed in cats in the last years. This increase could be due to the use of sandflies repellents in dogs, obligating the parasite to looking for other hosts. The role of cats in the epidemiology of this disease is unknown, although increase of prevalence of feline leishmaniosis has been observed in endemic areas in the last years. Diagnostic techniques and treatments in cats are not standardized, which makes it difficult to establish prevalence and epidemiology of feline leishmaniosis. Furthermore, the clinical signs and immune response against Leishmania in cats are different to those in dogs, with an observed increment of drug resistance. It is necessary to increase our knowledge about L. infantum infection in cats, including clinical signs, transmission, treatments, and the role of cats in the increasing of zoonoses. Finally, new alternative treatments are required for controlling the spread of this disease in all species of mammals.

Skin Lesions in Feline Leishmaniosis: A Systematic Review.

Feline leishmaniosis (FeL) is increasingly reported throughout the world and skin lesions predominate in the clinical picture. There are, however, few evidence-based data on cutaneous feline leishmaniosis and directions are strongly needed for a better management of the disease. In this study, we systematically reviewed what is currently known about the clinical dermatological presentation of FeL through analysis of the literature and, further, by adding unpublished cases managed by Italian veterinary dermatologists. Sixty-six feline cases of cutaneous leishmaniosis published in 33 articles between 1990 and 2020 met the inclusion criteria and were analyzed. Six unpublished cases of cutaneous FeL managed by Italian dermatologists were also reviewed. The majority of cases were reported from South America, followed by Europe and North America. Nodules were the most frequently reported clinical signs and the presence of Leishmania in lesioned skin was assessed mainly by cytology. A total of six Leishmania species have been identified as being responsible for skin lesions. Coinfections by FIV or FeLV were reported in 12.1% and 9.1% of the cases, respectively. Clinical data including treatment have been analyzed and discussed to provide directives for proper management of the disease for which cats may also serve as domestic reservoirs for human infections.

Leishmania infection in cats and feline leishmaniosis: An updated review with a proposal of a diagnosis algorithm and prevention guidelines.

Leishmaniosis is a vector-borne disease caused by protozoans of the genus Leishmania, which are transmitted to vertebrates, including cats, through the bites of female phlebotomine sand flies. An increasing number of epidemiological and experimental studies concerning Leishmania infection in cats, as well as case reports of clinical leishmaniosis in these felids, have been published in recent years. In the present study, a comprehensive review was made by sourcing the National Library of Medicine resources to provide updated data on epidemiology, immunopathogenesis, diagnosis, treatment, and prevention of feline leishmaniosis. Cats were found infected with Leishmania parasites worldwide, and feline leishmaniosis appears as an emergent disease mostly reported in countries surrounding the Mediterranean Sea and in Brazil. Cats with impaired immunocompetence seem to have a higher risk to develop clinical disease. The main clinical and clinicopathological findings are dermatological lesions and hypergammaglobulinemia, respectively. Diagnosis of feline leishmaniosis remains a challenge for veterinarians, in part due to the lack of diagnosis support systems. For this reason, a diagnostic algorithm for clinical decision support is herein proposed. No evidence-based treatment protocols are currently available, and these remain empirically based. Control measures are limited and scarce. Thus, a set of prevention guidelines are herein suggested.

Serum amyloid A levels and alpha 2 and gamma globulins on serum protein electrophoresis in cats exposed to and infected with Leishmania infantum.

BACKGROUND: Dogs are the main reservoir hosts of Leishmania infantum; nevertheless, recent investigations indicate a likely role for cats in the epidemiology of Leishmania infection. Feline leishmaniosis (FeL) remains poorly characterised, partly due to the lack of suitable diagnostic tools. This study aimed to compare serum amyloid A (SAA) levels and serum protein electrophoresis (SPE) profiles (specifically, alpha 2 and gamma globulins) in cats naturally exposed to or infected by L. infantum from southern Italy versus those of healthy controls and versus cats with neoplastic or inflammatory conditions from non-endemic areas. METHODS: Serum or plasma samples from four cohorts of cats were analysed for SAA levels and by SPE: (i) G1: healthy controls from Leishmania-non-endemic regions of Switzerland; (ii) G2: cats pre-diagnosed with neoplastic or inflammatory conditions available from the University of Cambridge sample archive; (iii) G3: L. infantum-seropositive, quantitative (q)PCR-negative cats from southern Italy; (iv) G4: L. infantum-seropositive and qPCR-positive cats from southern Italy. SAA data were assessed for normality and homoscedasticity using the Shapiro-Wilk and Levene's tests, respectively; the Kruskall-Wallis test, followed by Dunn's test with Bonferroni correction were subsequently used to compare SAA serum levels between groups. A weighted generalised linear model with a binomial distribution was used to assess statistically significant differences in the numbers of animals displaying elevated gamma globulins and increased alpha 2 globulins between groups. RESULTS: Overall, 68 samples were analysed (G1: n = 16, G2: n = 20, G3: n = 20, G4: n = 12). Cats suffering from neoplastic and inflammatory conditions (G2 ) showed significantly higher SAA levels than healthy controls (G1) (median values [interquartile range]: G1: 0.00 [0.00-0.00] mg/l versus G2: 0.85 [0.00-49.55] mg/l). G2, G3 and G4 cats showed higher percentages of individuals with increased alpha 2 globulins (percentages ± standard error: G1 = 20.0% ± 10.3, G2 = 80.0% ± 8.9, G3 = 70.0% ± 10.2, G4 = 75.0% ± 12.5) and gamma globulins (G1 = 0.0% ± 0, G2 = 65.0% ± 10.7, G3 = 50.0% ± 11.2, G4 = 58.3% ± 14.2) than healthy control cats (G1). For all three markers, no significant difference between cats within G2, G3 and G4 was recorded. CONCLUSIONS: This study indicates that the proportions of animals with elevated levels of alpha 2 and gamma globulins are significantly higher in cats exposed to and infected with L. infantum. Levels of SAA and alpha 2 and gamma globulins may not be used to differentiate between L. infantum infection or exposure, and neoplastic and/or inflammatory conditions.

Serological, molecular and clinicopathological findings associated with Leishmania infantum infection in cats in Northern Italy.

OBJECTIVES: The aims of this study were to investigate the prevalence of Leishmania species infection in cats in Northern Italy and to evaluate the associations between infection and signalment and clinicopathological data. METHODS: The study was carried out in a veterinary university hospital from June to November 2017. Blood, urine, conjunctival swabs and hair were collected from all randomly selected cats. Leishmania species infection was evaluated using the indirect fluorescent antibody test (IFAT), setting a cut-off value of 1:80, and using real-time PCR on blood, conjunctival and hair samples. A complete blood count, serum chemistry profile, serum electrophoresis and urinalysis were also carried out. The cats were grouped on the basis of the results of the diagnostic criteria adopted in positive, negative and unconfirmed Leishmania cases. Non-parametric variables and continuous data were compared among the study groups using the χ2 test and the Mann-Whitney U-test, respectively. RESULTS: One hundred and fifty-two cats were included. Nineteen of the 152 (12.5%) cats were positive (18/152 [11.8%] showed an IFAT titre of ⩾1:80 and 1/152 [0.7%] was real-time PCR-positive from a hair sample); 106/152 (69.7%) cats were negative; and 27/152 (17.8%) cats were unconfirmed for Leishmania species. Total proteins, beta2-globulin and gamma-globulin were significantly increased in the positive Leishmania group compared with the negative group. CONCLUSIONS AND RELEVANCE: The results of the present study demonstrated the spread of Leishmania infantum infection in cats in Northern Italy. Hyperproteinaemia and hypergammaglobulinaemia appeared to be significant clinicopathological abnormalities in this population of cats with L infantum infection.

Latest trends in L. infantum infection in dogs in Spain, Part II: current clinical management and control according to a national survey of veterinary practitioners.

BACKGROUND: Canine leishmaniosis (CanL) is a parasitic zoonotic disease, endemic in the Mediterranean basin including Spain. While knowledge about CanL, its management, treatment, prevention and control mounts, it remains unclear whether all clinical veterinarians follow the same international recommendations, such as those of the LeishVet group. This study was thus designed to assess recent trends in the clinical management of CanL in veterinary clinics across Spain through a questionnaire-based survey. Results were compared with those of a prior national multicenter questionnaire administered by our research team in 2005. METHODS: A questionnaire consisting of 28 questions about CanL was developed using Google Forms and distributed by email to 1428 veterinary clinics in Spain. Questions were designed to obtain data on common clinical signs, techniques and complementary exams used to diagnose the disease, and on its monitoring, treatment and control measures. Data were collected in a database for statistical analysis. RESULTS: Completed questionnaires were returned by 295 clinics. Compared to the situation in 2005, responses indicate that clinical signs of CanL have not changed significantly, cutaneous lesions being still the most prevalent sign observed by practitioners. Quantitative serological techniques are considered an adequate approach to diagnosis, provided their results are supported by the findings of a thorough physical exam, as well as complementary tests (complete blood count, biochemical profile, plasma protein electrophoretogram and complete urinalysis). Treatment protocols and check-ups follow international recommendations. Finally, a multimodal approach is being endorsed to adequately control CanL including preventive measures such as annual serological check-ups and the combination of repellents and vaccines. Additionally, owners are being better informed about CanL by veterinarians, which translates to the improved control of this zoonosis. CONCLUSIONS: The clinical management of CanL has recently undergone significant changes owing to improvements in clinical knowledge of the disease, more unified international criteria, improved diagnostic techniques and their adequate interpretation, as well as a greater awareness of the disease transmitted to owners.

Treatment and long-term follow-up of a cat with leishmaniosis.

BACKGROUND: Leishmania infection in cats is being increasingly reported in endemic areas. Nevertheless, only a few clinical cases have been described in cats, and even fewer have provided information on the response to treatment and a proper follow-up. Here we report a case of feline leishmaniosis not associated with any other disease or co-infection and document its response to allopurinol treatment and long-term follow-up data. RESULTS: A 6-year-old domestic shorthair female cat was referred for nodular blepharitis, mucocutaneous ulcerative lesions of the mouth and lymph node enlargement. The cat was moderately anaemic, hyperglobulinaemic and tested negative for feline leukaemia virus and feline immunodeficiency virus. Fine needle aspirates of nodules and mucocutaneous lesions showed the presence of numerous amastigote forms of Leishmania. Leishmania infection was further confirmed by serology (IFAT test, 1:640) and real-time PCR (RT-PCR) on blood and conjunctival swabs. The cat was treated with allopurinol (20 mg/kg SID), which was clinically effective, although the cat remained Leishmania-positive in serology and RT-PCR on blood and conjunctival swabs. Allopurinol treatment was interrupted after seven months because of the healing of all lesions and lack of compliance by the owner. After two years, the cat relapsed displaying almost the same clinical signs and clinicopathological alterations. On this occasion, the parasite was isolated by culture and identified as belonging to L. infantum. Allopurinol treatment was started again but was interrupted several times because of the itching side effect observed. The cat worsened progressively and died two months after the relapse without any chance to shift the treatment to another molecule (e.g. meglumineantimoniate or miltefosine). CONCLUSIONS: Out of all documented cases of feline leishmanosis, the present case has the longest follow-up period and it is one of the few in which the parasite was isolated and identified. It further confirms the potential progression of Leishmania infection to disease in cats even in the absence of comorbidities. Veterinarians practicing in endemic areas should be aware of this susceptibility, properly include feline leishmaniosis in the differential diagnosis and propose preventative measures to those cats at risk.

Feline leishmaniosis: Is the cat a small dog?

Leishmania infantum is a vector-borne zoonotic disease transmitted by phlebotomine sand flies and dogs are considered the main reservoir of the parasite. Feline leishmaniosis (FeL) caused by L. infantum is an emergent feline disease more and more frequently reported in endemic areas. This review summarizes current knowledge focusing similarities and differences with canine leishmaniosis (CanL). Cats are infected by the same Leishmania species than dogs but prevalence of the infection is lower and cases of disease are less frequently reported. Scarce information is available on adaptive immune response of cats naturally exposed to L. infantum infection and mechanisms responsible for susceptibility or resistance of feline hosts. However, about half of clinical cases of FeL are reported in cats with possible impaired immunocompetence. Coinfections or comorbidities are frequently detected in sick cats and they can contribute to a misrepresentation of clinical FeL albeit lesions associated with the presence of the parasite have been detected in skin, lymph nodes, spleen, bone marrow, liver, oral mucosa, stomach, large bowel, kidney, nasal exudate, lung, eye. As for dogs, skin or mucocutaneous lesions are the most common reason for veterinary consultation and finding on physical examination in cats with leishmaniosis. Molecular investigations of Leishmania DNA and anti- Leishmania antibody detection are largely used with the same methodologies for both CanL and FeL, however few information is available about their diagnostic performance in feline hosts. Treatment of cats with clinical FeL is still empirically based and off label by using the most common drugs prescribed to dogs. Life expectancy of cats with clinical FeL is usually good unless concurrent conditions or complications occur and prognosis does not seem significantly influenced by therapy or retroviral coinfection. According to current knowledge, cats can play a role as additional reservoir host of L. infantum and, in a « One Health ¼ perspective, preventative measures should be taken. In conclusion, albeit feline infection and the associated cat disease caused by L. infantum is increasingly reported in endemic areas and have many similarities with CanL, consolidated evidence-based knowledge is not available and we cannot exclude that important differences between dogs and cats exist about transmission, immunopathogenesis and best practice for management and prevention.

Feline leishmaniosis due to Leishmania infantum in Northwest Iran: The role of cats in endemic areas of visceral leishmaniosis.

Visceral leishmaniosis caused by Leishmania infantum (L. infantum) is an important zoonotic disease endemic in the Mediterranean region. Domestic dogs and other wild canines are the main reservoir hosts for the parasite, while domestic cats (Felis catus) may be carriers of L. infantum, and play a role as secondary reservoirs for the parasite. In the present study, serological (DAT), parasitological (microscopic smears and culture) and molecular methods (nested PCR) were used to evaluate infection with L. infantum in 103 stray cats collected from villages of Meshkin Shahr district, located in Ardabil province which is a well-known endemic region of human and canine visceral leishmaniosis in Iran. Overall, 25 out of 103 cats (24.27%) displayed anti-Leishmania antibodies with different titers. Amastigote forms of the parasite were detected in microscopic smears of the spleen of a cat with high anti-Leishmania antibodies using DAT. L. infantum was identified on microscopic slides by nested PCR, and the results were confirmed by sequence analysis. Based on the high rate of seropositive cats in this study, we conclude that cats may have an important role in the maintenance of L. infantum in the endemic areas of zoonotic visceral leishmaniosis in Iran.

Prevention of feline leishmaniosis with an imidacloprid 10%/flumethrin 4.5% polymer matrix collar.

BACKGROUND: Leishmaniosis caused by Leishmania infantum is one of the most important vector-borne diseases affecting animals and humans worldwide. Dogs are considered main reservoirs of the zoonotic forms, though in the last years the role of cats as reservoirs has been increasingly investigated. Feline leishmaniosis (FeL) occurs in endemic areas and no specific preventive measures have been investigated so far. In this study the efficacy of a 10% imidacloprid/4.5% flumethrin polymer matrix collar, licensed for tick and flea prevention, has been assessed against FeL in a longitudinal study on 204 privately owned cats from the Aeolian islands (Sicily), an area highly endemic for the disease. From March to May 2015 [Study Day 0 (SD 0)], cats negative for FeL were collared (G1, n = 104) or left untreated (G2, n = 100). Diagnosis consisted of serology and qPCR on blood and conjunctival swabs, which were collected at baseline (SD 0) and at the end of the study (SD 360). Interim clinical examinations were performed on SD 210 (when collars were replaced in G1) and SD 270. RESULTS: Of the 159 cats which completed the study, 5 in G1 and 20 in G2 were positive for L. infantum infection, in at least one of the diagnostic tests leading to a yearly crude incidence of 6.3% and 25.0% in G1 and G2, respectively (P = 0.0026). This translates into an efficacy of the collar of 75.0% in preventing feline Leishmania infection. The collar was generally well tolerated with no systemic adverse reactions and few local skin reactions were observed in the application area in four out of 104 treated cats (3.8%). CONCLUSIONS: The 10% imidacloprid/4.5% flumethrin collar significantly reduced the risk of L. infantum infection in cats. To our knowledge, this is the first study in which a preventative strategy against feline Leishmania infection is assessed under natural conditions. These findings close a gap in veterinary medicine, in that they confirm this collar as a tool in reducing the risk of Leishmania infection in cats. Such a preventative tool could contribute to the reduction of the risk of the disease in animal and in human populations when included in integrated leishmaniosis control programmes.

Feline leishmaniosis in Portugal: 3 cases (year 2014).

Leishmaniosis caused by Leishmania infantum is a zoonotic disease endemic in many countries of America, Asia and Europe, including Portugal. Dogs are the major reservoir of L. infantum, but domestic cats may also be infected. Three clinical cases of feline leishmaniosis are described, with ocular clinical signs as the only manifestation of the disease. A case had bilateral anterior uveitis and a granulomatous conjunctivitis, another one presented keratitis and the third case had a nodular blepharitis. All the affected cats had high serum titres of antibodies to L. infantum, while polymerase chain reaction results were positive in two of the cats. Although all cats in the present study improved after treatment with meglumine antimoniate and/or allopurinol, one of them died 6months later apparently due to a systemic L. infantum infection. The prevalence of disease may be underestimated in cats, because leishmaniosis is often not considered in the differential diagnosis of feline diseases. Feline leishmaniosis should be suspected in cats with ocular clinical signs and in those living in or traveling to areas where the zoonosis is endemic.