Microscopic detection of Leptospira bacterial organisms in urine sediment from a young dog with leptospirosis and a review of the pathobiology and diagnosis of canine leptospirosis.

Samples collected from an 11-month-old Dachshund-mix dog with a history of acute azotemia, fever, and enlarged and irregular kidneys were received at the Colorado State Veterinary Diagnostic Laboratory (CSU VDL). The submitting veterinarians were concerned about lymphoma versus acute nephritis/pyelonephritis. The CSU clinical pathology laboratory received urine for urinalysis and kidney aspirates for cytologic evaluation. Urine had also been submitted for aerobic culture and Leptospirosis PCR, and serum was submitted for Lepto-5 microscopic agglutination testing (MAT). Upon examination of a wet mount of the urine sediment, technical staff noted "vibrating" clumps of granular-appearing material throughout the slide, which prompted the preparation of a stained sediment slide for pathologist review. Very small, faintly staining organisms were observed, and an attempt was made to picture-match these with published reports of Leptospira in dog urine, but none could be found. In addition, some references claimed that Leptospira organisms are not seen in urine with light microscopy. The suspicion that these organisms were Leptospira sp. was supported by the MAT results and later confirmed by PCR. The organisms subsequently exhibited strong positive immunolabeling for the Leptospira antigen. This case report provides a searchable record of Leptospira organisms visualized by routine light microscopy in dog urine during natural infection and a review of canine leptospirosis pathobiology and diagnosis.

Functional characterization of galectin-3 from Nile tilapia (Oreochromis niloticus) and its regulatory role on monocytes/macrophages.

Galectin-3 is a kind of ß-galactoside-binding lectin involved in host defense against pathogen infection. However, the immune functions of fish galectin-3 remain poorly understood. In this study, the roles of a fish galectin-3 (OnGal-3) from Nile tilapia (Oreochromis niloticus) on the binding activity on bacterial pathogens or PAMPs, the agglutinating activity on bacterial pathogens and the regulatory effects on monocytes/macrophages activity were investigated. After in vitro challenge of Streptococcus agalactiae and Aeromonas hydrophila, OnGal-3 expressions were significantly up-regulated in monocytes/macrophages. In addition, recombinant OnGal-3(rOnGal-3) protein showed strong binding activity on bacterial pathogens or PAMPs. Also, rOnGal-3 agglutinated Gram-positive and Gram-negative bacteria. Moreover, rOnGal-3 could induce the inflammatory factors expressions in monocytes/macrophages and enhance phagocytosis and respiratory burst activity of monocytes/macrophages. These results suggest that fish galectin-3 participates in anti-bacterial immune response through recognizing pathogens and modulating monocytes/macrophages activity.

Isolation and Genetic Characterization of Toxoplasma gondii from Tissues of Wild Turkeys (Meleagris gallopavo) in Pennsylvania.

Toxoplasmosis in wild turkeys (Meleagris gallopavo) is of epidemiological interest because turkeys feed from the ground, and detection of infection in turkeys indicates contamination by oocysts in the environment. During the 2018 spring hunting season in Pennsylvania, fresh (unfixed, not frozen) samples were obtained from 20 harvested wild turkeys and tested for Toxoplasma gondii infection. Hearts from all wild turkeys and skeletal muscle from 1 were bioassayed for T. gondii by inoculation in outbred Swiss Webster (SW) and interferon-gamma gene knockout (KO) mice. Antibodies to T. gondii were detected in 1:5 dilution of neat serum from 5 of 15 wild turkeys and in fluid from the heart of 1 of 4 wild turkeys with the modified agglutination test (MAT); neat serum was not available from 4 wild turkeys. Viable T. gondii was isolated from hearts of 5 wild turkeys, 1 with MAT of 1:10, 1 with MAT of 1:5, and 3 seronegative (MAT < 1:5). Toxoplasma gondii was isolated from both heart and skeletal muscle in the 1 wild turkey that had skeletal muscle submitted. The KO mice inoculated with tissue from all 5 infected wild turkeys died or were euthanatized when ill, 7-21 days post-inoculation (PI). Tachyzoites were detected in lungs of all KO mice, and the T. gondii strains were successfully propagated in cell culture. The SW mice inoculated with tissues of wild turkeys remained asymptomatic, and tissue cysts were seen in the brains of infected mice when euthanatized in good health at 46 days PI; 1 of the 2 SW mice inoculated with the heart of 1 turkey died on day 26, and tachyzoites were detected in its lung. Genetic typing on DNA extracted from culture-derived tachyzoites using the PCR restriction fragment length polymorphism with 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) revealed that 4 isolates belonged to ToxoDB PCR-RFLP genotype #5 and 1 was genotype #216.

Soropositividade para Leptospira sp. e sorogrupos predominantes em cães do Pantanal Brasileiro / Seropositivity for Leptospira sp. and predominant serogroups in dogs from the Brazilian Pantanal

Em face à grande importância que a leptospirose possui no contexto sanitário mundial, tanto no aspecto humano como animal, este estudo teve por objetivo realizar a pesquisa de anticorpos anti-Leptospira sp. pela técnica de Soroaglutinação Microscópica (SAM) em 429 amostras de soros de cães provenientes de quatro municípios (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT e Corumbá/MS) localizados na região do Pantanal Brasileiro, bem como foram verificadas possíveis associações entre os resultados dos exames sorológicos e respostas aos questionários epidemiológicos aplicados aos proprietários. Do total de cães avaliados pela SAM (título ≥100), verificou-se que 34 (7,93%; IC 95%: 5,63%-11,00%) cães tinham anticorpos anti-Leptospira sp. Os títulos encontrados variaram entre 100 e 1600 e todos os municípios analisados tinham cães sororreagentes ao agente pesquisado. O sorogrupo reator mais frequente foi o Icterohaemorrhagiae, seguido pelo Australis. Por outro lado, foram observadas menores proporções de cães reagentes aos sorogrupos Tarassovi, Hebdomadis, Autumnalis e Grippotyphosa. As variáveis associadas com a ocorrência de leptospirose foram habitat rural (P<0,01) e área alagável (P=0,01). Estes resultados demonstram que os cães da região pantaneira tiveram contato com agentes do gênero Leptospira, o que representa uma informação relevante para a saúde pública local devido à importância zoonótica da doença.

Given the great importance that leptospirosis has the global health context, both in human and animal aspect, this study aimed to search for antibodies anti-Leptospira sp. by the technique of microscopic agglutination test (MAT) in 429 samples of sera from dogs from four municipalities (Poconé/MT, Santo Antônio de Leverger/MT, Barão de Melgaço/MT and Corumbá/MS) located in the Brazilian Pantanal region, in order for determine associations between the results of the serological tests and answers to epidemiological questionnaires applied to owners. Of the total dogs evaluated by MAT (titer ≥100), it was verified that 34 (7.93%, 95% CI: 5.63% -11.00%) dogs had antibodies against Leptospira sp. The titers found ranged from 100 to 1600 and all municipalities analyzed had seroreactive dogs for the investigated agent. The most frequent serogroup reactor was Icterohaemorrhagiae, followed by Australis. On the other hand, smaller proportions of reactive dogs were observed for serogroups Tarassovi, Hebdomadis, Autumnalis and Grippotyphosa. The variables associated with the occurrence of leptospirosis were rural habitat (P<0.01) and flooded area (P=0.01). These results demonstrate that dogs from the Pantanal region had contact with agents of the genus Leptospira, which represents information relevant to local public health due to the zoonotic importance of the disease.

First report on seroprevalence and risk factors of Toxoplasma gondii infection in sheep and goats in North Lebanon.

INTRODUCTION: Toxoplasmosis is of dual importance in both public and veterinary health due to the respective risk of transplacental transmission in primo-infected pregnant women and economic losses caused by abortions in mammals. One of the main routes of Toxoplasma gondii transmission to humans is the consumption of raw or undercooked meats containing parasitic cysts. Here, we performed the first epidemiological study to determine the seroprevalence and the risk factors of toxoplasmosis in livestock in Lebanon. METHODOLOGY: Using a modified agglutination test with a cut-off of 1:40, we tested the positivity rate of Immunoglobulin G antibodies in the sera of 100 sheep and 80 goats collected from 18 different livestock farms located in North Lebanon between March and June 2018. RESULTS: Anti-Toxoplasma gondii IgG antibodies were detected in 42% of sheep and 34% of goats. Adults (> 1 year) were significantly more infected by T. gondii than the lambs (< 1 year) in both species (p < 0.05). CONCLUSIONS: These findings indicated that food animals are highly exposed to T. gondii in Lebanon and could be potentially a major risk factor of T. gondii infection to humans. Consequently, national prophylactic strategies should be implemented to control and to prevent T. gondii transmission between animals and humans.

Detection of antibodies against Leptospira spp in free-living marsupials caught in the Eastern Amazon

Abstract INTRODUCTION: Serological surveys are important to assess the health status of wild animals. In this study, antibodies against Leptospira spp, causal agents of leptospirosis, were detected in free-living marsupials in the State of Pará, Brazil. METHODS: Nineteen blood samples collected from marsupials in the municipalities of Peixe-Boi, Viseu, and Castanhal were subjected to microscopic agglutination tests. RESULTS: In total, 36.8% (7/19) of samples were positive, and two exhibited co-agglutination. The most frequent serovars were Icterohaemorrhagiae (60%; 3/5), Panama (20%; 1/5), and Nupezo (20%; 1/5). CONCLUSIONS: Anti-Leptospira spp antibodies currently circulate in free-living marsupials in Northeastern Pará.

Serological and molecular detection of leptospira spp in dogs

Abstract INTRODUCTION: This study aimed to detect anti-Leptospira spp antibodies and Leptospira DNA in domestic dogs. METHODS: Blood and urine from 106 dogs were evaluated by microscopic agglutination test (MAT) and polymerase chain reaction (PCR), respectively. RESULTS: Six (5.7%) and one (1%) animals were positive by MAT and PCR, respectively. CONCLUSIONS: These results show a low prevalence of infection by Leptospira spp. The absence of positive results for the Icterohaemorrhagiae serogroup indicates the small relevance of these dogs as sources of human leptospirosis.

Prevalência de leptospirose em rebanhos bovinos no Pantanal de Mato Grosso do Sul / Prevalence of leptospirosis in cattle herds in the Pantanal of Mato Grosso do Sul

Foi realizado um estudo epidemiológico da leptospirose em fêmeas acima de 24 meses, provenientes de 246 rebanhos, e 2.766 animais amostrados aleatoriamente nos nove municípios que compõem a região do Pantanal de Mato Grosso do Sul, bem como identificados os fatores de risco associados à doença. As amostras de sangue foram coletadas no período de setembro a novembro de 2009 e examinadas pelo teste de aglutinação microscópica ante uma coleção de 24 antígenos vivos de Leptospira spp., representantes dos sorovares Australis, Bratislava, Autumnalis, Butembo, Castellonis, Batavie, Canicola, Whitcombi, Cynopteri, Grippotyphosa, Hebdomadis, Copenhageni, Icterohaemorrhagiae, Javanica, Panamá, Pomona, Pyrogenes, Hardjo, Wolffi, Shermani, Tarassovi, Sentot, Andamana e Patoc. Adicionalmente, representantes de doze estirpes de leptospiras isoladas no Brasil foram adicionados à coleção de antígenos do teste de soroaglutinação microscópica (SAM). A prevalência aparente foi de 66% e a prevalência real de animais infectados, de 79,80%, com intervalo de confiança (IC) de 95% (78,3-81,3) e 241 rebanhos apresentando pelo menos um animal reagente. Os sorovares mais prováveis foram o Hardjo seguido pelo Wolffi. Os resultados demonstram que a leptospirose bovina continua presente no Pantanal, com alta prevalência tanto em rebanhos quanto em indivíduos, sendo os principais fatores de risco para a doença o tipo de exploração e a raça.(AU)

This is an epidemiological study of leptospirosis in 24 month-old females from 246 herds. Two thousand, seven hundred and sixty six (2,766) animals were randomly sampled in the nine counties comprising the region of Pantanal of Mato Grosso do Sul, Brazil. The risk factors associated with the disease were also identified. Blood samples were collected from September to November 2009 and examined by the microscopic agglutination test (MAT) against a collection of 24 live antigens of Leptospira spp., representatives of serovars Australis, Bratislava, Autumnalis, Butembo, Castellonis, Batavie, Canicola Whitcombi, Cynopteri, Grippotyphosa, Hebdomadis, Copenhageni, Icterohaemorrhagiae, Javanica, Panama, Pomona, Pyrogenes, Hardjo, Wolffi, Shermani, Tarassovi, Sentot, Andamana, and Patoc. Additionally, twelve representatives of Leptospira strains isolated in Brazil were added to the collection of antigens for the microscopic agglutination test (MAT). The apparent prevalence was 66% and the actual prevalence of infected animals was 79.80%, with a confidence interval of 95% (78.3 to 81.3) and 241 herds having at least one reactive animal. The most likely serovars were Hardjo followed by Wolffi. Results show that bovine leptospirosis is still present in Pantanal, with high prevalence both in animals and herds, the main risk factors for the disease being the type of cattle farming and breeding.(AU)

Immune response of a commercial vaccine against Leptospira interrogans: Antibodies and cytokine levels.

Good vaccines should confer protection against specific pathogens in experimental and field conditions. However, some commercial vaccines are not capable to confer protection to animals, being inefficient in a bovine vaccine program. In this sense, the aim of this study was to evaluate the antibody levels involved in the immune response in vaccinated cows against leptospirosis, as well as acute phase protein and the immunological markers in a vaccine program in beef cattle. Twenty non-lactating cows, negative for leptospirosis and without vaccination against this disease were evaluated during five months. The herd was divided into two groups named as A (the control group) and B (the vaccinated group). Ten cows from the group B received an initial dose (5 mL) of vaccine on day 0 and one booster dose (5 mL) on day 29. In order to evaluate humoral response (MAT - titration 1:25), cytokine levels (tumor necrosis factor (TNF) and interleukin-10 (IL-10)), and C-reactive protein levels (an acute phase protein), blood samples were evaluated on days 0, 29, 40, 83 and 144 after vaccination. In none of the evaluated periods it was observed specific antibodies to any of the six serovars presents in the vaccine, as well as no difference between groups regarding cytokine and C-reactive protein levels. Therefore, the vaccine used did not stimulate the immune response of cattle, inferring the absence of protection against infections by L. interrogans.

Detection of Toxoplasma gondii in naturally infected domestic pigs in Northern Serbia.

Insufficiently cooked pork is considered as an important source of human infection with Toxoplasma gondii. The aim of our study was to investigate the presence of T. gondii in pigs intended for human consumption from Northern Serbia. Blood and diaphragm samples were collected from 182 naturally infected market-weight pigs, originating from both commercial farms and smallholdings. Sera were examined using modified agglutination test (MAT), and diaphragms from seropositive, as well as from some MAT-negative pigs, were bioassayed in mice. In addition, digests were examined for the presence of T. gondii DNA using a real-time polymerase chain reaction (qPCR) which was targeted at the 529 bp repetitive element of the T. gondii genome. The overall seroprevalence of T. gondii in pigs was 17% (31/182), with no difference between pigs from large commercial farms (17.8%) and those raised on smallholdings (16.3%). However, the seroprevalence in farm pigs was largely influenced by the findings on a single farm, where all examined animals tested positive. Parasites and/or parasite DNA were detected in the tissues of 15 of the 45 (25 seropositive and 20 seronegative) animals examined by either direct method. Tissue cysts were isolated in eight bioassays and an additional bioassay was positive by serology; all nine were confirmed positive by qPCR. All positive bioassays originated from seropositive pigs, but no correlation was observed between isolation rate and antibody titer. T. gondii DNA was detected in diaphragm tissues of eight pigs, of which three were seronegative. The results of our study provide further evidence for pork as a source of human T. gondii infection.

The Essential Oil of Hyptis mutabilis in Ichthyophthirius multifiliis Infection and its Effect on Hematological, Biochemical, and Immunological Parameters in Silver Catfish, Rhamdia quelen.

This study evaluated the activity of leaf essential oil (EO) from Hyptis mutabilis as well as its major constituent, (-)-globulol, in infections by the parasite Ichthyophthirius multifiliis (ich). Effects on hematological, biochemical, and immunological parameters of silver catfish, Rhamdia quelen, exposed to the same samples also were evaluated. In the first experiment, naturally infected fish were treated with EO (0, 10, and 20 mg · L-1) and ethanol, using several methods of exposure. Fish mortality and the number of trophonts per fish were assessed after 48 and 96 hr. Hour-long daily baths resulted in optimal survival, so this methodology was used for the second experiment, in which infected animals were exposed to (-)-globulol at 2.5 and 5 mg · L-1. The most effective concentrations in Experiments 1 and 2 were chosen for Experiment 3, in which healthy animals were subjected to hour-long daily baths with EO (20 mg · L-1) or (-)-globulol (2.5 mg · L-1). Additionally, an in vitro experiment was performed with EO and globulol at the same concentrations of the in vivo test. EO and (-)-globulol increased the survival of fish infected with ich and altered certain hematological and biochemical parameters. After 4 days, levels of hematocrit, erythrocytes, and leukocytes increased significantly in healthy animals exposed to EO. Exposure to (-)-globulol increased leukocyte number alone. No significant differences in nonspecific immunological parameters were detected when treated groups were compared to controls, but the leukocytosis observed in EO- and globulol-treated healthy animals indicates that EO and (-)-globulol increased innate immunity in these fish. An in vitro antiparasitic effect was observed in both samples.

Diagnostic utility of an immunochromatography test for the detection of Leptospira IgM antibodies in domestic dogs

A cross-sectional study using 99 serum samples of dogs from southern Chile was conducted to determine the diagnostic utility of a rapid immunochromatography assay for the detection of Leptospira specific IgM antibodies as screening test and as a potential aid in the diagnosis of leptospirosis in animals with and without clinical suspicion of the disease. The Microscopic Agglutination Test (MAT) was used as reference assay. Anti-Leptospira antibodies were detected in 37.3% of the dogs with MAT. Using the immunochromatography test, specific IgM antibodies were found in 13.1% of sampled dogs. The sensitivity of the rapid test as screening assay was 29.7% (95% Confidence Interval=16.4-47.2) and the specificity was 96.7% (95% Confidence Interval=87.8-99.4). 40.0% of the canines with clinical suspicion of leptospirosis and 37.1% of dogs without clinical signs were serological reactors to MAT, but none of MAT reactive dogs with clinical suspicion tested positive in the rapid test. Rapid and user-friendly diagnostic procedures for canine leptospirosis such as this immunochromatography assay could be important tools to use in clinical practice, however, further studies are needed to obtain more information about their utility, considering that diagnostic tests could not have similar performances in different geographic locations, clinical and epidemiological contexts.(AU)

Negative serosurvey of Toxoplasma gondii antibodies in Golden-headed Lion Tamarin (Leontopithecus chrysomelas) from Niterói/RJ, Brazil / Pesquisa sorológica de anticorpos anti-Toxoplasma gondii em Mico-leão-da-cara-dourada (Leontopithecus chrysomelas) de Niterói/RJ, Brasil

Abstract New World Nonhuman Primates are highly susceptible to clinical toxoplasmosis. Serum samples from 126 recently captured Leontopithecus chrysomelas, from an exotic and invasive population, were tested for Toxoplasma gondii antibodies by the modified agglutination test (MAT, cut-off 1:25); all were seronegative. The MAT is highly specific and is not species-specific. This is the first report of T. gondii survey in this tamarin in the wild. This result is consistent with prior reports that showed the high susceptibility of the species to infection by T. gondii usually with high mortality rates.

Resumo Primatas não humanos são extremamente susceptíveis a toxoplasmose. No presente estudo, 126 Leontopithecus chrysomelas foram capturados de uma população de vida livre, exótica e invasora, e amostras de soros foram testadas para a presença de anticorpos anti- Toxoplasma gondii pelo Teste de Aglutinação Modificado (MAT, ponto de corte 1:25). Todos os animais testados foram negativos. O MAT é um teste altamente específico e não é espécie-específico. Esse é o primeiro estudo de pesquisa por anticorpos anti- T. gondii nessa espécie em vida livre. O resultado corrobora com o conhecimento prévio sobre a susceptibilidade dessa espécie a infecção pelo parasite T. gondii.

Alloimmunisation in transfused patients: serial cross-matching in a population of hospitalised cats.

Objectives Cross-matching is currently recommended as part of pre-transfusion testing for repeat transfusions in cats 4 days after having received an initial transfusion. This prospective study determined when and if cats developed positive cross-match (CM) results after having been transfused with AB-compatible blood. Methods Donors were selected according to standard transfusion safety protocols. Twenty-one hospitalised anaemic recipients (blood type A: n = 20; blood type B: n = 1) received 1-4 (median 2) whole blood transfusions (WBTs) over 1-6 days (median 2) in 33 transfusion instances. The tube CM method, including major, minor and recipient control, was employed. Macroscopic and microscopic agglutination reactions were evaluated according to a predetermined scale. CM tests with a positive recipient control could not be evaluated. Results No signs of an acute transfusion reaction were observed. A total of 63 CMs were performed. In one cat with immune-mediated haemolytic anaemia the CM could not be evaluated (positive recipient control). The minor CM was negative in all cases. Fifteen of 20 cats had a negative major CM (MCM) 1-12 days (median 5) after their first transfusion. A positive MCM was observed in five cases after 2-10 days (median 5) post-first WBT. These five cats had received a total of 1-4 (median 2) WBTs. Cats with a negative MCM had received 1-3 (median 2) WBTs. In 51.5% (17/33) of transfusion instances, the cat's haematocrit increased as expected, with cats with a positive MCM at 40% (4/10) vs 56.5% (13/23) if MCM was negative. Conclusions and relevance Twenty-five percent (5/20) of the feline recipients likely developed alloantibodies against erythrocyte antigens outside of the AB system as early as 2 days post-first WBT. This adds data to the recommendation to include cross-matching in pre-transfusion screening tests.

Detection of Leptospira DNA in urine and presence of specific antibodies in outdoor cats in Germany.

Objectives Clinical manifestation of infection with Leptospira species in cats is rare. Nevertheless, cats can develop specific antibodies against the spirochetes after infection. In Canada, Taiwan and the USA it was recently demonstrated that naturally infected cats can also shed DNA from pathogenic Leptospira species in their urine, but the zoonotic potential of infected cats is still unclear. The objective of this study was to demonstrate if outdoor cats in Germany shed DNA from pathogenic Leptospira species in their urine. As a second aim, antibody prevalence was determined. Methods Two hundred and fifteen outdoor cats were prospectively recruited. Urine samples were tested by real-time PCR targeting the lipL32 gene of pathogenic Leptospira species. Antibody titres against eight serovars (Australis, Autumnalis, Bratislava, Canicola, Copenhageni, Grippotyphosa, Pomona, Saxkoebing) belonging to seven serogroups (Australis, Autumnalis, Canicola, Grippotyphosa, Icterohaemorrhagiae, Pomona, Sejroe) were determined by microscopic agglutination test. Results Urine samples from 7/215 cats (3.3%; 95% confidence interval [CI] 0.9-5.7) were PCR-positive. Specific antibodies were detected in 35/195 cats (17.9%; 95% CI: 12.5-23.3) with titres ranging from 1:100 to 1:6400. Australis, Bratislava and Grippotyphosa were the most common serovars. Conclusions and relevance Outdoor cats in Germany can shed DNA from pathogenic Leptospira species. Therefore, outdoor cats should be considered as a possible source of infection for dogs or humans. Further studies are needed to determine the role of Leptospira species as a cause of disease in cats.

A prototype of the direct agglutination test kit (DAT-Canis) for the serological diagnosis of canine visceral leishmaniasis.

This report describes the stege I/II development of a new direct agglutination test (DAT) for the diagnosis of canine visceral leishmaniasis (CVL) using freeze-dried antigen produced Coomassie blue-stained Leishmania (Leishmania) infantum promastigotes. In stage I, 16 canine serum samples, collected from eight dogs carrying CVL and eight healthy dogs, were assessed with the DAT using 2-mercaptoethanol (2-ME), N-acetyl-cysteine (NAC), kaolin or NAC plus urea (NAC+U) to improve the assay conditions. Stage II assessed the diagnostic accuracy with 100 serum samples collected from dogs with symptomatic CVL and clinically healthy dogs, comparing the four different sample diluents. The CVL-DAT prototype kit showed equivalent performances when 2-ME, NAC or NAC+U were used: 97.1% sensitivity (CI: 83-99.8%), 97% specificity (CI: 88.5-99.5%) and a 97% diagnostic accuracy (CI: 90.8-99.2). With kaolin, a 94.1% sensitivity (CI: 79-99%), 97% specificity (CI: 88.5-99.5%) and 96% diagnostic accuracy were observed (CI: 89.5-98.7), with no statistically significant differences among the four reagents (p=1.0). The NAC plus urea in sample diluent decreased non-specific agglutination, promoted a better defined sharp-edged blue spot and was thus chosen as a component for the new DAT prototype to diagnose canine VL, designated DAT-Canis.

Ocorrência de anticorpos anti-toxoplasma gondii em aves silvestres de três unidades de conservação federais da Paraíba e da Bahia / Occurrence of antibodies to toxoplasma gondii in wild birds from three federal conservation units of Paraíba and Bahia, Brazil

A vigilância e monitoramento de doenças em animais silvestres são imprescindíveis no contexto ambiental e de saúde pública, pois estes animais agem como sentinelas, refletindo alterações ambientais precocemente, o que proporciona maior eficácia no monitoramento ambiental e permite o acesso rápido a informações sobre as condições da área. Neste contexto, as aves são importantes no ciclo biológico do Toxoplasma gondii e na epidemiologia da toxoplasmose, principalmente porque seus tecidos representam importantes fontes de proteína na alimentação de felídeos e humanos. Objetivou-se detectar anticorpos anti-T. gondii, por meio do teste de aglutinação modificada em aves silvestres de três Unidades de Conservação (UC) Federais dos Estados da Paraíba e Bahia. No período de dezembro de 2011 a outubro de 2013 foram capturadas com redes de neblina 222 aves silvestres pertencentes a 67 espécies, 27 famílias e 12 ordens. Após a captura, foi colhido sangue de cada animal e separado o soro, que foi submetido ao Teste de Aglutinação Modificada (MAT≥1:25) utilizando taquizoítos inativados na formalina e 2-mercaptoetanol. Dentre as 222 amostras analisadas, três (1,3%) foram sororreagentes: 1 de 16 (6,2%) pipira-preta Tachyphonus rufus (título 50), 1 de 5 (20%) juriti-gemedeira Leptotila rufaxilla (título 50) e 1 de 1 (100%) caneleiro-enxofre Casiornis fuscus (título 25). Este é o primeiro relato da ocorrência de anticorpos anti-T. gondii nas referidas espécies de aves silvestres de vida livre nas duas UC Federais estudadas.

Surveillance and monitoring of wildlife pathogens are essential in the environmental context and human public health, as these animals act as sentinels, reflecting environmental changes early on, whath gives more efficient environmental monitoring and allows quick access to information on the conditions of area. Birds are important in the epidemiology and life cycle of Toxoplasma gondii, because their tissues are important source of protein in the diet of felids and humans. The objective was to determine antibodies to Toxoplasma gondii in wild birds from three Federal Conservation Units of the states of Paraíba and Bahia by Modified Agglutination Test (MAT). From December 2011 to October 2013, 222 wild birds of 67 species from 27 families and 12 Orders were captured with mist nets. Blood samples were then collected and the serum was separated by centrifugation. The sera were tested (MAT≥1:25) using formalin-fixed whole tachyzoites and 2-mercaptoethanol. Antibodies to T. gondii were found in 3 of 222 (1.3%) birds: in 1 of 16 (6.2%) white-lined tanager (Tachyphonus rufus, titer 50), in 1 of 5 (20%) gray-fronted dove (Leptotilla rufaxila, titer 50), and in 1 of 1 (100%) ashy-throated casiornis (Casiornis fuscus, titer 25). This is the first report of occurrence of antibodies to T. gondii in these tree bird species from two Federal Conservation Units.

Evaluation of the Leptospira species microscopic agglutination test in experimentally vaccinated cats and Leptospira species seropositivity in aged azotemic client-owned cats.

OBJECTIVES: The objectives of this study were to validate the microscopic agglutination test (MAT) using feline sera, determine cross-reactivity of Borrelia burgdorferi antibodies in the MAT, and evaluate if there is an association between Leptospira species seropositivity in aged (⩾10 years) client-owned cats with and without azotemia (creatinine >2 g/dl). METHODS: A four-serovar canine leptospiral vaccine was administered to two specific pathogen-free (SPF) cats on days 0 and 14. The MAT was performed intermittently until day 42 for the serovars Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona and Bratislava, with a cut-off value of ⩾1:100. Five purpose-bred cats were infested with wild-caught Ixodes scapularis adults with an average B burgdorferi infection rate of 50%, and tested for antibodies against B burgdorferi C6 peptide and DNA in skin biopsies, as well as by MAT. Sera from 66 azotemic and 75 non-azotemic cats ⩾10 years of age were tested for Leptospira species antibodies using the MAT and results were compared by the χ(2) test. RESULTS: Both SPF cats seroconverted by week 3 and formed antibodies against at least one serovar. There was no cross-reactivity in the MAT using samples from cats with antibodies to B burgdorferi. MAT results were positive for 4/66 azotemic cats and 8/75 non-azotemic cats; these results were not statistically different. CONCLUSIONS AND RELEVANCE: The MAT can be interpreted using feline serum and does not appear to cross-react in cats with B burgdorferi antibodies. There was no association between Leptospira species MAT results and azotemia in this group of aged client-owned cats but further studies are needed to determine if leptospirosis contributes to feline chronic kidney disease.

Toxoplasma gondii, Dirofilaria immitis, feline immunodeficiency virus (FIV), and feline leukemia virus (FeLV) infections in stray and pet cats (Felis catus) in northwest China: co-infections and risk factors.

This study was conducted to estimate the prevalence of Toxoplasma gondii, Dirofilaria immitis, feline immunodeficiency virus (FIV), and feline leukemia virus (FeLV) infections among stray and pet cats in Lanzhou, northwest China, and to identify the influence of age, gender, and regions on seropositivity. T. gondii antibodies were examined in cat sera by the modified agglutination test (MAT). The circulating antigens of D. immitis and FeLV and specific antibodies to FIV were examined using kits commercially available. The overall prevalence of T. gondii, FIV, FeLV, and D. immitis was 19.34, 9.12, 11.33, and 3.04 %, respectively. For the genetic characterization of T. gondii genotypes in cats, genomic DNA was extracted from the seropositive cats and the T. gondii B1 gene was amplified using a semi-nested PCR. DNA samples giving positive B1 amplification were then genotyped using multilocus PCR-RFLP. Two T. gondii genotypes (ToxoDB#9 and ToxoDB#1) were identified. Results of the multivariate logistic regression analysis showed that older cats are more likely to be seropositive than juveniles for T. gondii, FIV, FeLV, and D. immitis. This is the first report of T. gondii genotypes in cats in northwest China. Moreover, the present study is the first study of retrovirus and D. immitis seroprevalence in cats in China. The results revealed that T. gondii, FIV, and FeLV infections are common in stray and pet cats in northwest China.

Serological survey of Toxoplamosis in South American coatis (Nasua nasua) in Tietê Ecological Park, São Paulo, SP, Brazil / Inquérito sorológico da Toxoplasmose em quatis (Nasua nasua) do Parque Ecológico do Tietê, São Paulo, SP, Brasil

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis in humans, domestic animals and wild animals. Although there is serological evidence of T. gondii infection in wild animals, little is known about the role of wildlife in the epidemiological chain of this parasite. The South American coati (Nasua nasua) is an omnivorous species capable of adaptation to different environments. It can be found throughout Brazil, and in anthropized environments it can come into close contact with domestic animals. The present study found occurrences of T. gondii antibodies in South American coatis living in the Tietê Ecological Park, in the municipality of São Paulo, Brazil, through the modified agglutination test (MAT). In total, 99 samples were obtained and 70.70% (n = 70) were positive for anti-T. gondii antibodies, with titers that ranged from 50 to 3200. The data from this study indicate that South American coatis are exposed to this parasite.(AU)

Toxoplasma gondii é um protozoário parasita intracelular obrigatório que causa a toxoplasmose em humanos, animais domésticos e selvagens. Apesar das evidências sorológicas da infecção por T. gondii em animais selvagens, pouco se sabe sobre o papel da vida selvagem na cadeia epidemiológica deste parasito. Os quatis (Nasua nasua) são uma espécie onívora capaz de se adaptar em diferentes ambientes. Eles são encontrados em todo território brasileiro, e em ambientes antropizados podem apresentar um contato próximo com animais domésticos. O presente estudo verificou a ocorrência de anticorpos anti-T. gondii em quatis habitantes do Parque Ecológico do Tietê, São Paulo, Brasil, pelo Teste de Aglutinação Modificada (MAT). No total, 99 amostras foram obtidas das quais 70,70% (n = 70) foram positivas para anticorpos anti-T. gondii, com títulos de anticorpos que variaram de 50 a 3200. Os dados obtidos neste estudo indicam que quatis sul-americanos são expostos a este parasito.(AU)