Complete genome sequence and in silico analysis of L. interrogans Canicola strain DU114: A virulent Brazilian isolate phylogenetically related to serovar Linhai.

Canine leptospirosis is often caused by Leptospira interrogans serovar Canicola. Infected dogs may become asymptomatic carriers of the pathogen, which leads to many public health concerns. In this work, we present the complete genome sequencing and in silico analysis from a virulent Brazilian strain of L. interrogans serovar Canicola, previously isolated from a stray dog in Sao Paulo City. Comparative genomic analysis with a reference genome allowed identification of 1031 INDELs and several arrangement variations. Out of 35,361 SNPs identified, 6780 were missense mutations and 16,114 were synonymous mutations. The Gene Ontology terms more affected by mutations were described. Interestingly, phylogenetic analyses indicated a genetic relatedness of the isolate with serovar Linhai strain 56,609. In addition, we found several virulence-related genes and main outer membrane proteins associated with pathogenesis. This genomic information about canine isolates may help to elucidate the molecular diversity and mechanisms of Leptospira spp. pathogenicity.

Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples

Abstract A modified TaqMan real-time polymerase chain reaction targeting a 138 bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88-100%). The newly developed assay presented 91.6% (CI 95% 71.5-98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and sequencing), 100% (CI 95% 96.3-100%) relative specificity and 98.7% accuracy (CI 95% 94.8-100%). The lipl32 assay was able to detect and quantify at least 10 genome equivalents/reaction. DNA extracted from 17 pathogenic Leptospira spp., 8 intermediate/saprophytic strains and 21 different pathogenic microorganisms were also tested using the lipl32 assay, resulting in amplification exclusively for pathogenic leptospiral strains. The results also demonstrated high intra and inter-assay reproducibility (coefficient of variation 1.50 and 1.12, respectively), thereby qualifying the newly developed assay as a highly sensitive, specific and reliable diagnostic tool for leptospiral infection in dogs using urine specimens.

Prospective study of canine leptospirosis in shelter and stray dog populations: Identification of chronic carriers and different Leptospira species infecting dogs.

Dogs are highly susceptible to the leptospiral infection, notably stray and sheltered dogs. Unsanitary conditions often observed in dog shelters may predispose the introduction and spread of leptospires among sheltered populations, potentially increasing the chances for the inadvertent adoption of asymptomatically infected animals. The present work describes a longitudinal study using a multidisciplinary approach for the identification of chronically infected dogs and the characterization of potentially pathogenic strains circulating among stray and sheltered dog populations in São Paulo, Brazil. A total of 123 dogs from three populations were included. The initial evaluation consisted of blood and urine quantitative PCR testing (qPCR), the detection of specific antibodies by microscopic agglutination test (MAT), physical examination and hematological and serum biochemistry analyses. The qPCR-positive dogs were prospectively examined, and reevaluations also included culture from urine samples. Positive qPCR samples were subjected to 16S rRNA and secY gene phylogenetic analysis. The recovered strains were characterized by Multilocus Sequence Typing, polyclonal serogroup identification and virulence determination. Leptospiruria was detected in all populations studied (13/123), and phylogenetic analysis revealed that 10 dogs had L. interrogans infection. Three dogs (3/13) had L. santarosai infection. The secY phylogenetic analysis revealed that the L. santarosai sequences clustered separately from those obtained from other hosts. Ten leptospiruric dogs were reevaluated, and three dogs presented persistent leptospiruria, allowing culturing from two dogs. The strains were characterized as L. interrogans serogroup Canicola (virulent) and L. santarosai serogroup Sejroe (not virulent). Serum samples were retested by MAT using the DU92 and DU114 strains as antigens, and no increased seroreactivity was detected. Asymptomatic L. santarosai infection was observed in all populations studied, suggesting a possible role of dogs in the chain of transmission of this leptospiral species. The results suggest a genetic distinction between lineages of Brazilian L. santarosai maintained by dogs and other animal hosts. Our findings revealed that dogs could act as maintenance hosts for distinct pathogenic Leptospira, highlighting also that asymptomatically infected dogs can be inadvertently admitted and adopted in dog shelters, potentially increasing the risks of zoonotic transmission.

Diagnosis of acute canine leptospirosis using multiple laboratory tests and characterization of the isolated strains.

BACKGROUND: Dogs presenting with acute leptospirosis may present non-specific clinical and laboratory findings, and the definitive diagnosis may require additional confirmatory tests, including bacterial culture, for the direct or indirect identification of the pathogen. The present study describes the diagnosis of leptospirosis in suspected dogs based on the use of multiple diagnostic tests, including serological, molecular and bacteriological tests, along with the characterization of the recovered leptospiral strains. RESULTS: Urine, serum and blood samples were collected from 33 dogs with suspected clinical leptospirosis treated at the University of São Paulo Veterinary Hospital Service (Hovet FMVZ-USP) between 2013 and 2016. Only dogs with high blood urea nitrogen and creatinine levels in association with multiple clinical manifestations of the disease were included. Leptospiral culture, PCR and serology (Microscopic agglutination test - MAT) were performed in blood and urine samples taken from all suspected dogs at clinical presentation, and an additional prospective MAT titration was performed in seven dogs. Infection could be identified exclusively by PCR in 10 dogs (30.3%), exclusively by MAT in four dogs (12.1%) and by both tests in four dogs, totaling 18 dogs (54.5-95%CI: 37.6-71.5). Six out of eight MAT-confirmed cases presented with the highest titers against the Icterohaemorrhagiae serogroup. Leptospires were recovered from urine samples from two PCR-positive dogs, and both strains could be characterized by Multilocus Sequence Analysis and serogrouping as L. interrogans serogroup Icterohaemorrhagiae. Both isolates were shown to be pathogenic in the hamster model. CONCLUSIONS: The simultaneous use of MAT and PCR was able to increase the diagnosis of leptospirosis in clinically suspected cases. Despite the increasing incidence of new serovars affecting dogs being reported in different locations, our results suggest that leptospiral strains belonging to the Icterohaemorrhagiae serogroup are still a major causative agent of canine leptospirosis in São Paulo, Brazil.

Development and validation of a modified TaqMan based real-time PCR assay targeting the lipl32 gene for detection of pathogenic Leptospira in canine urine samples.

A modified TaqMan real-time polymerase chain reaction targeting a 138bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleotide sequencing analysis. The overall agreement between the assays was 94.8% (confidence interval [CI] 95% 88-100%). The newly developed assay presented 91.6% (CI 95% 71.5-98.5%) relative sensitivity (22[+] lipl32 PCR/24[+] 16S RNA and sequencing), 100% (CI 95% 96.3-100%) relative specificity and 98.7% accuracy (CI 95% 94.8-100%). The lipl32 assay was able to detect and quantify at least 10 genome equivalents/reaction. DNA extracted from 17 pathogenic Leptospira spp., 8 intermediate/saprophytic strains and 21 different pathogenic microorganisms were also tested using the lipl32 assay, resulting in amplification exclusively for pathogenic leptospiral strains. The results also demonstrated high intra and inter-assay reproducibility (coefficient of variation 1.50 and 1.12, respectively), thereby qualifying the newly developed assay as a highly sensitive, specific and reliable diagnostic tool for leptospiral infection in dogs using urine specimens.

Molecular and serological characterization of the first Leptospira santarosai strain isolated from a dog.

Leptospirosis is a zoonotic disease of global importance caused by pathogenic Leptospira species. Dogs can become asymptomatically infected, acting like reservoir hosts for pathogenic Leptospira, notably Leptospira interrogans serovar Canicola. Identification of such individuals and characterization of leptospires involved in chronic infections may unravel the role of dogs in the epidemiology of particular leptospiral strains. The aim of the present work was to describe the first Leptospira santarosai strain isolated from a dog. The dog was kept in a public shelter in São Paulo city, Brazil, and presented asymptomatic urinary shedding detected by PCR. Prospective evaluation was performed to fully characterize its chronic carrier state. The dog did not present anti-Leptospira titles or clinical/laboratorial abnormalities during the evaluations; nevertheless long-term urinary shedding was confirmed by PCR and leptospires were recovered from two occasions. The isolated strain was molecularly characterized by partial 16S rRNA and secY gene sequencing and MLST analysis. Serogroup identification was performed using polyclonal antibodies. The strain was identified as Leptospira santarosai, serogroup Sejroe. This is the first evidence in the literature of the isolation of L. santarosai in dogs. Our findings show that dogs can persistently harbor leptospires other than L. interrogans.

Hematological adaptation in Holstein calves during the neonatal period / Adaptação hematológica de bezerras Holandesas durante o período neonatal

The aim of this study was to evaluate the hematological profile of Holstein calves during the first month of life. Blood samples were harvested (n = 208) from 26 calves, from birth until 30 days of life. Hematologic values were determined by an automatic system associated with differential leukocyte count by manual methods. Variations in the erythrogram components were detected from birth up to the 30th day of life, except for hemoglobin (Hb) concentration and Mean Corpuscular Hemoglobin Concentration (MCHC). At birth, higher values were observed for hematocrit, Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) that decreased in subsequent moments. During the first days of life, leukocytosis was found due to neutrophilia and eosinopenia. A gradual increase of lymphocytes with the increase of age was also observed. Finally, the present research showed that the first month of life is a hematological adaptation period. Based on the results, it detected that blood component variations, characterized by hemoconcentration and leukocyte prolife compatible with glucocorticoids response up to the 4th day of life, were responsible for neutrophil lymphocyte ratio > 1.0 at birth.

Foi avaliado o perfil hematológico de bezerros da raça Holandesa durante o primeiro mês de vida. Foram colhidas 208 amostras de sangue total de 26 bezerras(os) do nascimento aos trinta dias de vida. Os valores hematológicos foram determinados por sistema automatizado associado à contagem diferencial dos leucócitos por metodologia manual. Foram detectadas variações nos componentes do hemograma do nascimento aos 30 dias de vida, exceto para os teores de hemoglobina (Hb) e concentração hemoglobínica corpuscular média (CHCM). Os maiores valores do hematócrito, volume corpuscular médio (VCM) e hemoglobina corpuscular média (HCM) foram observados ao nascimento, com decréscimo nos momentos subsequentes. Nos primeiros dias de vida foi observada leucocitose por neutrofilia e eosinopenia e com o avançar da idade houve aumento gradativo dos linfócitos. Com base nos resultados obtidos pode-se concluir que a adaptação dos bezerros no período pós-neonatal foi caracterizada por variações nos componentes do hemograma, observando-se hemoconcentração e padrão leucocitário compatível com resposta aos glicocorticoides até o 4º dia de vida, responsável pela relação neutrófilo-linfócito > 1,0 ao nascimento.

Estudo da ocorrência de litíase renal e ureteral em gatos com doença renal crônica / Study of nephrolithiasis and ureterolithiasis in cats with chronic kidney disease

Investigou-se a ocorrência de nefrolitíase e/ou ureterolitíase em 72 gatos portadores de doença renal crônica (DRC), classificados predominantemente no estágio II, segundo os critérios designados pela IRIS - International Renal Interest Society. Destes pacientes, 47 (65,27por cento) apresentaram litíase renal e ou ureteral. Não houve diferença estatística entre o grupo de estudo (DRC com cálculo) e o grupo controle (DRC sem cálculo) em relação à idade (p=0,274). Apesar disso, os pacientes portadores de nefrolitíase e/ou ureterolitíase apresentaram maiores indícios de lesão renal, caracterizados por diferenças estatisticamente relevantes da densidade urinária (p=0,013) e pelo menor tamanho dos rins direito (p=0,009) e esquerdo (p=0,048). Encontrou-se similaridade entre os grupos em relação a outros parâmetros, tais como as concentrações plasmáticas de cálcio total, cálcio ionizado, fósforo, sódio, potássio e paratormônio intacto (PTHi). Os valores das concentrações séricas de ureia e bicarbonato diferiram entre os grupos, com valores de p=0,039 e p=0,037, respectivamente. Além disso, foi mensurada a pressão arterial, que se manteve inalterada na comparação entre o grupo de estudo e o grupo controle. Os resultados obtidos reforçam a necessidade de acompanhamento ultrassonográfico de todos os pacientes portadores de DRC, mesmo daqueles assintomáticos ou em estágios iniciais da doença.

Nephrolithiasis and/or ureterolithiasis were investigated by means of ultrasonography in 72 cats with chronic kidney disease (CKD), predominantly classified in stage II, according to IRIS - International Renal Interest Society criteria. Of these patients, 47 (65.27 percent) had nephrolithiasis and/or ureterolithiasis. There was no statistical difference between the study group (CKD with calculi) and control group (CKD without calculi) regarding age (p=0.274). Nevertheless, patients with nephrolithiasis and/or ureterolithiasis had greater evidence of renal injury, characterized by statistically significant differences in the urinary density (p=0.013) and the smaller size of the right kidney (p=0.009) and left kidney (p=0.048), measured in the longitudinal plane. There were no difference between groups in the other parameters investigated such as plasmatic total calcium, ionized calcium, phosphorus, sodium, potassium and intact parathyroid hormone concentrations. The values of serum urea and bicarbonate differ between groups with p=0.039 and p=0.037, respectively. Furthermore, arterial blood pressure was measured, remaining unchanged between the groups. One can conclude that nephrolithiasis and/or ureterolithiasis are common findings in cats with CKD and these results reinforce the need to perform image investigation in cats with CKD even in the asymptomatic ones, or those in the early stages of the disease.

Risk factors for feline leukemia virus (FeLV) infection in cats in São Paulo, Brazil / Fatores de risco da leucemia viral felina em São Paulo, Brazil

The aim of this study was to identify risk factors for feline leukemia virus (FeLV) infection in cats (n = 812) from the city of São Paulo. Information on age, gender, outdoor access, reproductive status, origin and number of potential contacts were obtained for each animal. Direct immunofluorescence test was used to identify the infected cats. Fifty cats (6.2 %) were positive for FeLV infection. The risk factors identified were “outdoor access” (OR = 47.2; p < 0.001), “having been rescued from the street” (OR = 3.221; p = 0.008) or being three to six year old (OR = 3.046; p = 0.009); the most important risk factor was free outdoor access. A predictive model for FeLV infection was built based on the results of the multivariate analysis. Cats with free outdoor access are more predisposed to infection, with 18% more chances of becoming infected. If the animal is one to three year old, the probability increases, reaching 34%. If the cat is exposed to these three risk factors, the probability of infection is even higher (63%). When analyzed together or as isolated risk factors, the age and being rescued from the street represent less impact on the spreading of the FeLV, as the probability of infection for cats exposed to each of these risk factors is 1 and 4%, respectively. Thus, free roaming and outdoor access are the most important risk factors associated to FeLV infection among cats in São Paulo city and must be taken in consideration in the prevention of this retrovirus infection.

O objetivo deste estudo foi o de identificar os fatores de risco para a leucemia viral felina (FeLV) em uma amostragem de 812 gatos da cidade de São Paulo, Brasil. Informações sobre idade, sexo, acesso à rua, estado reprodutivo, origem e número de contactantes foram obtidos para cada animal. Teste de imunofluorescência direto para a pesquisa de antígeno viral em esfregaço de sangue periférico foi utilizado para a identificação dos felinos infectados. Foram encontrados 51 gatos (6,2%) positivos para a infecção. Os fatores de risco identificados foram “acesso a rua” (OR = 47,2; p < 0,001), “ter sido resgatado da rua” (OR = 3,221; p = 0,008) e “ter idade entre 3 e 6 anos” (OR = 3,046; p = 0,009), sendo o “livre acesso à rua” o fator de risco mais importante. Foi elaborado um modelo preditivo para a infecção pelo FeLV, baseandose nos resultados da análise multivariada. Gatos com acesso livre à rua apresentam 18% mais chances de se infectarem e se estiverem na faixa etária entre 3 e 6 anos de idade o risco aumenta para 34%. Se o gato estiver exposto aos três fatores de risco, a probabilidade de infecção é ainda mais alta, de 63%. Quando analisados em conjunto ou como fatores de risco isolados, a idade e o fato de “ter sido resgatado da rua” possuem menor impacto na disseminação de FeLV, já que a probabilidade de infecção para os gatos expostos a cada um desses fatores é de 1% e de 4%, respectivamente. Assim, o acesso livre a rua e ter sido resgatado da rua foram os fatores de risco mais importantes associados com a infecção pelo FeLV na cidade de São Paulo e devem ser levados em consideração na profilaxia da leucemia viral felina.

Diagnóstico sorológico de erliquiose canina com antígeno brasileiro de Ehrlichia canis / Serological diagnosis of canine monocytic ehrlichiosis with Brazilian antigen of Ehrlichia canis

O presente trabalho relata o isolamento de Ehrlichia canis em cultivo de células DH82 e posterior padronização da Reação de Imunofluorescência Indireta (RIFI). Leucócitos de uma cadela experimentalmente infectada com o isolado Jaboticabal de E. canis foram inoculados em cultivo de células DH82. A inoculação foi monitorada após a segunda semana, a cada 5-6 dias, através de exames citológicos e pela amplificação de um fragmento do gene dsb de Ehrlichia pela Reação em Cadeia pela Polimerase (PCR) para confirmação da infecção. A cultura apresentou-se positiva aos 27 dias pós-inoculação pela PCR e aos 28 dias pela citologia. No 33o dia pós-inoculação, observou-se 20 por cento de células infectadas e, aos 53 dias, 60 por cento de infecção. Atualmente, o isolado encontra-se estabelecido em células DH82, com várias passagens atingindo 90-100 por cento de células infectadas entre 7-10 dias após a inoculação. Após o seqüenciamento do produto de PCR, o isolado apresentou-se 100 por cento similar à seqüência correspondente de E. canis depositada no GenBank. As células infectadas foram utilizadas como antígeno para a padronização da RIFI para detecção da infecção em cães.

The present study describes a successful isolation of Ehrlichia canis and its establishment in DH82 cells, followed by the development of an Indirect Fluorescent Antibodies Test (IFAT). Leukocytes collected from an experimentally infected dog with the Jaboticabal strain of E. canis were used to inoculate a DH82 cell monolayer. Two weeks later, the inoculated culture was checked for infectivity, every 5-6 days by both cytological staining and PCR, targeting a fragment of the dsb gene. The cell culture showed to be infected by Ehrlichia on day 27 by PCR and on day 28 by cytological staining. By the day 33, the infection rate reached 20 percent and on day 53, 60 percent. Currently, the isolate is established in DH82 cells, with several passages reaching 90-100 percent of infected cells, within 7 to 10 days post inoculation. After sequencing, the amplicon was identical to other E. canis corresponding sequences available in the GenBank. DH82 infected cells were used to standardize an IFAT for the diagnosis of canine ehrlichiosis.

Concentrações séricas de proteína total, albumina e gamaglobulinas em gatos infectados pelo vírus da imunodeficiência felina / Serum protein, albumin and gamaglobulin concentrations in cats infected with feline immunodeficiency virus

A infecção pelo vírus da imunodeficiência dos felinos (VIF) apresenta um curso prolongado, caracterizado por uma fase aguda, em que ocorre a replicação viral no organismo hospedeiro, seguida de um período de menor replicação, no qual o animal é praticamente assintomático. Anos depois, no estádio final da infecção, desenvolve-se a síndrome da imunodeficiência dos felinos. Alguns animais infectados podem desenvolver hipergamaglobulinemia do tipo policlonal, principalmente na fase crônica ou final da infecção. Este fato tem sido atribuído a um distúrbio na produção de citocinas, causado pela infecção viral de linfócitos T CD4+. Não obstante, pouco se sabe a respeito das concentrações de proteínas séricas, especificamente gamaglobulinas, na fase aguda da infecção pelo VIF. Objetivando esclarecer isto, procedeu-se à determinação das proteínas séricas de dez felinos, SRD de ambos os sexos, infectados aos 7 meses de idade com o VIF (clade B), antes da infecção e 4, 8 e 12 meses após. A infecção pelo VIF foi confirmada pela soroconversão, com a presença de anticorpos específicos, pesquisados por meio da técnica de imunoadsorção enzimática (ELISA) e pela demonstração de material genético do vírus (PCR). Outros dez felinos VIF-, da mesma faixa etária foram mantidos como controle. Previamente à infecção experimental, todos os felinos eram negativos ao VIF, fato comprovado pela ausência de anticorpos específicos. A proteína sérica total foi determinada pelo método do buireto e as frações protéicas foram obtidas por eletroforese em tiras de acetato de celulose lidas por densitometria. Verificou-se aumento de gamaglobulinas (2,01 ±0,27g dL-1, P<0,0001) quatro meses após a infecção nos animais infectados. Doze meses após a infecção, não foram observadas diferenças nas concentrações das frações protéicas entre os felinos infectados e os controles.

Feline immunodeficiency virus (FIV) infection is known as a lifelong infection of cats. The acute phase corresponds to the period of viral replication in the host organism, followed by a period of lower replication when the animal is asymptomatic. Some years later, at the final stage, an immunodeficiency syndrome subsides. Hypergammaglobulinemia or a polyclonal gammapathy might be seen in some, but not all affected cats, mainly in the chronic phase of infection. This is thought to be due to a disturbance of cytokines production as a result of T CD4 + cells infection. Nonetheless, little is known regarding serum protein, specifically, gammaglobulin concentration in the early phase of FIV infection. Aiming at clarifing this, serum protein concentrations were determined before and 4, 8 and 12 months after FIV infection of ten domestic short-haired, male or female, 7 months old cats (group I). Ten cats of same age were maintained as controls (group II). All cats were FIV-naive cats, as confirmed by ELISA test. After infection, both, ELISA and PCR became positive only for the cats belonging to the group I. Serum protein concentration was measured by biuret method and protein fractions were obtained by electrophoresis on cellulose acetate strips. An increase in the gammaglobulin concentration (2.01 ± 0.27g/dL, P<0.0001) could be observed 4 months p.i. in the group I. Twelve months after infection, no differences could be found on protein concentrations between both groups of cats.

Fragilidade osmótica eritrocitária em gatos acometidos por hepatopatias e gatos com insuficiência renal / Erythrocyte osmotic fragility in cats with hepatic disease and cats with renal failure

A fragilidade osmótica eritrocitária (FOE) avalia a resistência osmótica das células vermelhas em concentrações decrescentes de solução salina. A resistência depende da forma, volume, tamanho, conteúdo de hemoglobina e meia vida dos eritrócitos, e pode ser alterada por vários fatores fisiológicos ou patológicos. Existem poucos relatos sobre a FOE em gatos. O objetivo deste estudo foi avaliar a FOE em gatos hígidos, com enfermidades hepáticas e naqueles com insuficiência renal. Foram estudados 27 animais divididos em três grupos. A hemólise 50 por cento foi notada de forma similar em todos os grupos. Em 100 por cento de hemólise, os eritrócitos de gatos com insuficiência renal e doença hepática mostraram maior fragilidade osmótica, embora esses animais apresentassem maiores níveis de colesterol sérico quando comparados aos gatos sadios do grupo controle. O achado de maior FOE mesmo com níveis elevados de colesterol pode estar associado a características metabólicas da espécie.

Anemia hemolítica imunomediada não regenerativa em um cão / Nonregenerative immune-mediated hemolytic anemia in a dog

Quadros hemolíticos não eritrorregenerativos são descritos em cães e podem ser decorrentes de doença medular primária, bem como, da destruição dos precursores eritróides medulares por imunoglobulinas. Um cão macho, de três anos de idade, sem raça definida, foi atendido no Hospital Veterinário da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo apresentando sinais de anemia hemolítica aguda arregenerativa. Após a instituição de terapia imunossupressora (prednisona), houve remissão da hemólise sem, no entanto, sinais de eritrorregeneração. No décimo dia de tratamento, o mielograma demonstrou discreta hipoplasia e displasia eritróide, descartando a possibilidade de aplasia medular. Associou-se ciclofosfamida e azatioprina ao tratamento, tendo havido resposta eritrorregenerativa e recuperação dos valores hematológicos. A ocorrência deste caso de anemia hemolítica não eritrorregenerativa deve servir como alerta para a ocorrência desta condição mórbida, como também, da importância da utilização do mielograma como método auxiliar no diagnóstico de anemias arregenerativas.

Semelhanças entre a doença idiopática do trato urinário inferior dos felinos e a cistite intersticial humana / Similarities between the feline idiopathic lower urinary tract disease and human interstitial cystitis

A doença idiopática do trato urinário inferior dos felinos (DITUIF) continua sendo um grande desafio para o clínico veterinário uma vez que, apesar dos avanços nas técnicas diagnósticas, sua etiologia continua por ser determinada. O objetivo do presente trabalho é demonstrar as principais semelhanças entre a doença urinária felina e a cistite intersticial humana (CI), principalmente no que se refere à comprovaçäo do caráter neurogênico da inflamaçäo vesical, o papel do estresse na gênese e/ou exacerbaçäo dos sinais clínicos da doença, e os principais achados cistoscópicos e histopatológicos em ambas as síndromes urinárias. Conclui-se, portanto, que além da dieta comercial, podem existir outros fatores com participaçäo na inflamaçäo vesical de felinos com DITUI.

Borrelia burgdorferi antibodies in dogs from Cotia county, Säo Paulo State, Brazil

Dogs sera samples collected from Cotia County, Säo Paulo were tested using indirect immunoenzymatic test (ELISA) in order to study Lyme disease serology in dogs. ELISA method was standardized and G39/40 North American strain of Borrelia burgdorferi was used as antigen. Positive results were confirmed employing the Western blotting technique. Because of the possibility of cross-reactions, sera were also tested for different serological strains of Leptospira interrogans and L. biflexa using microscopic sera agglutination test. Twenty-three of 237 (9.7 percent) serum samples were positive in the ELISA; 20 of them (86.9 percent) were confirmed by the Western blotting, what suggests that Cotia may be a risk area for Lyme disease. Although 4 samples (1.7 percent) were positive for Lyme disease and leptospirosis, no correlation was found between the results (X² = 0.725; p = 0.394) what suggests absence of serological cross reactivity

Experimental infection and horizontal transmission of Bartonella henselae in domestic cats

In order to study B. henselae transmission among cats, five young cats were kept in confinement for two years, one of them being inoculated by SC route with B. henselae (10(5) UFC). Only occasional contact among cats occurred but the presence of fleas was observed in all animals throughout the period. Blood culture for isolation of bacteria, PCR-HSP and FTSZ (gender specific), and BH-PCR (species-specific), as well as indirect immunofluorescence method for anti-B. henselae antibodies were performed to confirm the infection of the inoculated cat as well as the other naive cats. Considering the inoculated animal, B. henselae was first isolated by blood culture two months after inoculation, bacteremia last for four months, the specific antibodies being detected by IFI during the entire period. All contacting animals presented with bacteremia 6 months after experimental inoculation but IFI did not detect seroconversion in these animals. All the isolates from these cats were characterized as Bartonella (HSP and FTSZ-PCR), henselae (BH-PCR). However, DNA of B. henselae could not be amplified directly from peripheral blood by the PCR protocols used. Isolation of bacteria by blood culture was the most efficient method to diagnose infection compared to PCR or IFI. The role of fleas in the epidemiology of B. henselae infection in cats is discussed

Avaliaçäo da resposta imune humoral em cäes jovens imunizados contra a cinomose com vacina de vírus atenuado / Evaluation of humoral immune response in puppies immunized to canine distemper with attenuated virus vaccine

Este estudo foi realizado com o objetivo de se investigar a cinética da resposta imune em cäes jovens vacinados contra cinomose, com vírus atenuado, por meio do teste de soroneutralizaçäo. Onze cäes sadios da raça Beagle, observados do nascimento até 30 meses de idade, isolados de outros cäes, receberam vacina monovalente viva atenuada de cinomose canina aos 75, 105 e 135 dias de idade e doze meses após a terceira dose vacinal. O desenvolvimento de anticorpos foi mensurado pela reaçäo de soroneutralizaçäo imediatamente antes da vacinaçäo, trinta dias após a administraçäo da primeira e terceira doses, noventa dias após a administraçäo da terceira dose, nove e doze meses após a administraçäo da terceira dose, trinta dias, seis e doze meses após a dose anual. Näo havia títulos detectáveis de anticorpos contra cinomose canina antes da primovacinaçäo. O título de anticorpos variou de 4,047 a 4,880 (em logaritmos na base 10), trinta dias após a administraçäo da primeira dose e 6 meses após a dose anual, respectivamente. Todos os cäes apresentaram uma resposta similar, embora com variaçöes, com a produçäo de anticorpos com títulos maiores do que 2, considerado o título mínimo protetor. Doze meses após a dose anual, a maioria dos cäes apresentou alto título de anticorpos, sugerindo que a revacinaçäo anual poderia ser desnecessária

Variaçöes dos níveis séricos de sódio, potássio e glicose de cäes em choque séptico / Sodium, potassium and glucose serum levels changes in dogs with endotoxic shock

Foram avaliados os níveis séricos de sódio, potássio e glicose em trinta e cinco cäes, machos e fêmeas, que se apresentaram em estado de choque séptico decorrente da gastrenterite hemorrágica, antes e após terapia sintomática (Ringer com lactato de sódio e glicose), com a finalidade de se derterminar a melhora clínica obtida. Observou-se que 74,2 por cento dos animais apresentavam hipocalemia, 57,1 por cento hipoglicemia e 60 por cento discreta hiponatremia, quando da primeira colheita. Após 2 horas da instituiçäo da terapia sintomática, observou-se que a hipopotassemia ainda persistia, porém a hipoglicemia fora suficientemente corrigida (apenas 5,8 por cento mantiveram-se hipoglicêmicos). Conclui-se que a terapia sintomática é efetiva para a correçäo dos valores glicêmicos, porém näo é capaz de repor a deficiência de potássio apresentada por estes animais, devendo ser entäo acrescida deste íon.

Toxoplasma gondii infection in Brazilian domestic outpatient cats

The occurrence of Toxoplasma antibodies in domestic outpatient cats in the city of São Paulo was evaluated using the indirect immunofluorescence assay. A total of 248 blood samples obtained from male and female cats seen at the Veterinary Teaching Hospital at the University of São Paulo between February 1996 and January 1997 were tested. Of these, 17.7 percent were positive, with a 64 titer being detected in most animals. The frequence of Toxoplasma antibodies was significantly higher in older cats, those fed raw meat and those with free access to the outdoor environment. There was no significant difference in reactivity between males and females. We conclude that diet and free access to the outdoor environment were equally important as predisposing factors to the risk of Toxoplasma infection