Isolation of potential zoonotic Mycobacterium spp. from diseased freshwater angelfish (Pterophyllum scalare) from an aquarium / Isolamento de Mycobacterium spp. potencialmente zoonótica de peixes anjo de água doce (Pterophyllum scalare) doentes de um aquário

Nontuberculous mycobacteria infection is one of the most common chronic bacterial diseases in ornamental aquarium fish and appears to be directly related to stressful husbandry practices. Furthermore, it also represents zoonotic potential. Here we present the isolation and characterization of non-tuberculous mycobacteria from diseased freshwater angelfish (Pterophyllum scalare) in São Paulo, Brazil. Nine discarded breeding females with signs of disease were evaluated. The fish exhibited lethargy, loss of appetite, cachexia, skin ulcers, and exophthalmia. At necropsy, four fishes presented macroscopic granulomas in the spleen. Mycobacterium chelonae, M. fortuitum, M. gordonae, M. intracellulare and M. peregrinum were isolated and identified by hsp65 PCR restriction analysis. Histopathological analysis revealed microscopic lesions compatible with mycobacteriosis, and Mycobacterium bacillus were observed by Ziehl-Neelsen stain. Notably, all Mycobacterium species identified in this study have already been reported in human patients; therefore, diseased animals may be a source of infection for people who handle fish and aquariums.

A infecção por micobactérias não tuberculosas é uma das doenças bacterianas crônicas mais comuns em peixes ornamentais de aquário e parece estar diretamente relacionada a práticas de manejo estressantes. Além disso, também representa potencial zoonótico. Aqui, apresentamos o isolamento e a caracterização de micobactérias não tuberculosas de peixes-anjo de água doce doentes (Pterophyllum scalare) em São Paulo, Brasil. Foram avaliadas nove fêmeas reprodutoras descartadas com sinais de doença. Os peixes exibiam letargia, perda de apetite, caquexia, úlceras cutâneas e exoftalmia. Na necropsia, quatro peixes apresentaram granulomas macroscópicos no baço. Mycobacterium chelonae, M. fortuitum, M. gordonae, M. intracellulare e M. peregrinum foram isolados e identificados por análise de restrição por PCR de hsp65. A análise histopatológica revelou lesões microscópicas compatíveis com micobacteriose, e o bacilo de Mycobacterium foi observado pela coloração de Ziehl-Neelsen. Notavelmente, todas as espécies de Mycobacterium identificadas neste estudo já foram relatadas em pacientes humanos; portanto, animais doentes podem ser uma fonte de infecção para pessoas que manipulam peixes e aquários.

Isolation of potential zoonotic Mycobacterium spp. from diseased freshwater angelfish (Pterophyllum scalare) from an aquarium.

Nontuberculous mycobacteria infection is one of the most common chronic bacterial diseases in ornamental aquarium fish and appears to be directly related to stressful husbandry practices. Furthermore, it also represents zoonotic potential. Here we present the isolation and characterization of non-tuberculous mycobacteria from diseased freshwater angelfish (Pterophyllum scalare) in São Paulo, Brazil. Nine discarded breeding females with signs of disease were evaluated. The fish exhibited lethargy, loss of appetite, cachexia, skin ulcers, and exophthalmia. At necropsy, four fishes presented macroscopic granulomas in the spleen. Mycobacterium chelonae, M. fortuitum, M. gordonae, M. intracellulare and M. peregrinum were isolated and identified by hsp65 PCR restriction analysis. Histopathological analysis revealed microscopic lesions compatible with mycobacteriosis, and Mycobacterium bacillus were observed by Ziehl-Neelsen stain. Notably, all Mycobacterium species identified in this study have already been reported in human patients; therefore, diseased animals may be a source of infection for people who handle fish and aquariums.

Genetic and antigenic characterization of Brazilian SRLV strains: Natural small ruminant interspecies transmission from mixed herds.

Cross-species transmission events and mixed infection of small ruminant lentiviruses (SRLVs) were studied in seven goats and two sheep from three small ruminant mixed flocks from Northeast and Southeast Brazil. Genetic and antigenic analyses with gag/env genes and ELISA multiepitope SU1/SU5 recombinant antigens were carried out, respectively. The genetic analysis of gag and env sequences showed high viral diversity in both species, MVV-like (subtype A1) and CAEV-like B1 in goats, and CAEV-like (subtype B1) in sheep, revealing SRLV interspecies transmission from sheep to goats and vice versa in Brazilian farms. Two Brazilian caprine lentiviruses were segregated in two new genetic clades based on gag analyses, which suggests a new classification into heterogenic genotype A. Furthermore, goat isolates were grouped into subtype A1 and B1 clusters. Cross-reactive antibodies were detected in goats using ELISA with a recombinant antigen carrying SU1 and SU5 immunodominant epitopes; the results showed anti-CAEV and MVV antibodies in goats and anti-CAEV antibodies in sheep. This result can be associated with the high divergence in the V4 region due to SRLV variability. All results confirm cross-species infection of SRLV in Brazilian mixed herds.

High genetic diversity of hepatitis E virus in swine in São Paulo State, Brazil / Diversidade genética do vírus da hepatite E em suínos no estado de São Paulo, Brasil

A hepatite E é uma zoonose emergente que afeta diversas espécies de mamíferos, inclusive o ser humano. É ocasionada por um vírus da espécie Orthohepevirus A que possui diversos genótipos e subgenótipos. No Brasil é descrito o genótipo HEV-3, cujo principal reservatório é o porco doméstico. Testes moleculares e sorológicos demonstram o HEV-3 em diferentes estados, tanto em animais quanto em humanos. No estado de São Paulo, existem diversos estudos sobre a epidemiologia da hepatite E em humanos, mas faltam informações sobre o HEV-3 em suínos. Assim, o objetivo deste trabalho foi verificar a ocorrência de HEV por meio da técnica de RT-PCR e posterior sequenciamento em um banco de amostras de fezes de suínos colhidas entre 2008 e 2009, na região metropolitana de Campinas. Das 89 amostras analisadas, foi possível detectar o HEV-3 em sete e, pela reconstrução filogenética, foram encontrados os subgenótipos HEV-3b, HEV-3h, e HEV-3j. Uma amostra disponível no GenBank, proveniente de São Paulo, que ainda não havia sido subgenotipada, foi agrupada ao HEV-3i. Os subgenótipos HEV-3j e HEV-3i ainda não tinham sido relatados no país. O estudo demonstra uma grande diversidade genética do HEV no estado de São Paulo e reforça o caráter zoonótico da HEV-3.(AU)

Lymphocyte proliferative responses in dairy cows supplemented with an immunomodulatory feed additive and administered polyvalent vaccination / [Proliferação de linfócitos em vacas leiteiras suplementadas com um aditivo alimentar imunomodulador e vacinadas com vacina polivalente]

O presente estudo avaliou o efeito da suplementação com OmniGen-AF® na proliferação de linfócitos e títulos de anticorpos após vacinação em bovinos leiteiros. Amostras de sangue periférico foram coletadas de 32 vacas leiteiras para quantificação dos títulos de anticorpos anti-Leptospira, e amostras de sangue periférico de 16 vacas leiteiras foram também coletadas para avaliação da proliferação de linfócitos. Observou-se que a suplementação com OmniGen-AF® aumentou a proliferação basal de linfócitos (sem estímulos) 21 dias após a vacinação (P=0,03), apesar de reduzir a proliferação de linfócitos B quando estimulada com Leptospira borgpetersenii serovar Hardjo inativada pelo calor (P=0,03). Ademais, nenhum efeito da suplementação sobre a proliferação de linfócitos no momento imediatamente anterior à vacinação e nos títulos de anticorpos anti-Leptospira foi encontrado. Além disso, a proliferação de linfócitos estimulada com lipopolissacarídeos foi maior em vacas multíparas que em primíparas 21 dias após a vacinação (P=0,03). Desse modo, o presente estudo demonstra que a suplementação com OmniGen-AF® não afetou de forma robusta a proliferação de linfócitos e os títulos de anticorpos anti-Leptospira após vacinação em vacas leiteiras sadias.(AU)

Adenocarcinoma pulmonar associado à infecção por Mycobacterium sp. em vaca - relato de caso / [Pulmonary adenocarcinoma associated with infection by Mycobacterium sp. in a cow - case report]

Descrevem-se os aspectos clínicos, anatomopatológicos, imuno-histoquímicos, microbiológicos e moleculares de um caso de adenocarcinoma pulmonar associado à infecção por Mycobacterium sp. em uma vaca. O animal apresentou hiporexia, emagrecimento, vocalizações, postura ortopneica, ingurgitamento da jugular, estase venosa positiva, gemido expiratório e morte. Na necropsia, os pulmões estavam aumentados e apresentavam, na superfície pleural, nódulos branco-amarelados, firmes, multifocais a coalescentes, interpostos por áreas avermelhadas. Ao corte, os nódulos aprofundavam-se ao parênquima e possuíam múltiplos focos de aspecto caseoso e friável e áreas de mineralização. O saco pericárdico e os linfonodos traqueobrônquicos, ilíacos, lombares aórticos e mamários apresentavam lesões semelhantes. Histologicamente, observou-se neoformação carcinomatosa associada a áreas multifocais de necrose e mineralização. As células neoplásicas foram fortemente imunomarcadas pelo anticorpo antipancitoqueratina AE1/AE3. Na cultura microbiológica de fragmentos dos pulmões, houve crescimento de colônias bacterianas compatíveis com micobactérias atípicas. O sequenciamento molecular submetido ao BLASTn identificou o Mycobacterium sp. WCM 7299 (ID: gb|KJ873243.1|).(AU)

The clinical, anatomopathological, immunohistochemical, microbiological and molecular aspects of a case of pulmonary adenocarcinoma associated with infection by Mycobacterium sp. in a cow are described. The animal presented hyporexia, weight loss, vocalizations, orthopneic posture, jugular engorgement, positive venous stasis, expiratory groaning and death. At necropsy, the lungs were enlarged and presented firm, multifocal to coalescent yellowish nodules, interposed by reddish areas on the pleural surface. At cut, the nodules deepened to the parenchyma and had multiple foci of caseous and friable appearance and areas of mineralization. The pericardial sac and tracheobronchial, iliac, aortic lumbar and mammary lymph nodes showed similar lesions. Histologically, a carcinomatous neoformation, associated with multifocal areas of necrosis and mineralization, was observed. Neoplastic cells were strongly immunolabelled by anti-PanCytokeratin antibody AE1/AE3. Microbiological culture of lung fragments showed growth of bacterial colonies compatible with atypical mycobacteria. Molecular sequencing submitted to BLASTn identified the Mycobacterium sp. WCM 7299 (ID: gb|KJ873243.1|).(AU)

Características epizootiológicas da infecção natural pelo vírus da cinomose canina em Belo Horizonte / Natural canine distemper virus infection epizootiology characteristics in Belo Horizonte

O perfil epizootiológico da cinomose canina em Belo Horizonte é desatualizado e não alberga algumas características relevantes. Uma análise recente da distribuição do vírus em relação às características do hospedeiro e do meio ambiente associada aos principais sinais clínicos e achados laboratoriais são importantes para se adotarem medidas estratégicas para o controle da enfermidade. Objetivou-se, assim, determinar as características epizootiológicas da infecção pelo vírus da cinomose canina associada à variedade de sinais clínico-neurológicos e laboratoriais em Belo Horizonte, auxiliando no diagnóstico precoce da infecção e na diminuição das taxas de morbidade e mortalidade da doença. A avaliação do perfil epizootiológico de 90 cães revelou que a doença é mais frequente em animais adultos (um a seis anos de idade) e que não receberam vacinas conforme recomendado pelos protocolos. Os sinais clínicos extraneurais e neurais foram variados, com predomínio para manifestações gastrentérica e respiratória, mioclonia e déficit motor, respectivamente. O exame do fluido cérebro-espinhal demonstrou predomínio de proteinorraquia associada à pleocitose linfocítica. O teste de imunocromatografia para pesquisa de antígeno com amostras do fluido cerebroespinhal foi eficaz para identificar a doença em pacientes com sinais neurológicos, diferentemente das amostras do swab conjuntival, que não devem ser utilizadas.(AU)

The epizootiology profile of canine distemper in Belo Horizonte is outdated and does not harbor some important characteristics. A recent analysis of the virus distribution in relation to host and environmental characteristics associated with the main clinical signs and laboratory findings are important for adopting strategic measures to control the disease. The aim of this study was to determine the epizootiology characteristics of canine distemper virus infection associated with a variety of clinical and neurologic signs and laboratory findings in Belo Horizonte, helping to detect early infection and reduce morbidity and mortality rates. The evaluation of the epizootiology profile of 90 dogs revealed that the disease is more frequent in adult animals (1-6 years of age) and did not receive vaccines as recommended by the protocols. Extra neural and neural clinical signs were varied, with predominance for gastrointestinal and respiratory manifestations and myoclonus and motor deficit, respectively. Examination of the cerebrospinal fluid of 16 dogs showed a predominance of increase protein associated with lymphocytic pleocytosis. The immunochromatography test for antigen screening with samples of cerebrospinal fluid in 76 animals with neurological signs was effective in identifying the disease, unlike conjunctival swab samples, which should not be used.(AU)

Características epizootiológicas da infecção natural pelo vírus da cinomose canina em Belo Horizonte / Natural canine distemper virus infection epizootiology characteristics in Belo Horizonte

O perfil epizootiológico da cinomose canina em Belo Horizonte é desatualizado e não alberga algumas características relevantes. Uma análise recente da distribuição do vírus em relação às características do hospedeiro e do meio ambiente associada aos principais sinais clínicos e achados laboratoriais são importantes para se adotarem medidas estratégicas para o controle da enfermidade. Objetivou-se, assim, determinar as características epizootiológicas da infecção pelo vírus da cinomose canina associada à variedade de sinais clínico-neurológicos e laboratoriais em Belo Horizonte, auxiliando no diagnóstico precoce da infecção e na diminuição das taxas de morbidade e mortalidade da doença. A avaliação do perfil epizootiológico de 90 cães revelou que a doença é mais frequente em animais adultos (um a seis anos de idade) e que não receberam vacinas conforme recomendado pelos protocolos. Os sinais clínicos extraneurais e neurais foram variados, com predomínio para manifestações gastrentérica e respiratória, mioclonia e déficit motor, respectivamente. O exame do fluido cérebro-espinhal demonstrou predomínio de proteinorraquia associada à pleocitose linfocítica. O teste de imunocromatografia para pesquisa de antígeno com amostras do fluido cerebroespinhal foi eficaz para identificar a doença em pacientes com sinais neurológicos, diferentemente das amostras do swab conjuntival, que não devem ser utilizadas.(AU)

The epizootiology profile of canine distemper in Belo Horizonte is outdated and does not harbor some important characteristics. A recent analysis of the virus distribution in relation to host and environmental characteristics associated with the main clinical signs and laboratory findings are important for adopting strategic measures to control the disease. The aim of this study was to determine the epizootiology characteristics of canine distemper virus infection associated with a variety of clinical and neurologic signs and laboratory findings in Belo Horizonte, helping to detect early infection and reduce morbidity and mortality rates. The evaluation of the epizootiology profile of 90 dogs revealed that the disease is more frequent in adult animals (1-6 years of age) and did not receive vaccines as recommended by the protocols. Extra neural and neural clinical signs were varied, with predominance for gastrointestinal and respiratory manifestations and myoclonus and motor deficit, respectively. Examination of the cerebrospinal fluid of 16 dogs showed a predominance of increase protein associated with lymphocytic pleocytosis. The immunochromatography test for antigen screening with samples of cerebrospinal fluid in 76 animals with neurological signs was effective in identifying the disease, unlike conjunctival swab samples, which should not be used.(AU)

Identification of a novel protein in the genome sequences of Leptospira interrogans with the ability to interact with host's components.

BACKGROUND: Leptospirosis is an infectious disease that affects humans and animals worldwide. The etiological agents of this disease are the pathogenic species of the genus Leptospira. The mechanisms involved in the leptospiral pathogenesis are not full understood. The elucidation of novel mediators of host-pathogen interaction is important in the detection of virulence factors involved in the pathogenesis of leptospirosis. OBJECTIVE: This work focused on identification and characterization of a hypothetical protein of Leptospira encoded by the gene LIC10920. METHODS: The protein of unknown function was predicted to be surface exposed. Therefore, the LIC10920 gene was cloned and the protein expressed in Escherichia coli BL21 (DE3) Star pLysS strain. The recombinant protein was purified by metal affinity chromatography and evaluated with leptospirosis human serum samples. The interaction with host components was also performed. RESULTS: The recombinant protein was recognized by antibodies present in leptopsirosis human serum, suggesting its expression during infection. Immunofluorescence and intact bacteria assays indicated that the bacterial protein is surface-exposed. The recombinant protein interacted with human laminin, in a dose-dependent and saturable manner and was named Lsa24.9, for Leptospiral surface adhesin, followed by its molecular mass. Lsa24.9 also binds plasminogen (PLG) in a dose-dependent and saturable fashion, fulfilling receptor ligand interaction. Moreover, Lsa24.9 has the ability to acquire PLG from normal human serum, exhibiting similar profile as observed with the human purified component. PLG bound Lsa24.9 was able of generating plasmin, which could increase the proteolytic power of the bacteria. CONCLUSIONS: This novel leptospiral protein may function as an adhesin at the colonization steps and may help the invasion process by plasmin generation at the bacterial cell surface.

Identification of a novel protein in the genome sequences of Leptospira interrogans with the ability to interact with host's components

Background Leptospirosis is an infectious disease that affects humans and animals worldwide. The etiological agents of this disease are the pathogenic species of the genus Leptospira. The mechanisms involved in the leptospiral pathogenesis are not full understood. The elucidation of novel mediators of host-pathogen interaction is important in the detection of virulence factors involved in the pathogenesis of leptospirosis. Objective This work focused on identification and characterization of a hypothetical protein of Leptospira encoded by the gene LIC10920. Methods The protein of unknown function was predicted to be surface exposed. Therefore, the LIC10920 gene was cloned and the protein expressed in Escherichia coli BL21 (DE3) Star pLysS strain. The recombinant protein was purified by metal affinity chromatography and evaluated with leptospirosis human serum samples. The interaction with host components was also performed. Results The recombinant protein was recognized by antibodies present in leptopsirosis human serum, suggesting its expression during infection. Immunofluorescence and intact bacteria assays indicated that the bacterial protein is surface-exposed. The recombinant protein interacted with human laminin, in a dose-dependent and saturable manner and was named Lsa24.9, for Leptospiral surface adhesin, followed by its molecular mass. Lsa24.9 also binds plasminogen (PLG) in a dose-dependent and saturable fashion, fulfilling receptor ligand interaction. Moreover, Lsa24.9 has the ability to acquire PLG from normal human serum, exhibiting similar profile as observed with the human purified component. PLG bound Lsa24.9 was able of generating plasmin, which could increase the proteolytic power of the bacteria. Conclusions This novel leptospiral protein may function as an adhesin at the colonization steps and may help the invasion process by plasmin generation at the bacterial cell surface.

Inhibition of the growth of major mastitis-causing pathogens by non-aureus Staphylococcus isolates using the cross-streaking method / Comunicação: Inibição do crescimento dos principais patógenos causadores de mastite por isolados de estafilococos não aureus pelo método cross-streaking

O objetivo do presente estudo foi avaliar a capacidade de estafilococos não aureus (NAS) isolados de diferentes nichos ecológicos (leite, ambiente e ápice do teto), associados a vacas leiteiras, de inibir os principais agentes etiológicos da mastite bovina (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis e Escherichia coli). Neste estudo, 38 isolados NAS de diferentes nichos ecológicos foram avaliados quanto à capacidade de inibir o crescimento in vitro de importantes patógenos causadores de mastite pelo método cross-streaking. No total, 19 (50%) isolados de NAS (oito isolados de S. chromogenes, 10 de S. fleurettii e um de S. haemolyticus) apresentaram inibição contra os principais patógenos causadores de mastite. No entanto, a inibição dos patógenos causadores da mastite bovina por isolados de NAS foi maior contra bactérias Gram-positivas. Além disso, o presente estudo não sugeriu que os nichos ecológicos influenciam a capacidade do NAS de inibir os principais patógenos causadores da mastite bovina. Com base nesses resultados, concluiu-se que certos isolados de NAS apresentam potencial efeito protetor contra os principais patógenos da mastite, pelo menos in vitro.(AU)

Inhibition of the growth of major mastitis-causing pathogens by non-aureus Staphylococcus isolates using the cross-streaking method / Comunicação: Inibição do crescimento dos principais patógenos causadores de mastite por isolados de estafilococos não aureus pelo método cross-streaking

O objetivo do presente estudo foi avaliar a capacidade de estafilococos não aureus (NAS) isolados de diferentes nichos ecológicos (leite, ambiente e ápice do teto), associados a vacas leiteiras, de inibir os principais agentes etiológicos da mastite bovina (Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis e Escherichia coli). Neste estudo, 38 isolados NAS de diferentes nichos ecológicos foram avaliados quanto à capacidade de inibir o crescimento in vitro de importantes patógenos causadores de mastite pelo método cross-streaking. No total, 19 (50%) isolados de NAS (oito isolados de S. chromogenes, 10 de S. fleurettii e um de S. haemolyticus) apresentaram inibição contra os principais patógenos causadores de mastite. No entanto, a inibição dos patógenos causadores da mastite bovina por isolados de NAS foi maior contra bactérias Gram-positivas. Além disso, o presente estudo não sugeriu que os nichos ecológicos influenciam a capacidade do NAS de inibir os principais patógenos causadores da mastite bovina. Com base nesses resultados, concluiu-se que certos isolados de NAS apresentam potencial efeito protetor contra os principais patógenos da mastite, pelo menos in vitro.(AU)

Evaluation of antibodies against Toxoplasma gondii and Leptospira spp. in Magellanic penguins (Spheniscus magellanicus) on Magdalena Island, Chile.

Toxoplasmosis has been reported in many avian species, but little information is available from wild penguin populations. Leptospira can infects domestic and wild animals. Spheniscus magellanicus belong to the order Sphenisciformes, family Spheniscidae, and are colonial birds. These seabirds live in temperate waters along the Atlantic shores of South America, and their total population has been estimated to be 1,300,000 breeding pairs. Magdalena Island (Chile) hosts an important breeding colony but, over recent decades, a marked decline in the number of birds has been seen. The objective of this study was to determine occurrences of antibodies against T. gondii and Leptospira spp. in penguins (Spheniscus magellanicus) on Magdalena Island, from where no previous data on these agents were available. Serum samples were collected from 132 penguins on Magdalena Island. Antibodies against Toxoplasma gondii were detected using the modified agglutination test (Titer ≥20), and anti-Leptospira spp. antibodies were detected using the microscopic agglutination test (Titer ≥100). T. gondii antibodies were detected in 57 (43.18%) of the 132 serum samples, with titers that ranged from 20 to 320. None of the penguins in this study was reactive to anti-Leptospira spp. antibodies. This is the first report of T. gondii seropositivity in free-living Magellanic penguins in Chile.

Occurrence of anti-Leptospira antibodies in dogs in the southwestern region of the state of São Paulo / Ocorrência de anticorpos anti-Leptospira em cães na região sudeste do estado de São Paulo

Introduction: Leptospirosis is an emerging disease with different prevalence in dog populations. Dogs are crucial in the disease epidemiology, acting as accidental or maintenance hosts. Infective serovars present different geographic distribution among these populations, depending on exposure to hosts from infected wild or domestic animal reservoirs. The most common serovars that infect dogs prior to the introduction of the vaccines against leptospirosis were Icterohaemorrhagiae and Canicola. Objective: To analyze the occurrence of anti-leptospira antibodies in dogs from southwestern region of the state of São Paulo, using the microscopic agglutination test (MAT).(AU)

Occurrence of anti-Leptospira antibodies in dogs in the southwestern region of the state of São Paulo / Ocorrência de anticorpos anti-Leptospira em cães na região sudeste do estado de São Paulo

Introduction: Leptospirosis is an emerging disease with different prevalence in dog populations. Dogs are crucial in the disease epidemiology, acting as accidental or maintenance hosts. Infective serovars present different geographic distribution among these populations, depending on exposure to hosts from infected wild or domestic animal reservoirs. The most common serovars that infect dogs – prior to the introduction of the vaccines against leptospirosis – were Icterohaemorrhagiae and Canicola. Objective: To analyze the occurrence of anti-leptospira antibodies in dogs from southwestern region of the state of São Paulo, using the microscopic agglutination test (MAT).

Respiratory signs, fever and lymphopenia in calves inoculated with Brazilian HoBi-like pestiviruses.

Hobi-like viruses (HobiPeV) comprise a novel, recently classified species of bovine pestiviruses, originally identified in commercial fetal bovine serum of Brazilian origin and, subsequently, isolated from diseased animals in several countries. Although frequently isolated from clinical cases, most HobiPeV isolates failed to reproduce overt disease in cattle upon experimental inoculation. Herein, we describe the outcome of experimental infection of four to six months-old seronegative calves with two Brazilian HobiPeV isolates. Calves inoculated intranasally with isolate SV478/07 developed viremia between days 2 and 9 post-inoculation (pi) and shed virus in nasal secretions up to day 11pi. These animals presented hyperthermia (day 7 to 10-11 pi) and lymphopenia from days 4 to 8pi. Clinically, all four calves developed varied degrees of apathy, anorexia, mild to moderate respiratory signs (nasal secretion, hyperemia), ocular discharge and pasty diarrhea in the days following virus inoculation. In contrast, calves inoculated with isolate SV757/15 presented only hyperthermia (days 3 to 10-11 pi) and lymphopenia (days 4-8 pi), without other apparent clinical signs. In these animals, viremia was detected up to day 9 pi and virus shedding in nasal secretions lasted up to day 12-14 pi. Both groups seroconverted to the inoculated viruses, developing virus neutralizing (VN) titers from 320 to 5120 at day 28pi. These results extend previous findings that experimental infections of calves with HobiPeV are predominantly mild, yet they also indicate that field isolates may differ in their ability to cause disease in susceptible animals.

Antimicrobial susceptibility patterns of Escherichia coli phylogenetic groups isolated from bovine clinical mastitis.

Determination of antimicrobial susceptibility (AMS) of Escherichia coli causing clinical mastitis (CM) according to the phylogenetic groups and its association with descriptors at the cow and herd level may help improve specific strategies for treatment and control of this pathogen in dairy herds. The aims of the present study were to (a) determine the frequency of phylogenetic groups of E. coli isolated from CM in dairy cows, and its association with cow-level descriptors (parity, lactation stage, CM severity, and affected quarter position), housing system, and season; and (b) determine and compare AMS among E. coli phylogenetic groups. A quadruplex PCR method was used to classify E. coli isolates into 1 of the 7 phylogenetic groups. Minimal inhibitory concentrations were determined for 10 antimicrobials, and survival analysis was performed to evaluate the AMS differences among E. coli phylogroups. Most E. coli isolates belonged to phylogroups A (52%) and B1 (38%). None of the cow- and herd-level descriptors were associated with the E. coli phylogenetic groups. Overall, E. coli isolates were mostly susceptible to ceftiofur (96.8%), sulfadimethoxine (75.5%), and cephalothin (74.5%). Based on the survival analysis, differences in AMS between phylogenetic groups of E. coli was observed only for cephalothin, in which strains of phylogroup A were inhibited at lower minimum inhibitory concentration than strains of phylogroup B1. Results of this study indicated low susceptibility of E. coli isolates identified from CM to most antimicrobials. In addition, differences in AMS can occur among E. coli phylogenetic groups, although they may be uncommon as they were limited to only one antimicrobial (i.e., cephalothin).

Mastitis in the transition period: identification of potential blood markers / Mastite no período de transição: identificação de potenciais marcadores no sangue

This study identified potential blood markers associated with mastitis in dairy cows under different dry therapies during the transition period, using a logistic regression model. Thirty-four Holstein dairy cows were divided into three groups: untreated controls (13 cows, 42 quarters); animals that received an antimicrobial at drying-off (10 cows, 40 quarters); and animals that were administered an internal teat sealant at drying-off (11 cow, 44 quarters). Blood and quarter milk samples were collected 60 days before the expected day of calving, on the day of calving, and three, seven, 15, 21, and 30 days after calving. Milk samples were submitted for bacteriological analysis and somatic cell count. Blood samples were collected for analyses of the following: the erythrogram and leucogram; plasm fibrinogen concentration; hepatic and renal functions; metabolic profile; serum calcium and phosphorous levels; total serum protein and albumin concentrations. The concentration of total serum proteins was associated with a high somatic cell count. Similarly, the concentrations of total serum proteins and triglycerides were associated to milk bacteriological positive samples during the transition period. Thus, the occurrence of mastitis in dairy cows during the transition period was greater in animals that showed higher concentrations of serum total proteins and triglycerides, in contrast to the use of dry cow therapy.(AU)

O presente estudo identificou potenciais marcadores sanguíneos associados tanto à mastite, durante o período de transição em vacas leiteiras submetidas à antimicrobianoterapia, quanto à utilização do selante interno de teto na secagem por modelo de regressão logística. Trinta e quatro vacas da raça Holandesa foram divididas em três grupos experimentais, a saber: 13 animais (42 quartos mamários) que não receberam tratamento na secagem; 10 animais (40 quartos mamários) tratados por via intramamária com uma bisnaga do antimicrobiano para vaca seca à base de cefalônio anidro após a última ordenha em cada quarto mamário; e 11 animais (44 quartos mamários) que receberam, por via intramamária, uma bisnaga de selante à base de subnitrato de bismuto após a última ordenha em cada quarto mamário. As amostras de leite e sangue foram coletadas 60 dias antes da data prevista do parto, na secagem, e no dia do parto e após três, sete, 15 e 21 dias após o parto. As amostras de leite foram utilizadas para o exame microbiológico e a determinação da contagem de células somáticas. As amostras de sangue foram utilizadas para determinação do eritrograma, leucograma, fibrinogênio plasmático, funções hepáticas e renais, perfil metabólico, proteína total e albuminas séricas e concentração sérica de cálcio e fósforo. A concentração total de proteínas séricas foi associada à alta contagem de células somáticas. Similarmente, a concentração total de proteínas séricas e triglicérides foi associada ao isolamento de patógenos causadores de mastite nas amostras de leite durante o período de transição. Desse modo, conclui-se que vacas leiteiras com concentrações séricas maiores de proteína total e triglicérides têm maior chance de apresentar mastite durante o período de transição; em contraste, o uso da antimicrobianoterapia de vaca seca reduz esse risco.(AU)

Mastitis in the transition period: identification of potential blood markers / Mastite no período de transição: identificação de potenciais marcadores no sangue

This study identified potential blood markers associated with mastitis in dairy cows under different dry therapies during the transition period, using a logistic regression model. Thirty-four Holstein dairy cows were divided into three groups: untreated controls (13 cows, 42 quarters); animals that received an antimicrobial at drying-off (10 cows, 40 quarters); and animals that were administered an internal teat sealant at drying-off (11 cow, 44 quarters). Blood and quarter milk samples were collected 60 days before the expected day of calving, on the day of calving, and three, seven, 15, 21, and 30 days after calving. Milk samples were submitted for bacteriological analysis and somatic cell count. Blood samples were collected for analyses of the following: the erythrogram and leucogram; plasm fibrinogen concentration; hepatic and renal functions; metabolic profile; serum calcium and phosphorous levels; total serum protein and albumin concentrations. The concentration of total serum proteins was associated with a high somatic cell count. Similarly, the concentrations of total serum proteins and triglycerides were associated to milk bacteriological positive samples during the transition period. Thus, the occurrence of mastitis in dairy cows during the transition period was greater in animals that showed higher concentrations of serum total proteins and triglycerides, in contrast to the use of dry cow therapy.(AU)

O presente estudo identificou potenciais marcadores sanguíneos associados tanto à mastite, durante o período de transição em vacas leiteiras submetidas à antimicrobianoterapia, quanto à utilização do selante interno de teto na secagem por modelo de regressão logística. Trinta e quatro vacas da raça Holandesa foram divididas em três grupos experimentais, a saber: 13 animais (42 quartos mamários) que não receberam tratamento na secagem; 10 animais (40 quartos mamários) tratados por via intramamária com uma bisnaga do antimicrobiano para vaca seca à base de cefalônio anidro após a última ordenha em cada quarto mamário; e 11 animais (44 quartos mamários) que receberam, por via intramamária, uma bisnaga de selante à base de subnitrato de bismuto após a última ordenha em cada quarto mamário. As amostras de leite e sangue foram coletadas 60 dias antes da data prevista do parto, na secagem, e no dia do parto e após três, sete, 15 e 21 dias após o parto. As amostras de leite foram utilizadas para o exame microbiológico e a determinação da contagem de células somáticas. As amostras de sangue foram utilizadas para determinação do eritrograma, leucograma, fibrinogênio plasmático, funções hepáticas e renais, perfil metabólico, proteína total e albuminas séricas e concentração sérica de cálcio e fósforo. A concentração total de proteínas séricas foi associada à alta contagem de células somáticas. Similarmente, a concentração total de proteínas séricas e triglicérides foi associada ao isolamento de patógenos causadores de mastite nas amostras de leite durante o período de transição. Desse modo, conclui-se que vacas leiteiras com concentrações séricas maiores de proteína total e triglicérides têm maior chance de apresentar mastite durante o período de transição; em contraste, o uso da antimicrobianoterapia de vaca seca reduz esse risco.(AU)

Epidemiological Profile of Wild Rabies in Brazil (2002-2012).

Rabies is one of the most important zoonosis in the world with high impact on public health. Studies report the presence of Lyssavirus in reservoirs of the wild cycle, highlighting the role of wild canines, marmosets, and vampire and non-vampire bats as potential vectors of the disease to domestic animals and human beings. Therefore, the reintroduction of rabies in urban environments from reservoirs of the wild cycle is a matter of concern. This study describes the profile of rabies cases documented in Brazil from 2002 to 2012, with emphasis on the wild transmission cycle of the disease. We carried out a descriptive study using records with information on the time of infection, persons with infection and location of confirmed cases of rabies in humans and animals, as well as data on anti-rabies treatments obtained from the Information System of Notifiable Diseases (Sinan) database. Within the study period, 82 cases of rabies transmitted by wild animals to humans were reported, predominantly in rural areas of the northern and north-eastern regions. Of the cases in humans, 72% did not receive post-exposure prophylaxis. Among wild mammals, vampire bats were the most frequent vectors of the disease. In the north-east region, 460 terrestrial wild mammals were reported with confirmed rabies. Over the study period, 1703 bats were reported to carry the rabies virus. In the south-east region, the most frequently reported carriers of the virus were non-vampire bats. The midwest and northern regions presented a lower number of records of rabies cases among terrestrial wild mammals. However, the high number of rabies cases among bovines reflects the role of the vampire bat as a maintainer of the rabies virus in the rural cycle. The present results are key to adjust the planning of rabies control in Brazil to the current epidemiological trends.