Seroepidemiology of Neospora caninum among goats (Capra hircus) in the state of Paraíba, northeastern Brazil / Soroepidemiologia de Neospora caninum em caprinos (Capra hircus) do estado da Paraíba, Nordeste do Brasil

Although goat dairy farms in Brazil may have a higher risk of infection by Neospora caninum than beef farms, risk factor evaluation on a representative population remains to be fully established in Brazil. Accordingly, this study aimed to establish the occurrence of anti-N. caninum antibodies and factors associated with exposure in 406 blood samples from five dairy and three beef goat farms in the state of Paraíba, northeastern Brazil. Anti-N. caninum antibodies were detected by indirect immunofluorescence assay (IFA), with samples considered positive when reacting with dilution ≥ 1:50. A total of 106/406 goats (26.11%; 95% CI: 21.96-30.72%) were seroreactive comprising 2/61 (3.28%), 10/45 (22.22%), 13/50 (26.00%), 17/51 (33.33%) to 29/46 (63.04%) in dairy farms, and from 3/54 (5.56%), 12/50 (24.00%) to 20/49 (40.82%) on the beef farms. No significant associations were found in relation to age, gender, dairy versus beef farms, occurrence of abortions or mummified fetuses, and seroreactivity to N. caninum (P>0.05). In conclusion, goat farms in the state of Paraíba showed the highest occurrence of anti-N. caninum antibodies to date in Brazil.(AU)

Embora as criações caprinas de leite no Brasil possam ter maior probabilidade de risco de infecção por Neospora caninum do que as de carne, a avaliação dos fatores de risco em uma população representativa ainda não está totalmente estabelecida no Brasil. Dessa forma, este estudo teve por objetivo estabelecer a soroprevalência de N. caninum e seus fatores associados à exposição em 406 amostras de sangue de cinco fazendas de leite e três de corte provenientes do estado da Paraíba, região Nordeste do Brasil. A detecção de anticorpos anti-N. caninum foi realizada utilizando-se a reação de imunofluorescência indireta (RIFI), com as amostras consideradas positivas na diluição ≥ 1:50. No total, 106/406 (26,11%; IC 95%: 21,96-30,72%) caprinos foram sororreagentes, variando de 2/61 (3,28%), 10/45 (22,22%), 13/50 (26,00%), 17/51 (33,33%) a 29/46 (63,04%) em fazendas de leite, e de 3/54 (5,56%), 12/50 (24,00%) a 20/49 (40,82%) em fazendas de corte. Não foram observadas associações significativas entre idade, sexo, criação de leite e carne, ocorrência de abortamentos ou fetos mumificados e sororreatividade para N. caninum (P>0,05). Em conclusão, fazendas de caprinos da Paraíba mostraram as mais altas ocorrências de anticorpos anti-N. caninum até o momento no Brasil.(AU)

Bronchoalveolar lavage (BAL) in tapirs (Tapirus terrestris) with Mycobacterium tuberculosis / Lavado broncoalveolar (LBA) em antas (Tapirus terrestris) com Mycobacterium tuberculosis

This paper describes the in vivo Bronchoalveolar lavage (BAL) technique by endoscopy in tapirs (Tapirus terrestris) with clinical signs of tuberculosis. The technique was performed in two tapirs, male and female, from Curitiba Zoo, Paraná, Brazil. A flexible endoscope and a polyethylene catheter were used after the chemical restraint of the animals. For BAL technique, 60mL of saline 0.9% were infused with a polyethylene catheter, introduced by the endoscope's working channel, and 15mL of BAL were recovered, analyzed and submitted to cytocentrifugation. Slides were stained by Papanicolaou, periodic acid-Schiff (PAS) and Ziehl-Neelsen methods contained high quantity of inflammatory cells on light microscopy (macrophages 27.5%, lymphocytes 0.5%, neutrophis 67% and eosinophis 5%). BAL samples were submitted to culture, bacilloscopy and PCR and were negative for both animals. Based on this study, it was concluded that the bronchoalveolar lavage technique in tapirs is feasible, simple, noninvasive, practical and fast, providing an important clinical information in vivo regarding the functional status of the lower respiratory tract...

O presente trabalho descreve a técnica de lavado broncoalveolar (LBA) por endoscopia em antas (Tapirus terrestris) in vivo com sinais clínicos de tuberculose. A técnica foi realizada em duas antas, um macho e uma fêmea, provenientes do Zoológico de Curitiba, Paraná, Brasil, utilizando-se endoscópio flexível e sonda de polietileno, após a contenção química desses animais. Para o LBA, 60mL de solução fisiológica 0,9% foram infundidos com auxílio de cateter de polietileno, introduzido pelo canal de trabalho do endoscópio, e, aproximadamente, 15mL de LBA foram recuperados, acondicionados, analisados e submetidos à citocentrifugação. As lâminas foram coradas pelas técnicas de Papanicolau, ácido periódico de Schiff (PAS) e Ziehl- Neelsen, método que contém altas quantidades de células inflamatórias em microscopia (macrófagos 27,5%, linfócitos 0,5%, neutrófilos 67% e eosinófilos 5%). Amostras de LBA foram submetidas a cultura, baciloscopia e PCR e foram negativas em ambos os animais. Concluiu-se, baseado no presente trabalho, que técnica de lavado broncoalveolar é simples, não invasiva, funcional e rápida. Pode fornecer ao clínico importantes informações acerca do estado de funcionamento do aparelho respiratório in vivo...

Cardiac lesions in 30 dogs naturally infected with Leishmania infantum chagasi.

The hearts of 30 dogs naturally infected with Leishmania infantum chagasi were evaluated histologically and immunohistochemically. Myocardial lesions were detected in all dogs, including lymphoplasmacytic myocarditis (27/30), myonecrosis (24/30), increased interstitial collagen (22/30), lepromatous-type granulomatous myocarditis (7/30), fibrinoid vascular change (3/30), and vasculitis (1/30). The parasite was detected in the hearts of 20 of 30 dogs. The number of parasitized cells correlated with the intensity of the inflammation and with the number of granulomas. The results indicate that cardiac lesions are prevalent in dogs with naturally occurring leishmaniasis even in the absence of clinical signs of cardiac disease.

Serology for Brucella abortus in cart horses from an urban area in Brazil / Sorologia para Brucella abortus em cavalos de carroça de área urbana do Brasil

Avaliou-se a infecção por Brucella abortus em cavalos de carroça de Curitiba e São José dos Pinhais-PR. Um total de 123 amostras foi submetido ao teste do antígeno tamponado acidificado (ATA), soroaglutinação lenta em tubos (SAL) e prova do 2-mercaptoetanol (2-ME) para confirmação dos resultados. Oito (6,5%) equinos foram positivos para o ATA e um animal permaneceu positivo ao teste confirmatório. Existem evidências da presença de brucelose entre os cavalos de carroça.

Prevention educational program of human rabies transmitted by bats in rain forest preserved area of southern Brazilian coast.

Guaraqueçaba city is a rain forest environmental protected area located on the southern coast of Brazil. Recently, the local Animal Health Service has noticed haematophagous bats feeding from humans and domestic animals, as well as bat colonies located in houses and public schools. In 2007, two non-haematophagous bats were tested positive by direct immunofluorescence for rabies in a nearby city. Native fauna and environmental laws protect non-haematophagous bats in Brazilian preserved areas such as Guaraqueçaba, making non-haematophagous bat population control almost impossible. Accordingly, the aim of this study was to evaluate a simple and feasible educational protocol applied by a multi-institutional task force in local elementary schools to prevent rabies transmitted by bats. Information was transmitted to children by video, lectures and oral question-answer section; evaluation was made by written questionnaires to teachers and students. Interinstitutional task force included public and animal health public services, a federal university and the city secretary of environment, of education, of agriculture and of animal health, and also participation of local community. Information was effectively absorbed by children when evaluated just after being given. As important, questionnaires showed that handling and playing with bats at day time was common in several elementary school students, exposing themselves to what may represent higher risk of rabies transmission than haematophagous bat feeding directly from humans. Training of teachers and students may effectively prevent rabies by bats in such communities. Insertion of this subject into science content of local elementary school educational programme was proposed in order to establish a continuing education programme on rabies transmitted by bats in environmental preserved areas.

A quantitative TaqMan PCR assay for the detection of Mycoplasma suis.

AIM: To develop a TaqMan probe-based, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of Mycoplasma suis in the blood of pigs. METHODS AND RESULTS: Primers and probes specific to Myc. suis 16S rRNA gene were designed. The qPCR assay's specificity, detection limit, intra- and inter-assay variability were evaluated and its performance was compared with a Myc. suis conventional PCR assay (cPCR). Blood of two experimentally infected pigs, 40 Indiana pigs, 40 Brazilian sows and 28 peccaries were tested. The assay detected as few as ten copies of Myc. suis plasmids and was 100-fold more sensitive than the cPCR. No cross-reactivity with nontarget pig mycoplasmas was observed. An average of 1·62 × 10(11) and 2·75 × 10(8) target copies ml(-1) of blood were detected in the acutely and chronically infected pigs, respectively. Three (7·5%) pigs and 32 (80·0%) sows were positive while all peccaries were negative for Myc. suis. CONCLUSION: The developed qPCR assay is highly sensitive and specific for Myc. suis detection and quantification. SIGNIFICANCE AND IMPACT OF THE STUDY: TaqMan qPCR is an accurate and quick test for detection of Myc. suis infected pigs, which can be used on varied instrumentation platforms.

Validation of a Leishmania infantum ELISA rapid test for serological diagnosis of Leishmania chagasi in dogs.

Canine visceral leishmaniasis (CVL) is caused by Leishmania donovani complex parasites including L. donovani, Leishmania infantum and Leishmania chagasi. As some studies suggest that L. chagasi and L. infantum may be very similar or even the same species, the aim of the present study was to evaluate a commercial rapid ELISA test, originally designed for L. infantum, in the diagnosis of CVL in dogs naturally infected by L. chagasi. A total of 400 serum canine samples, including 283 positive dogs for CVL from an endemic area, 86 clinically healthy dogs from a non-endemic area and 31 dogs seropositive for confounding infectious agents (Trypanosoma cruzi, Toxoplasma gondii, Neospora caninum, Babesia canis and Ehrlichia canis) were used for test validation. An overall sensitivity of 94.7% (95% CI=91.41-97.01%) and specificity of 90.6% (95% CI=83.80-95.21%) was found, with a high degree of agreement (k=0.8445) to the indirect ELISA. When confounding infectious diseases were excluded, specificity increased to 100% (95% CI=95.8-100%), with a higher degree of agreement (k=0.8928). In conclusion, the commercial kit designed for L. infantum was a highly sensitive and specific device for detection of L. chagasi infection in dogs, which indicates high immunoreactivity similarities between L. infantum and L. chagasi.

Soroprevalência de anticorpos para o Vírus da Leucose Enzoótica em bovinos criados na região metropolitana de Curitiba, Paraná / Seroprevalence for Bovine Leukosis Virus (BLV) in dairy cattle of Curitiba and the surrounding area, Paraná, Brazil

A Leucose Enzoótica Bovina (LEB) é uma doença infecto-contagiosa causada pelo Vírus da Leucemia Bovina (VLB), um retrovírus oncogênico da família Retroviridae. O presente estudo teve como objetivo avaliar a soroprevalência e a respectiva influência de fatores etários da LEB em bovinos leiteiros das raças Holandesa Preta e Branca, Jersey, Pardo-Suíço e mestiços, criados na região metropolitana de Curitiba, PR. Foram testadas 268 amostras de soros sanguíneos pela prova de Imunodifusão em gel de Agar (IDGA), colhidas em cinco propriedades situadas nos municípios de São José dos Pinhais, Campina Grande do Sul, Pinhais e Fazenda Rio Grande. Foram encontrados 151/268 (56,34%) animais positivos e 117/268 (43,66%) negativos. Animais mais velhos mostraram um aumento estatisticamente significativo de soropositividade. Pode-se concluir que a LEB está amplamente disseminada em bovinos leiteiros da região metropolitana de Curitiba e há necessidade de adequada aplicação de medidas de controle e prevenção da LEB.

Enzootic bovine leukosis (EBL) is aninfectious and contagious disease caused by bovine leukemia virus (BLV), an oncogenic retrovirus of the family Retroviridae. The present study aimed to evaluate the seroprevalence and respective age influence of BLV in Holstein, Jersey, Brown Swiss and mixed-breed dairy cattle raised in Curitiba and the surrounding area, Paraná, Brazil. A total of 268 samples of bovine serum from five different herds in the counties of São José dos Pinhais, Campina Grande do Sul, Pinhais and Fazenda Rio Grande were tested with immunodiffusion (ID). A total of 151/268 (56.34%) testedseropositive, while 117/268 (43.66%) were considered seronegative. Older animals prsented a significant rise in seropositivity. In conclusion, BLV is widely distributed among dairy cattle of Curitiba and the surrounding area, present in all tested herds, thus requiring adequate application of measures in EBL control and prevention.

Mapping risk of bovine fasciolosis in the south of Brazil using Geographic Information Systems.

Fasciolosis, caused by Fasciola hepatica, is an endemic disease of ruminants that occurs in several countries of South America where it can lead to decreased production and fertility and, in severe cases, animal death. Although very prevalent, information on the epidemiology of the disease is incomplete in Brazil. The objective of the present study was to define the prevalence of F. hepatica in the livers of cattle from slaughterhouses and correlate the data with the animal's origin (climate and altitude) using a Geographic Information System (GIS). The data was used to create an epidemiological map of fasciolosis by state (Rio Grande do Sul, Santa Catarina, Paraná), by municipality (n=530) and by year (2003-2008). Information was analyzed using a databank from slaughterhouses with Federal Inspection Services of the Ministry of Agriculture. The highest cattle infection rate was found in the two most Southern states of Rio Grande do Sul (18.7%) and Santa Catarina (10.1%). Animals from the Campanha region of Rio Grande do Sul and from the central coast area of Santa Catarina had prevalences of greater than 40%. Cattle from low altitudes municipalities were significantly more likely to have the disease (p<0.05). No significant differences were found between high or low prevalence and ambient temperatures. Risk maps resulting from this study provide information on the epidemiology and transmission of F. hepatica in Southern Brazil needed for design of appropriate control measures to control economic impacts. F. hepatica may represent an important source of zoonotic infection of humans as well; therefore these findings may be complemented by future studies on human infections in high risk areas.

Detection of Bartonella spp. in neotropical felids and evaluation of risk factors and hematological abnormalities associated with infection.

Although antibodies to Bartonella henselae have been described in all neotropical felid species, DNA has been detected in only one species, Leopardus wiedii. The aim of this study was to determine whether DNA of Bartonella spp. could be detected in blood of other captive neotropical felids and evaluate risk factors and hematological findings associated with infection. Blood samples were collected from 57 small felids, including 1 Leopardus geoffroyi, 17 L. wiedii, 22 Leopardus tigrinus, 14 Leopardus pardalis, and 3 Puma yagouaroundi; 10 blood samples from Panthera onca were retrieved from blood banks. Complete blood counts were performed on blood samples from small felids, while all samples were evaluated by PCR. DNA extraction was confirmed by amplification of the cat GAPDH gene. Bartonella spp. were assessed by amplifying a fragment of their 16S-23S rRNA intergenic spacer region; PCR products were purified and sequenced. For the small neotropical felids, risk factors [origin (wild-caught or zoo-born), gender, felid species, and flea exposure] were evaluated using exact multiple logistic regression. Hematological findings (anemia, polycythemia/hyperproteinemia, leukocytosis and leukopenia) were tested for association with infection using Fisher's exact test. The 635bp product amplified from 10 samples (10/67=14.92%) was identified as B. henselae by sequencing. Small neotropical felid males were more likely to be positive than females (95% CI=0.00-0.451, p=0.0028), however other analyzed variables were not considered risk factors (p>0.05). Hematological abnormalities were not associated with infection (p>0.05). This is the first report documenting B. henselae detection by PCR in several species of neotropical felids.

Survey of owned feline and canine populations in apartments from a neighbourhood in Curitiba, Brazil.

Animal population estimates are essential for public health services to ensure the success of zoonoses control programmes. Canine and feline populations vary among different regions mainly because of local human income status and type of human residence. Accordingly, the present study estimated the pet population size living in apartments of a vertical neighbourhood in Curitiba, Brazil. We chose a neighbourhood with a predominance of apartment buildings. All apartment buildings were visited, and questionnaires were completed by doormen or residents. Data were obtained from 120 of 173 apartment buildings. Survey questions included the number of apartments, residents, dogs and cats. Two thousand nine hundred and sixty six apartments with a total of 7429 residents were surveyed. The number of dogs and cats was 569 and 86 respectively. Thus, the human:dog and human:cat ratios were 13.05:1 and 86.38:1. These ratios were higher than those observed in other neighbourhoods in Curitiba. The present study indicates that the number of pets from apartments may be different from houses, and different among distinct areas within the same city.

Exploratory study of Mycoplasma suis (Eperythrozoon suis) on four commercial pig farms in southern Brazil.

Mycoplasma suis (Eperythrozoon suis) was detected by PCR and Southern blot in 186 pigs (121 sows, 61 piglets and four boars) on four farms in southern Brazil. DNA was extracted from blood samples and a 16S rRNA gene fragment of M suis was amplified by PCR; Southern blot analysis was then performed on all the samples. Twenty-two of the sows (18.2 per cent) were positive by PCR, and 40 (33.1 per cent) were positive by Southern blot; only one piglet and one boar were positive. The packed cell volume and total plasma protein of the pigs and their PCR and Southern blot results were not significantly different on the four farms, but higher proportions of the pigs were positive by Southern blot than by PCR (P<0.05). The packed cell volume and total plasma protein concentrations of the M suis positive and negative sows were not significantly different.

Imunologia tumoral como ferramenta na oncologia veterinária / Tumoral immunology as a tool for veterinary oncology / Immunologìa tumoral como herramienta en la oncología veterinaria

A busca de métodos terapêuticos e profiláticos mais eficazes contra o câncer resultou em grande impacto no entendimento das bases imunológicas da oncologia. A descoberta dos antígenos tumorais e dos mecanismos de escape tumoral da vigilância imunológica revolucionaram a compreensão da atuação do sistema imune na gênese e desenvolvimento tumoral. As proteínas parecem ser a chave do processo tumoral, permitindo um melhor entendimento do sistema imune como um regulador final da sua origem, desenvolvimento e/ou destruição das neoplasias. Sendo assim, as proteínas tumorais oferecem uma via potencial para a compreensão dos mecanismos de interação e evasão celular, busca de novos marcadores tumorais, e a produção de uma imunoterapia eficaz e irrestrita. Esta revisão tem como objetivo principal discutir a atuação do sistema imune no desenvolvimento neoplásico no sentido de produzir protocolos de tratamento e prevenção eficientes como futuras ferramentas na oncologia veterinária.

The search for more effective therapeutic and prophylactic methods against cancer resulted in a great impact on the understanding of the immunological basis of oncology. The discovery of tumoral antigens and tumoral escape mechanisms from immunological surveillance revolutionized the comprehension of the role of the immune system on the tumoral genesis and its development. Proteins seem to be the key for the tumoral process, allowing a better understanding of the immune system as a final check point of the neoplasic beginning, development and/or destruction. Therefore, tumoral proteins provide a potential pathway for the comprehension of its interaction and cellular evasion mechanisms, the search for new tumoral markers, and the production of an effective and non-restricted immunotherapy. This review has as its main objective to discuss the role of the immune system regarding the neoplasic development towards the production of an effective therapy and prophylaxis protocols as future tools for veterinary oncology.

La búsqueda por métodos terapéuticos y profilácticos más eficazes contra el cáncer resultó en gran impacto en el entendimiento de las bases inmunológicas de la oncología. El descubrimiento de los antígenos tumorales y de los mecanismos de escape tumoral de la vigilancia inmunológica revolucionaron la comprensión de la actuación del sistema inmune y en el génesis y desarrollo tumoral. Las proteínas parecen ser la llave del proceso tumoral, permitiendo un mejor entendimiento del sistema inmune como un regulador final de su nacimiento, desarrollo y/o destrucción de las neoplasias. Así pues, las proteínas tumorales ofrecen un potencial camino para la comprensión de los mecanismos de interacción y evasión celular, búsqueda de nuevos marcadores tumorales, y la producción de una inmunoterapia eficaz y sin restricción. Esta revisión tiene como objetivo principal discutir la actuación del sistema inmune en el desarrollo neoplásico con objeto de producir protocolos de tratamiento y prevención eficientes como futuras herramientas en la oncología veterinaria.

Immunohistochemistry of atrial and brain natriuretic peptides in control cats and cats with hypertrophic cardiomyopathy.

Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are cardiac hormones involved in electrolyte and fluid homeostasis. Our laboratory has investigated the use of ANP and BNP as diagnostic markers of cardiac disease in cats. We hypothesize that the cardiac distribution of ANP and BNP increases in cats with hypertrophic cardiomyopathy (HCM). Accordingly, we evaluated the immunohistochemical distribution of ANP and BNP in hearts of four cats with naturally occurring HCM relative to five healthy controls. Indirect immunoperoxidase was performed with polyclonal immunoglobulin G against feline ANP (1-28) and proBNP (43-56). In control cats, ANP and BNP immunoreactivity was restricted to the atria. Staining for both peptides was most intense adjacent to the endocardial surface. Auricles stained more diffusely than atria for both peptides. The interstitial capillaries and nerve fibers within the heart were positive only for BNP. Atrial immunoreactivity for ANP and BNP was more diffuse and had a less distinctly layered pattern in HCM than in control cats. Ventricular cardiomyocytes of HCM cats were negative for ANP but stained lightly and diffusely for BNP. The capillaries and nerve fibers remained positive for BNP. We conclude that in cats with HCM, the cardiac distribution of ANP and BNP is more diffuse in the atria and that novel expression of BNP in the ventricular cardiomyocytes occurs.

Canine hepatozoonosis in Brazil: description of eight naturally occurring cases.

Eight cases of canine hepatozoonosis were diagnosed at the Veterinary Hospital (Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista, Campus de Botucatu), between October 1993 and April 1994. Clinical signs included anorexia, pale mucous membranes, weight loss, pain, diarrhoea, vomit, gait abnormalities, fever, polyuria and polydipsia. Haematologic findings revealed anaemia in seven cases, leucocytosis with neutrophilia in three cases, lymphopenia in three cases and monocytosis in four cases. Serum biochemistries included alterations in many parameters. The micrometry of Hepatozoon canis gametocytes ranged from 6.8 x 4.0 microns to 7.5 x 4.5 microns. Parasitaemia ranged from less than 0.5% to 2%. In all the cases reported other concurrent diseases were present. Diagnosis of canine hepatozoonosis was made by identifying H. canis gametocytes within leucocytes in stained blood smears.