Tuberculosis determined by Mycobacterium bovis in captive waterbucks (Kobus ellipsiprymnus) in SÃo paulo, Brazil.

Two waterbucks from São Paulo Zoo Foundation exhibited respiratory symptoms in July 2004. After euthanasia, granulommas in lungs and mediastinic lymph nodes were observed. Acid-fast bacilli isolated were identified as Mycobacterium bovis spoligotype SB0121 by PRA and spoligotyping. They were born and kept in the same enclosure with the same group, without any contact to other species housed in the zoo. This is the first detailed description of M. bovis infection in Kobus ellipsiprymnus.

Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308.

The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (p< 0.0001) and GT (p=0.0004). There was no difference between the PK and GT protocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and vice-versa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol.

Economic analysis of vaccination to control bovine brucellosis in the States of Sao Paulo and Mato Grosso, Brazil.

Brucellosis is a zoonotic disease that causes important economic losses in Brazil, and the country has therefore established a national program for its control and eradication. Using data generated in the last national brucellosis survey, we conducted an economic analysis in two Brazilian States with different brucellosis status, Mato Grosso (with high prevalence) and Sao Paulo (with low prevalence). The economic analysis was based on the calculation of the additional benefits and costs of controlling bovine brucellosis through the vaccination of heifers aged between 3 and 8 months with S19 vaccine, considering maximal and minimal impacts of the disease. The analysis showed that vaccinating 90% of the replacement heifers aged 3-8 months of age offers the best economic performance in a vaccination program against bovine brucellosis if compared to vaccination rates of 70% and 80%. Moreover, regions with higher prevalences of bovine brucellosis would experience significant economic advantages when implementing a vaccination strategy to control the disease. This economic analysis will allow decision makers to plan more economically effective vaccination programs.

Ticks (Acari: Ixodidae) associated with rural dogs in Uruará, eastern Amazon, Brazil.

A survey of 124 dogs for ticks was conducted from August to November 1998 in 68 small farms of Uruará municipality in Eastern Amazon-Brazil. We collected 55 ticks from 18 dogs (14.5%). Amblyomma oblongoguttatum Koch, Amblyomma ovale Koch, and Amblyomma cajennense (F.) were found on nine, six, and one dog, respectively. Fourteen small farms (20.5%) had at least one dog with ticks.

A case of gynandromorphism in Amblyomma oblongoguttatum (Acari: Ixodidae).

During a survey conducted from August to November 1998 to ticks associated with dogs in the rural area of Uruará, in the state of Para, Eastern Amazon region of Brazil, we collected one gynandromorph specimen out of 18 Amblyomma oblongoguttatum Koch specimens collected from dogs. It is a perfect bipartite gynandromorph, reported in this species for the first time.

Seroprevalence of Toxoplasma gondii in captive neotropical felids from Brazil.

Seroprevalence of Toxoplasma gondii was determined in 865 captive neotropical felids from 20 states from Brazil, sampled from September 1995 to April 1997. Sera were tested by the modified agglutination test (MAT) using formalin-fixed whole tachyzoites and mercaptoethanol. Antibodies (MAT> or =1:20) to T. gondii were found in 472 of 865 (54.6%) cats: in 45 of 99 (45.9%) jaguarundis (Herpailurus yagouaroundi), in 97 of 168 (57.7%) ocelots (Leopardus pardalis), in 68 of 131 (51.9%) oncillas (L. tigrinus), in 35 of 63 (55.5%) margays (L. wiedii), in 1 of 8 (12.5%) Pampas-cat (Oncifelis colocolo), in 9 of 12 (75.0%) Geoffroys-cat (O. geoffroyi), in 134 of 212 (63.2%) jaguars (Panthera onca), and in 83 of 172 (48.2%) pumas (Puma concolor). Antibody titers were: 1:20 in 27 felids, 1:25 in 142 felids, 1:40 in 6 felids, 1:50 in 292 felids, and > or =1:500 in 5 felids. The high seroprevalence of T. gondii antibodies found in the present study suggested a widespread exposure of neotropical cats to T. gondii in zoos in Brazil. The results warrant an investigation on the mode of exposure and oocyst shedding by neotropical cats.

Leptospira interrogans serovar icterohaemorrhagiae seropositivity and the reproductive performance of sows.

The reproductive performance of 28 sows seropositive to Leptospira interrogans serovar icterohaemorrhagiae was compared with that of 87 Leptospira sp. seronegative dams belonging to the same herd. Sows were sampled during 1988 to 1993. During this period the herd was not submitted to any kind of intervention (antibiotic therapy, immunoprophylaxis or rodent control). Relative risks (RR) of return to heat, mummified fetuses, stillbirth, and weak newborn piglets for infected sows were assessed and the differences in means of total piglets born per litter, piglets born alive, piglets effectively housed, weaned piglets, stillbirths, mummified fetuses, weak newborn piglets, weight at birth of the piglets effectively housed, weight at 21 days of life and weight at weaning were evaluated. Seropositive dams had a greater risk of having weak newborn piglets (RR = 1.67, 1.02 < or = CI 95% < or = 2.72) and also of having more weak newborn piglets per litter (P = 0.01). Other variables examined were not different (P > 0.05).

Prevalence of Toxoplasma gondii antibodies in sera of domestic cats from Guarulhos and São Paulo, Brazil.

Antibodies to Toxoplasma gondii were determined in serum samples of 502 domestic cats from Brazil by the modified agglutination test (MAT), using formalin-fixed whole tachyzoites and mercaptoethanol. Antibodies (MAT > or = 1:20) were found in 132 (26.3%) of 502 cats. With respect to origin, antibodies were found in 26.7% of 430 stray cats from São Paulo, 10% of 40 stray cats from Guarulhos, and 40.6% of 32 cats from a cat breeder in São Paulo. Antibody titers were: 1:20 in 10 cats, 1:25 in 40 cats, 1:50 in 73 cats, and > or = 1:500 in 9 cats. Exposure rates of T. gondii in cats from São Paulo, Brazil are similar to that in domestic cats in North America.

[Epidemiologic study of bovine and human leptospirosis in eastern Brazilian Amazon]. / Estudo epidemiológico da leptospirose bovina e humana na Amazônia oriental brasileira.

The seroprevalence study for leptospirosis in bovines and humans was realized in family holder farms along the Transamazon Highway. The prevalence of bovine leptospirosis was 97% [90.9 - 99.5%] of farms with at least one positive animal according to microscopic agglutination test for the leptospirosis diagnostic. In 61.2% of the tested herds, the serovar hardjo was the most common, followed by the serovar bratislava (9%) and the serovar shermani (4.5%). The serologic prevalence of leptospirosis in humans was 32.8% [23.4 - 43.5%] in family groups with at least one positive individual according to microscopic agglutination test for the leptospirosis diagnostic. In 9% of family groups, the serovar bratislava was the most common, while serovar hardjo and grippotyphosa accounted for 6% and 4.5%, respectively. The impact of these results is discussed in relation to animal production and public health. Suggestions have been proposed in order to improve the situation in the region.

[Tuberculosis lymphadenitis in slaughtered swine from the State of São Paulo, Brazil: microscopic histopathology and demonstration of mycobacteria]. / Linfadenites tuberculóides em suínos abatidos no estado de São Paulo, Brasil: aspectos macroscópicos histopatológicos e pesquisa de micobactérias.

INTRODUCTION: As the occurrence of tuberculosis lymphadenitis in swine constitutes a public health risk, especially in immunosuppressed individuals, the distribution of tuberculoid lesions and the presence of microbacteria in lymph nodes and hepatic and muscular tissue in swine for slaughter, in the State of S. Paulo, SP (Brazil), in the period from 1993 to 1994, were studied. MATERIAL E METHOD: Tuberculous lesions in 60 carcasses of swine, slaughtered at abattoirs during the period 1993-1994, were studied. When macroscopic lesions were observed, a representative sampling of lymph nodes, hepatic and muscular tissues (masseter and diaphragm) were examined using histopathological and microbacterial isolation techniques. RESULTS: The macroscopic lesions were found predominantly in mesenteric lymph nodes. The MAI Complex (M.avium-intracellulare) was isolated mainly in the carcass group that showed lesions and was not found in the control group. Microorganisms of the MAI Complex were not isolated from hepatic and muscular tissues. CONCLUSION: No clear relationship between the type of mycobacteria isolated and the macroscopic lesions observed during the carcass inspection was found.

Toxoplasma gondii antibodies in exotic wild felids from Brazilian zoos.

Serum samples from 37 captive exotic felids in 12 zoos from six Brazilian states were assayed for antibodies to Toxoplasma gondii by the modified agglutination test using formalin-fixed whole tachyzoites. Titers greater than or equal to 1:20 were considered positive. Antibodies to T. gondii were found in 24 of 37 (64.9%) felids, including one European lynx (Lynx lynx), two jungle cats (Felis chaus), two servals (Leptailurus serval), two tigers (Panthera tigris), three leopards (Panthera pardus), and 14 of 27 lions (Panthera leo). This is the first serologic analysis for T. gondii infection in exotic wild felids from Brazilian zoos.

Seroprevalence and risk factors of bovine herpesvirus 1 infection in cattle herds in the state of Paraná, Brazil.

Epidemiological data describing bovine herpesvirus 1 (BoHV-1) infection in Brazilian cattle herds are scarce. A cross-sectional study was conducted in the state of Paraná between December 2001 and July 2002 with the objective of estimating the apparent prevalence of BoHV-1-seropositive herds and animals and identifying the potential risk factors for infection in farms with breeding animals in the state of Paraná in Southern Brazil. The state was divided into seven regions based on the livestock production dynamics of the different areas. Sampling was performed in two stages. Initially, herds were randomly selected, followed by a random selection of animals within the selected herds. Blood samples were collected from 14803 females, aged ≥24 months, from 2018 BoHV-1 non-vaccinated herds. Serum samples were tested for antibodies against BoHV-1 using an indirect enzyme-linked immunosorbent assay (ELISA). The apparent prevalence of seropositivity in the herds and animals in Paraná was 71.3% (95% CI: 69.3-73.3) and 59.0% (95% CI: 56.2-61.8), respectively. Multiple logistic regression analysis revealed that risk factors associated with the presence of the infection were as follows: beef herds [OR = 1.58 (1.12-2.23)], natural service [OR = 1.48 (1.02-2.14)], purchase of animals [OR = 1.90 (1.52-2.37)], pasture rental [OR = 2.24 (1.51-3.33)], existence of calving pens [OR = 1.56 (1.20-2.03)] and records of abortion in the last 12 months [OR = 1.45 (1.08-1.95)]. These results indicate that BoHV-1 infection is widespread in the state of Paraná.

Evaluation of four DNA extraction protocols for Brucella abortus detection by PCR in tissues from experimentally infected cows with the 2308 strain.

This study compared 4 protocols for DNA extraction from homogenates of 6 different organs of cows infected with the Brucella abortus 2308 strain. The extraction protocols compared were as follows: GT (guanidine isothiocyanate lysis), Boom (GT lysis with the carrying suspension diatomaceous earth), PK (proteinase K lysis), and Santos (lysis by boiling and freezing with liquid nitrogen). Positive and negative gold standard reference groups were generated by classical bacteriological methods. All samples were processed with the 4 DNA extraction protocols and amplified with the B4 and B5 primers. The number of positive samples in the placental cotyledons was higher than that in the other organs. The cumulated results showed that the Santos protocol was more sensitive than the Boom (p=0.003) and GT (p=0.0506) methods and was similar to the PK method (p=0.2969). All of the DNA extraction protocols resulted in false-negative results for PCR. In conclusion, despite the disadvantages of classical bacteriological methods, the best approach for direct diagnosis of B. abortus in organs of infected cows includes the isolation associated with PCR of DNA extracted from the cotyledon by the Santos or PK methods.

Tuberculosis determined by Mycobacterium bovis in captive waterbucks (Kobus ellipsiprymnus) in São Paulo, Brazil

Two waterbucks from São Paulo Zoo Foundation exhibited respiratory symptoms in July 2004. After euthanasia, granulommas in lungs and mediastinic lymph nodes were observed. Acid-fast bacilli isolated were identified as Mycobacterium bovis spoligotype SB0121 by PRA and spoligotyping. They were born and kept in the same enclosure with the same group, without any contact to other species housed in the zoo. This is the first detailed description of M. bovis infection in Kobus ellipsiprymnus.

Avaliação de testes sorológicos de triagem para diagnóstico da brucelose bubalina / Evaluation of screening serological tests for brucellosis diagnosis in buffaloes

The study compared the performance of three screening serological tests: buffered plate antigen (BPA), Rose-Bengal produced with 1119-3 Brucella abortus strain (RB1119-3), and Rose-Bengal produced with 99 Brucella abortus strain (RB99). Sera from 696 adult female animals were submitted to BPA, RB1119-3, RB99, 2-mercaptoethanol test (ME), and complement fixation test (FC). The gold standard was the combination of CF and ME. The Kappa values for BPA, RB99, and RB1119-3 were 0.82, 0.74, and 0.70, respectively. The relative sensitivity and specificity for the same tests were 0.98 and 0.96, 0.92 and 0.94, and 0.95 and 0.92, respectively. These results indicate that BPA is a better screening test than RB for buffalo, regardless of the B. abortus strain in RB.

Tipificação genética, através da técnica de spoligotyping, de isolados de Mycobacterium bovis em animais abatidos na região metropolitana de Salvador, Bahia, Brasil / Use of spoligotyping for genetic typing of Mycobacterium bovis isolates from slaughtered animals in the metropolitan area of Salvador, Bahia, Brazil

A tuberculose é uma enfermidade infecciosa crônica, que afeta mamíferos e aves e constitui um sério problema de saúde pública e animal. Objetivando realizar um levantamento molecular da enfermidade em bovinos abatidos em matadouros frigoríficos no Estado da Bahia, Brasil, foram analisadas as lesões pulmonares e de linfonodos mediastínicos de 43 carcaças de animais abatidos em três matadouros-frigoríficos localizados na Região Metropolitana de Salvador, Bahia. Sete isolados de Mycobacterium bovis foram identificados, através da técnica do spolygotyping, e discriminados em três diferentes espoligotipos (SB1055, SB0120 e SB0268) descritos no Brasil e em diversas áreas do mundo. Os resultados indicam que o método de diagnóstico utilizado pode contribuir para a criação de uma base de dados para o estudo epidemiológico da tuberculose bovina no Estado da Bahia.

Tuberculosis is an infectious chronic disease that affects mammals and birds and constitutes a serious problem for public and animal health. Pulmonary and mediastinic lymph node lesions of 43 animals slaughtered in 3 slaughterhouses in the metropolitan region of the city of Salvador, Bahia, Brazil, were analyzed with the objective of obtaining a molecular survey of the disease in bovines slaughtered in slaughterhouses in the state. Seven isolates ofMycobacterium bovis were identified through the spoligotyping technique and classified into 3 different spoligotypes (SB1055, SB0120, SB0268), described in Brazil and in many areas worldwide. The results indicate that the diagnostic method utilized may contribute to the creation of a database for the epidemiologic study of bovine tuberculosis in the state of Bahia.

Modelagem matemática do controle de brucelose bovina por vacinação / Mathematical modeling of bovine brucellosis control by vaccination

As fêmeas bovinas, por sua importância na transmissão e na manutenção da brucelose, constituíram o alvo dos inquéritos do Programa Nacional de Controle e Erradicação da Brucelose e da Tuberculose Animal. Com base em informações obtidas em unidades federativas onde foram realizados inquéritos sorológicos e observadas prevalências de animais acima de 2 por cento, elaborou-se um modelo para simular a dinâmica da brucelose em rebanhos bovinos formados exclusivamente por fêmeas, analisando o efeito de estratégias de vacinação. Para baixa cobertura vacinal, da ordem de 30 por cento, o tempo para reduzir a prevalência a 2 por cento, valor adotado como referência, pode ser longo, aproximando-se do dobro do tempo necessário para uma cobertura mais alta, de 90 por cento. De acordo com o modelo, o tempo para reduzir a prevalência a 1 por cento ou 2 por cento, que permitam passar à fase de erradicação, pode chegar a uma década. Recomenda-se a intensificação do esforço para a vacinação de fêmeas, procurando atingir alta cobertura vacinal.

Due to the important role played by female bovines in the transmission and maintenance of brucellosis, they were the target of the serological surveys of the Brazilian National Program for the Control and Eradication of Bovine Brucellosis and Tuberculosis. Based on the information obtained in Brazilian states, where the serological surveys were carried out and prevalences higher than 2 percent were observed, a model to simulate the dynamics of brucellosis in herds of female bovines was developed to analyze the effects of vaccination strategies. For low vaccination coverage (around 30 percent), the time to reduce the prevalence to 2 percent, adopted as a reference, may be long, approximately twice as long as the time observed for a higher coverage (90 percent). According to the model, the time to reduce the prevalence to levels of 1 percent or 2 percent, adequate to start the eradication phase, may reach a decade. It is recommended an intensification of the effort to vaccinate females, aiming at attaining high vaccination coverage.

Identification of two novel Mycobacterium avium allelic variants in pig and human isolates from Brazil by PCR-restriction enzyme analysis.

Mycobacterium avium complex (MAC) is composed of environmental mycobacteria found widely in soil, water, and aerosols that can cause disease in animals and humans, especially disseminated infections in AIDS patients. MAC consists of two closely related species, M. avium and M. intracellulare, and may also include other, less-defined groups. The precise differentiation of MAC species is a fundamental step in epidemiological studies and for the evaluation of possible reservoirs for MAC infection in humans and animals. In this study, which included 111 pig and 26 clinical MAC isolates, two novel allelic M. avium PCR-restriction enzyme analysis (PRA) variants were identified, differing from the M. avium PRA prototype in the HaeIII digestion pattern. Mutations in HaeIII sites were confirmed by DNA sequencing. Identification of these isolates as M. avium was confirmed by PCR with DT1-DT6 and IS1245 primers, nucleic acid hybridization with the AccuProbe system, 16S ribosomal DNA sequencing, and biochemical tests. The characterization of M. avium PRA variants can be useful in the elucidation of factors involved in mycobacterial virulence and routes of infection and also has diagnostic significance, since they can be misidentified as M. simiae II and M. kansasii I if the PRA method is used in the clinical laboratory for identification of mycobacteria.