Abstract in English:
ABSTRACT.- Souza F.A.L., Braga J.F.V., Pires L.V., Carvalho C.J.S., Costa E.A., Ribeiro M.F.B., Santos R.L. & Silva S.M.M.S. 2013. Babesiosis and anaplasmosis in dairy cattle in Northeastern Brazil. Pesquisa Veterinária Brasileira 33(9):1057-1061. Departamento de Clínica e Cirurgia Veterinária, Centro de Ciências Agrárias, Universidade Federal do Piauí, Av. Universitária s/n, Teresina, PI 64049-550, Brazil. E-mail: chicoleite@hotmail.com
The goal of this study was to characterize the epidemiological situation and the factors involved in the prevalence of babesiosis and anaplasmosis in cattle in the dairy basin of Parnaíba, Piauí, Brazil. The study was conducted in 22 farms, and collected blood samples from 202 cattle to study serological, molecular and determination of the packed cell volume (PCV). On the farms were applied surveys involving epidemiological aspects. Seroprevalence rates were: Babesia bigemina 52.5%, B. bovis 68.8%, and Anaplasma marginale 89.1%. Of the samples analyzed, 73.3% were reactive for Babesia spp. and A. marginale, showing co-infection. In PCR, B. bigemina and B. bovis were positive in 52.0% and 33.2% respectively, and A. marginale in 76.2%. Of these, 51.5% amplified DNA of Babesia spp. and A. marginale. The semi-intensive management predominated in 68.0% of the farms studied. The clinical history of babesiosis and anaplasmosis, was reported from 73% of the farms. There was no significant difference (p>0.05) between age groups and for the PCV of positive compared with negative animals. The study indicates that in this region is enzootic instability for babesiosis and enzootic stability for anaplasmosis, reinforcing the fact that in Brazil there are areas of enzootic instability, even in tropical regions of the country. The PCR technique was a valuable tool for the diagnosis of these diseases and may be used to characterize a geographic region.
Abstract in Portuguese:
RESUMO.- Souza F.A.L., Braga J.F.V., Pires L.V., Carvalho C.J.S., Costa E.A., Ribeiro M.F.B., Santos R.L. & Silva S.M.M.S. 2013. Babesiosis and anaplasmosis in dairy cattle in Northeastern Brazil. [Babesiose e anaplasmose em bovinos leiteiros no Nordeste do Brasil.] Pesquisa Veterinária Brasileira 33(9):1057-1061. Departamento de Clínica e Cirurgia Veterinária, Centro de Ciências Agrárias, Universidade Federal do Piauí, Av. Universitária s/n, Teresina, PI 64049-550, Brazil. E-mail: chicoleite@hotmail.com
O objetivo deste estudo foi caracterizar a situação epidemiológica e os fatores envolvidos na prevalência da babesiose e anaplasmose em bovinos da bacia leiteira de Parnaíba, Piauí, Brasil. O estudo foi realizado em 22 propriedades, sendo coletadas amostras de sangue de 202 bovinos para estudos sorológicos, moleculares e determinação do volume globular (VG). Nas propriedades foram aplicadas inquéritos envolvendo aspectos epidemiológicos. As taxas de soroprevalência foram: 52,5% para Babesia bigemina, 68,8% B. bovis, e 89,1% para Anaplasma marginale. Das amostras analisadas, 73,3% foram reagentes para Babesia spp. e A. marginale, demostrando co-infecção. Na PCR, B. bigemina e B. bovis foram positivas em 52,0% e 33,2% respectivamente, e A. marginale em 76,2%. Destes, 51,5% amplificaram DNA de Babesia spp. e A. marginale. O manejo semi-intensivo predominou em 68,0% das propriedades estudadas. O histórico clínico de babesiose e anaplasmose foi relatado em 73% das propriedades. Não houve diferença significativa (p>0,05) entre as faixas etárias e para o VG de animais positivos comparados com os negativos. O estudo indica que nesta região há instabilidade enzoótica para babesiose e estabilidade enzoótica para anaplasmose, reforçando o fato de que, no Brasil, existem áreas de instabilidade enzoótica, mesmo em regiões tropicais do país. A técnica de PCR demonstrou ser uma ferramenta valiosa para o diagnóstico destas doenças e pode ser utilizada para caracterizar uma região geográfica.
Abstract in English:
ABSTRACT.- Silva J.B., Lopes C.T.A., Pinheiro C.P., Lima D.H.S., Silva R.S.L., Fonseca A.H., Araújo F.R. & Barbosa-Neto J.D. 2013. [Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes (Bubalus bubalis) on Marajo Island, State of Pará, Brazil.] Prevalência sorológica e molecular de Babesia bovis e Babesia bigemina em búfalos (Bubalus bubalis) na Ilha de Marajó, Pará. Pesquisa Veterinária Brasileira 33(7):847-850. Departamento de Epidemiologia e Saúde Pública, Universidade Federal Rural do Rio de Janeiro, BR 465 Km 7, Seropédica, RJ 23890-000, Brazil. E-mail: jenevaldo@hotmail.com
The aim of the study was to estimate the prevalence of Babesia bovis and Babesia bigemina in water buffaloes of the Marajó Island, State of Pará, Brazil. We used an indirect enzyme-linked immunosorbent assay (iELISA), with total antigen containing proteins outer surface, and polymerase chain reaction (qPCR), involving the use of SYBR Green based on amplification of a small fragment of the cytochrome b gene. The prevalence of positive animals in iELISA to B. bovis B. bigemina and mixed infection was 24.87% (199/800), 20.75% (166/800) and 18.75% (150/800), respectively. Using the PCR, the presence of B. bovis was detected in 15% (18/199) and B. bigemina in 16% (19/199) of animals, and of these, 58% (11/19) presented co-infected by the two agents. The results show a low prevalence of antibodies anti-B. bovis and anti-B. bigemina in water buffaloes from Marajó Island. However, it was observed that the agents of bovine babesiosis circulate in buffaloes, and these may act as reservoirs.
Abstract in Portuguese:
RESUMO.- Silva J.B., Lopes C.T.A., Pinheiro C.P., Lima D.H.S., Silva R.S.L., Fonseca A.H., Araújo F.R. & Barbosa-Neto J.D. 2013. [Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes (Bubalus bubalis) on Marajo Island, State of Pará, Brazil.] Prevalência sorológica e molecular de Babesia bovis e Babesia bigemina em búfalos (Bubalus bubalis) na Ilha de Marajó, Pará. Pesquisa Veterinária Brasileira 33(7):847-850. Departamento de Epidemiologia e Saúde Pública, Universidade Federal Rural do Rio de Janeiro, BR 465 Km 7, Seropédica, RJ 23890-000, Brazil. E-mail: jenevaldo@hotmail.com
O objetivo do estudo foi testar a prevalência sorológica e molecular de Babesia bovis e Babesia bigemina em búfalos da Ilha de Marajó, Pará. Foi utilizado ensaio de imunoadsorção enzimático indireto (iELISA) com antígeno total contendo proteínas de superfície externa e reação em cadeia da polimerase (qPCR), envolvendo o uso de SYBR Green com base na amplificação de um pequeno fragmento de gene do citocromo b. A prevalência de animais positivos no ELISA para B. bovis, B. bigemina e para infecção mista foi de 24.87% (199/800), 20.75% (166/800) e 18.75% (150/800), respectivamente. Na PCR foi detectado a presença de B. bovis em 15% (18/199) e de B. bigemina em 16% (19/199) dos animais, sendo que destes, 58% (11/19) apresentavam-se co-infectados pelos dois agentes. Os resultados mostram uma baixa prevalência de anticorpos anti-B. bovis e anti-B. bigemina em búfalos da Ilha do Marajó. Porém, observou-se que os agentes da babesiose bovina circulam em búfalos, podendo estes atuar como reservatórios.
Abstract in English:
ABSTRACT.- Tembue A.A.M., Silva F.J.M., Silva J.B., Santos T.M., Santos H.A., Soares C.O. & Fonseca A.H. 2011. Risk factors associated with the frequency of antibodies against Babesia bovis and Babesia bigemina in cattle in southern Mozambique. Pesquisa Veterinária Brasileira 31(8):663-666. Laboratório de Doenças Parasitárias, Departamento de Parasitologia Animal, Universidade Federal Rural de Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br
The study aimed to evaluate the risk factors associated with the frequency of IgG antibodies against Babesia bovis and B. bigemina in cattle in southern Mozambique. Eight hundred and nine serum samples were collected from cattle in three provinces namely Maputo, Gaza and Inhambane, and tested by indirect enzyme-linked immunosorbent assay (i-ELISA) to assess the humoral immune response towards B. bovis and B. bigemina. The chi-square test at 5% significance was used to determine whether there was an association between gender, age and geographic origin of seropositive animals. The overall prevalence was 78.8% (548/695) for B. bovis and 76.0% (528/695) for B. bigemina. The origin of the animals showed a significant association (p<0.05) with seropositivity to both agents, while gender and age was not associated (p>0.05). Maputo province had the highest rate of positive animals, with 93.7% (118/126) for B. bovis and 97.6% (123/126) for B. bigemina. In Gaza province 77.3% (321/415) of the animals were positive for B. bovis and 67.5% (280/415) for B. bigemina, while in the province of Inhambane the levels of seropositivity were 70.8% (109/154) and 81.2% (125/154) for B. bovis and B. bigemina respectively. In the present study, the frequency of cattle positive for B. bovis and B. bigemina was shown to increase among older age groups, suggesting that infection and re-infection persisted even after the primary infection. Thus, this region is considered to be in a state of enzootic stability with regards to B. bovis and B. bigemina.
Abstract in Portuguese:
RESUMO.- Tembue A.A.M., Silva F.J.M., Silva J.B., Santos T.M., Santos H.A., Soares C.O. & Fonseca A.H. 2011. Risk factors associated with the frequency of antibodies against Babesia bovis and Babesia bigemina in cattle in southern Mozambique. [Fatores de risco associados à frequência de anticorpos contra Babesia bovis e B. bigemina em bovinos da Região Sul de Moçambique.] Pesquisa Veterinária Brasileira 31(8):663-666. Laboratório de Doenças Parasitárias, Departamento de Parasitologia Animal, Universidade Federal Rural de Rio de Janeiro, Seropédica, RJ 23890-000, Brazil. E-mail: adivaldo@ufrrj.br
Foram avaliados os fatores de risco associados a frequência de anticorpos da classe IgG contra Babesia bovis e B. bigemina em bovinos da região sul de Moçambique. Oitocentos e nove amostras de soros foram coletadas de bovinos em três províncias nomeadamente Maputo, Gaza e Inhambane e testados por ensaio de imunoadsorção enzimático indireto (i-ELISA) para avaliar a resposta imune humoral contra B. bovis e B. bigemina. O teste de Qui-quadrado a 5% de significância foi utilizado para verificar a associação entre as variáveis sexo, faixa etária e origem geográfica com a soropositividade dos animais. A prevalência geral foi de 78,8% (548/695) para B. bovis e 76,0% (528/695) para B. bigemina. A origem dos animais apresentou associação (p<0,05) com a soropositividade a ambos os agentes, enquanto variável sexo não apresentou associação (p>0,05). A província de Maputo apresentou a maior taxa de animais positivos, com 93,7% (118/126) para B. bovis e 97,6% (123/126) para B. bigemina. Na província de Gaza a soropositividade foi de 67,5% (280/415) para B. bigemina e 77,3% (321/415) para B. bovis enquanto que na província de Inhambane a positividade foi de 81,2% (125/257) e 70,8% (109/257) para B. bigemina e B. bovis, respectivamente. Na presente pesquisa, a freqüência de bovinos positivos para B. bovis e B. bigemina aumentou nas faixas etárias superiores, sugerindo que as infecções e as re-infecções persistem mesmo após primo-infecção. A região estudada apresenta-se na condição de estabilidade enzoótica para os agentes estudados.
Abstract in English:
ABSTRACT.- Baldani C.D., Machado R.Z., Raso T.F. & Pinto A.A. 2007. Serodiagnosis of Babesia equi in horses submitted to exercise stress. Pesquisa Veterinária Brasileira27(4):179-183. Departamento de Patologia Veterinária, Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP 14870-000, Brazil. E-mail: zacarias@fcav.unesp.br
A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.
Abstract in Portuguese:
ABSTRACT.- Baldani C.D., Machado R.Z., Raso T.F. & Pinto A.A. 2007. Serodiagnosis of Babesia equi in horses submitted to exercise stress. Pesquisa Veterinária Brasileira27(4):179-183. Departamento de Patologia Veterinária, Universidade Estadual Paulista, Faculdade de Ciências Agrárias e Veterinárias, Via de Acesso Prof. Paulo Donato Castellane s/n, Jaboticabal, SP 14870-000, Brazil. E-mail: zacarias@fcav.unesp.br
A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.
Abstract in English:
ABSTRACT.- Almeida M.B., Tortelli F.P., Riet-Correa B., Ferreira J.L.M., Soares M.P., Farias N.A.R., Riet-Correa F. & Schild A.L. 2006. [Tick fever in southern Brazil: a retrospective study of 1978-2005.] Tristeza parasitária bovina na região sul do Rio Grande do Sul: estudo retrospectivo de 1978-2005. Pesquisa Veterinária Brasileira 26(4):236-242. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br
A retrospective study of tick fever was made, which occurred from 1978-2005 in southern Rio Grande do Sul in the influence area of the Regional Diagnostic Laboratory of the Federal University of Pelotas. From 4,884 cattle specimens, sent by practitioners or which were from necropsies performed at the Diagnostic Laboratory, 231 (4.7%) were diagnosed as tick fever. Data from 221 of those outbreaks were analyzed. Ninety one (41.1%) outbreaks were caused by Babesia bovis, 11 (4.9%) by Babesia bigemina, and 65 (29.41%) by Anaplasma marginale. In other 33 (14.93%) outbreaks of babesiosis there is no information if the disease was caused by B. bovis or B. bigemina, and 21 (9.5%) outbreaks were caused by mixed infection of A. marginale and B. bovis or B. bigemina. Mean morbidity, mortality, and letality rates in 149 outbreaks were 11.17%, 6.81%, and 70.04%, respectively. Most outbreaks occurred during summer (January-March) and autumn (April-June), mainly in 1 to 3-year-old cattle. Clinical signs were depression, weakness, fallen ears, fever, and weight loss. Low packed cell volume values were always found. Hemoglobinury was observed in babesiosis. Neurological signs characterized by gait alterations, muscular tremors, aggressiveness and falling down with tonic and clonic convulsions were observed in babesiosis by B. bovis. The main gross lesions were anemia, jaundice, splenomegaly, hepatomegaly, yellow liver and cardiac hemorrhages. Hemoglobinury was observed in babesiosis, and congestion of the cerebral cortex in babesiosis by B. bovis. It is concluded that B. bovis is the main agent causing thick fever in southern Rio Grande do Sul. In that region with a cattle population of 2,630,000 heads the annual losses due to tick fever can be estimated in 6,220 cattle or US$ 1,623,000.00. Preventive measures to diminish tick fever losses in the region are necessary.
Abstract in Portuguese:
ABSTRACT.- Almeida M.B., Tortelli F.P., Riet-Correa B., Ferreira J.L.M., Soares M.P., Farias N.A.R., Riet-Correa F. & Schild A.L. 2006. [Tick fever in southern Brazil: a retrospective study of 1978-2005.] Tristeza parasitária bovina na região sul do Rio Grande do Sul: estudo retrospectivo de 1978-2005. Pesquisa Veterinária Brasileira 26(4):236-242. Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas, Campus Universitário s/n, Pelotas, RS 96010-900, Brazil. E-mail: alschild@terra.com.br
A retrospective study of tick fever was made, which occurred from 1978-2005 in southern Rio Grande do Sul in the influence area of the Regional Diagnostic Laboratory of the Federal University of Pelotas. From 4,884 cattle specimens, sent by practitioners or which were from necropsies performed at the Diagnostic Laboratory, 231 (4.7%) were diagnosed as tick fever. Data from 221 of those outbreaks were analyzed. Ninety one (41.1%) outbreaks were caused by Babesia bovis, 11 (4.9%) by Babesia bigemina, and 65 (29.41%) by Anaplasma marginale. In other 33 (14.93%) outbreaks of babesiosis there is no information if the disease was caused by B. bovis or B. bigemina, and 21 (9.5%) outbreaks were caused by mixed infection of A. marginale and B. bovis or B. bigemina. Mean morbidity, mortality, and letality rates in 149 outbreaks were 11.17%, 6.81%, and 70.04%, respectively. Most outbreaks occurred during summer (January-March) and autumn (April-June), mainly in 1 to 3-year-old cattle. Clinical signs were depression, weakness, fallen ears, fever, and weight loss. Low packed cell volume values were always found. Hemoglobinury was observed in babesiosis. Neurological signs characterized by gait alterations, muscular tremors, aggressiveness and falling down with tonic and clonic convulsions were observed in babesiosis by B. bovis. The main gross lesions were anemia, jaundice, splenomegaly, hepatomegaly, yellow liver and cardiac hemorrhages. Hemoglobinury was observed in babesiosis, and congestion of the cerebral cortex in babesiosis by B. bovis. It is concluded that B. bovis is the main agent causing thick fever in southern Rio Grande do Sul. In that region with a cattle population of 2,630,000 heads the annual losses due to tick fever can be estimated in 6,220 cattle or US$ 1,623,000.00. Preventive measures to diminish tick fever losses in the region are necessary.
Abstract in English:
ABSTRACT.- D’Andrea L.A.Z., Sartor I.F., Madruga C.R., Freitas S.B.Z., Kroll L.B. & Kronka S.N. 2005. [Immunological condition of cattle in Holstein and Nelore breed in regard to Babesia bovis and B. bigemina in two regions of the State of São Paulo.] Condição imunológica de bovinos das raças Holandesa e Nelore frente a Babesia bovis e B. bigemina em duas regiões do Estado de São Paulo. Pesquisa Veterinária Brasileira 26(2):74-78.. Seção de Biologia Médica do Instituto Adolfo Lutz, Laboratório Regional de Presidente Prudente, Av. Cel José Soares Marcondes 2357, Presidente Prudente, SP 19013-050, Brazil. E-mail: zampieri@ial.sp.gov.br
The immunological reply of a population to an infectious agent can vary between races and handling of this population. Regional research becomes important, in order to know the interrelation between the agent and its host. In this way, the occurrence of immunoglobulins of class G, anti-Babesia bovis and anti-Babesia bigemina in the Nelore (Bos indicus) and Hostein breed (Bos taurus), was investigated in two regions of the State of São Paulo, 300 km distant from each other. For the indirect method of ELISA, 1,161 bovine serum samples were tested. The medium frequencies of antibodies showed that in the two regions exists an enzootic stability for B. bovis in both breeds studied; even so there was a tendency of marginal area for the Nelore breed in one of the regions. Regarding B. bigemina, in both regions exists enzootic stability for the Hostein and enzootic instability for the Nelore breed. Therefore, acute cases of the disease or specific outbreaks by B. bigemina infection in the Nelore breed may occur in these regions.
Abstract in Portuguese:
ABSTRACT.- D’Andrea L.A.Z., Sartor I.F., Madruga C.R., Freitas S.B.Z., Kroll L.B. & Kronka S.N. 2005. [Immunological condition of cattle in Holstein and Nelore breed in regard to Babesia bovis and B. bigemina in two regions of the State of São Paulo.] Condição imunológica de bovinos das raças Holandesa e Nelore frente a Babesia bovis e B. bigemina em duas regiões do Estado de São Paulo. Pesquisa Veterinária Brasileira 26(2):74-78.. Seção de Biologia Médica do Instituto Adolfo Lutz, Laboratório Regional de Presidente Prudente, Av. Cel José Soares Marcondes 2357, Presidente Prudente, SP 19013-050, Brazil. E-mail: zampieri@ial.sp.gov.br
The immunological reply of a population to an infectious agent can vary between races and handling of this population. Regional research becomes important, in order to know the interrelation between the agent and its host. In this way, the occurrence of immunoglobulins of class G, anti-Babesia bovis and anti-Babesia bigemina in the Nelore (Bos indicus) and Hostein breed (Bos taurus), was investigated in two regions of the State of São Paulo, 300 km distant from each other. For the indirect method of ELISA, 1,161 bovine serum samples were tested. The medium frequencies of antibodies showed that in the two regions exists an enzootic stability for B. bovis in both breeds studied; even so there was a tendency of marginal area for the Nelore breed in one of the regions. Regarding B. bigemina, in both regions exists enzootic stability for the Hostein and enzootic instability for the Nelore breed. Therefore, acute cases of the disease or specific outbreaks by B. bigemina infection in the Nelore breed may occur in these regions.
Abstract in English:
ABSTRACT:
Guimarães A.M., Lima J.D. & Ribeiro M.EB. 2003. Ultrastructure of Babesia equitrophozoites isolated in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 23(3):101-104. [Ultra-estrutura de trofozoítos de Babesia equi isolados em Minas Gerais, Brasil.] Departamento de Medicina Veterinária, Universidade Federal de Lavras, Cx. Postal 37, Lavras, MG 37200-000, Brazil. E-mail: amg@ufla.br
A transmission electron microscope study was carried out on Babesia equi obtained from a splenectomized horse, from the municipality of Santa Luzia, Minas Gerais, Brazil. The isolate was inoculated into two splenectomized foals (1.05 x 1010 parasitized erythrocytes by B. equi). Trophozoites have a single membrane in direct contact with the cytoplasm of the red blood cells, a prominent nucleus, well-developed rough and smooth endoplasmic reticulum, numerous free ribosomes and small food vacuóles. B. equi trophozoites have a cytostome and a long tubular feeding structure in direct contact with the blood plasmá.
Abstract in Portuguese:
RESUMO:
Guimarães A.M., Lima J.D. & Ribeiro M.EB. 2003. Ultrastructure of Babesia equitrophozoites isolated in Minas Gerais, Brazil. Pesquisa Veterinária Brasileira 23(3):101-104. [Ultra-estrutura de trofozoítos de Babesia equi isolados em Minas Gerais, Brasil.] Departamento de Medicina Veterinária, Universidade Federal de Lavras, Cx. Postal 37, Lavras, MG 37200-000, Brazil. E-mail: amg@ufla.br
Neste estudo de microsco-pia eletrônica de transmissão utilizou-se um isolado de Babesia equi obtido a partir de um eqüino esplenectomizado, oriundo do município de Santa Luzia, Minas Gerais, Brasil. O isolado foi inoculado em dois potros esplenectomizados (1,05 x 1010 eritrócitos parasitados com B. equi). Os trofozoítos apresentaram uma membrana simples em· contato direto com o citoplasma das· hemácias, núcleo proeminente, retículo endoplasmático liso e rugoso bem desenvolvidos, numerosos ribosomos livres e pequenos vacúolos alimentares. Em trofozoítos de B. equi foi observado citostoma e uma longa estrutura tubular de alimentação em contato direto com o plasma sangüíneo.
Abstract in English:
ABSTRACT.- Mendonça C.L., Vieira D., Kohayagawa A., Schenk M.A.M., Madruga C.R. & Afonso J.A.B. 2003. [Clinical and hematological evaluation of Nelore calves experimentally infected with isolates of Babesia bigemina from the Southeastern, Northeastern and Northern regions of Brazil.] Avaliação clínica e hematológica em bezerros Nelore infectados experimentalmente com isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil. Pesquisa Veterinária Brasileira 23(2):52-60. Clínica de Bovinos, Campus Garanhuns, Univ. Fed. Rural de Pernambuco, Cx. Postal 152, Garanhuns, PE 55292-901, Brazil.
A comparative study was made regarding the clinical and hematological alterations caused by isolates of Babesia bigemina from southeastern, northeastern and northern Brazil in experimentally infected Nelore calves. Eighteen calves between 7 and 9 months of age, without antibodies against Babesia sp and raised free from ticks, were used. Three animais were previously inoculated with 2.0x109 parasitic erythrocytes (PE) for each stabilate. The other 15 calves were subdivided into three groups, with five animais each, that were subinoculated with 1.0x1010 PE of the respective isolates. The clinical and hematological alterations were evaluated by the determination of parasitaemia, haemogram, plasmatic fibrinogen,reticulocyte count, descriptive analysis of the bone marrow and erythrocytic osmotic fragility, for 30 days, totalizing seven moments of observation. The follow-up of the immunological response by the indirect fluorescent antibody test was carried out daily until the 10th day after inoculation (DAI) and after that, on the 151 20th, 25th and 30th DAI. A mild clinical manifestation of the disease was observed. The laboratory findings revealed low leveis of parasitaemia; decrease of the erythrogram values; absence of reticulocytes, initial decrease in the total count of leukocytes, neutrophils and lymphocytes with a posterior elevation of the number of these cells; hypercellularity of the erythrocytic series and decrease of the myeloid: erythroid relation which was more accentuated between the 8th and 12th DAI, and an increase of the erythrocytic osmotic fragility in the groups inoculated with the Southeast and Northeast isolates. None of the three isolates of B. bigemina gave rise to the clinical characteristic form of the disease, although they induced an humoral immune response.
Abstract in Portuguese:
RESUMO.- Mendonça C.L., Vieira D., Kohayagawa A., Schenk M.A.M., Madruga C.R. & Afonso J.A.B. 2003. [Clinical and hematological evaluation of Nelore calves experimentally infected with isolates of Babesia bigemina from the Southeastern, Northeastern and Northern regions of Brazil.] Avaliação clínica e hematológica em bezerros Nelore infectados experimentalmente com isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil. Pesquisa Veterinária Brasileira 23(2):52-60. Clínica de Bovinos, Campus Garanhuns, Univ. Fed. Rural de Pernambuco, Cx. Postal 152, Garanhuns, PE 55292-901, Brazil.
O presente trabalho teve por objetivo estudar comparativamente as alterações clínicas e hematológicas desencadeadas por isolados de Babesia bigemina das regiões Sudeste, Nordeste e Norte do Brasil em bezerros Nelore infectados experimentalmente. Foram utilizados 18 bezerros com idade entre sete e nove meses, isentos de anticorpos contra Babesia sp. e criados livres de carrapatos. Três animais foram previamente inoculados com 2,0x109 eritrócitos parasitados (EP) para cada isolado. Os outros 15 bezerros foram subdivididos em três grupos de cinco animais, que foram subinoculados com 1,0x1010 EP dos respectivos isolados. Foram avaliadas as alterações clínicas e hematológica por meio da determinação da parasitemia, do hemograma, do fibrinogênio plasmático, da contagem de reticulócitos, da análise descritiva da medula óssea e da fragilidade osmótica eritrocitária, no decorrer de 30 dias, perfazendo um total de sete momentos de observação. O acompanhamento da resposta imunológica pelo teste de imunofluorescência indireta foi realizado diariamente até o 10º dia pós-inoculação (DPI) e posteriormente no 15º, 20º, 25º e 30º DPI. Clinicamente, observou-se uma manifestação muito branda da doença. Os achados laboratoriais revelaram baixos níveis de parasitemia; decréscimo nos valores do eritrograma; ausência de reticulócitos; diminuição inicial na contagem total dos leucócitos, neutrófilos e linfócitos com posterior elevação do número destas células; hipercelularidade da série eritrocítica e decréscimo da relação mielóide:eritróide mais acentuada entre o 8º e 12º DPI e um aumento da fragilidade osmótica eritrocitária nos grupos inoculados com os isolados sudeste e nordeste. Nenhum dos três isolados de B. bigemina desencadeou a forma clínica característica da enfermidade, apesar de induzirem uma resposta imune humoral.
Abstract in English:
ABSTRACT.- Madruga C.R., Leal C.R.B., Ferreira A.M.T., Araújo F.R., Bonato A.L.V., Kessler R.H., Schenk M.A.M. & Soares C.O. 2002. Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil. Pesquisa Veterinária Brasileira 22(4):153- 160. [Análise genética e antigênica de isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil.] Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil.
A molecular epidemiological study was performed with Babesia bigemina isolates from five geographical regions of Brazil. The genetic analysis was done with random amplification of polymorphic DNA (RAPD), repetitive extragenic palindromic elements-polymerase chain reaction (REP-PCR) and enterobacterial repetitive intergenic consensus sequences-polymerase chain reaction (ERIC-PCR) that showed genetic polymorphism between these isolates and generated fingerprinting. In RAPD, IL0872 and IL0876 primers were able to detect at least one fingerprinting for each B. bigemina isolate. The amplification of B. bigemina DNA fragments by REP-PCR and ERIC-PCR gave evidence for the presence in this haemoprotozoan of the sequences described previously in microorganisms of the bacterial kingdom. For the first time it was demonstrated that both techniques can be used for genetic analysis of a protozoan parasite, although the ERIC-PCR was more discriminatory than REP-PCR. Toe dendogram with similarity coeficiente among isolates showed two clusters and one subcluster. The Northeastern and Mid-Westem isolates showed the greatest genetic diversity, while the Southeastem and Southem isolates were the closest. Toe antigenic analysis was done through indirect fluorescent antibodytechnique and Westem blotting using a panei of monoclonal antibodies directed against epitopes on the merozoite membrane surface, rhoptries and membrane of infected erythrocytes. As expected, the merozoite variable surface antigens, major surface antigen (MSA)-1 and MSA-2 showed antigenic diversity. However, B cell epitopes on rhoptries and infected erythrocytes were conserved among all isolates studied. In this study it was possible to identify variable and conserved antigens, which had already been described as potential immunogens. Considering that an attenuated Babesia clone used as immunogen selected populations capable of evading the immunity induced by this vaccine, it is necessary to evaluate more deeply the cross-protection conferred by genetically more distant Brazilian B. bigemina isolates and make an evaluation of the polymorphism degree of variable antigens such as MSA-1 and MSA-2.
Abstract in Portuguese:
RESUMO.- Madruga C.R., Leal C.R.B., Ferreira A.M.T., Araújo F.R., Bonato A.L.V., Kessler R.H., Schenk M.A.M. & Soares C.O. 2002. Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil. Pesquisa Veterinária Brasileira 22(4):153- 160. [Análise genética e antigênica de isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil.] Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil.
Um estudo de epidemiologia molecular foi executado com isolados de Babesia bigemina das cinco regiões fisiográficas do Brasil. A análise genética foi feita com amplificação aleatória de DNA polimórfico (RAPD), reação da polimerase em cadeia com seqüências de elementos extragênicos repetitivos palindrômicos (REP-PCR) e reação da polimerase em cadeia com seqüências repetitivas enterobacterianas intergênicas de consenso (ERIC-PCR) que apresentaram polimorfismo genético entre os isolados e geraram marcadores. No RAPD com os oligonucleotídeos iniciadores IL0872 e IL0876, foi possível detectar pelo menos um marcador por isolado de B. bigemina. A amplificação de fragmentos de DNA de B. bigemina por REPPCR e ERIC-PCR demonstrou a presença dessas seqüências, descritas anteriormente somente em microrganismos bacterianos, nesse hemoprotozoârio, e, pela primeira vez, foi verificado que podem ser utilizadas para análise genética de um protozoário. O ERIC-PCR foi mais discriminatório que o REP-PCR. O dendograma formado com o coeficiente de similaridade entre os isolados evidenciou dois agrupamentos e um subgrupo. Os isolados do Nordeste e Centro-Oeste demonstraram maior diversidade genética, enquanto que os isolados do Sudeste e Sul foram os mais próximos. A análise antigênica foi executada por meio de imunofluorescência indireta e Western blotting usando um painel de anticorpos monoclonais direcionados a epitopos B na membrana dos merozoítos, roptries e membrana de eritrócitos infectados. Os antígenos variáveis da superfície dos merozoítos, antígeno principal da superfície do merozoíto (APSM)-1 e APSM-2 apresentaram diversidade antigênica. Entretanto, os epítopos de células B nas roptries e nos eritrócitos infectados foram conservados em todos os isolados. Nesse estudo foi possível identificar antígenos variáveis e conservados que anteriormente haviam sido descritos como potenciais imunógenos. Considerando que um clone atenuado de Babesia utilizado para imunização selecionou populações capazes de evadir a resposta imune à vacina, torna-se necessário avaliar mais detalhadamente a imunidade cruzada existente entre os isolados brasileiros mais distantes geneticamente e realizar uma avaliação do grau de polimorfismo dos antígenos variáveis APSM-1 e APSM-2.
Abstract in English:
ABSTRACT.- Madruga C.R., Marques A.P.C., Araújo F.R., Miguita M., Carvalho C.M.E., Araújo F.S., Umaki A.C.S., Crocci A.J. & Queiróz R.A. 2001. Evaluation of an ELISA for detection of antibodies to Babesia bigemina in cattle and it's application in an epidemiological survey in Brazil. [Avaliação de um ELISA para detecção de anticorpos contra Babesia bigemina em bovinos e sua aplicação em um inquérito sorológico no Brasil.] Pesquisa Veterinária Brasileira 21 (2):72-76. Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil.
An indirect enzyme-linked immunosorbent assay (ELISA) using a crude antigen was evaluated for its performance to detect Babesia bigemina antibodies. The sensitivity and specificity were 98.0% and 99.0%, respectively. ln agreement with the high specificity, no cross-reactions were verified with sera from calves inoculated three times with 107 Babesia bovis organisms. With regard to the comparison of ELISA and indirect fluorescent antibody test (lFAT) in detecting antibodies against B. bigemina in calves experimentally infected with five Brazilian geographical isolates of this hemoparasite, lFAT was able to detect antibodies one day earlier in most of the calves' sera. There was a good agreement between results shown by ELISA and IFAT with sera from an enzootically stable area (k=0.61). However, there was no agreement between these serological tests with sera from an enzootically unstable area (k=0.33). The ELISA was employed in an epidemiological survey using with 1,367 sera from four counties in the Pantanal of Mato Grosso do Sul and characterized this region as an enzootically stable area, since the prevalence ranged from 87.7 to 98.9%. Therefore, this ELISA with high sensitivity, specificity and performance similar to lFAT can be employed in serological diagnosis of B. bigemina.
Abstract in Portuguese:
RESUMO.- Madruga C.R., Marques A.P.C., Araújo F.R., Miguita M., Carvalho C.M.E., Araújo F.S., Umaki A.C.S., Crocci A.J. & Queiróz R.A. 2001. Evaluation of an ELISA for detection of antibodies to Babesia bigemina in cattle and it's application in an epidemiological survey in Brazil. [Avaliação de um ELISA para detecção de anticorpos contra Babesia bigemina em bovinos e sua aplicação em um inquérito sorológico no Brasil.] Pesquisa Veterinária Brasileira 21 (2):72-76. Embrapa Gado de Corte, Rodovia BR 262 Km 4, Cx. Postal 154, Campo Grande, MS 79002-970, Brazil.
Um ensaio de imunoadsorção enzimática (ELISA) baseado em antígeno bruto foi avaliado na detecção de anticorpos contra Babesia bigemina. A sensibilidade e a especificidade do teste foram de 98,0% e 99,0%, respectivamente. Concordando com a alta especificidade do teste, não foram verificadas reações cruzadas com soros de bezerros inoculados três vezes com 107 merozoítos de Babesia bovis. Com relação à comparação do ELISA com a imunofluorescência indireta (IFAT) na detecção de anticorpos contra B. bigemina em bezerros experimentalmente infectados com cinco isolados brasileiros geograficamente distintos deste hemoparasito, o IFAT foi capaz de detectar anticorpos um dia antes do ELISA na maioria dos soros dos animais. Houve urna boa concordância entre os resultados encontrados no ELISA e no IFAT com soros de bovinos de região de estabilidade enzoótica (k=0.61 ). No entanto, não houve concordância entre os testes sorológicos com soros de animais de área de instabilidade enzoótica (k=0.33). O ELISA foi empregado em um inquérito epidemiológico com 1.367 soros de quatro municípios do Pantanal de Mato Grosso do Sul e caracterizou esta região corno urna área de estabilidade enzoótica, urna vez que as prevalências variaram de 87, 7 a 98,9%. Dessa forma, este ELISA, que apresentou alta sensibilidade, especificidade e desempenho similar ao IFAT, pode ser utilizado no diagnóstico sorológico de B. bigemina.