Histopathological and molecular methods as complementary diagnostic in case of lymphadenopathies suggestive of bovine tuberculosis.

Bovine tuberculosis (TB) is a chronically evolving zoonotic infectious disease caused by Mycobacterium bovis. Anatomopathological examination during post mortem inspection in bovines is the main resource engaged in sanitary slaughter; however, it is very troublesome since many granulomatous inflammatory processes have similar morphological characteristics. Thus, this study aims to use complementary diagnosis methods (histopathological and polymerase chain reaction - PCR assays) to confirm the macroscopic assessment of lymphadenopathies indicative of tuberculosis in bovines slaughtered in a refrigerated slaughterhouse in Tailândia city, PA, Brazil. Fifty­one samples were collected from lesions indicative of tuberculosis in pre­scapular and pre­pectoral lymph nodes (or different lymphadenitis) in condemned carcasses. Histological processing employed routine techniques carried out at the Laboratory of Animal Pathology of the Federal Rural University of the Amazon, while the PCR assay was performed at the Bacteriology Laboratory of the Evandro Chagas Institute. Results showed that 1.96% of the histopathology samples corresponded to inflammatory processes typical of TB and that, in PCR, 4.25% of the samples had the amplification profile of the M. bovis species. These results indicate the importance of adding complementary methods to assist the sanitary inspection line and make inspection more efficient in its decisions.

Low transmission rates of Equine infectious anemia virus (EIAV) in foals born to seropositive feral mares inhabiting the Amazon delta region despite climatic conditions supporting high insect vector populations.

BACKGROUND: Marajó Island, within in the Amazon River Delta, supports numerous bands of feral equids including the genetically distinct Marajoara horses. Approximately 40% of the equids on the island are infected with Equine infectious anemia virus (EIAV). This high seropositivity rate coupled with the need to preserve rare breeds such as the Marajoara horse precludes euthanasia as the primary means for controlling EIAV in this region. In the absence of iatrogenic transmission, spread of this lentivirus is mediated primarily by hematophagous insects, whose year-round prevalence on the island is supported by favorable climatic conditions. In addition, cases of vertical EIAV transmission have been observed suggesting inclusion of seropositive mares in restorative breeding programs could result in their progeny becoming infected with this virus either pre-parturition or post-partum via hematophagous insects. Therefore, the aim of this study was to evaluate EIAV vertical and post-partum insect-mediated transmission rates among foals born to seropositive feral mares until natural weaning. Serum samples from foals born to seropositive feral mares within the Soure municipality, of Marajó Island, were collected to investigate their serological status, using an indirect ELISApgp45, with positive samples confirmed using the classical agar gel immunodiffusion (AGID) assay. RESULTS: The serological status of 28 foals were monitored over a 2-year period with some subjects, depending on their date of birth, being sampled up to six times. All foals remained with their respective mares until fully weaned at approximately 10 months of age. Only 2 foals (7.14%) in the study group became seropositive against EIAV. CONCLUSION: The results demonstrate that in most cases it is possible to obtain seronegative foals born to and eventually weaned by EIA positive mares, even in equatorial regions where substantial rainfall and high temperatures favor the proliferation of insect vectors.

Glanders and brucellosis in equids from the Amazon region, Brazil.

Glanders and brucellosis are zoonotic infectious diseases that affect equids in several countries worldwide. On Marajó Island (Amazon region of Brazil), Marajoara and Puruca horses, which are well adapted to the climatic and territorial adversities of the region, play a fundamental role in the local economy and in the sociocultural lives of the population. However, these animals have undergone a drastic reduction in number, markedly due to precarious veterinary care, unknown causes of morbidity and mortality, and disordered crossing with other breeds introduced to the island. Thus, this study aimed to investigate the occurrence of glanders and brucellosis in equids on a property located in the municipality of Soure, Marajó Island (Brazil). Serum samples were collected from 388 animals (357 horses and 31 mules), maintained in an extensive breeding system, in a property that was also extensively breeding buffaloes, goats, and sheep, with contact among species. The sera were tested for glanders using an indirect ELISA (ELISAi), and the results were confirmed by immunoblotting. The diagnosis of brucellosis was made using the Rose Bengal test (RBT) and confirmed through the Serum Agglutination test (SAT) and 2-mercaptoethanol test. In the case of glanders, 2.31% (9/388) of animals were positive in ELISAi test, of which eight had results confirmed by immunoblotting, representing 2.06% seropositivity in the entire herd. For brucellosis, serum samples from 6.7% (26/388) horses were reactive in the RBT, of which 4.12% (18/388) had a titer ≥50 and 2.06% (8/388) had a titer ≥100 in the SAT. This is the first study to report the occurrence of glanders and equine brucellosis in the municipality of Soure/Marajó Island. Monitoring the occurrence of such diseases is extremely important since they affect the herds economically and zootechnically, in addition to their high zoonotic potential. The number of animals sampled in this study, as well as the way they are raised and managed, is representative of the total equid population of the island. These results, combined with previous studies on buffaloes, indicate that these diseases are endemic in the Marajo Island.

"Which Factors Affect Pregnancy Until Calving and Pregnancy Loss in Buffalo Recipients of in vitro Produced Embryos?"

In vitro embryo production and embryo transfer (ET) in buffaloes has been developed for decades. However, most studies are focused on the donor or laboratory improvements, and there is a lack of reports regarding the recipients. Therefore, our aim was to investigate factors associated to pregnancy (P/ET), pregnancy loss (PL), and calving rates in buffalo recipients. The studied factors were season, recipient parity, the synchronization protocol, the CL diameter, asynchrony between the embryo and the recipient, the day of the recipient estrous cycle, the embryo (fresh vs. vitrified), the day of embryo development, and the embryo stage. These retrospective data, from a program of in vitro produced embryos, were analyzed by logistic regression, and the odds ratio was also estimated. Two factors were related to P/ET and the calving rate: (1) progesterone associated to estradiol plus eCG protocol for fixed time ET tended to affect positively P/ET on day 30 (41.9 vs. 36.1%, respectively; P = 0.07; AOR = 1.28) and P/ET on day 60 (37.8 vs. 36.1%, respectively; P = 0.09; AOR = 1.08) compared to the Ovsynch protocol; and (2) the CL diameter (≥14.5 mm) at transfer increased P/ET on day 30 (47.4 vs. 32.5%; P < 0.01; AOR = 1.87) and on day 60 (45.3 vs. 27.7%; P < 0.01; AOR = 2.16), and also the calving rate (37.9 vs. 21.7%; P < 0.01; AOR = 2.20). PL was greater when ET was done in the nonbreeding season compared to the breeding season (PL 30-60: 12.8 vs. 0.0%, P = 0.01; AOR > 999.99; PL 60-calving: 26.8 vs. 3.6%, P = 0.03; AOR = 9.90; and PL 30-calving: 36.2 vs. 3.6%, P = 0.01; AOR = 15.30). In conclusion, the data of our study indicated that the synchronization protocol, the CL diameter, and ET during the breeding season impacted the reproductive efficiency of buffalo recipients.

High positivity values for bovine leukemia virus in human breast cancer cases from Minas Gerais, Brazil.

Bovine leukemia virus (BLV) is a retrovirus that causes lymphoma in cattle worldwide and has also been associated with breast cancer in humans. The mechanism of BLV infection in humans and its implication as a primary cause of cancer in women are not known yet. BLV infection in humans may be caused by the consumption of milk and milk-products or meat from infected animals. Breast cancer incidence rates in Brazil are high, corresponding to 29.5% a year of cancer cases among women. In 2020, an estimated 66,280 new cases of breast cancer are expected, whereas in 2018 breast cancer has led to 17,572 deaths, the highest incidence and lethality among cancers in women in this country that year. BLV infection occurrence ranges from 60 to 95% in dairy herds. In addition, there are some regions, such as the Minas Gerais State, southeastern Brazil, where the population traditionally consume unpasteurized dairy products. Taken together, this study aimed to verify if there is a higher association between breast cancer and the presence of BLV genome in breast tissue samples within this population that consumes raw milk from animals with high rates of BLV infection. A molecular study of two BLV genes was carried out in 88 breast parenchyma samples, between tumors and controls. The amplified fragment was subjected to BLV proviral sequencing and its identity was confirmed using GenBank. BLV proviral genes were amplified from tumor breast parenchyma samples and healthy tissue control samples from women, revealing a 95.9% (47/49) and 59% (23/39) positivity, respectively. Our results show the highest correlation of BLV and human breast cancer found in the world to date within the population of Minas Gerais, Brazil.

Genetic material from buffalo and cattle: crucial importance in the formalization of bilateral trade between India and Brazil.

The trade in live animals between India and Brazil dates from the late nineteenth century when European travellers traded animals of Indian origin for display in zoos. Considering the origin of coffee and sugar cane, as well as the expertise related to mineral evaluation, we need to consider that India was involved in important economic cycles of Brazil, even indirectly. This virtuous flow of trade has been maintained and intensified throughout modern history, especially after these two nations gained political independence from their colonisers, thereby becoming independent in mercantile affairs. This paper addresses the main points related to the use of animals of Indian origin in Brazil. We revisit some of the historical aspects of the process of colonisation of Brazil, as well as the importation of animals from India. The restrictions imposed on this process due to the occurrence of diseases in cattle and buffalo in India will be examined. At the end of the text, emphasis will be given to the risks of introducing exotic diseases into Brazil.

Evaluation of post-mortem diagnostic tests' sensitivity and specificity for bovine tuberculosis using Bayesian latent class analysis.

This study aimed to evaluate the performance of real-time PCR (qPCR), ELISA IDEXX™, and bacterial isolation as post-mortem diagnostic tests in animals with lesions compatible with bovine tuberculosis detected by Brazilian Federal Inspection Service as part of the bovine tuberculosis active surveillance. Bayesian latent class models were used to estimate diagnostic tests' sensitivity, specificity, correlations, predictive values and frequency of infected animals. Samples of tuberculosis-suggestive lesions collected by FIS sanitary inspection routine in slaughterhouses from 11 Brazilian states were analyzed. Isolation was the most sensitive technique, 94.54% (95% Credible Interval (CrI) 90.09%-97.65%), qPCR was 64.69% (95% CrI 54.41%-74.15%) sensitive and ELISA IDEXX™ 26.74% (95% CrI 22.82%-30.97%). Tests' specificities were 98.19% (95% CrI 95.75%-99.45%), 93.49% (95% CrI 79.28%-99.66%), 95.53% (95% CrI 91.71%-98.02%) respectively. Despite its low sensitivity, ELISA IDEXX™ was able to identify positive samples that were not detected by the other techniques. These samples had high probability to be true positives given ELISA's positive predictive value. The correlations between qPCR and isolation were neither biologically nor statistically significant. The low sensitivity of the qPCR is a limiting factor to its use as a post-mortem diagnosis in bovine tuberculosis suggestive lesions. Its use could be recommended in situations of high prevalence, or in parallel association with other tests, such as ELISA IDEXX™. ELISA IDDEX™ should not be used as a unique test, or in substitution of the other tests, for the post-mortem diagnosis of bovine tuberculosis due to its sensitivity.

Prevalence and risk factors for bovine tuberculosis in Minas Gerais State, Brazil.

The aim of the present study was to estimate the prevalence and regional spread of bovine tuberculosis in the state of Minas Gerais, Brazil, to identify herd-level risk factors, and to provide guidance for disease control and mitigation of its impact in public health. The study comprised a large-scale random sample survey of 22,990 animals and 1586 herds, distributed in seven regions. A questionnaire was applied on each farm to collect epidemiological and herd management data. Overall, 5.04 % of herds and 0.81 % of animals were positive for bovine tuberculosis. The highest herd prevalence was found in Alto Paranaíba, an expanding dairy region. The more technologically advanced dairy herds showed a prevalence ratio of 2.83 compared to others and are obvious candidates for risk-based surveillance and herd accreditation schemes. Small farms cannot be left out of disease control efforts because they are the vast majority of herds, albeit with lower individual risk. With regard to public health, there is widespread practice of producing homemade fresh cheese with raw milk and of slaughtering culled cows in places without sanitary inspection. This poses a risk to consumers and limits the efficacy of surveillance at slaughter.

Molecular detection of bovine immunodeficiency virus in water buffaloes (Bubalus bubalis) from the Amazon region, Brazil.

Bovine immunodeficiency is a chronic progressive disease caused by a lentivirus that affects cattle and buffaloes. Although the infection has been described in cattle in some countries, including in Brazil, there are only two reports of infection in buffaloes: one in Pakistan and one in Cambodia. The aim of the present study was to survey the occurrence of bovine immunodeficiency virus (BIV) in water buffaloes from the Amazon region, Pará state, Brazil. BIV proviral DNA was surveyed in 607 whole blood samples of water buffaloes from 10 farms located in the state of Pará using semi-nested polymerase chain reaction (PCR) (PCR-SN) to amplify the pol region of the viral genome. Of the 607 samples tested, 27 (4.4 %) were positive for BIV proviral DNA. The amplified fragments were confirmed by sequence analysis after cloning and nucleotide sequencing. The sequence obtained had 99 % similarity to the reference strain (R-29). The present study provides important epidemiological data because BIV was detected for the first time in water buffaloes in Brazil. Further, the results suggest the possibility of the virus being a risk factor for herd health because it may be a potential causal agent of chronic disease and, also may be associated to other infectious diseases.

Comparison of the tuberculin test, histopathological examination, and bacterial culture for the diagnosis of tuberculosis (Mycobacterium bovis) in buffaloes (Bubalus bubalis) in Brazil.

Tuberculosis is a disease with a great zoonotic potential. It is considered a major obstacle to cattle production and is responsible for severe losses in several production systems. A comparative cervical test (CCT) was performed in 1140 buffaloes from different mesoregions of the state of Pará, Brazil, with the aim of comparing the sensitivity and specificity of CCT with histopathological examination and bacterial culture. Of the animals tested using CCT, 4.65% (53/1140) were positive, 2.98% (34/1140) were inconclusive, and 92.36% (1053/1140) were negative. Among the 168 sacrificed animals, 33 were positive, 18 were inconclusive, and 117 were negative by CCT, and samples from the sacrificed animals were collected for histopathological examination and bacterial culture. A qualitative evaluation of the tuberculin test was performed by comparing the test results with the histopathological and bacteriological results. The latter two tests yielded a prevalence of 4.16%, a sensitivity of 71.43%, and a specificity of 82.61%. Based on these results, we concluded that CCT yielded satisfactory results and can be applied in diagnostic studies in buffaloes. The prevalence rate obtained using three distinct diagnostic methods suggests that Mycobacterium bovis was present in a few animals in the population evaluated.

Bovine herpesvirus 6 in buffaloes (Bubalus bulalis) from the Amazon region, Brazil.

This study presents the first description of Bovine herpesvirus 6 (BoHV-6) that was isolated from buffaloes of Amazon region in Brazil. Phylogenetic analysis showed that the BoHV-6 Brazilian strains clustered with the sequence of BoHV-6 from elsewhere available at the GenBank. It was observed in some buffaloes with lymphoproliferative disease in one herd, thus the animals were also tested for Bovine leukemia virus (BLV), which has been associated to lymphoma in bovines. All animals were negative to BLV. These results indicate that BoHV-6 is present in buffaloes in Brazil, but the importance and impact of this infection and its association with any illness is still undefined.

Inquérito sorológico da infecção pelos vírus da encefalomielite equina no estado de Minas Gerais, Brasil / Serological survey of equine encephalomyelitis virus infection in the state of Minas Gerais, Brazil

Foi investigada a ocorrência da infecção pelos vírus da Encelafalomielite Equina do Leste (EEE), Encefalomielite Equina do Oeste (WEE) e Encefalomielite Equina Venezuelana (VEE) em equídeos não vacinados contra tais agentes, criados em dez delegacias regionais do estado de Minas Gerais (Almenara, Bambuí, Curvelo, Governador Valadares, Montes Claros, Oliveira, São Gonçalo do Sapucaí, Teófilo Otoni, Unaí e Viçosa), empregando-se a técnica de soroneutralização em microplacas. Dos 826 animais examinados, 30,2% ((250/826) foram soropositivos para EEE e 1,9% (16/826) para o zuelano de Encefalomielite Equina circulam na população equina do estado de Minas Gerais.

The occurrence of Equine Eastern Encephalomyelitis (EEE), Equine Western Encephalomyelitis (WEE) and Equine Venezuelan Encephalomyelitis (VEE) virus infection was investigated in equids not vaccinated against these viruses. The animals were distributed in ten regional districts of the state of Minas Gerais (Almenara, Bambuí, Curvelo, Governador Valadares, Montes Claros, Oliveira, São Gonçalo do Sapucaí, Teófilo Otoni, Unaí e Viçosa). Microplate serum neutralization test was used to detect antibodies against encephalitis virus. Two hundred and fifty animals (30.2%, 250/826) were EEE-seropositive, while 1.9% of them (16/826) were VEE-seropositive. No animals were found to be seropositive for WEE. In conclusion, either EEE or VEE viruses circulate in the equid population of the state of Minas Gerais.

Quill mites in Brazilian psittacine birds (Aves: Psittaciformes).

The primary and secondary feathers of 170 Brazilian psittacine birds (Aves: Psittaciformes) were examined in order to identify feather quill mite fauna. Birds were held captive in two locations in the state of Minas Gerais (MG), and two in the state of Espirito Santo (ES). The quills were cut longitudinally and were examined under optical microscopy. The genus of quill mites most frequently found was Paralgopsis (Astigmata: Pyrogliphidae), followed by Cystoidosoma (Astigmata: Syringobiidae). Astigmata: Syringophilidae mites were sporadically observed. After analyzing the data using logistic regression models, it was determined that there was higher infestation risk for psittacines in ES state, as compared with those in MG, and a significant increase in risk depending on the psittacine host species. However, the location of captivity did not have a significant effect. Lesions were observed in infested feathers. Cystoidosoma sp. and Paralgopsis sp. were always observed together, with parts of Paralgopsis found inside Cystoidosoma sp., suggesting thanatochresis or predation.

Development and validation of a method for purification of mallein for the diagnosis of glanders in equines.

BACKGROUND: The allergic test of mallein is one of the most frequently used tests, together with the Complement Fixation Test (CFT), for the diagnosis of glanders in endemic areas. Mallein, a purified protein derivative (PPD), is produced similarly to PPD tuberculin and the end product is a primarily proteic antigen, which is only poorly purified. The immuno-allergic activity of mallein is believed to be due to a high molecular weight group of proteins present in the antigen. To improve the quality of the antigen, in terms of sensitivity and specificity, a new method of mallein production was developed, in which purification was accomplished by ultrafiltration in a Tangential Flow Filtration system (TFF). RESULTS: The TFF methodology efficiently separated the high and low molecular weight protein groups of mallein. The five TFF-purified malleins, produced from Burkholderia mallei strains isolated from clinical cases of glanders in Brazil, proved to be more potent than standard mallein in the induction of an allergic reaction in sensitized animals. Regarding specificity, two of the purified malleins were equivalent to the standard and three were less specific. CONCLUSION: Some of the TFF-purified malleins showed considerable potential to be used as an auxiliary test in the diagnosis of glanders.

Prevalence and risk factors for brucellosis in Tocantins and Brazilian National Program to fight this disease / Prevalência e Fatores de Risco para Brucelose no Tocantins e o Programa Nacional Brasileiro de combate à doença

Em 2010 foram analisados 1.818 soros de vacas com 24 ou mais meses de idade. Estes animais estavam distribuídos por 102 rebanhos em 16 dos 17 municípios da microrregião de Araguaína, Estado do Tocantins. Foram pesquisadas imunoglobulinas contra Brucella abortus. Os donos ou responsáveis pelas propriedades responderam a um questionário sobre fatores de risco para a brucelose. Os soros positivos no teste do antígeno acidificado e tamponado (AAT) foram submetidos à prova do 2-Mercaptoetanol (2-ME). As prevalências de rebanho e de animal foram de 43,5por cento (42,3-44,8por cento) e de 6,2por cento (6,1-6,2por cento), respectivamente. Excetuando uma pequena região, ao Norte daárea estudada, a prevalência de rebanho deste trabalho é maior (p menor que 0,05) do que aquela encontrada sete anos antes em todas as outras regiões do Estado. Não se verificou diferença na prevalência dadoença em vacas com e sem vacinação (OR igual 1,52; [0,73 a 3,54]). Fazem-se necessários estímulospara remoção das fontes de infecção dos rebanhos e para boas práticas de manejo sanitário. São também necessárias normas mais consistentes para realização e monitoramento da vacinação e para controle do trânsito animal. Programas estaduais próprios, para combate à brucelose, devem serincentivados em respeito às diferenças regionais.

Curva de anticorpos pós-vacinais em ovinos imunizados com uma ou duas doses de bacterina oleosa anti-leptospirose, produzida com a sorovariedade Hardjo, tipo Hardjoprajitno, estirpe Norma, isolada no Brasil / Post vaccinal antibodies profile of sheep immunized with one or two doses of an oil emulsified anti leptospirosis bacterin produced with serovar Hardjo, type Hardjoprajitno, strain Norma, isolated in Brazil

Foi comparado o nível de anticorpos de ovelhas imunizadas com uma ou duas doses de bacterina oleosa produzida com a sorovariedade Hardjo, tipo Hardjoprajitno, estirpe Norma, isolada da urina de bovino no Brasil. Culturas de 2x10(8) leptospiras/mL foram inativadas com formalina a 0,3 por cento, à concentração final e emulsionada em óleo Emulsigen® 12 por cento. A dose da vacina foi padronizada para a concentração de 1x10(8) leptospiras/mL. Quarenta ovinos adultos, da raça Santa Inês, de um rebanho livre de leptospirose por exames clínicos e sorológicos durante um ano foram escolhidos para o experimento. O grupo A (n=15) recebeu duas doses de 3,0mL da vacina por via subcutânea, com intervalo de 30 dias. O grupo B (n=15) recebeu dose única de 3,0mL, via subcutânea e o grupo C (controle) recebeu uma dose subcutânea de 3,0mL de solução 0,85 por cento de cloreto de sódio. Os títulos de anticorpos pós-vacinação foram mensurados pelo teste de soroaglutinação microscópica (SAM) e um teste imunoenzimático (ELISA) a cada 30 dias durante 120 dias. Os títulos dos grupos A e B na primeira colheita variaram de 80 a 160. No grupo A, após a segunda dose, os títulos aumentaram duas a quatro vezes, até 3.200, enquanto no grupo B os títulos de aglutininas foram menores que 160 e diminuíram uma a duas vezes após 60 dias da vacinação. Utilizando-se dose única, os anticorpos persistiram por somente 30 dias e, com duas doses, com 30 dias de intervalo, os anticorpos foram detectáveis por 60 dias por meio do teste de SAM e 120 dias no teste de ELISA. Assim, o teste de SAM detectou títulos de IgM vacinal somente por 60 dias, enquanto o teste de ELISA foi capaz de detectar anticorpos durante os 120 dias. No grupo controle negativo, ocorreram no ELISA reações inespecíficas de títulos até 80, porém no SAM os títulos dos mesmos animais se mantiveram em zero. O teste de ELISA pode ser utilizado para medir anticorpos vacinais para a sorovariedade Hardjo, tipo Hardjoprajitno, estirpe Norma em ovinos.

It was compared the level of antibodies of sheep immunized with one or two doses of an oil bacterin produced with serovar Hardjo, type Hardjoprajitno, strain Norma, isolated from cattle urine in Brazil. Cultures of 2x10(8) leptospires/mL were inactivated with formalin 0.3 percent, final concentration and emulsified in oil Emulsigen® 12 percent. The vaccine dose was standardized to the concentration of 1x10(8) leptospires/mL. Forty adult sheep, Santa Inês breed from a herd free of leptospirosis by clinical and serological examinations during one year were chosen for the experiment. Group A (n=15) received two subcutaneous 3.0 mL vaccine dose, interval of 30 days. Group B (n=15) received one subcutaneous 3.0 mL vaccine dose. Group C (control) received one subcutaneous dose of 3.0 mL of 0.85 percent sodium chloride solution. Post vaccination antibody titers were measured by microscopic agglutination test (MAT) and an in house enzyme linked immunosorbent assay (ELISA) every 30 days during 120 days. At 30 days post-vaccination, the titers of the groups A and B ranged from 80 to 160. In group A, after the booster dose, the titers increased two to four times until 3,200, while in the group B the titers were lower than 160 and decreased by one to two times after 60 days after vaccination. Using a single dose, the antibodies persisted for only 30 days, and with two doses with 30 days of interval, the antibodies were detectable for 60 days through the MAT test and 120 days through the ELISA. The MAT test detected IgM titers of vaccine only for 60 days, while the ELISA was able to detect antibodies during the 120 days. In the negative control group, nonspecific reactions occurred in the ELISA up to titer 80, however titers in the MAT of the same animals remained at zero. The ELISA test can be used to assess anti leptospire vaccinal antibody level to the serovar Hardjo, type Hardjoprajitno, strain Norma in sheep.

Genotyping of the pseudorabies virus by multiplex PCR followed by restriction enzyme analysis.

Suid herpesvirus 1 (SuHV-1) is the causative agent of Aujeszky's disease. The infectious agent has only one serotype, but it was classified by restriction enzyme analysis of the whole genome into four genotypes, named I to IV. The aim of this study was to standardize a rapid method for genotyping SuHV-1 without virus isolation, using a multiplex-PCR followed by enzymatic restriction analysis. The complete genome of the virus was analyzed in silico to determine the restriction sites for the enzyme BamHI. Primers were designed to flank sites with emphasis on certain points of differentiation of genotypes. The standard PCRs were able to detect the SuHV-1 and also to differentiate genotypes from brain tissue of infected pigs. The BamHI-PCR is a rapid, practical, and sensitive way to genotype SuHV-1.

Principais aspectos da infecção por Leptospira sp em ovinos: [revisão] / Main aspects of Leptospira sp infection in sheep: [review]

Este trabalho tem como objetivo revisar as infecções por Leptospira sp em ovinos. São abordados os aspectos epidemiológicos, incluindo a ocorrência no Brasil e as formas de transmissão, os sinais clínicos e as lesões, o diagnóstico e as medidas de prevenção e controle.

An updated review of Leptopspira sp infection in sheep is presented emphasizing some epidemiological aspects including the occurrence of the disease in Brazil and mechanisms of transmission, clinical signs and lesions, diagnosis, prevention and control measures.

Diagnóstico e genotipagem do vírus da pseudoraiva por nested-PCR e análise de restrição enzimática / Diagnosis and genotyping of pseudorabies virus by nested-PCR and restriction enzyme analysis

A pseudoraiva (PR) é uma enfermidade viral responsável por consideráveis perdas econômicas na indústria de suínos. O vírus da pseudoraiva (PrV) apresenta apenas um sorotipo, mas, por análise de restrição enzimática, foi classificado em quatro genótipos denominados I, II, III e IV. Os métodos usados para genotipagem dependem do isolamento do vírus, da purificação do DNA viral, da restrição enzimática do genoma completo e da visualização após eletroforese. O objetivo deste trabalho foi estabelecer um método mais rápido e sensível para detectar e genotipar o PrV por nested-PCR e análise de restrição enzimática. Vinte isolados do PrV das regiões Sul e Sudeste do Brasil e a estirpe padrão Shope foram replicadas em células PK-15 e submetidas à nested-PCR para o gene da glicoproteína E. Além desses vírus previamente isolados, foram avaliadas 75 amostras clínicas de cérebro de suíno em um total de 25 animais positivos para a PR no isolamento e na soroneutralização viral e 50 amostras negativas provenientes de animais negativos na soroneutralização viral e de granjas sem histórico de PR. Todas as amostras clínicas tiveram resultados compatíveis com o isolamento e a soroneutralização, e a totalidade das amostras positivas foi classificada como genótipo II. A sensibilidade analítica da nested-PCR foi de 10-1,3 TCID50 mL-1. A combinação da nested-PCR e da restrição enzimática foi capaz de detectar e genotipar o vírus com resultados em um a dois dias, sendo mais rápida que os métodos convencionais de restrição do genoma completo que podem demorar até sete dias.

Pseudorabies is a disease caused by Suid herpesvirus 1 (PrV) and is responsible for considerable economic losses in the swine industry. The PrV has only one serotype, but based on RFLP (restriction fragment length polymorphism) the virus was divided into four genotypes named I, II, III, IV. The classical methods for PrV genotyping usually require virus isolation, DNA purification enzyme restriction analysis and a long electrophoresis. The aim of this research was to describe a faster and more sensitive method to detect and genotype PrV based on nested-PCR and restriction enzyme analysis. Twenty PrV isolates from south and southeast regions of Brazil, and the standard strain Shope were grown in PK-15 cells and submitted to PCR for glycoprotein E gene amplification. Additionally were tested 75 clinical samples (swine brain), with 25 positives for virus isolation and seroneutralization, and 50 negatives from a flock free PR with negative results in seroneutralization test. There was 100 percent of agreement between results of nested-PCR and virus isolation and seroneutralization and all samples detected were classified as genotype II. The nested-PCR, combined with restriction enzyme analysis, was able to detect and genotype PrV in 1-2 days with a sensitivity of 10-1,3 TCID50 mL-1. It was faster than classical methods described in the literature that require at least 7 days to be completed.

Evaluation in vitro of the infection times of engorged females of Rhipicephalus (Boophilus) microplus by the entomopathogenic nematode Steinernema glaseri CCA strain / Avaliação in vitro dos tempos de infecção de fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus pelo nematoide entomopatogênico Steinernema glaseri estirpe CCA

Studies have shown that ticks are susceptible to infection by entomopathogenic nematodes. These studies indicate different susceptibilities of ticks to infection by these fungi, depending on the tick species, development phase, entomopathogenic nematodes species and strains and the time the ticks are exposed to them. Usually this period ranges from 24 to 72 hours. The aim of this study was to evaluate the infection times in vitro of engorged Rhipicephalus (Boophilus) microplus females by the entomopathogenic nematodes Steinernema glaseri CCA strain, by analysis of the ticks' biological parameters. The results show that a 2-hour exposure time was sufficient for the engorged R. microplus females to be infected by S. glaseri CCA, but that a minimum exposure time of 24 hours was necessary to generate treatment efficacy above 90 percent.

Os carrapatos são susceptíveis à infecção por nematoides entomopatogênicos. Essa susceptibilidade diverge quanto às espécies de carrapato estudadas, à fase evolutiva, às espécies e estirpes dos nematoides e ao tempo ao qual os carrapatos ficam expostos a estes. O presente trabalho teve como objetivo avaliar os tempos de infecção in vitro de fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus pelo nematoide entomopatogênico Steinernema glaseri estirpe CCA, pela análise dos parâmetros biológicos do carrapato. Os resultados obtidos demonstraram que um período de duas horas de exposição foi suficiente para que fêmeas ingurgitadas de R. microplus fossem infectadas por S. glaseri CCA e que um período de exposição mínimo de 24h foi necessário para que houvesse infecção de fêmeas ingurgitadas de R. microplus por S. glaseri estirpe CCA, capaz de gerar, in vitro, eficácia no tratamento superior a 90 por cento.