High frequency of genital carriers of Leptospira sp. in sheep slaughtered in the semi-arid region of northeastern Brazil.

Although some studies in sheep have indicated leptospire colonization of the genital tract, further studies are needed to clarify the role of genital carriers in this species. Thus, this study aimed to evaluate the colonization of pathogenic leptospires in the genital and urinary tract of slaughtered sheep. Fifty-seven adult, female woolless sheep destined for slaughter were used. Renal (n = 57), bladder (n = 57), ovary (n = 34), uterine tube (n = 44), and uterus (n = 33) samples were collected for molecular detection of Leptospira sp. DNA, and blood samples (n = 57) for serological testing. The molecular testing was performed using polymerase chain reaction (PCR), and the serological testing was performed using microscopic serum agglutination test (MAT). Samples with amplifying DNA were subjected to genetic sequencing. In total, leptospiral DNA was found in the tissues of 44 (77.2%) sheep, whereas only nine animals were positive on both PCR and MAT; there was slight agreement between PCR and MAT techniques (k = 0.0268; p = 0.684). In 61 (54.9%) genital tract and in five (4.4%) urinary tract samples, the leptospiral DNA was detected, with significant difference (p < 0.001). The genes of one sample from the uterine tube and another from the bladder were sequenced and demonstrated 99% similarity to Leptospira interrogans. Anti-Leptospira antibodies were detected in 11 (19.3%) of the tested animals. The results reinforce the importance of the genital tract as an extra-renal site of colonization, suggesting the possibility of venereal transmission in sheep.

Use of chitosan and ß-tricalcium phosphate, alone and in combination, for bone healing in rabbits.

The aim of this research was to evaluate the process of bone regeneration in rabbits, using chitosan and beta-tricalcium phosphate (ß-TCP) independently and in combination. A total of 12 New Zealand rabbits of both sexes, with average weight of 3.0 ± 0.57 kg were used. Animals were randomly divided into two experimental time points, with six animals euthanized 45 days after surgery and six euthanized 90 days after surgery. We performed two osteotomies in each tibia. The left tibia was used for the chitosan (QUI) and control groups, and the right tibia was used for the ß-TCP alone and in combination with chitosan (QUI+TCP) groups. Tomographic evaluation showed no statistically significant difference among groups; however radiopacity was higher in the treated groups. Comparative descriptive histological evaluation found that treatment groups stimulated a more pronounced tissue repair reaction and accelerated bone repair. Morphometric analysis showed that treatment groups presented statistically higher bone formation compared with the control group.

Excretion of Brucella abortus vaccine B19 strain during a reproductive cycle in dairy cows.

This paper aimed to determine the excretion period of B19 vaccine strain during a complete reproductive cycle (from estrus synchronization, artificial insemination, pregnancy and until 30 days after parturition) of dairy cows from 3 to 9 years old that were previously vaccinated from 3 to 8 months. Three groups were monitored with monthly milk and urine collection during 12 months: G1 with seven cows from 3 to 4 years old; G2 with three cows from 5 to 6 years old; and G3 with four cows from 7 to 9 years old. Urine and milk samples were submitted to bacteriological culture and urine and PCR reactions for detection of Brucella spp. and PCR-multiplex for B19 strain identification. Ring test (RT) was also performed in the milk samples, and serum samples were tested by buffered acidified plate antigen test (BAPA). All animals were serologically negative at BAPA and Brucella spp. was not isolated from both urine and milk samples. RT revealed 13/210 (6.2%) positive milk samples. PCR reactions detected DNA of Brucella spp. in 86/420 (20.5%) samples. In urine it was found a significantly higher frequency (35.2%; 74/210) than in milk (5.7%; 12/210), more frequently from the estrus to 150 days of pregnancy and after parturition (6.7%; 10/150), and from 150 days of pregnancy to parturition (3.4%; 2/60), and they were all identified as B19 strain. In three groups, intermittent excretion of B19 strain was detected mainly in urine samples, which confirmed its multiplication and persistence in cows for until 9 years.

Isolation of Toxoplasma gondii from goats from Brazil.

Goats are economically important in many countries, and little is known of caprine toxoplasmosis in Brazil. Antibodies to Toxoplasma gondii were assayed in the sera of 143 goats from 3 Brazilian states, using modified agglutination test (MAT titer > or = 1:25); 46 (32.2%) tested positive. Samples of brain, heart, diaphragm, and masseter of seropositive animals were pooled, digested in pepsin, and bioassayed in mice. Viable T. gondii specimens were isolated from tissue homogenates of 12 goats; the isolates were designated TgGtBr1-12. Ten of the 12 isolates killed 100% of infected mice, indicating that goats can harbor mouse-virulent T. gondii and, hence, can serve as a source of infection for humans.

Toxoplasma gondii isolates from free-range chickens from the northeast region of Brazil.

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in 152 free-range chickens (Gallus domesticus) from 22 municipalities in 7 northeastern states (Pernambuco, Rio Grande do Norte, Maranhão, Bahia, Ceará, Sergipe, and Alagoas) of Brazil was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT); 81 (53.3 %) chickens had titers of 1:5 in 26, 1:10 in 9, 1:20 in 4, 1:40 in 1, 1:80 in 6, 1:160 in 6, 1:320 in 13, 1:640 in 6, 1:1,280 in 3, 1:2,560 in 6, and 1:5,120 or higher in 1. Hearts and brains of 81 seropositive chickens were bioassayed individually in mice. Toxoplasma gondii was isolated from 23 chickens with MAT titers of 1:5 or higher; the isolates were designated TgCKBr165-187. Five isolates killed all infected mice. Results indicate widespread contamination of rural environment in Brazil with T. gondii oocysts.

Evaluation of efficacy and safety of zinc gluconate associated with dimethyl sulphoxide for sexually mature canine males chemical neutering.

The aim of this research was to evaluate the efficacy of zinc gluconate associated with dimethyl sulphoxide (DMSO) for chemical neutering in canine males. Fifteen sexually mature male dogs were divided in two groups, named control and treated. An injection was administered to both testicles, at a concentration of 26.2 mg zinc gluconate per ml and 0.5% DMSO in the treated group (11 dogs). The control group was given injections of saline solution (four dogs). Clinical examination and blood collection for a haemogram were done both before and after drug injection. There were 12 spermograms performed to analyse sperm motility, sperm vigour, ejaculate volume, testicle size, pathology and sperm concentrations. Libido was also measured. An ultrasound examination and histopathology were performed at the end of the experiment. Dogs' libido after chemical injection was reduced by over 50%. The spermogram analysis showed final mean results of 14.54% for sperm motility, 0.72 of sperm vigour and 37,150 per million spermatozoa per millilitre, values considered below the necessary levels at which fertilization can occur. Ultrasound and histopathology analyses of testicles for the treated group revealed more intense injuries when compared with the control group, with compromised testicular parenchyma and a decrease of germ cell number leading to total atrophy, indicating that the treatment reduced the fertilizing potential of male dogs, promoting a possible subfertile status.

A randomized controlled trial of homeopathic treatment of weaned piglets in a commercial swine herd.

AIM: To evaluate the zootechnic performance and occurrence of diarrhea in piglets in the week post-weaning comparing supplementation with sucrose saline which contained or did not contain added homeopathic medicine. METHOD: Animals were randomly divided into three groups of 24 piglets each. The control group did not receive any treatment. Another group received sucrose saline, and the third group received sucrose saline with homeopathic medicine added, in the period of zero to seven days post-weaning. The homeopathic treatment consisted of Echinacea angustifolia, Avena sativa, Ignatia amara, Calcarea carbonica, all 6cH. Piglets were weighed daily for weight gain or loss, and observed for diarrhea and feed intake. RESULTS: Animals receiving sucrose saline alone and sucrose saline with homeopathy had less weight loss than control (p=0.017, p=0.0001 respectively). There was no statistical difference in relation to overall incidence of diarrhea or food consumption. These data suggest that the supplementation with sucrose saline with added homeopathic medicine in the first seven days post-weaning may be an useful option to reduce weight loss in weaned piglets.

Characterization of dog serum virome from Northeastern Brazil.

Domestic dogs share habitats with human, a fact that makes them a potential source of zoonotic viruses. Moreover, knowledge regarding possible bloodborne pathogens is important due to the increasing application of blood transfusion in dogs. In the present study, we evaluated the serum virome of 520 dogs using throughput sequencing (HTS). The serum samples were pooled and sequenced using an Illumina MiSeq platform. Our unbiased method identified prevalent canine pathogens as canine protoparvovirus 1 (canine parvovirus 2), undersearched agents as canine bocaparvovirus 1 (minute virus of canines) and canine circovirus, circular viruses closely related to viruses recently found in human samples, and new parvovirus and anelloviruses. The dog virome described in the present work furthers the knowledge concerning the viral population in domestic animals. The present data includes information regarding viral agents that are potentially transmitted through blood transfusion among dogs.

Seroepidemiology of Toxoplasma gondii and Neospora caninum in dogs from the state of Paraíba, Northeast region of Brazil.

A cross-sectional study was conducted to determine the seroprevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies and to investigate the risk factors related to antibodies against T. gondii and N. caninum in dogs of the city of Campina Grande, state of Paraiba, Northeast region of Brazil. For this purpose, 286 blood samples were collected from dogs during the rabies vaccination campaign, in September 2003, and on this occasion questionnaires addressing epidemiological aspects of the infections were given to each dog owner. The sera were analyzed for anti-T. gondii and anti-N. caninum antibodies by indirect fluorescent antibody tests. Of the total of 286 dogs, 129 were positive for T. gondii (titer16) with a prevalence value of 45.1% (95% CI=39.24-51.07%). For N. caninum, 24 dogs were positive (titer50), with a prevalence value of 8.4% (95% CI=5.45-12.23%). Antibodies to T. gondii and N. caninum were found simultaneously in 14 dogs (4.9%; 95% CI=2.7-8.08%). For T. gondii infection, the risk factors associated with seroprevalence was the age of the animals, with dogs older than one year presenting higher values of odds ratio, and co-habitation of cats in the household. For N. caninum infection, dogs that have street contact had higher odds of seropositivity than dogs that remained exclusively in a domestic environment.

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

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

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

Ehrlichia canis in dogs in a semiarid region of Northeastern Brazil: serology, molecular detection and associated factors.

This study investigated infection by Ehrlichia spp. agents by PCR, immunofluorescence assay test (IFAT), and by Giemsa-stained blood smears in 108 dogs from a semiarid region of the state of Paraíba, Northeastern Brazil. Seventy-five (69.4%) of the 108 dogs were found to be seropositive to Ehrlichia canis, while only four dogs (3.7%) were positive in real-time PCR for E. canis. In six dogs (5.6%) E. canis-like morulae were observed in monocytes. Animals that stayed in environment whose floor was dried dirt, and dogs whose owners reported low frequency of cleaning the dog environment had higher (P<0.05) PCR positivity for E. canis. Increasing seropositivity was found in older dogs (P=0.012). This study provides the first molecular detection of E. canis in the semiarid region of Northeastern Brazil.

Effect of piglet castration with nonsurgical sterilant on the zootechnical performance and pork carcass quality / Efeito da castração de leitões com esterilizante não cirúrgico no desempenho zootécnico e na qualidade da carcaça suína

A non-surgical sterilant (NSS) was used with the objective of assessing the zootechnical performance, carcass and meat quality, and hormone levels. 90 male piglets were selected with birthweights varying between 1.5kg to 2.0kg. The control group was constituted of 45 males castrated by the conventional surgical method, at the seventh day of age, and the treated group was constituted of 45 males castrated with NSS (active principle of zinc gluconate in the concentration of 26.2mg/mL associated to the dimethyl sulfoxide at 0.5%), with the application of the first dose on the seventh day of age and the second dose on the fourteenth day of age. The zootechnical assessments were carried out on the farm and consisted: weight gain in the periods per animal and feed conversion. The slaughter of the animals and the assessment of the carcass and meat quality and hormone levels were carried out in a slaughterhouse and the municipality of Campinas (SP). The study demonstrated that the use of NSS was a viable alternative in relation to the zootechnical performance and the carcass quality. In relation to the meat quality there was no significant difference in the majority of the assessed parameters.(AU)

Foi utilizado esterilizante não cirúrgico (ENC) com o objetivo de se avaliar o desempenho zootécnico e a qualidade de carcaça, de carne e de níveis hormonais. Foram selecionados 90 leitões machos de peso entre 1,5kg e 2,0kg ao nascimento. O grupo controle foi constituído por 45 machos castrados pelo método cirúrgico convencional, ao sétimo dia de idade, e o grupo tratado foi constituído por 45 machos castrados com ENC (princípio ativo de gluconato de zinco na concentração de 26,2mg/mL associado ao dimetil sulfóxido a 0,5%), com primeira dose de aplicação ao sétimo dia de idade e a segunda dose ao 14° dia de idade. As avaliações consistiram de: ganho de peso nos períodos por animal e conversão alimentar. O abate dos animais e a avaliação da carne e de níveis hormonais foram realizados em um frigorífico, na cidade de Campinas (SP). O uso de ENC foi uma alternativa viável em relação ao desempenho zootécnico e à qualidade da carcaça. Em relação à qualidade da carne, não houve diferença significativa na maioria dos parâmetros avaliados.(AU)

Risk factors associated with Neospora caninum seropositivity in sheep from the State of Alagoas, in the northeast region of Brazil.

The aim of this study was to identify risk factors associated with Neospora caninum seropositivity in sheep from the State of Alagoas, in the northeast region of Brazil. Twenty-six herds were selected, and blood samples were collected from 343 animals > 6-mo-old. The indirect fluorescent antibody test (IFAT) was used for serological diagnosis of N. caninum infection. Epidemiological questionnaires were applied for each herd to identify the risk factors associated with infection. Thirty-three (9.6%) animals tested positive for anti- N. caninum IgG antibodies, with titers ranging from 1 ratio 50 to 1 ratio 1,600. Fourteen herds (53.8%) presented at least 1 seropositive animal. Risk factors identified were property size

Toxoplasma gondii and Neospora caninum in domestic cats from the Brazilian semi-arid: seroprevalence and risk factors / Toxoplasma gondii e Neospora canimum em gatos no semiárido nordestino: soroprevalência e fatores de risco

In the present study we aimed to establish the seroprevalence of T. gondii and N. caninum in stray and domiciled cats from the municipality of Patos, Paraíba state, Brazil. Blood samples were collected from 201 animals: 132 domiciled cats and 69 stray cats. An epidemiological questionnaire was conducted with all cat owners. Indirect immunofluorescent antibody test (IFAT) was performed at cut-offs of 1:16 and 1:50 for T. gondii and N. caninum, respectively. Overall prevalence of seroreagent cats for T. gondii was 43.8 percent. We found a prevalence of 47.7percent in domiciled cats and 36.2 percent in stray cats. Antibody titers ranged from 1:16 (cut-off) to 1:8192; 1:128 was the most frequent titer. No statistical difference was observed between domiciled cats and stray cats. Correlation was verified between seroreagent for T. gondii and age and hunting habit (P<0.05). No animals tested seroreagent for N. caninum. It was possible to conclude that there is high prevalence of cat seroreagent for T. gondii and that N. caninum is not present in cats from the area studied...

Este trabalho teve como objetivo determinar frequência de Toxoplasma gondii e Neospora caninum em gatos domiciliados e errantes do município de Patos, mesorregião do sertão paraibano. Foram coletadas amostras de sangue provenientes de 201 gatos, 132 domiciliados e 69 errantes. Nos domiciliados, foi aplicado um questionário epidemiológico aos proprietários. A reação de imunofluorescência indireta (RIFI) foi realizada levando-se em consideração os pontos de corte 1:16 e 1:50 para T. gondii e N. caninum, respectivamente. Foi obtida uma prevalência total de 43.8 por cento de gatos sororreagentes para T. gondii. Nos domiciliados, a prevalência foi de 47,7 por cento (63/132), e nos errantes de 36,2 por cento (25/69), com títulos variando de 1:16 a 1:8192, sendo a titulação 1:128 a mais frequente. Não se observou diferença estatística entre animais domiciliados e errantes. Verificou-se correlação entre sororreagentes a T. gondii e idade e hábito de caçar (P<0.05). Nenhum animal foi sororreagente para N. caninum. Concluiu-se que é alta a prevalência de gatos sororreagentes para T. gondii e que o N. caninum não está presente em gatos na área estudada...

Interrelação entre frequência de anticorpos anti-leptospira Spp. e exames histopatológicos (hematoxilina-eosina e Warthin Starry) em suínos abatidos no semiárido paraibano / Interrelation between frequency of anti-leptospira spp. and histopathological examination (hematoxylin-eosin and Warthin-Starry) in pigs slaughtered in the semiarid of Paraiba

Este trabalho foi realizado em suínos abatidos no Município de Patos, Estado da Paraíba, Brasil,com o objetivo de determinar a frequência de anticorpos anti-Leptospira spp., comparando os achados sorológicos com exames histopatológicos de rim, fígado, ovário e útero. A soroaglutinação microscópica foi realizada em 126 animais. Os exames histopatológicos realizados em cortes de fígado, rim, ovário e útero, corados pela hematoxilina-eosina (HE), foram realizados em 20 animaisescolhidos aleatoriamente, sendo 10 do grupo com títulos ≥ 100 e 10 do grupo com títulos < 100. Paralelamente, foi realizada pesquisa direta de leptospiras pela técnica de Warthin-Starry em amostras de rim de todos os animais soropositivos e nos 10 animais soronegativos submetidos à HE. Dos 126 animais examinados, 18 (14,6%) foram soropositivos, com predominância de reações para o sorovar Autumnalis (11 animais; 8,73%). Quatro animais soropositivos e dois animais soronegativosapresentaram infiltrado inflamatório e necrose de graus variados em um dos rins e no fígado. Os ovários e úteros examinados não apresentaram lesões. A pesquisa direta de leptospiras pela técnica de Warthin-Starry não revelou animais positivos em nenhuma amostra testada. Em face da soropositividade encontrada (14,6%), sugere-se a importância da conscientização por parte dos produtores acerca da implantação de medidas de prevenção adequadas com o objetivo de impedir, ou pelo menos diminuir, a disseminação das leptospiras em suínos e, consequentemente, bloquear a possível transmissão do agente para os seres humanos...

Interrelation between frequency of anti-leptospira spp. Antibodies and findings of histopathological examinations (hematoxylin-eosin and warthin-starry) in pigs slaughtered in the semiarid of paraiba state, northeastern brazil. This work was conducted in slaughtered pigs in Patos County, Paraiba State, Brazil, with the aim to determine the frequency of anti-leptospira antibodies and to compare the serological findings with the histopathological findings of kidney, liver, ovary and uterus. The microscopic agglutination test (MAT) was performed on 126 animals. The histopathological examination performed in sections of livers, kidneys, ovaries and uteruses stained with hematoxylin-eosin (HE) was conducted in 20 randomly chosen animals (10 from group with serological titer ≥ 100 and 10 from group < 100). Parallel to this, a direct search for leptospires was carried out by Warthin-Starry technique in kidney samples from all seropositive animals and in the 10 seronegative animals submitted to HE. In the 126 animals examined, 18 (14.6%) were seropositive, with prevalence of reactions to Autumnalis serovar (11 animals; 8.73%). Four seropositive and two seronegative animals showed different degrees of inflammatory infiltrate and necrosis in liver or kidney. The ovaries and uterus no showed lesions. Direct analysis of leptospires by the Warthin-Starry technique did not reveal positive animals in any sample tested. The rate of seropositivity found (14.6%) suggests the importance of awareness by producers about the implementation of preventive measures aimed at preventing, or at least reducing the spread of leptospiras in pigs and thereby blocking the possible transmission of the agent to humans...

Validação de nova proposta de espermocultura quantitativa aplicada a sêmen industrializado de touros / Validation of a new proposal of quantitative sperm culture for industrialized bull semen

O Ministério da Agricultura, Pecuária e Abastecimento (MAPA) tem discutido a obrigatoriedade da realização de espermocultura em sêmen industrializado não somente para garantia da biossegurança mas também por falhas na fertilização in vitro pela contaminação dos ovócitos com bactérias ubiquitárias e oportunistas da microbiota prepucial. A técnica quantitativa de Pour Plate, preconizada pela Organização Internacional de Epizootias, é operacionalmente difícil e de custo elevado para a rotina em centrais de inseminação artificial (CIAs). Desse modo, avaliou-se a técnica de contagem de bactérias viáveis por superfície (CBVS) em UFC/mL de sêmen industrializado de touros de CIAs, comparando-a à técnica de Pour Plate, com intuito de validação. Foram empregadas palhetas de sêmen de touros do Projeto Hungria do MAPA. A partir da diluição seriada de 10-1 a 10-5 do sêmem, foram empregadas, paralelamente, as técnicas de Pour Plate e CBVS. A média ou mediana de UFC/mL obtida em cada diluição foi comparada entre as técnicas, e não foram observadas diferenças estatísticas entre as duas técnicas quanto à quantificação de bactérias em UFC/mL, sugerindo a possibilidade de substituição da técnica de Pour Plate pela de CBVS, com vantagens de praticidade e menor custo.

The Brazilian Ministry of Agriculture (MAPA) has discussed the mandatory culture of industrialized semen, both to ensure biosefety, and to prevent in vitro fertilization problems caused by oocyte contamination with ubiquitous and opportunistic bacteria from preputial microbiota. Pour plate, a quantitative technique recommended by the World Organization for Animal Health (OIE), is operationally difficult and costly for routine analysis in Artificial Insemination Centers (AICs). The objectives of this study were to evaluate and validate viable superficial bacteria counts (VSBC), in CFU/mL, compared with pour plate counts, in industrialized bull semen samples from AICs. Semen straws from Projeto Hungria - MAPA bulls were used. VSBC and pour plate were carried out in parallel in serial dilutions of the samples, from 10-1 to 10-5. CFU/mL means or medians recorded in each dilution and technique were compared, and no statistical differences were observed between the two techniques regarding the quantification of bacteria in CFU/mL, suggesting that it may be possible to replace pour plate for CBSV, a cheaper and more practical technique.

Excretion of Brucella abortus vaccine B19 strain during a reproductive cycle in dairy cows

This paper aimed to determine the excretion period of B19 vaccine strain during a complete reproductive cycle (from estrus synchronization, artificial insemination, pregnancy and until 30 days after parturition) of dairy cows from 3 to 9 years old that were previously vaccinated from 3 to 8 months. Three groups were monitored with monthly milk and urine collection during 12 months: G1 with seven cows from 3 to 4 years old; G2 with three cows from 5 to 6 years old; and G3 with four cows from 7 to 9 years old. Urine and milk samples were submitted to bacteriological culture and urine and PCR reactions for detection of Brucella spp. and PCR-multiplex for B19 strain identification. Ring test (RT) was also performed in the milk samples, and serum samples were tested by buffered acidified plate antigen test (BAPA). All animals were serologically negative at BAPA and Brucella spp. was not isolated from both urine and milk samples. RT revealed 13/210 (6.2%) positive milk samples. PCR reactions detected DNA of Brucella spp. in 86/420 (20.5%) samples. In urine it was found a significantly higher frequency (35.2%; 74/210) than in milk (5.7%; 12/210), more frequently from the estrus to 150 days of pregnancy and after parturition (6.7%; 10/150), and from 150 days of pregnancy to parturition (3.4%; 2/60), and they were all identified as B19 strain. In three groups, intermittent excretion of B19 strain was detected mainly in urine samples, which confirmed its multiplication and persistence in cows for until 9 years.

Brucelose bovina no estado da Paraíba: estudo retrospectivo / Bovine brucelosis in the state of Paraíba: a retrospective survey

O objetivo do presente trabalho foi determinar a frequência de propriedades positivas (focos) e de animais soropositivos para a brucelose bovina no Estado da Paraíba. Foram utilizados dados da Agência de Defesa Agropecuária do Estado, coletados de suas 23 microrregiões, durante o período de janeiro de 2008 a julho de 2009. Durante esse período, foram examinadas 11.149 propriedades, e 55.691 soros de bovinos foram submetidos ao diagnóstico de brucelose. Para o diagnóstico sorológico, foi utilizado o teste do antígeno acidificado tamponado (AAT). Uma propriedade foi considerada foco quando apresentou pelo menos um animal soropositivo. Das propriedades investigadas, 104 (0,93%) apresentaram pelo menos um animal soropositivo e, dos animais analisados, 199 (0,36%) foram soropositivos. Houve diferença significativa (p < 0,001) na proporção de fêmeas (0,47%) e machos (0,04%) soropositivos.

The aim of this study was to determine the frequency of positive herds (foci) and seropositive animals for bovine brucellosis in the state of Paraíba, Northeast region of Brazil. Data from the Agency of Agricultural Protection in the state, collected from its 23 microregions, during the period from January 2008 to July 2009, were used. During this period, 11,149 herds were examined, and 55,691 cattle sera were submitted to the diagnosis of brucellosis. For serological diagnosis the rose bengal test was used. A herd was considered a focus when it presented at least one seropositive animal. Of the herds investigated, 104 (0.93%) had at least one seropositive animal, and of the animals examined, 199 (0.36%) were seropositive. There was a significant difference (p < 0.001) in the proportion of seropositivity for females (0.47%) and males (0.04%).

Detecção dos genes da toxina citoletal distensiva em estirpes de Campylobacter jejuni isoladas de carcaças de frangos / Detection of cytolethal distending toxin genes in strains of Campylobacter jejuni isolated from broiler carcasses

Foram analisadas 80 amostras de sobrecoxas de frangos de corte resfriados provenientes de feiras livres e hipermercados do município de São Paulo, SP. Treze estirpes de Campylobacter spp. foram isoladas em 10 (12,5 por cento) sobrecoxas, sendo cinco amostras originárias de feiras livres e cinco de hipermercados. Onze estirpes foram identificadas como Campylobacter jejuni e duas como Campylobacter coli. As 11 estirpes foram confirmadas como C. jejuni pela PCR do gene da hipuricase (hip), e destas, quatro (36,4 por cento) apresentaram os três genes (cdtA, cdtB e cdtC) codificantes da toxina citoletal distensiva pela multiplex-PCR, sendo três estirpes provenientes de hipermercados e uma de feira livre. Observou-se a presença de estirpes virulentas de C. jejuni, portadoras do complexo de genes cdt, nas amostras de frango resfriado, não só na linha de abate, mas até o ponto final da cadeia de distribuição, nos dois principais centros de venda a varejo.

Eighty samples of refrigerated broiler thighs purchased in street markets and supermarkets in the city of São Paulo, SP, were analyzed. Thirteen Campylobacter spp. strains were isolated in 10 (12.5 percent) thighs, five of them from street market samples and other five from supermarkets. Eleven strains were identified as Campylobacter jejuni and two of them as Campylobacter coli. The 11 strains were confirmed to be C. jejuni using PCR for hippuricase (hip) gene. From these, multiplex-PCR showed that four (36.4 percent) strains presented the three genes (cdtA, cdtB, and cdtC) encoding cytolethal distending toxin: three strains from supermarket and one from street market samples. These results are important, because they demonstrate the presence of virulent C. jejuni strains in refrigerated broiler thigh samples, not only in the slaughterhouse but in the final point of the distribution chain, at the two most important food retail commercer.

Influência das condições ambientais na transmissão da leptospirose entre criações de ovinos e bovinos da região de Sorocaba, SP / The influence of environment conditions on the transmission of leptospirosis between sheep and cattle herds of Sorocaba region, SP, Brazil

Leptospiras excretadas pela urina podem sobreviver por longos períodos em águas de superfície e solos, na dependência do pH e teor de umidade e de matéria orgânica. Investigou-se a influência do meio ambiente na transmissão da leptospirose em dois rebanhos exclusivos de ovinos (A e C) e dois de ovinos consorciados com bovinos (F e H) da região de Sorocaba, SP, no período de dezembro de 2007 a setembro de 2008. Foram examinadas amostras de soro pela reação de soroaglutinação microscópica; de urina, água e solo pelo cultivo para leptospiras e urina de ovinos pela PCR. Condições edafoclimáticas, pH das águas de superfície e solo, granulometria e permeabilidade do solo foram analisadas. Todos os rebanhos apresentaram pelo menos um animal sororeagente para Leptospira spp. Apenas a PCR de um pool de urina de ovinos (H) foi positiva. Leptospira spp. foi isolada do lago de F. O pH das águas de superfície variou entre 6,0-7,0; e nos solos entre 4,5 e 6,8. Os índices de matéria orgânica em A, C e H variaram de 24 a 35 g/dm3, e 63 g/dm3 em F. A composição do solo de A e F mostrou-se franco-argiloarenosa, C argilosa e H franco-siltosa; como texturas mistas são capazes de manter a umidade, principalmente devido a argila. Diante da presença de animais sororeatores e portanto da circulação de Leptospira spp. nos rebanhos, conclui-se que o ciclo de transmissão é dependente da interação sinérgica e antagônica de muitas variáveis; onde o pastejo num habitat com alto teor de umidade parece ser limitante.

Leptospires excreted by urine are able to survive for long periods in surface water and soil depending on the pH, humidity and organic matter presence. This paper reported the influence of environment conditions on the transmission of leptospirosis in two sheep-only farms (A and C) and two cattle-sheep farms (F and H) from December 2007 to September 2008. Serum samples were examined by microscopic agglutination test; urine, surface water and soil samples were cultured for leptospires, and ovine urine pools were analyzed by PCR. Regional edaphoclimatic conditions, pH of surface water and soil, granulometry and permeability of soil were analyzed. All herds presented at least one reactor to Leptospira spp. Only the PCR of an ovine urine pool of herd H was positive and Leptospira spp. was isolated from the F lake. The pH of water samples ranged from 6.0 to 7.0; while in soil it was around from 4.5 to 6.8. Soil organic matter were 24 to 35 g/dm3 in A, C e H, and 63 g/dm3 in F. Soil samples of A and F showed loamy-clay texture; C had clay soil, and H loamy-silt soil; as mixed compositions are able to maintain the humidity, mainly where clay is present. As the presence of reactors in all herds indicated the contact with Leptospira spp., it was concluded that the cycle of transmission is dependent on the synergistic and antagonistic interaction of many variables; but the close contact of animals grazing in a high humidity habitat seems to be limiting.