Isolamento e caracterização molecular de Dichelobacter nodosus isolados de ovinos no estado da Bahia, Brasil / Isolation and molecular characterization of Dichelobacter nodosus isolated from sheep in Bahia, Brazil

Background: Pododermatitis or footrot is an infectious disease that affects the hoof and interdigital tissue of sheep causing lameness. The disease is caused by the interaction of the agent Dichelobacter nodosus and symbiotic bacteria in the complex environment of the epidermal tissues of the hoof and host immune system. D. nodosus is not able to invadehealthy hooves, so the infection is preceded by colonization of the interdigital skin by Fuso bacterium necrophorum. The aim of this research was to perform the isolation andcharacterization of D. nodosus in sheep farms of different municipalities of Bahia, obtaining the serogroups present in each herd.Materials, Methods & Results: The study was carried out in nine sheep farms from eight municipalities in the state of Bahia. All farms presented history of foot diseases. A total of 620 animals were observed, 140 of which were examined for lameness. To collect the contents of the lesions, sterile swabs were introduced into tubes containing sterile Thorley transport medium under refrigeration at 8°C and sent for laboratory analysis. Subsequently, each swab collected was seeded in two Petri dishes containing 4% hoof agar medium and incubated in anaerobic at 37°C for 96 hours. The purified samples were seeded on 2% hoof agar and incubated under the same conditions as above. The colonies were identified by the morphological characteristic and Gram staining. The DNA was extracted and stored at -20°C until its use in PCR, for identification and classification of D. nodosus in serogroups (A-I). In the nine farms visited were found animals with clinical signs of infectious pododermatitis. After processing, there was success of isolation in 39 samples (41%), confirming the presence of D. nodosus in all municipalities evaluated.[...]

Isolamento e caracterização molecular de Dichelobacter nodosus isolados de ovinos no estado da Bahia, Brasil / Isolation and molecular characterization of Dichelobacter nodosus isolated from sheep in Bahia, Brazil

Background: Pododermatitis or footrot is an infectious disease that affects the hoof and interdigital tissue of sheep causing lameness. The disease is caused by the interaction of the agent Dichelobacter nodosus and symbiotic bacteria in the complex environment of the epidermal tissues of the hoof and host immune system. D. nodosus is not able to invadehealthy hooves, so the infection is preceded by colonization of the interdigital skin by Fuso bacterium necrophorum. The aim of this research was to perform the isolation andcharacterization of D. nodosus in sheep farms of different municipalities of Bahia, obtaining the serogroups present in each herd.Materials, Methods & Results: The study was carried out in nine sheep farms from eight municipalities in the state of Bahia. All farms presented history of foot diseases. A total of 620 animals were observed, 140 of which were examined for lameness. To collect the contents of the lesions, sterile swabs were introduced into tubes containing sterile Thorley transport medium under refrigeration at 8°C and sent for laboratory analysis. Subsequently, each swab collected was seeded in two Petri dishes containing 4% hoof agar medium and incubated in anaerobic at 37°C for 96 hours. The purified samples were seeded on 2% hoof agar and incubated under the same conditions as above. The colonies were identified by the morphological characteristic and Gram staining. The DNA was extracted and stored at -20°C until its use in PCR, for identification and classification of D. nodosus in serogroups (A-I). In the nine farms visited were found animals with clinical signs of infectious pododermatitis. After processing, there was success of isolation in 39 samples (41%), confirming the presence of D. nodosus in all municipalities evaluated.[...](AU)

Afecções podais com ênfase em pododermatite infecciosa em ovinos no estado da Bahia

O presente trabalho foi dividido em dois experimentos visando determinar a ocorrência e diagnosticar as doenças podais que acometem rebanhos ovinos em diferentes municípios baianos, incluindo a caracterização clínica e sorológica nos casos de pododermatite infecciosa. O primeiro trabalho objetivou o isolamento e caracterização do Dichelobacter nodosus em fazendas de criação de ovinos, durante surtos de pododermatite em oito municípios da Bahia. Durante as visitas, 620 animais foram inspecionados, procedendo-se exame podal completo em 140 deles, sendo 95 destinados a coleta por swab para bacteriologia. Após cultura, foi possível isolar 39 amostras, obtendo-se identificação do D. nodosus em todos os municípios visitados. As amostras positivas encaminhadas para sorotipagem por PCR revelaram a presença de sete sorogrupos (A, B, D, E, F, H, I), sendo o D (59%) e H (17%) os mais prevalentes e o sorogrupo I detectado pela primeira vez no país. Do total de amostras, 11,5% apresentaram infecções mistas com dois ou três sorogrupos por animal. A detecção de um novo sorogrupo e de infecções mistas apontaram para a necessidade de ampliar o foco da investigação e das medidas de diagnóstico, profilaxia e controle da pododermatite infecciosa nos ovinos do Brasil. O segundo trabalho objetivou determinar a ocorrência e caracterizar as doenças podais encontradas em criações de ovinos de oito municípios da Bahia. Em cada visita, realizou-se a inspeção do rebanho, questionário e caracterização das doenças encontradas. A ocorrência de animais com doenças podais foi de 23,3% (140/600) variando de 12,8 a 55,5% por propriedade. Em cada fazenda, foi possível observar ao menos três tipos de enfermidades podais. Ao todo foram registrados 220 lesões envolvendo nove doenças. A pododermatite infecciosa foi a enfermidade mais diagnosticada, totalizando 51,3% dos casos (113/220), seguida da dermatite interdigital, com 27,7% (61/220). Dentre os 113 casos de pododermatite infecciosa, 70% correspondiam a estágios avançados e terminais da doença indicando que a forma virulenta era a prevalente nos rebanhos consultados. As outras doenças identificadas foram: crescimento excessivo do casco com 8,6% dos casos (19/220), doença da linha branca com 3,2% (7/220), sola dupla com 3,2% (7/220), granuloma do dígito com 2,3% (5/220), hiperplasia interdigital com 2,3% (5/220), úlcera de sola com 0,9% (2/220) e abscesso de casco com 0,5% (1/220). Quanto ao membro afetado, não houve diferença significativa entre os membros torácicos e pélvicos. Quanto a quantidade de membros acometidos, 20% dos animais (28/140) possuíam lesões em somente um membro, 32,1%(45/140) em dois membros, 20,7% (29/140) em três membros e 27,2% (38/140) apresentaram lesões nos quatro membros. Avaliando-se a quantidade de doenças por animal, observou-se que 53,6% (75/140) dos animais foram acometidos por um único tipo de enfermidade podal, 35,7% (50/140) por dois tipos, 9,3% (13/140) por três tipos e 1,4% (2/140) apresentaram quatro tipos de doenças podais. A claudicação foi observada em todos os animais, porém seu grau variava conforme a gravidade da doença e quantidade de cascos afetados. As condições ambientais e de manejo foram determinantes para o surgimento de lesões podais entre os animais avaliados. Além da influência dos diversos fatores predisponentes encontrados, observou-se que o diagnóstico precoce e profilaxia não são realizados na maioria dos casos, o que contribuiu para a elevada ocorrência e diversidade de doenças podais diagnosticadas.

This study was divided into two experiments to determine the occurrence and diagnose foot diseases that affect sheep flocks in different towns in Bahia, including the clinical and serological characterization of foot rot cases. The first paper aimed the isolation and characterization of Dichelobacter nodosus in sheep farms during outbreaks in eight towns of Bahia. During the visits 620 animals were inspected, proceeding complete foot examination at 140 of them and 95 for collection by swab for bacteriology. After culture was possible to isolate 39 samples, obtaining identification Dichelobacter nodosus in all farms visited. Positive samples sent for serotyping PCR revealed the presence of seven serogroups (A, B, D, E, F, H, I), D (59%) and H (17%) were the most prevalent and serogroup I first reported in the country. Of the total sample, 11.5% had mixed infections with two or three serogroups per animal. The detection of a new serogroup, and mixed infections point to the need to expand the focus of research and the diagnostic, prevention and control measures of footrot in Brazil. The second paper aimed to determine the occurrence and characterize foot diseases found in sheep farms of eight towns in Bahia. In each visit, inspection of the herd, survey questionnaire and characterization of diseases were proceeded. The occurrence of foot diseases was 23.3% (140/600) ranging from 12.8 to 55.5% by farm. In each farm was observed at least three types of foot diseases. Altogether 220 cases were recorded involving nine diseases. Foot rot was the most diagnosed disease totaling 51.3% of the cases (113/220) followed by Interdigital dermatitis with 27.7% (61/220). Among the 113 cases of footrot, 70% were advanced or terminal stages of the disease indicating that the virulent form was prevalent in herds consulted. Other diseases have been identified: excessive growth of the hoof with 8.6% (19/220), white line disease with 3.2% (7/220), double sole with 3.2% (7/220), toe granuloma with 2.3% (5/220), interdigital hyperplasia with 2.3% (5/220), sole ulcer with 0.9% (2/220) and hoof abscess with 0.5% (1/220). As for the affected limb, there was no significant difference between the fore and hindlimbs. As the number of affected limbs, 20% of animals (28/140) had lesions only one limb, 32.1% (45/140) in two limbs 20.7% (29/140) in three limbs and 27.2% (38 / 140) had injuries in all limbs. Evaluating the amount of diseases per animal, it was observed that 53.6% (75/140) of the animals were affected by a single type of foot disease, 35.7% (50/140) of two types, 9.3% (13/140) of three types and 1.4% (2/140) four types of feet diseases. Lameness was observed in all animals, but its level varied according to the severity of the condition and quantity of affected hooves. Environmental conditions and management were crucial to the development of foot lesions in evaluated animals. Besides the influence of various predisposing factors, it was observed that the early diagnosis and prophylaxis are not performed in most cases, which contributed to the occurrence and diversity of diseases diagnosed.

Microbiota anaeróbia isolada de bovinos com pododermatite

The objective of this study was to isolate and identify the anaerobic bacteria species in bovine with different degrees of pododermatitis. Sixty bovines were divided into four groups of 15 as follows: group I included healthy animals, used as control; group II with animals in the initial phase of the process; group III, with animals displaying interdigital skin hyperplasia and group IV with animals with footrot. Interdigital tissue fragments were collected and processed for isolation and identification of anaerobic agents. The bacteria species isolated were: Dichelobacter nodosus in groups II, III and IV and Fusobacterium necrophorum only in groups III and IV, with frequencies of 26.7%, 6.7%, and 20.0% for the former and 6.7% and 13.3% for the latter, respectively. In addition, Fusobacterium symbiosum was present in 40.0% of group I, 6.7% of group II, 13.3% of group III and 13.3% of group IV; Bacteroides sp. in 6.7% of groups I and IV; Bacteroides ruminatus in 33.3% of group I, 6.7% of group II, 33.3% of group III and 13.3% of group IV; Bacteroides oralis in 6.7% of group III, and Fusobacterium mortiferum in 6.7% of group IV.

O presente trabalho teve como objetivo isolar e identificar espécies bacterianas anaeróbias presentes nos pés de bovinos portadores de vários graus de pododermatite. Foram utilizados 60 bovinos, distribuídos em quatro grupos de 15. O grupo I foi constituído por animais saudáveis e serviu de controle; o grupo II, por bovinos na fase inicial do processo; o grupo III, por animais portadores de pododermatite interdigital vegetativa e o grupo IV, por bovinos portadores de pododermatite necrosante. Foram colhidos fragmentos de tecido interdigital para cultura e as principais espécies bacterianas isoladas foram: Dichelobacter nodosus nos grupos II, III e IV e Fusobacterium necrophorum nos grupos III e IV, com freqüências de 26,7%, 6,7%, 20,0%, 6,7% e de 13,3%, respectivamente. Encontraram-se também Fusobacterium symbiosum em 40,0% no gb>rupo I, 6,7% no grupo II, 13,3% no grupo III e 13,3% no grupo IV, Bacteroides sp. em 6,7% nos grupos I e IV, Bacteroides ruminatus em 33,3% no grupo I, 6,7% no grupo II, 33,3% no grupo III e 13,3% no grupo IV, Bacteroides oralis em 6,7% no grupo III e Fusobacterium mortiferum em 6,7% no grupo IV.