Unidade Acadêmica de Medicina Veterinária Centro de Saúde e … · 2017-06-06 · Slide 15 Patogenia Penetração mucosas oral, nasal, ocular, prepucial, retal, vaginal e uterina

Slide 1

Universidade Federal de Campina GrandeCentro de Saúde e Tecnologia Rural

Unidade Acadêmica de Medicina Veterinária

Brucelose Ovina

PROF. TITULAR CLEBERT JOSE ALVES 1

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Slide 2

Brucelose Ovina

A brucelose ovina é caracterizada por um processo infeccioso clínico ou subclínico e de tendência à cronicidade;

Presença de lesões genitais de epididimite no macho e placentite nas fêmeas;

Casos de abortamento, levando a mortalidade de recém-nascidos e elevada frequência de nascidos com baixo peso e baixa viabilidade condicionando à elevada mortalidade em cordeiros

Sendo uma das principais causas de perdas econômicas no rebanho

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Slide 3

Agente EtiológicoAs brucelas são coco-bacilos gram-negativos que normalmente aparecem isolados, embora, às vezes, aos pares e até em pequenos grupos;

Variam de 0,6 a 1,5 μm de comprimento por 0,5 a 0,7 μm de diâmetro;

São imóveis, não formam esporos e não apresentam cápsula;

Crescem em meios de cultura comuns, enriquecidos com soro normal equino, bovino ou ovino, em uma concentração final de 5% a 10%. Podendo-se utilizar o sangue desfibrinado de ovino como enriquecedor.

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Slide 4

Agente EtiológicoAs colônias em meio sólido são visíveis em três a cinco dias de incubação, apresentando-se pequenas, circulares, de bordos regulares, opacas, superfície granular e com cor variando do branco ao marrom.

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Slide 5

Agente Etiológico

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Slide 6

Importância EconômicaOs prejuízos econômicos decorrentes da brucelose ovina devem-se:

*diminuição da fertilidade dos carneiros;

* abortos em ovelhas;

* elevadas perdas perinatais de cordeiros;

•* descarte e substituição de reprodutores;

* problemas de manejo na estação reprodutiva. (ROBLES et al.,

1993; BAIGÚN et al., 2000)

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Slide 7

HospedeirosA Brucella ovis infecta de forma natural exclusivamente à espécie ovina (HOMSE et al., 1995);

Macho mais suscetível que a fêmea (TAMAYO et al., 1989);

Existe ainda diferença de suscetibilidade entre raças importadas e nativas (FICAPAL et al., 1998).

Em caprinos, foi demostrado experimentalmente que a infecção por B. ovis é leve e de curta duração, mostrando que eles são menos suscetíveis que os ovinos

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Slide 8

HospedeirosSendo desconhecido se a infecção se transmite do ovino ao caprino em condições de campo (ESTEIN, 1999)

Burgess et al. (1985), após Infecção experimental de caprinos com B. ovis, demonstraram que eles reagem sorologicamente e também excretam a bactéria no sêmen, porém não relataram a ocorrência de infecção natural.

Outras espécies de Brucella podem infectar ovinos, como a B. melitensis, especialmente se caprinos e ovinos são criados juntos

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Slide 9

Vias de Eliminação*Sêmen constitui a mais importante via de eliminação deste agente;

*Eliminação ocorre de forma intermitente;

* Por períodos prolongados, chegando a 80 semanas pós

Infecção;

*sendo isolado, inclusive, do sêmen de carneiros soronegativos

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Slide 10

Vias de EliminaçãoOvelhas, naturalmente infectadas a bactéria foi isolada do útero;

Tanto em casos de infecção natural como em infecção experimental, os envoltórios fetais e o feto constituem vias de eliminação da B. ovis, uma vez que a bactéria foi isolada a partir do feto e placenta;

A urina também é uma via de eliminação da B. ovis, uma vez que esta bactéria foi isolada da urina de carneiros experimentalmente infectadas

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Slide 11

Porta de EntradaFoi demonstrado que a B. ovis penetra no organismo através das:

* Mucosas conjuntival;

* Cervico-vaginal;

* Prepucial;

* Retal;

* Nasal;

* Pele lesada

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Slide 12

Vias de transmissão* A transmissão de B. ovis pode ocorrer por via venérea

* via vaginal, com sêmen contaminado;

* ovelhas infectadas excretando B. ovis via vaginal, há alta

possibilidade de transmissão ativa para o carneiro durante a cópula;

* comportamento homossexual - sendo a única via de transmissão quando o rebanho é composto só por machos;

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Slide 13

Vias de transmissão*Como o sêmen é a principal via de excreção da B. ovis e as mucosas vaginal e cervico-uterina são importantes portas de entrada do agente a transmissão da B. ovis do carneiro para a ovelha na cópula contribui para a manutenção da infecção;

*A persistência da infecção por B. ovis em rebanhos, apesar da eliminação dos carneiros reatores sorologicamente, pode ser o resultado da presença de carneiros soronegativos que mantêm a infecção e excretam a bactéria, infectando assim os carneiros sadios

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Slide 14

Vias de transmissão*A brucelose ovina causa o nascimento de cordeiros prematuros ou débeis e os que sobrevivem podem ser um potencial portador podendo desenvolver a doença ao atingir a puberdade

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Slide 15

Patogenia

Penetração mucosas oral, nasal,

ocular, prepucial, retal, vaginal e uterina

Permanecer nelas por um mês, devido a

propriedade de

resistir à destruição intrafagocitária, multiplicando-se

lentamente

Multiplicação nos gânglios regionais, as brucelas invadem os vasos linfáticos regionais e daí o ducto torácico e acorrente

sanguínea. O estágio bacterêmico parece cessar aproximadamente no final do segundo mês de infecção

tropismo pelos órgãos genitais sendo isolada da ampola dos ductos deferentes, vesículas seminais, glândulas

bulbo uretrais, epidídimos. testículo,

onde provoca hiperplasia epitelial e

formação de cistos

A

localização das brucelas no epitélio

da cauda dos epidídimos produz

alterações degenerativas.

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LesõesMacroscópicas:

*Aderência testicular entre a túnica albugínea e a túnica vaginal;

*Atrofia testicular, onde testículo afetado se apresenta pequeno e de consistência firme devido a fibrose ou mineralização;

*Orquite com exsudato purulento;

*Lesões mais consistentes localizam-se nos epidídimos,

sendo unilaterais em mais de 77% dos casos

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Lesões* O epidídimo se apresenta aumentado de tamanho, principalmente a cauda, de consistência firme e, ao corte pode-se observar um ou vários pequenos cistos ou nódulos variando de 0,7 a 1,0 centímetro de diâmetro. Estes cistos ou nódulos contêm um fluido verde-amarelado a esbranquiçado;

*As ovelhas que abortam apresentam a placenta edematosa e coberta com exsudato fibrinoso amarelado, principalmente nas áreas intercotiledonárias;

*Feto abortado encontra-se, autolisado, com exsudato supurativo nos brônquios e pulmões, e exsudato fibrinoso na cavidade peritoneal

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LesõesMicroscópicas: Lesões histológicas encontradas com frequência:

* Na cauda do epidídimo caracterizada por uma epididimite onde o tecido intersticial e os espaços perivasculares adjacentes à camada basal do epitélio tubular, apresentam uma infiltração moderada de células mononucleares, sobretudo linfócitos;

* Moderada hiperplasia epitelial afetando grupos de túbulos de algumas regiões da cauda dos epidídimos;

* Degeneração hidrópica do epitélio dos túbulos do epidídimo, levando a extensiva formação de cistos intra-epiteliais que ficam repletos de agrupamentos de neutrófilos e leucócitos necróticos;

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Lesões* O lúmen de vários túbulos epididimários pode conter um exsudato com abundantes neutrófilos e algumas células epiteliais descamadas;

* testículos pode-se encontrar moderado edema intertubular e alterações degenerativas dos túbulos seminíferos, atrofia e mineralização;

* A vesiculite seminal caracteriza-se por densa infiltração focal com muitos linfócitos e poucas células plasmáticas e neutrófilos ocasionais que distribuem-se no tecido intersticial

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Slide 20

Lesões* Nas ovelhas, observa-se uma severa endometrite, caracterizada por infiltração difusa de macrófagos, linfócitos e plasmócitos, envolvendo todas as camadas da mucosa, particularmente os espaços perivasculares e periglandulares;

*Endometrite necrótica com focos de desnudação da mucosa;

* Na placenta estão presentes alterações na área intercotiledonária incluindo necrose de trofoblastos, edema e placentite supurativa multifocal;

*Alterações histológicas no feto abortado incluem a presença de exsudato supurativo nos brônquios, bronquíolos e pulmões

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Slide 21

Sinais e Sintomas* Os sinais clínicos inerentes à infecção por B. ovis estão relacionados à esfera reprodutiva;

*Lesões palpadas no conteúdo escrotal de carneiros adultos;

*Epididimite ou orquite palpável quatro a 12 semanas pós-infecção

*Carneiros jovens podem apresentar epididimite com lesões, principalmente na cabeça do epidídimo, causadas por outros organismos pleomórficos gram-negativos como Histophilus ovis e Actinobacillus seminis;

*Os testículos apresentam-se deformados, aumentados de tamanho e consistência suave, no entanto, em alguns testículos observa-se diminuição do tamanho e consistência firme devido a atrofia e frequente mineralização

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Sinais e Sintomas* Alterações na qualidade do ejaculado;

*Presença de células inflamatórias no sêmen;

*Redução na performance reprodutiva dos animais afetados e até infertilidade;

* Abortos no final da gestação e nascimentos de cordeiros fracos ou débeis

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Slide 23

Sinais e Sintomas

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DiagnosticoO diagnóstico da epididimite infecciosa dos carneiros deve basear-se na combinação do exame clínico e sua confirmação pelo isolamento da B. ovis do sêmen e/ou resultados positivos na provas sorológicas, devendo-se levar em consideração os dados epidemiológicos do rebanho.

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Slide 25

Diagnostico

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Slide 26

DiagnosticoDiagnostico Clinico:

* Observação e palpação dos testículos e epidídimos devem ser realizadas para identificar a presença de lesões aparentes, devendo-se atentar para alterações como volume dos testículos e epidídimos, consistência, presença ou ausência de dor e se as lesões são uni ou bilaterais;

*

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Slide 27

DiagnosticoDiagnostico Laboratorial:

Métodos diretos

* Cultivo em meios seletivos para isolamento da bactéria a partir do sêmen de carneiros, descargas vaginais;

* Após a necropsia, os órgãos preferidos em termos de probabilidade de isolamento do agente são epidídimo, vesícula seminal, ampola, glândula bulbouretral e linfonodos inguinais em carneiros;

* B. ovis pode ser isolada em meios não seletivos, como ágar base enriquecido com 10% de soro ovino ou bovino estéreis, ou em meio ágar sangue com 5 a 10% de sangue ovino desfibrinado

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Slide 28

Diagnostico* A identificação da bactéria é feita pela morfologia da colônia, teste positivo da acriflavina, reação de catalase positiva, oxidase negativa, não produz H2S

* Métodos Moleculares:

* Reação em cadeia de polimerase (PCR) para o diagnóstico da infecção por B. ovis em amostras de sêmen de carneiros;

* PCR em paralelo com a cultura bacteriológica;

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Slide 29

DiagnosticoDiagnostico Laboratorial:

Métodos Indiretos

*Provas sorológicas as mais utilizadas são:

* Reação de Fixação de Complemento (FC);

*Imunodifusão em Gel de Agarose (IDGA);

*Ensaio Imunoenzimático (ELISA).

*O método de IDGA é o teste mais utilizado para o diagnóstico de triagem de rebanhos.

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Slide 30

Diagnostico*Estudos comparativos demonstraram que o teste de ELISA é mais sensível (97,6%) que o IDGA (96,4%) e FC (92,7%);

*A combinação de IDGA e ELISA apresentou 100% de sensibilidade contra 96,4% obtidos com a combinação de IDGA e FC.

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Slide 31

Controle e Profilaxia* Identificação e eliminação dos carneiros soropositivos e/ou com lesões genitais devem ser a base para impedir a perpetuação da doença no rebanho;

* A identificação dos animais infectados e/ou doentes pode ser feita através de métodos sorológicos como a IDGA, RFC’ e ELISA;

* Histórico do rebanho com relação à introdução de animais de áreas onde a doença é frequente, problemas reprodutivos e alterações na qualidade do sêmen;

* O mais recomendado é associação de vários métodos de diagnóstico, uma vez que o uso de um método isolado apresenta limitações como, por exemplo: as lesões clínicas dos órgãos genitais só são apresentadas em cerca de 50% dos carneiros infectados

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Slide 32

Controle Profilaxia* Programa Nacional de Sanidade de Caprinos e Ovinos (PNSCO) do MAPA – Brasil

*Não prevê o uso de vacinação para o controle da brucelose ovina;

* Comentário: Amostra Rev. 1 de B. melitensis é provavelmente a melhor vacina disponível para a profilaxia da infecção por B. ovis – proibida a comercialização no Brasil

* Certificação voluntária de propriedades como livres do microrganismo;

* Estabelecimentos serão examinados todos os carneiros não castrados com idade acima de seis meses

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Slide 33

Controle Profilaxia* Animais soropositivos serão separados e sacrificados;

* Estabelecimento obter três testes sorológicos negativos com intervalos semestrais para obter a certificação;

* A renovação do certificado será anual

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Slide 34

Epidemiologia e Dados Regionais

ALVES, F. S. F.; PINHEIRO, R. R.; ANDRIOLI, A.; YOSHIDA FACCIOLI, P. Y. F.; VESCHI, J. L. A.; ALVES, C. A.; SANTOS, F. A. ConsideraçõesEpidemiológicas e do Diagnóstico Sorológico da Brucelose Ovina no Brasil. 2016. Embrapa Ovinos e Caprinos. Sobral – CE, Nota Técnica.10p.

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Slide 35

Inquéritos sorológicos

35

Clementino et al. (2007) prev. 5,57%(28/498); Lima (2015) prev. 5,4%(13/241); Alves et al.(2010) prev. 7,5%(6/80); Santos et al. (2013) prev. 5,20%(59/1134)

Chaves et al. (2002) prev. 5,6(12/216)

Silva et al. (2009) prev. 3,28%(6/183); Souza et al. (2012) prev. 0,72% (5/694); Araújo et al. (2013) prev.6,94%(55/793)

Silva et al. (2003) prev. 34%(103/290); Lima (2015) prev. 7,65% (36/470); Azevedo et al. (2004)prev.11,3%

Lima (2015) 3,5%(21/593)

Pinheiro Junior et al. (2009) prev.3,1% (18/579)

Batista (2012) prev. 8,54%(40/468)

Coleto et al. (2003) prev. 16,25%(26/160)

Batista (2012) prev. 7,81%(30/384)

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Slide 36

Epidemiologia

ALVES, F. S. F.; PINHEIRO, R. R.; ANDRIOLI, A.; YOSHIDA FACCIOLI, P. Y. F.; VESCHI, J. L. A.; ALVES, C. A.; SANTOS, F. A. ConsideraçõesEpidemiológicas e do Diagnóstico Sorológico da Brucelose Ovina no Brasil. 2016. Embrapa Ovinos e Caprinos. Sobral – CE, Nota Técnica.10p.

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Slide 37

Epidemiologia

ALVES, F. S. F.; PINHEIRO, R. R.; ANDRIOLI, A.; YOSHIDA FACCIOLI, P. Y. F.; VESCHI, J. L. A.; ALVES, C. A.; SANTOS, F. A.Considerações Epidemiológicas e do Diagnóstico Sorológico da Brucelose Ovina no Brasil. 2016. Embrapa Ovinos e Caprinos.Sobral – CE, Nota Técnica. 10p.

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Slide 38

Inquéritos sorológicos

Costa et al. (2016) prev. 2,4%(3/124); Salaberry et al. (2011) prev.0%(0/334); Marques (2006) prev. 5,3%(44/833)

Schäfer et al. (1997) prev. 0%(0/69)

Reis (2005) prev. 0,9%

(1/109); Cunha Filho et al. (2007) prev. 1,4%(3/213)

Gonçalves (2005) prev. 27,6% (29/105)

Juliano et al. (2011) prev. 12,7% (153/1198)

Marinho & Mathias (1996) prev. 0%(0/850); Rizzo et

al. (2009) prev. 1,96% (4/204); Nozaki et al. (2004)

prev. 12% (124/1033)Machado et al. (2015) prev.2,9%(52/1800); Magalhães Neto & Gil-Turnes

(1996) prev. 13,43%(220/1638); Ramos (1966) prev. 6,5%(216/3317)

Lima et al. (2007) prev. 3,64%(13/357)

Martins et al. (2013) prev. 31,6%(45/142)

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Slide 39

Isolamentos

Alves et al, 2010

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Pesq. Vet. Bras. 33(4):459-463, abril 2013

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Slide 58


Soroprevalência e comparação de técnicas sorológicas utilizadas no diagnostico da Brucelose Ovina no Brasil

Trabalho submetido à Revista Semina: Ciências Agrária – PR. Qualis B1-JCR 0,229. Manuscrito em português.

Fabrine Alexandre dos Santos, Francisco Selmo Fernandes Alves,Raimundo Rizaldo Pinheiro, Alice Andrioli, Patrícia Yoshida FaccioliMartins, Ana Milena Cesar Lima, Luis Antônio Mathias, GlaucenyraCecília Pinheiro da Silva, Sérgio Santos de Azevedo, Clebert José Alves.

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Slide 59

Epidemiologia e Dados Regionais Resultados

EstadosAnimais Propriedades

Nº total Positivos (%) Nº total Positivos (%)

Paraíba 240 13 (5,4% IC 3,19 – 9,05) 24 9 (37,50% IC 21,16 – 57,29)

Rio Grande do Norte 470 35 (7,4% IC 5,40 – 10,18) 47 23 ( 48,94% IC 35,28 – 62,76)

Ceará 412 31 (7,5% IC 5,35 - 10,48) 34 19 (55,88% IC 39,45 – 71,12)

Piauí 750 50 (6,6% IC 5,09-8,68) 63 27 (42,86% IC 31,40 – 55,14)

Sergipe 593 20 (3,3% IC 2,19 –5,15) 50 16 (38 % IC 25,86 – 51,85)

Total 2465 149 (6,04% IC 5,17% -7,06%) 218 94 (43,11% IC 36,72 – 49,76)

Tabela 1. Prevalência de propriedades positivas (focos) e animais para a infecção por Brucella ovisem cinco Estados ( Piauí, Ceará, Rio Grande do Norte, Paraíba e Sergipe), da região do Nordeste doBrasil. entre os anos de 2010 a 2012

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Tabela 2- Resultados da Sorologia para Brucelose ovina por diferentes técnicas diagnosticas.

Amostras IDGA FC

ELISA

P S N

Banco Soros CNPC/ Sobral 130/130 0/130 2 6 122

Banco Soros LDT 41/41 0/41 2 14 25

Total 171/171 0/171 4 20 147

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Tabela 3- Resultados da Sorologia para Brucelose ovina em diferentes Estados da região doNordeste em função das técnicas diagnosticas.

Amostras IDGA FCELISA

P S N

Paraíba 13/13 0/13 0/13 1/13 12/13

Rio Grande do

Norte35/35 0/35 1/35 1/35 33/35

Sergipe 20/20 0/20 1/20 2/20 17/20

Piauí 33/33 0/33 0/33 2/33 31/33

Ceará 29/29 0/29 0/29 0/29 29/29

Total 130/130 0/130 2/130 6/130 122/130

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Documents

Unidade Acadêmica de Medicina Veterinária Centro de Saúde e … · 2017-06-06 · Slide 15 Patogenia Penetração mucosas oral, nasal, ocular, prepucial, retal, vaginal e uterina