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Previous issue date: 2008-04-01 / Funda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / This study had the objective of evaluating the capacity of Stomoxys calcitrans fly to vehicular
bacterial agents causing bovine mastitis. Just as evaluate the enterobacterian microbiota on the
external surface, mouth parts and abdominal digestive tract of the stable fly. And also realize
a survey of the bacterial microbiota of the cases of bovine mastitis on the visited properties.
There were taken mastitis milk samples and 20 specimens of stable flies on 10 visited
properties in the municipatility of Barra Mansa and Resende, Rio de Janeiro State. The
collected milk was submitted to bacterial isolation in the laboratory of Bacteriology UFRRJ.
Samples were subcultures on MacConkey (MC) agar, Brain-Heart Infusion agar (BHI) and
Manitol-Phenol Red agar. The flies were individually washed in BHI broth, sterilized and had
its mouth parts and abdominal intestinal content dissected under stereoscope microscope.
These structures were macerated in BHI broth and, after 24 hours at 37 ?C, subcultured at MC
agar, Agar Eosin Methilen Blue (EMB) and enriched in Sodium Tetrathionate Broth after
incubation, and were subcultured in Salmonella-Shigella agar. After this stage, each isolated
colony was observed for differences in morphology as size and pigment production
characteristics to be identified through biochemical specific tests. It was also realized an
antimicrobial resistance test to verify to which antibiotics the isolated bacteria were sensible.
When were coincidences between the enterobacterian species in the milk and on the flies
evaluated segments, was realized a Random Amplified Polymorphic DNA (RAPD-PCR) as
an instrument of evaluating the genetic diversity and the eletrophoretic profile similarity the
bacterial subpopulations. In agreement with the obtained results, was verified that the mastitis
milk had as etiological agents specially the species Staphylococcus aureus, coagulasenegative
staphylococci and Escherichia coli. The antimicrobials with less resistance rate were
Amoxilin-Clavulanic Acid and Norfloxacin. Regarding to the enterobacterian microbiota
verified on S. calcitrans segments, Escherichia coli, Enterobacter agglomerans, Enterobacter
cloacae e Salmonella spp were the most prevalent bacteria. The segment with the higher
number of isolations was the external surface, with 73 (45.91%) distinct colonies isolated.
The mouth parts had 46 (28.93%) colonies and the abdominal digestive tract 40 (25.16%)
isolated colonies. The eletrophoretic profile similarity happened just on E. coli subpopulations
on milk samples and S. calcitrans segments of the same property. This way, was possible to
observe that the flies do not act as a vehicle of any bovine mastitis causative E. coli.
Regarding to the identified E. coli, was verified that 13.79% were Shiga Toxin-Producing,
and the genes stx1, stx2 and eae were identified. On the present study, was verified that this
fly has the capacity to act as vehicle to enterobacterian, on its external surface as well as
inside its body. The stable fly control may contribute to the improvement on animal
productivity and sanity. / Este estudo teve o objetivo de avaliar a capacidade da mosca Stomoxys calcitrans em veicular
agentes bacterianos causadores de mastite bacteriana bovina, bem como avaliar a microbiota
enterobacteriana na superf?cie externa, aparelho bucal e trato digest?rio abdominal da mosca
dos est?bulos; e fazer um levantamento da microbiota bacteriana nos casos de mastite bovina
das propriedades visitadas. Foram coletadas amostras de leite com mastite e 20 esp?cimes da
mosca dos est?bulos em 10 propriedades visitadas nos munic?pios de Barra Mansa e Resende,
Estado do Rio de Janeiro. No Laborat?rio de Bacteriologia da UFRRJ foi realizado o
isolamento bacteriano do leite coletado em Agar MacConkey (MC), Agar Infuso C?rebro
Cora??o (BHI) e Manitol Vermelho de Fenol. As moscas foram lavadas individualmente em
Caldo BHI, esterilizadas e, sob a luz de um microsc?pio estereosc?pio, tinham seu aparelho
bucal e conte?do intestinal abdominal dissecado. Estas estruturas foram maceradas em Caldo
BHI e ap?s 24 horas a 37 ?C repicados em Agar MC, Agar Eosina Azul de Metileno (EMB) e
enriquecidos em Caldo Tetrationato de S?dio que ap?s incuba??o, era repicado em Agar
Salmonella-Shigella. Ap?s esta etapa, cada col?nia isolada era avaliada quanto ?s suas
caracter?sticas morfo-tintoriais para serem identificadas atrav?s de testes bioqu?micos
espec?ficos. Tamb?m foi realizado o teste de resist?ncia antimicrobiana, com a finalidade de
se verificar, as quais antibi?ticos as bact?rias isoladas foram sens?veis. Quando havia
coincid?ncia de esp?cies enterobacterianas no leite e nos segmentos avaliados das moscas, se
realizava a t?cnica de an?lise do polimorfismo dos segmentos de DNA obtidos por
amplifica??o rand?mica (RAPD-PCR) como instrumento de avalia??o da diversidade gen?tica
e das rela??es de clonalidade entre estas subpopula??es bacterianas. De acordo com os
resultados obtidos, foi verificado que o leite com mastite, teve como principais agentes
etiol?gicos as esp?cies Staphylococcus aureus, Estafilococos Coagulase Negativa (ECN) e
Escherichia coli. Os antimicrobianos com menor taxa de resist?ncia foram Amoxicilina/?cido
Clavul?nico e Norfloxacina. Com rela??o ? microbiota enterobacteriana verificada nos
segmentos de S. calcitrans, foi observado que Escherichia coli, Enterobacter agglomerans,
Enterobacter cloacae e Salmonella spp foram as bact?rias mais prevalentes. O segmento com
o maior n?mero de isolados foi a superf?cie externa, onde foram isoladas 73 col?nias distintas
(45,91%), seguida pelo aparelho bucal com 46 col?nias (28,93%), e pelo trato digest?rio
abdominal com 40 col?nias isoladas (25,16%). Houve clonalidade apenas de sub-popula??es
de E. coli entre as amostras de leite e de segmentos de S. calcitrans de uma mesma
propriedade. Desta forma, foi poss?vel observar que as moscas n?o veiculavam nenhuma E.
coli causadora de mastite bovina. No que se refere ?s E. coli identificadas, foi verificado que
13,79% eram Shiga-Toxig?nicos, sendo identificados os genes stx1, stx2 e eaeA. No presente
estudo, verificou-se que esta mosca tem a capacidade de veicular enterobact?rias, tanto em
sua superf?cie externa como no interior de seu corpo. O controle da mosca dos est?bulos
poder? contribuir para melhora na sanidade e produtividade animal.
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:tede/828 |
| Date | 01 April 2008 |
| Creators | Castro, Bruno Gomes de |
| Contributors | Bittencourt, Avelino Jos?, Souza, Miliane Moreira Soares de |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Curso de P?s-Gradua??o em Ci?ncias Veterin?rias, UFRRJ, Brasil, Parasitologia Veterin?ria |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/doctoralThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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