Biofilm represents a way of life that allows greater survival of microorganisms in hostile habitats. Campylobacter jejuni is able to form biofilms in vitro and on surfaces at several points in the poultry production chain. Genetic determinants related to their formation are expressed differently between strains and external conditions are decisive in this respect. Our approach combines phylogenetic analysis and the presence of seven specific genes linked to biofilm formation in association with traditional microbiology techniques, using Mueller Hinton and chicken juice as substrates in order to quantify, classify, determine the composition and morphology of the biomass of simple and mixed biofilms of 30 C. jejuni strains. It also evaluates the inhibition of its formation by biocides commonly used in industry and also by zinc oxide nanoparticles. Genetic analysis showed high heterogeneity with the identification of 23 pulsotypes. Despite the diversity, the presence of flaA, cadF, luxS, dnaJ, htrA, cbrA, and sodB genes in all strains shows the high potential for biofilm formation. This ability was only expressed in chicken juice, where they presented phenotype of a strong biofilm producer, with a mean count of 7.37 log CFU/mL and an ultrastructure characteristic of mature biofilm. The composition of simple and mixed biofilms was predominantly composed by proteins. The exceptions were found in mixed biofilms with Pseudomonas aeruginosa, which includes a carbohydrate-rich matrix, lower ability to sessile form in chicken juice and compact architecture of the biofilm, this aspects are intrinsic to this species. Hypochlorite, chlorhexidine, and peracetic acid were more effective in controlling viable cells of C. jejuni in biofilm, but the existence of tolerant strains indicates exposure to sublethal concentrations and development of adaptation mechanisms. This study shows that in chicken juice C. jejuni presents greater potential in producing mature biofilms.
RESUMO.-[Ocorrência e caracterização de isolados de Campylobacter spp. em cães, gatos e crianças.] Com o objetivo de melhorar o entendimento das infecções porCampylobacter spp. em cães, gatos e crianças no Brasil, foram avaliadas 160 amostras fecais de crianças e 120 swabs retais de pets (103 cães e 17 gatos). Do total das amostras das crianças, 6,87% foram positivas para Campylobacter spp. e em cães e gatos a positividade foi de 18,3%. Das 33 amostras positivas para Campylobacter spp., 57,6% foram identificadas como C. jejuni e 33,4% foram identificadas como C. coli. Mais de 50% das amostras isoladas de pets foram resistentes a ceftiofur, sulphazotrim, norfloxacina e tetraciclina. Em crianças, a maioria das amostras foi resistente a amoxilina, cefazolina, ceftiofur, eritromicina e norfloxacina. De 19 isolados de C. jejuni, 11 isolados de crianças e cinco (5) de cães tinham dois (2) dos quatro (4) genes de virulência flaA, pldA, cadF or ciaB. Associação positiva entre a presença de Campylobacter spp. e diarreia em cães e gatos foi observada em animais desverminados e com hemograma sugestivo de infecção bacteriana. Também houve associação positiva entre a presença dos genes de virulên-cia e a ocorrência de diarreia, e entre o uso de antibióticos e a positividade para Campylobacter spp. em suabes fecais de pets. To improve the understanding of implications of Campylobacter spp. infections in pets and children of different environments were analysed 160 faecal samples from children and 120 from pets (103 dogs and 17 cats). Campylobacter spp. were detected in 6.87% of the children and in 18.3% of the dogs and cats. From 33 stool samples positive for Campylobacter spp., 57.6% were identified as C. jejuni, and 33.4% were identified as C. coli. More than 50% of the isolates in pets were resistant to ceftiofur, sulphazotrim, norfloxacin and tetracycline. In humans, most of the isolates were resistant to amoxicillin, cefazolin, ceftiofur, erythromycin and norfloxacin. From 19 isolates of C. jejuni, 11 isolates from children and 5 from dogs contained two to four of the virulence genes flaA, pldA, cadF or ciaB. We found an association between the presence of virulence genes and diarrhoea. Furthermore, an association was observed between the presence of Campylobacter spp. and diarrhoea in dewormed pets with blood picture suggestive of bacterial infection, and the therapeutic use of antibiotics was associated with more positive detection of Campylobacter spp. in the faeces of pets. Our data indicate that virulent strains of Campylobacter spp. can be risk factor to diarrhoea in animals, and that high resistance to antimicrobial agents is common in pets. de Campylobacter spp. são fatores de risco para diarreia em cães e a resistência antimicrobiana é comum em isolados de cães.
-It was studied, in this work, the effects of supplementation of probiotics on the performance, on the pH of crop content, on the morphometry of the ileum and on the number of enterobacteria in the cecum of the broilers.Out of the 120 studied birds, 60 belonged to the test group and 60 to the control group. At 1, 7, 12, 18, 23 and 28 days of age, the chickens and rations were weighed and mortality and morbidity were determined. Then, the birds were sacrificed and the pH of the crop content was measured, ileum fragments were collected for morphometric analysis and the enterobacteria in the cecal content were quantified. The mean pH was lower in the group that was given probiotics at 1, 7 and 18 days of age. There were no differences on ration consumption, weight, mortality and morbidity rates. The counts of enterobacteria were lower in the supplemented animals at the ages of 7, 18 and 28 days. The measure of villus of the ileum was higher for the group treated with probiotics at all ages, except for 1 day of age in comparison to the control group.This work proves that supplementation with probiotics reduces the pH of the crop content at the ages of 1, 7 and 18 days and it can contribute to the reduction of pathogenic bacteria on the first days of life by reducing the quantity of enterobacteria in the cecum. In addition, probiotics increase the height of villi in the ileum of the birds.Key Words: body weights, competitive exclusion, enterobacteria, intestinal villi, ration Microbiota cecal, morfometria do íleo, pH do inglúvio e desempenho zootécnico de frangos de corte sob suplementação com probióticos RESUMO -Neste trabalho foram estudados os efeitos da suplementação de probióticos sobre o desempenho, o pH do conteúdo do papo, a morfometria do íleo e o número de enterobactérias no ceco de frangos de corte. Das 120 aves estudadas, 60 eram pertencentes ao grupo teste e 60 ao grupo controle. Nas idades de 1, 7, 12, 18, 23 e 28 dias, os frangos e a ração foram pesados e a mortalidade e morbidade foram determinadas. As aves foram, então sacrificadas, o pH do papo foi medido, os fragmentos do íleo foram coletados para análise da morfometria e as enterobactérias do ceco foram quantificadas. A média do pH foi menor no grupo que recebeu probióticos nas idades de 1, 7 e 18 dias. Não houve diferença no consumo de ração, no peso das aves, e nas taxas de mortalidade e morbidade. A contagem de enterobactérias foi menor nos animais sob suplementação com probióticos nas idades de 7, 18 e 28 dias. A medida das vilosidades do íleo foi maior no grupo tratado com probióticos em todas as idades, à exceção de 1 dia em relação ao grupo controle. Este trabalho comprova que a suplementação com probióticos diminui o pH do conteúdo do papo nas idades de 1, 7 e 18 dias, e pode contribuir na redução de bactérias patogênicas nos primeiros dias de vida, diminuindo a quantidade de enterobactérias no ceco. Além disso, os probióticos aumentaram a altura dos vilos do íleo das aves.Palavras-chave: enterobactérias, exclusão competitiva, pes...
Campylobacter coli is an important species involved in human cases of enteritis, and chickens are carriers of the pathogen mainly in developing country. The current study aimed to evaluate the transmission of C. coli and its pathogenic effects in chicken embryos. Breeder hens were inoculated intra-esophageally with C. coli isolated from chickens, and their eggs and embryos were analyzed for the presence of bacteria using real-time PCR and plate culture. The viability of embryos was verified. In parallel, SPF eggs were inoculated with C. coli in the air sac; after incubation, the embryos were submitted to the same analysis as the embryos from breeder hens. In embryos and fertile eggs from breeder hens, the bacterium was only identified by molecular methods; in the SPF eggs, however, the bacterium was detected by both techniques. The results showed no relationship between embryo mortality and positivity for C. coli in the embryos from breeder hens. However, the presence of bacteria is a cause of precocious mortality for SPF embryos. This study revealed that although the vertical transmission is a possible event, the bacteria can not grow in embryonic field samples.
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