Daise Aparecida Rossi

Campylobacter sp in eggs from cloacal swab positive breeder hens

Campylobacter sp is a microaerophilic, thermotolerating Gram negative bacterium, known to be one of the main causes of food-borne human infections. Among the foods that carry these microorganisms, the chicken is outstanding. In Brazil, a large chicken exporting country, few researches are conducted about their prevalence in breeder hens and the transmission through eggs. The aim of this research was to verify the presence of Campylobacter sp in the shells and within the eggs from positive cloacal swab breeder hens. Microbiological analyses were made on cloacal swabs of 140 weighed breeder hens. The positive breeder hens were set aside and in a total of 244 of their eggs, Campylobacter sp was present in macerated shells and yolk contents during 7 weeks. Out of the 140 researched breeder hens, 25 (17.8%) were positive from cloacal swabs, however the eggs were not positive. The physiological characteristics of the birds, their eggs and Campylobacter sp favor the bacterium entering and surviving in the eggs, but in this study, no positive result was found in macerated shells or in the yolks, indicating that vertical transmission is probably an unusual event.

Transfer, viability and colonisation of Campylobacter jejuni in the chicken vitellus and in embryos

1. The objective of this study was to evaluate the ability of Campylobacter jejuni to penetrate and colonise eggs from specific-pathogen-free (SPF) and heavy breeder hens, and to determine its effects on the viability of SPF embryos. 2. We detected C. jejuni in 10% of breeder hens and 20% of SPF eggs, which demonstrates the ability of the bacteria to go through the pores of eggs and contaminate the vitellus after 3 h of contact. These results indicate that there is a risk of contamination under commercial production conditions, where, after oviposition, there is contact between the egg and organic material such as faeces and blood. 3. We observed that in 80% of SPF eggs analysed, C. jejuni survived the 21-d incubation period. This positive result suggests that this microorganism was also responsible for early embryonic mortality. 4. The ability of C. jejuni to penetrate the eggs in this study suggests that serious problems may occur under natural field conditions, which may cause significant problems for producers.

Microbiota of the cecum, ileum morphometry, pH of the crop and performance of broiler chickens supplemented with probiotics

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.

Characterization of the virulence, growth temperature and antibiotic resistance of the Campylobacter jejuni IAL 2383 strain isolated from humans

The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.

Occurrence and characterization of Campylobacter spp.isolates in dogs, cats and children

Com o objetivo de melhorar o entendimento das infeccoes por Campylobacterspp. em caes, gatos e criancas no Brasil, foram avaliadas 160 amostras fecais de criancas e 120 swabs retais de pets (103 caes e 17 gatos). Do total das amostras das criancas, 6,87% foram positivas para Campylobacter spp. e em caes e gatos a positividade foi de 18,3%. Das 33 amostras positivas para Campylobacter spp., 57,6% foram identificadas como C. jejunie 33,4% foram identificadas como C. coli. Mais de 50% das amostras isoladas de pets foram resistentes a ceftiofur, sulphazotrim, norfloxacina e tetraciclina. Em criancas, a maioria das amostras foi resistente a amoxilina, cefazolina, ceftiofur, eritromicina e norfloxacina. De 19 isolados de C. jejuni, 11 isolados de criancas e cinco (5) de caes tinham dois (2) dos quatro (4) genes de virulencia flaA, pldA, cadFor ciaB. Associacao positiva entre a presenca de Campylobacter spp. e diarreia em caes e gatos foi observada em animais desverminados e com hemograma sugestivo de infeccao bacteriana. Tambem houve associacao positiva entre a presenca dos genes de virulencia e a ocorrencia de diarreia, e entre o uso de antibioticos e a positividade para Campylobacterspp. em suabes fecais de pets. Os dados desse trabalho indicam que cepas virulentas de Campylobacter spp. sao fatores de risco para diarreia em caes e a resistencia antimicrobiana e comum em isolados de caes

Identificação sorológica e relação filogenética de Salmonella spp. de origem suína

Salmonella spp. is an important zoonotic pathogen that can spread along the production chain of swines. The objective was to evaluate the incidence of Salmonella spp. in feces of swines in termination phase in the farm, in the pre-slaughter and environmental samples, identify the serotypes and establish a phylogenetic relationship among the isolates. Three collections were done in different batches of pigs housed in the termination pen and in the same animals after transport to the slaughterhouse totaling 90 plots and 9 environmental samples. The transport does not influenced the percentage of isolation of the microorganism (p>0.05). Of the total of 99 samples, 50 (50.5%) were identified as Salmonella spp., and was identified a variety of serovars: Agona (30%), Typhimurium (26%), Minnesota (24%), Infantis (18%) and Panama (2%). Dendrograms showed homology among isolates of different serovars grouped into clusters. The similarity was independent of the local of isolation, indicating the presence of several clones. The main sources of infection were cross-contamination between animals and environment and the consumption of contaminated feed. The diversity of strains and homology among the isolates indicates a common origin, demonstrating a need for monitoring of zoonotic bacterias and the deployment of more effective control measures for Salmonella spp. in swines.

Campylobacter spp. and Related Organisms in Poultry

Members of the Campylobacter genus are Gram-negative, microaerophilic, small bacilli with polar flagellation. Bacterial movement has a typical corkscrew characteristic. There are many species associated with different diseases. This chapter describes the taxonomy, genetic structure, and general characteristics of the Campylobacter genus focusing on thermophilic species and/or those that colonize the intestinal tract of birds.

Campylobacter jejuni in commercial eggs

This study evaluated the ability of Campylobacter jejuni to penetrate through the pores of the shells of commercial eggs and colonize the interior of these eggs, which may become a risk factor for human infection. Furthermore, this study assessed the survival and viability of the bacteria in commercial eggs. The eggs were placed in contact with wood shavings infected with C. jejuni to check the passage of the bacteria. In parallel, the bacteria were inoculated directly into the air chamber to assess the viability in the egg yolk. To determine whether the albumen and egg fertility interferes with the entry and survival of bacteria, we used varying concentrations of albumen and SPF and commercial eggs. C. jejuni was recovered in SPF eggs (fertile) after three hours in contact with contaminated wood shavings but not in infertile commercial eggs. The colonies isolated in the SPF eggs were identified by multiplex PCR and the similarity between strains verified by RAPD-PCR. The bacteria grew in different concentrations of albumen in commercial and SPF eggs. We did not find C. jejuni in commercial eggs inoculated directly into the air chamber, but the bacteria were viable during all periods tested in the wood shavings. This study shows that consumption of commercial eggs infected with C. jejuni does not represent a potential risk to human health.

Antimicrobial effect of turmeric (Curcuma longa) on chicken breast meat contamination

The aim of this study was to evaluate the efficacy of turmeric (Curcuma longa), also known in Brazil as saffron, on the reduction of Staphylococcus aureus and Escherichia coli counts in chicken meat. Forty breast meat samples were divided in two groups (A and B). In group A, 103-104E. coli (ATCC 25922) cells were inoculated and group B samples were inoculated with 104-105S. aureus (ATCC 9801) cells, after which each group was divided in three samples. The first sample was analyzed immediately after inoculation. The second sample (control group) was stored at 4 oC for 48 hours and turmeric at 1% (w/w) was added to the third sample, which was homogenized and then stored under the same conditions as the second sample. E. coli and S. aureus were enumerated in all samples. Mean bacterial counts determined for the control samples and for the samples with turmeric addition after 48h of storage were 1.83 x 104 CFU g-1 and 1.80 x 104 CFU g-1 for S. aureus, and 9.36 x 103 CFU g-1 and 7.25 x 103 CFU g-1 for E. coli, respectively. The results showed that there was no significant reduction in bacterial counts with the addition of 1% turmeric to chicken breast meat.

Campylobacter sp in organs and meconium of day-old broiler chicks derived from naturally infected breeder hens

Campylobacter sp is an important agent that causes foodborne infection, particularly in food of poultry origin. Therefore, the efficient control of the transmission routes in chicken farms is of outmost importance to prevent it from spreading. In chicken farms, the main transmission route of this microorganism is horizontal, as the vertical route continues to be the object of inconclusive researches. The objective of this study was to verify the presence of Campylobacter sp in breeder hens, meconium, and other organs of day-old chicks derived from these breeders in order to obtain information on vertical transmission. Microbiological analyses were performed, using cloacal swabs from 279 breeder hens. Positive breeders were segregated, and the presence of Campylobacter sp. was verified in their progeny by analyzing 117 meconium samples; 36 heart, liver and spleen samples (pool of 3 day- old chicks per sample), and 34 intestine samples (pool of 3 chickens in each sample). The analysis of the 279 breeder hens showed that 39 (13.97%) were positive for Campylobacter sp, using the cloacal swab method. The meconium and the organs of day-old chicks were not positive. The physiological characteristics of breeder hens, of eggs, and of Campylobacter sp favor the entrance of bacteria and their survival inside the eggs, and therefore, this probably is the contamination route of day-old chicks. However, chick meconium and organs were negative in the present experiment, indicating that the vertical way of transmission is a rare event.