A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

Meeting abstrac

Transference in vitro of the resistance to the antimicrobials between Escherichia coli, Lactobacillus spp. and Salmonella enteritidis isolated from chickens

O objetivo deste trabalho foi verificar a possibilidade de transferência de resistência aos antimicrobianos entre bactérias normais da microbiota de frangos e Salmonella Enteritidis. Utilizamos amostras de Lactobacillus spp. (L. spp.), Salmonella Enteritidis (SE) e Escherichia coli (E. coli) previamente isolados de frangos, selecionados após prova de sensibilidade antimicrobiana in vitro conforme metodologia padrão (Comitê Nacional para Padrões Clínicos de Laboratório). Utilizamos aqueles com resistência e sensibilidade aos antimicrobianos indutores, chamados de bactérias doadoras e receptoras, respectivamente. Os antimicrobianos indutores foram utilizados para estimular a transferência de resistência aos antimicrobianos entre as bactérias. A possibilidade de transferência foi verificada da E. coli resistente para a SE e L. spp. Também foi verificada a transferência de uma amostra de L. spp resistente aos antimicrobianos indutores para a SE. Só foi possível verificar a transferência da resistência aos antimicrobianos indutores quando a bactéria doadora foi a E. coli e a bactéria receptora foi a SE. No presente estudo concluímos que a transferência de resistência aos antimicrobianos entre bactérias é possível, mas nem todas as bactérias participam desse evento, não transmitindo e nem adquirindo esta resistência.The objective of this work was to verify the possibility of transference of resistance to the antimicrobials between bacteria that are in the present normal microbiota of chickens and Salmonella Enteritidis. Samples of Lactobacillus spp. (L. spp.), Salmonella Enteritidis (SE) and Escherichia coli (E. coli) previously isolated from chickens, selected after the test of sensitivity antimicrobial in vitro according the standard method (National Committee for Clinical Laboratory Standards) utilizing those with resistance and sensibility to the antimicrobials inductors, named donor and receptor bacteria, respectively were used. Antimicrobials inductors were utilized to stimulate the transference of resistance to the antimicrobials between the bacteria. The possibility of transference was verified from the E. coli resistant to the SE and L. spp. Transference of a sample of L. spp resistant to the antimicrobials inductors to the SE was also verified. It was only possible to verify the transference of the resistance to the antimicrobials inductor when the donor bacteria was the E. coli and the bacteria receptor was SE. In the present study we conclude that the transference of resistance to the antimicrobials between bacteria is possible, however, not all bacteria participate in that trial, not transmitting and neither acquiring this resistance

Evaluation in vitro of the antagonistic substances produced by Lactobacillus spp. isolated from chickens

To determine the inhibitory capacity of lactic acid bacteria due to the action of antagonistic substances, we tested 474 isolates of Lactobacillus from the crop and cecum of chickens against gram-positive and gram-negative indicator microorganisms by the spot-on-the-lawn and well-diffusion antagonism methods. Of the 474 isolates, 265 demonstrated antimicrobial activity against the indicator microorganisms. Isolates identified as L. reuteri, L. salivarius, or Lactobacillus spp. inhibited Enterococcus faecalis, E. faecium, Listeria monocytogenes, and Salmonella spp. but not L. casei, L. delbrueckii, L. fermentum, or L. helveticus by the well-diffusion simultaneous antagonism method under anaerobic incubation conditions. The antagonistic substances produced by some of the Lactobacillus isolates were inactivated after treatment by proteolytic enzymes, which suggested that the substances could be antimicrobial peptides or bacteriocins

Avaliação dos efeitos de 5-hidroxitriptofano em-hidroxibenzilhidrazine associados a Lactobacillus spp. na morfometria intestinal e imunomarcação de serotonina em frangos de corte desafiados com Salmonella Enteridis

Resumo As células enterocromafins são um dos componentes da mucosa intestinal que liberam serotonina para o lúmen, promovendo atividades secretórias e crescimento celular de vários tecidos, incluindo vilosidades intestinais. O presente estudo avaliou as influências do 5-hidroxitriptofano (5HTP) e do m-hidroxibenzilhidrazine (NSD1015), associados a Lactobacillus spp., sobre o peso corporal e o desenvolvimento das vilosidades intestinais na porção proximal do duodeno de frangos de corte desafiados com Salmonella Enteritidis. Verificou-se também se a presença de Lactobacillus spp. e Salmonella Enteritidis influenciaram a imunomarcação de serotonina no duodeno e, para isso, o estudo foi dividido em dois experimentos, com e sem desafio por S. Enteritidis. No Experimento 1, em aves sem desafio, os pesos corporais não diferiram significantemente (p>0,05) e, no Experimento 2, aves com desafio, os tratamentos com o precursor isolado e associado a Lactobacillus spp. determinaram maior peso corporal das aves. Nos dois experimentos, as aves tratadas com 5HTP apresentaram aumento na densidade e altura das vilosidades no duodeno, sugerindo a atuação de 5HTP como um agente trófico. A administração de Lactobacillus spp. também determinou altura maior de vilosidades duodenais. Quanto a imunomarcação de serotonina, as aves tratadas com Lactobacillus spp. no Experimento 1 e as aves tratadas com Lactobacillus spp. e desafiadas com S. Enteritidis no Experimento 2, apresentaram valores superiores aos demais tratamentos, sugerindo que a presença destas bactérias promove maior liberação de serotonina para o duodeno, porém o mecanismo exato de como este processo ocorre necessita ser mais elucidado

Comparison of three diagnostic methods for Salmonella enterica serovars detection in chicken rinse

ABSTRACT: Salmonella detection is a key point in food safety testing, because of the frequent association of this pathogen with food poisoning in humans. The standard bacteriological tests currently used for Salmonella-detection are time-consuming; therefore, there is a need to develop alternative methods to accelerate the detection. In order to accelerate Salmonella diagnosis, we used the immunomagnetic separation assay associated with bacteriophage P22 for the rapid detection of the following Salmonella serovars in chicken rinses of drumsticks, artificially contaminated with 5, 10, and 100 CFU/25mL of bacteria: Salmonella enterica subsp. enterica serovar Heidelberg (S. Heidelberg), Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) and Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium). The efficiency of the technique, represented by the time required for detection of positive and negative samples, was compared with that of the standard diagnostic tests used for this pathogen, the bacteriological assay and the polymerase chain reaction (PCR)-based test. This study confirmed the ability of the bacteriophage-associated immunomagnetic separation assay to identify 99.6% of Salmonella-positive samples of the three serovars tested. In contrast, the bacteriological assay and PCR-based test detected 95.1% and 98.5% of the Salmonella-positive samples respectively

Evaluation of in vitro and in vivo adhesion and immunomodulatory effect of Lactobacillus species strains isolated from chickens

This study aimed to characterize the in vitro and in vivo adhesion and immunomodulatory effect of Lactobacillus strains isolated from chickens. Lactobacillus samples isolated from 65-wk-old birds were identified by PCR; their adhesion was evaluated in vitro via basement membrane-type cell matrix and in vivo through carboxyfluorescein succinimidyl amino ester staining inoculation in 1-d-old birds and duodenum, jejunum, ileum, and cecum collections at 1, 4, 12, and 24 h after inoculation. The 5 best adhesive samples at the in vitro test formed a pool for total IgA and IgG measurement in sera and intestinal fluid. The birds were divided into groups by inoculation scheme: group 1 was treated with a pool of Lactobacillus spp. at 2-d-old and challenged 1 d later with Salmonella Enteritidis and then treated again with a pool of Lactobacillus spp. at 4 d of age; group 2 was treated with a pool of Lactobacillus spp. at 2 and 4 d of age; group 3 was challenged with Salmonella Enteritidis at 3 d of age; and group 4 was a negative control. Collections were taken at 7, 14, 21, 28, and 35 d after the first inoculation. The results suggest that basement membrane matrix use represents an important technique for triage of samples for subsequent in vivo evaluation and that carboxyfluorescein succinimidyl amino ester staining is efficient for identifying this bacterial characteristic. The Lactobacillus treated groups (1 and 2) presented the highest IgA concentrations at the end of the experiment (12,054.6 and 10,568.4 ng/mL, respectively). The group 2 IgG values in intestinal fluid exceeded those of the other 3 groups (P 0.05), whereas group 3 presented the highest concentration of this antibody. It is concluded that there was greater adhesion of strains in the cecum and an important correlation between in vitro and in vivo results. These results also suggest the immunomodulatory action of Lactobacillus spp. in the chicken.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Evaluation of Predisposing Factors of Necrotic Enteritis in Experimentally Challenged Broiler Chickens

Clostridium perfringens is the etiological agent of NE, a disease that greatly affects the poultry industry. Experiments on the induction of NE are difficult to carry out, as it is a multifactorial disease, and thus different predisposing factors have been used. This study evaluated the effect of the Gumboro disease vaccine virus vaccine (IBDV-vac) associated or not with infection by Eimeria spp. in broilers, as a predisposing factor for NE. Broilers (n = 99) were divided into groups (11) challenged with IBDV-vac, Eimeria spp. CP type G (CP13, CP14 and CP03) or both. The macroscopic evaluation revealed that the highest average (3.45) of injury occurred for the CP13 + IBDV-vac group. The microscopic analysis showed that Eimeria spp. increased the population of intraepithelial lymphocytes and reduced the villus/crypt ratio in duodenum and jejunum when associated with CP13 or CP14. There was a synergistic effect between the CP strain used and the predisposing factors; nevertheless, it was not clear which was the most effective predisposing factor to potentiate the lesions, suggesting that the association of the strain with the factors should first be evaluated for each experimental protocol