Evaluation of propanediol and cobalamin metabolism in the intestinal colonization and systemic invasion of Salmonella Enteritidis in laying hens / [Avaliação do metabolismo do propanodiol e da cobalamina na colonização intestinal e na infecção sistêmica de Salmonella Enteritidis em poedeiras]

Embora Salmonella Enteritidis (SE) seja capaz de metabolizar 1,2-propanodiol (1,2-Pd), utilizado como fonte de carbono e de energia ao longo de uma rota dependente de vitamina B12, a importância deste composto na infeção de Gallus gallus domesticus por SE permanece desconhecida. No presente estudo, foram construídos um mutante de SE sem os genes pduCDE, que codifica a propanodiol desidratase (Pdu), e outro contendo as deleções no pduCDE e também nos genes cobS e cbiA, responsáveis pela síntese de vitamina B12. Em seguida, avaliou-se a importância do metabolismo do 1,2-Pd em SE para colonização intestinal de infecção sistêmica de poedeiras comerciais. As estirpes mutantes de SE foram capazes de colonizar o intestino, de serem excretadas nas fezes e de invadir o baço e o fígado na mesma intensidade que a estirpe selvagem, o que sugere que os produtos dos genes pduC, pduD, pduE, cobS e cbiA não são essenciais durante infecção por Salmonella Enteritidis nessa espécie.(AU)

Safety and efficacy of a salmonella gallinarum ΔcobSΔcbiA strain with potential to prevent chicken infections by salmonella gallinarum and salmonella enteritidis

New vaccine design techniques have allowed the development of effective vaccine strains against Salmonella infections inwhich the risks of reversion to the wild type and virulence is null. The mutant strain Salmonella Gallinarum ΔcobSΔcbiA was previously shown to be avirulent in chickens. In this study, this strain was tested as a vaccine against Salmonella Gallinarum (SG) and S. Enteritidis (SE) infections, and its protection levels, safety and possible risks of reversion to virulence after vaccination of layers were evaluated. Birds were vaccinated at five days of age or at five and 25 days of age. At 45 days of age, brown and white layers were challenged with SG and SE wild strains, respectively. Two assays to test the possibility of reversion to virulence were performed. Five successive bacterial passages in brown layers were carried out in the first assay. In the second assay, brown layers received a ten-fold concentrated inoculum of the SGΔcobSΔcbiA strain and were evaluated for clinical signs and mortality. In both experiments, no birds that received the inoculation of the attenuated strain died. Additionally, the use of the mutant strain as a vaccine provided good protection levels against both challenge strains.(AU)

The contribution of genes required for anaerobic respiration to the virulence of Salmonella enterica serovar Gallinarum for chickens

Salmonella enterica serovar Gallinarum (SG) is an intracellular pathogen of chickens. To survive, to invade and to multiply in the intestinal tract and intracellularly it depends on its ability to produce energy in anaerobic conditions. The fumarate reductase (frdABCD), dimethyl sulfoxide (DMSO)-trimethylamine N-oxide (TMAO) reductase (dmsABC), and nitrate reductase (narGHIJ) operons in Salmonella Typhimurium (STM) encode enzymes involved in anaerobic respiration to the electron acceptors fumarate, DMSO, TMAO, and nitrate, respectively. They are regulated in response to nitrate and oxygen availability and changes in cell growth rate. In this study mortality rates of chickens challenged with mutants of Salmonella Gallinarum, which were defective in utilising anaerobic electron acceptors, were assessed in comparison to group of bird challenged with wild strain. The greatest degree of attenuation was observed with mutations affecting nitrate reductase (napA, narG) with additional attenuations induced by a mutation affecting fumarate reductase (frdA) and a double mutant (dmsA torC) affecting DMSO and TMAO reductase.

Isolamento de salmonella: comparação das etapas de pré- enriquecimento e enriquecimento direto de amostras de fezes armazenadas por 24 e 96 horas

ABSTRACT This work was carried out to assess the preenrichment (PE) and enrichment (DE) steps for isolating Salmonella serotypes Enteritidis (SE) and Typhimurium (STM) from chicken feces kept at 4° C for 24 and 96h. The samples were artificially contaminated and kept in 1% peptone water at 4° C for 24 or 96h. After that, part of them was incubated at 37° C/24h and part was inoculated into enrichment broth, selenite broth plus novobiocin (SN) and tetrathionate broth plus novobiocin (TN) incubated at 37°C/24h. The PE culture was inoculated in SN, TN and Rapapport-Vassiliadis novobiocin (RVN), also incubated at 37° C/24h. The enrichment broth was plated on brilliant green agar (BGA), MacConkey agar (MCA), Hektoen agar (HEA), Salmonella-Shigella agar (SSA), xylose-lysine desoxicholate agar (XLDA) and xylose-lysine tergitol 4 (XLT4), which were incubated at 37° C/24h. Salmonella-like colonies were submitted to TSI agar and LIA agar, and incubated at 37° C/24h, as well as to slide agglutination tested with poly O and poly H Salmonella antiserum. When the samples were stored for 24h there was no difference between PE and DE (p > 0.05). However after 96h the PE was superior to DE (p 0.05). For enrichment, better results were seen with RVN broth (p 0.05). The XLD yielded

RESUMO Este trabalho foi desenvolvido para avaliar comparativamente o isolamento de Salmonella sorotipos Enteritidis (SE) e Typhimurium (STM) a partir do enriquecimento direto (ED) ou processamento com pré-enriquecimento (PE) de amostras de fezes de aves adultas, armazenadas em água peptonada tamponada a 1% (APT) por 24 ou 96h a 4º C. Utilizou-se os caldos de enriquecimento Rapapport-Vassiliadis novobiocina (RVN), tetrationato-novobiocina (TN) e selenitonovobiocina (SN) e os meios para plaqueamento ágar verde brilhante (VB), ágar de MacConkey (MC), ágar de Hektoen (HE), ágar Salmonella-Shigella (SS), ágar xilose lisina desoxicolato (XLD) e ágar xilose lisina tergitol 4 (XLT4). O procedimento bacteriológico incluiu as etapas de pré-enriquecimento, enriquecimento em caldo seletivo, plaqueamento, testes bioquímicos presuntivos e confirmação sorológica com utilização de soros polivalentes anti-antígenos somáticos e anti-antígenos flagelares de Salmonella. Não houve diferença estatisticamente significativa (p > 0,05) para as amostras armazenadas por 24h submetidas tanto ao PE quanto ao ED. Entretanto, em armazenagem por 96h o número de isolamentos nas amostras submetidas ao PE foi estatisticamente superior às submetidas ao ED (p 0,05). Quanto aos caldos enriquecedores, não houve diferença estatística de número de isolamentos (p > 0,05) entre os caldos SN e TN, mas o caldo RVN mostrou-se estatisticamente superior aos demais (p 0,05). Para os meios de plaqueamento, o XLD destacou-se por promover maior número de recuperações, embora sem significado estatístico (p > 0,05) para as amostras estocadas por 24h. Entre os dois sorotipos de Salmonella (SE e STM) não houve diferença estatística no número de recuperações (p > 0,05).