Inactivation of phoPQ genes attenuates Salmonella Gallinarum biovar Gallinarum to susceptible chickens

Abstract Salmonella Gallinarum is a host-restrict pathogen that causes fowl typhoid, a severe systemic disease that is one of the major concerns to the poultry industry worldwide. When infecting the bird, SG makes use of evasion mechanisms to survive and to replicate within macrophages. In this context, phoPQ genes encode a two-component regulatory system (PhoPQ) that regulates virulence genes responsible for adaptation of Salmonella spp. to antimicrobial factors such as low pH, antimicrobial peptides and deprivation of bivalent cations. The role of the mentioned genes to SG remains to be investigated. In the present study a phoPQ-depleted SG strain (SG ΔphoPQ) was constructed and its virulence assessed in twenty-day-old laying hens susceptible to fowl typhoid. SG ΔphoPQ did cause neither clinical signs nor mortality in birds orally challenged, being non-pathogenic. Furthermore, this strain was not recovered from livers or spleens. On the other hand, chickens challenged subcutaneously with the mutant strain had discreet to moderate pathological changes and also low bacterial counts in liver and spleen tissues. These findings show that SG ΔphoPQ is attenuated to susceptible chickens and suggest that these genes are important during chicken infection by SG.

Contribution of flagella and motility to gut colonisation and pathogenicity of Salmonella Enteritidis in the chicken

ABSTRACT Salmonella Enteritidis causes fowl paratyphoid in poultry and is frequently associated to outbreaks of food-borne diseases in humans. The role of flagella and flagella-mediated motility into host-pathogen interplay is not fully understood and requires further investigation. In this study, one-day-old chickens were challenged orally with a wild-type strain Salmonella Enteritidis, a non-motile but fully flagellated (SE ΔmotB) or non-flagellated (SE ΔfliC) strain to evaluate their ability to colonise the intestine and spread systemically and also of eliciting gross and histopathological changes. SE ΔmotB and SE ΔfliC were recovered in significantly lower numbers from caecal contents in comparison with Salmonella Enteritidis at early stages of infection (3 and 5 dpi). The SE ΔmotB strain, which synthesises paralysed flagella, showed poorer intestinal colonisation ability than the non-flagellated SE ΔfliC. Histopathological analyses demonstrated that the flagellated strains induced more intense lymphoid reactivity in liver, ileum and caeca. Thus, in the present study the flagellar structure and motility seemed to play a role in the early stages of the intestinal colonisation by Salmonella Enteritidis in the chicken.

Protection of chickens against fowl typhoid using field vaccine programs formulated with the live attenuated strain Salmonella Gallinarum ΔcobSΔcbiA / Proteção de aves contra o tifo aviário utilizando a cepa atenuada Salmonella Gallinarum ΔcobSΔcbiA em diferentes programas vacinais de campo

Salmonella enterica serovar Gallinarum biovar Gallinarum (SG) is a host-specific bacteria that causes the fowl typhoid (FT). This disease is highly pathogenic to commercial chickens, specially brown layers and breeders, causing acute septicemia followed by high morbidity and mortality. Vaccination is extensively adopted in the fields as a biosafety tool for prevention of isolated infections and outbreaks in commercial poultry flocks. The present study evaluated the use of an attenuated SG with deletions on genes cobS and cbiA (SGΔcobSΔcbiA) as a live vaccine, using vaccination schemes adjusted for field conditions. To this end, brown layers were used in two different experiments, to evaluate the long-term protection, necessary in the fields. The vaccination scheme on the first experiment consisted of two doses, the first at 4 th week-of-age and the booster dose at 8 th week-of-age with challenge at 16 th week-of-age with wild SG strain. On the second experiment, the vaccination was carried out by different routes using three doses of the live vaccine, at 4 th , 8 th and 12 th weeks-of-age, and the challenge was done at 20 th weeks-of-age. After the challenge, the mortality was recorded during 28 days, and the egg production (experiment 2) was evaluated and compared with the group of unvaccinated layers. In both experiments, the mortality was significantly reduced, and the egg production was not affected in vaccinated layer-hens. In summary, this study shows the efficacy and the protection of different vaccination schemes against FT that can be applied under field conditions in commercial poultry farms.(AU)

Salmonella enterica sorovar Gallinarum biovar Gallinarum (SG) é uma bactéria hospedeira específica que causa o tifo aviário (TA). Essa doença é altamente patogênica em aves comerciais, especialmente galinhas poedeiras de linhagem vermelha e aves reprodutoras pesadas, causando septicemia aguda, e consequentemente, alta morbidade e mortalidade. A vacinação é amplamente utilizada no campo como uma ferramenta de biossegurança para a prevenção de infecções isoladas e surtos nas granjas avícolas comerciais. O atual estudo avaliou o potencial vacinal de uma cepa viva atenuada de SG com deleções nos genes cobS e cbiA (SGΔcobSΔcbiA), utilizando esquemas de vacinação formulados para utilização em campo. Para isso, as galinhas poedeiras de linhagem vermelha foram utilizadas em dois experimentos diferentes, para avaliar a proteção a longo prazo, necessária no campo. O esquema de vacinação no primeiro experimento consistiu em duas doses, a primeira na quarta semana de vida e a dose de reforço na oitava, e o desafio na 16ª semana com a estirpe selvagem SG. No segundo experimento, a vacinação foi realizada por diferentes rotas usando três doses da vacina viva, na quarta, na oitava e na décima segunda semana de vida, e o desafio foi feito na 20ª semana de vida. Após o desafio, a mortalidade foi acompanhada por 28 dias, e no experimento 2 a produção de ovos também foi avaliada e comparada com o grupo de galinhas não vacinadas. Em ambos os experimentos, a mortalidade foi significativamente reduzida, e a produção de ovos não foi afetada nos grupos de galinhas poedeiras vacinadas. Este estudo mostra a eficácia da proteção dos diferentes programas de vacinação contra o TA, que podem ser aplicados em granjas comerciais em condições de campo.(AU)