Utilização de um mutante atenuado de Salmonella enterica subesp. enterica sorovar GALLINARUM cobS cbiA para proteção de aves contra a infecção por Salmonella enterica subesp. enterica sorovares GALLINARUM E ENTERITIDIS

Salmonella Gallinarum (SG) causa o Tifo Aviário, uma doença caracterizada por alta mortalidade e morbidade em aves. Salmonella Enteritidis (SE) é uma salmonela paratífica, capaz de acometer aves e mamíferos. Carne de aves e ovos são as principais fontes de transmissão de SE, causando doença transmitida por alimentos em humanos. O controle destas bactérias nas criações avícolas é feito através da combinação de medidas sanitárias e a vacinação das aves. As vacinas vivas mostram-se melhores do que as inativadas para combater esses sorotipos, no entanto, a proteção contra SE, é parcial. A cepa SG cobS cbiA é incapaz de sintetizar vitamina B12. A ausência dessa substância prejudicou a sobrevivência da bactéria no organismo parasitado, causando atenuação na virulência. Neste trabalho, foram avaliadas a mortalidade e a infecção sistêmica causadas por SG cobS cbiA em comparação com uma cepa selvagem de SG. Também foi avaliado seu potencial vacinal contra SG e SE em aves de postura e sua utilização para impedir a colonização cecal por SE em pintinhos de corte de um dia. As aves de postura foram vacinadas com uma dose aos cinco dias, ou com duas doses aos cinco e vinte e cinco dias de vida e foram desafiadas com as respectivas cepas de SG e SE aos 45 dias de vida. As aves que receberam tanto uma quanto duas doses da cepa vacinal foram parcialmente protegidas contra SG. A proteção contra SE foi significativa no grupo de aves que recebeu duas doses de SG cobS cbiA. Porém, a inoculação de SG cobS cbiA em pintinhos com um dia de vida não impediu a colonização do ceco por SE. A cepa SG cobS cbiA demonstrou ter potencial vacinal contra SG e SE.Salmonella Gallinarum (SG) causes Fowl Typhoid, a disease caracterized by high mortality and morbidity rates in poultry. Salmonella Enteritidis (SE) is a paratyphoid salmonelae that infects birds and mammals. Poultry meat and eggs are the main sources of transmission of SE, causing foodborne disease in humans. The control of these bacterias inside the avian breeding sites is done by the combination of sanitary measures and poultry vaccination. Live vaccines have demonstrated greater efficacy on the protection of poultry against these serovars than the killed vaccines, though, protection against SE is still partial. The strain SG cobS cbiA is uncapable to synthesize vitamin B12. The absence of this substance harmed the bacterial survival in the infected organism, causing attenuation of the virulence. On this work, the mortality and the sistemic infection by SG cobS cbiA were measured and compared with a wild strain of SG. It was also evaluated the potential use of the attenuated strain as a vaccine against SG and SE in laying hens and its utilization to prevent the colonization of the ceca by SE on 1-day-old chickens. The laying hens were vaccinated with one dose at 5- day-old, or with two doses at 5 and 25-day-old and were challenged at 45-day-old with the respective strain of SG or SE. The vaccination either with one or two doses was able to protect the birds against challenge with SG. The administration of two doses of SG cobS cbiA protected the laying hens against SE. Although, the inoculation of SG cobS cbiA by oral gavage on newly hatched chicks was unable to prevent the colonization of the ceca by SE. The protection confered by the strain SG cobS cbiA demonstrated its potential to be used as a vaccine.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Requirement for cobalamin by Salmonella enterica serovars Typhimurium, Pullorum, Gallinarum and Enteritidis during infection in chickens

Salmonella enterica serovar Typhimurium synthesizes cobalamin (vitamin B12) only during anaerobiosis. Two percent of the S. Typhimurium genome is devoted to the synthesis and uptake of vitamin B12 and to B12-dependent reactions. To understand the requirement for cobalamin synthesis better, we constructed mutants of Salmonella serovars Enteritidis and Pullorum that are double-defective in cobalamin biosynthesis (ΔcobSΔcbiA). We compared the virulence of these mutants to that of their respective wild type strains and found no impairment in their ability to cause disease in chickens. We then assessed B12 production in these mutants and their respective wild type strains, as well as in S. Typhimurium ΔcobSΔcbiA, Salmonella Gallinarum ΔcobSΔcbiA, and their respective wild type strains. None of the mutants was able to produce detectable B12. B12 was detectable in S. Enteritidis, S. Pullorum and S. Typhimurium wild type strains but not in S. Gallinarum. In conclusion, the production of vitamin B12 in vitro differed across the tested Salmonella serotypes and the deletion of the cbiA and cobS genes resulted in different levels of alteration in the host parasite interaction according to Salmonella serotype tested.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

IncIl/ST113 and IncIl/ST114 conjugative plasmids carrying bla(CTX-M-8) in Escherichia coli isolated from poultry in Brazil

Extended-spectrum beta-lactamase-producing Escherichia coli isolated from poultry in Brazil showed bla(CTX-M-8) gene. IS10 was found upstream of bla(CTX-M-8), harbored on plasmids Inch, ST113/ST114 subtypes. Genomic relationship revealed a heterogeneous E. coli population. The gene bla(CTX-M-8) is established in South America in food-producing animals, which represent risk of dissemination for other countries.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Protection of chickens against fowl typhoid using field vaccine programs formulated with the live attenuated strain Salmonella Gallinarum ΔcobSΔcbiA

<div><p>ABSTRACT: 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.</p></div

Antimicrobial susceptibility of Salmonella Gallinarum and Salmonella Pullorum isolated from ill poultry in Brazil

ABSTRACT: Salmonella Gallinarum (S. Gallinarum) and Salmonella Pullorum (S. Pullorum) are poultry host-specific, agents of fowl typhoid and pullorum disease, respectively. These biovars cause septicemic infections, resulting in high mortality. Outbreaks are frequently reported worldwide, causing losses due to the elimination of infected flocks and treatments. The use of antimicrobial agents is frequent in poultry farms to prevent or treat gastrointestinal infections. In the present research it was evaluated the antimicrobial susceptibility of 50 S. Gallinarum and S. Pullorum isolates, from outbreaks that occurred between 1987 to 1991 and 2006 to 2013. The comparison of the susceptibility profiles showed that all isolates were susceptible to β-lactams. All isolates from 1987-1991 were susceptible to all antibiotics tested except NAL and CIP (78%). The susceptibility profile of S. Gallinarum (2006 - 2013 period) was the following NAL (58%), CIP (63%), ENR (67%), TET (92%), FFC (96%) and SXT (96%). S. Pullorum isolates (2006 - 2013 period) showed the following susceptibility rates to NAL (65%), CIP (71%), ENR (94%) and TET (94%). All isolates were susceptible to β-lactams tested, however, resistance to quinolones and fluoroquinolones increased over time. Furthermore, low levels of resistance to other antibiotics were found in recent isolates, such as tetracyclines

Immunomodulatory activity and control of Salmonella Enteritidis colonization in the intestinal tract of chickens by Lactobacillus based probiotic

Lactobacillus-based probiotics (LBP) are used as competitive exclusion to control pathogenic enterobacterial infections and improve the weight gain in broiler chickens. This study assessed the inhibition of Salmonella Enteritidis (SE) infection in one-week-old broiler chicks, using an experimental LBP containing four Lactobacillus strains isolated from chickens (L. acidophilus, L. fermentum, L. reuteri, L. salivarius). The immunomodulatory effects of this treatment were evaluated, through the analysis of cytokines and influx of macrophages, γδ, CD4(+) and CD8(+) T cells in the gut. The intestinal colonization by SE was reduced by 1.8CFU/g (log10) in chicks treated with LBP (p<0.05). The levels of pro-inflammatory cytokines (IL-1β, LITAF) were significantly reduced in treated chicks (p<0.05), whilst untreated chicks showed elevated inflammatory stimulus and an increased population of CD8(+) T cells in the intestinal mucosa after challenge (p<0.05). Additionally, the LBP stimulated TLR2 expression in caecal tonsils. The adjuvant property of the Lactobacillus cell wall (LCW) was evaluated, demonstrating good capability to stimulate T helper 2 (Th2) cell proliferation. Pretreatment of chicks with LBP decreased the intestinal colonization by SE, minimizing the tissue lesions and inflammation after challenge and showed a potential use as adjuvant with injectable killed vaccines.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES