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A comparison on the prevalence of Salmonella infection in layer hens from commercial layer farms with high and low rodent densities was investigated. Out of 280 laying hens sampled from three commercial layer farms with high rodent densities, Salmonella enterica subsp. enterica serovar Enteritidis (Salmonella Enteritidis) was isolated from 20 (7.14%) hens and Salmonella enterica subsp. enterica serovar Infantis (Salmonella Infantis) from three (1.07%) hens. In contrast, layer hens sampled from four commercial layer farms with low rodent densities were negative for any salmonellae. Significant differences (P < 0.05) in the isolation rates of Salmonella from various organs of infected layer hens were also noted. For Salmonella Enteritidis, liver (55.0%) and the oviduct (55.0%) had the highest isolation rates while all Salmonella Infantis isolates were from the oviduct. Pulsed field gel electrophoresis (PFGE) analysis of BlnI-digested chromosomal DNA of Salmonella Enteritidis isolated from layer hens and rodents showed similar patterns. PFGE analysis of Salmonella Infantis isolated from layer hens, rodents, eggs, and the environment yielded identical patterns. In this study, the significantly higher prevalence rate (P < 0.05) of Salmonella Enteritidis and Salmonella Infantis in layer hens from high rodent density farms could be attributed to the high rodent population density. The persistent Salmonella Enteritidis and Salmonella Infantis infection inside layer houses may have been amplified by the increasing numbers in the rodent population over the years, which increased the opportunity for environment-rodent-chicken interaction and the transmission of salmonellae to chickens. Monitoring of salmonellae from rodents inside poultry premises is recommended to be an effective additional tool in the assessment of the Salmonella status of layer flocks.  相似文献   

3.
The ability of Salmonella enterica serotype Enteritidis isolates to adhere to immobilized secretions of the isthmus of the laying hen was determined in an ELISA-type assay. One-third of the 56 isolates tested in the logarithmic growth phase, adhered to the isthmal secretions. Using a binding assay of the isolates to thin paraffin sections of the oviduct, we demonstrated that the receptor of the adhesion was localized inside the tubular gland cells of the isthmus. The adhesion to immobilized isthmal secretions as well as to the paraffin sections was blocked by the addition of mannose. A fimD mutant of S. Enteritidis, lacking type 1 fimbriae, did not adhere, confirming that the adhesion was mediated by type 1 fimbriae. Mannosylated glycoproteins were demonstrated in the isthmus glandular cells using confocal laser scanning microscopy by FITC-labelled Lens culinaris lectins. It is hypothesized that the binding of S. Enteritidis to isthmal secretions could play a role in the contamination of eggs through incorporation of the bacteria in the shell membranes.  相似文献   

4.
In the chicken, maternal antibodies are transferred into the egg and subsequently transported into the developing embryo. IgG (called IgY) is the primary immunoglobulin isotype of the egg yolk. Their level in serum depends on the correct function of immunological system in laying hens. Many factors have a direct or indirect influence on antibody level in fowl. One of them is a commonly used antibiotic, but its influence on avian immune system is still unknown. The objective of the study was to determine the effect of enrofloxacin and chloramphenicol on the level of IgY antibody in serum and egg yolk after immunostimulation of hens with living cells of Salmonella enterica subsp. enterica serovar Enteritidis and lipopolisaccharide. Forty adult egg-laying Arbor Acres and Isa 215 hens (32 and 50 weeks old) from the reproductive flocks and 1640 of their eggs were used for the investigation. No clinical symptoms of any diseases were observed in birds during the entire breeding period. Additionally the birds were checked as free from Salmonella spp. in the beginning of the experiment. The birds were divided into 6 experimental and 2 control groups (5 birds in one group). The hens in the experimental groups were immunized with S. Enteritidis antigens: living bacteria and lipopolisaccharide and treated with enrofloxacin or chloramphenicol. Antibiotics were administered in drinking water for 10 days (from 3rd to 13th day of experiment). To indicate anti-S. Enteritidis, antibodies in sera and egg yolk were used indirectly on ELISA based on lipopolisaccharide from S. Enteritidis. As conjugate these were applied anti-chicken IgY with horseradish peroxidase and ABTS with H2O2 as obtained. Additionally, to detect antibody in serum, a rapid slide test was used with Pullognost and Enterognost standard antigens made in the laboratory. The study revealed that both antibiotics tested decreased the level of specific IgY in laying hens immunized with living bacteria and lipopolisaccharide. It seems that antibiotics have a suppressive effect on the immunological system. The strongest immunosuppressive effect was exerted by chloramphenicol.  相似文献   

5.
Three mature hens were immunized with an Aro- mutant of Salmonella typhimurium beginning with a subcutaneous dose in adjuvant followed by two oral boosters. Isotype-specific antibodies were measured in the white and yolk eggs collected weekly over a period of 230 days. Two hens showed a memory response to the first oral booster, with large increases in egg yolk IgG and smaller increases in IgA and IgM antibodies in egg whites. Smaller amounts of IgA and IgM antibodies were found in egg yolks, and a slight increase in IgG occurred in the whites. One hen showed an increase in serum titers of all isotypes against S. typhimurium. The second hen had high serum titers before immunization was started which did not change. The third hen had a high level of IgM in the white of eggs before immunization was started. This hen showed erratic responses in egg white antibodies following immunization, no increase in IgA or IgM in yolks and only a slight increase in IgG, no increase in serum IgG, and was the only hen with a high level of IgM antibody against S. typhimurium in the bile, conditions reflecting a state of oral tolerance. With the exception of this hen, the results showed that IgA and IgM antibodies were aroused in hens by immunization with an avirulent mutant of S. typhimurium, and that these antibodies were present in the white of eggs from immunized hens.  相似文献   

6.
In order to determine the epidemiological link between the Salmonella Enteritidis contamination in a rat-infested chicken layer farm, an attached egg processing facility and liquid egg samples, several S. Enteritidis isolates were analyzed by pulsed-field gel electrophoresis (PFGE) and bacteriophage typing. A total of 33 S. Enteritidis strains were isolated from a total of 4,081 samples. Similar pulsed-field patterns were generated by S. Enteritidis isolates from liquid eggs, rats and effluent water. Additionally, only two phage types were detected among the S. Enteritidis isolates, PT 1b and PT 6. These results suggest that S. Enteritidis isolates from rats, egg processing facility, and liquid eggs are genetically related. Furthermore, S. Enteritidis infection in rats in layer farms poses a serious public health concern and should be included in future epidemiological studies.  相似文献   

7.
Eighteen chickens were immunized subcutaneously with purified type 1 fimbriae from Salmonella enterica serotype Enteritidis at 18 and 21 weeks of age. Evidence of IgG and IgA responses was found in the eggs and in the sera of the immunized hens. Three weeks later, immunized and non-immunized chickens (n=18) were challenged intravenously with 2x10(7) live Salmonella enterica serotype Enteritidis. There was no significant difference in the numbers of eggs laid by immunized and non-immunized birds. The percentage of Salmonella contaminated eggs was significantly higher in the non-immunized group than in the immunized group due to a higher percentage of contamination of the externally disinfected egg shells. There were no statistical differences in the percentages of contaminated yolks and egg whites between control and immunized birds. No differences in the number of colonizing bacteria could be found in the spleen nor in the liver between the immunized and the control groups throughout the experiment. Salmonella was cleared from the ovary of the immunized birds in the second week p.i., in contrast to the control birds where Salmonella was isolated till the third week after infection. Oviducts were significantly more infected in the control group than in the immunized group. Salmonella was cleared from the oviducts at 3 weeks p.i. in the immunized hens but not in the control hens. In conclusion, we demonstrated that the immunization of laying hens with type 1 fimbriae reduced the number of contaminated eggs and reduced the colonization of the reproductive organs.  相似文献   

8.
Internal contamination of eggs by Salmonella Enteritidis has been a significant source of human illness for several decades and is the focus of a recently proposed U.S. Food and Drug Administration regulatory plan. Salmonella Heidelberg has also been identified as an egg-transmitted human pathogen. The deposition of Salmonella strains inside eggs is apparently a consequence of reproductive tissue colonization in infected laying hens, but the relationship between colonization of specific regions of the reproductive tract and deposition in different locations within eggs is not well documented. In the present study, groups of laying hens were experimentally infected with large oral doses of Salmonella Heidelberg, Salmonella Enteritidis phage type 13a, or Salmonella Enteritidis phage type 14b. For all of these isolates, the overall frequency of ovarian colonization (34.0%) was significantly higher than the frequency of recovery from either the upper (22.9%) or lower (18.1%) regions of the oviduct. No significant differences were observed between the frequencies of Salmonella isolation from egg yolk and albumen (4.0% and 3.3%, respectively). Some significant differences between Salmonella isolates were observed in the frequency of recovery from eggs, but not in the frequency or patterns of recovery from reproductive organs. Accordingly, although the ability of these Salmonella isolates to colonize different regions of the reproductive tract in laying hens was reflected in deposition in both yolk and albumen, there was no indication that any specific affinity of individual isolates for particular regions of this tract produced distinctive patterns of deposition in eggs.  相似文献   

9.
Using a deletion mutant in the regulator of SPI-2, ssrA, we investigated the role of SPI-2 in invasion, intestinal colonization and reproductive tract infection of chickens by Salmonella Enteritidis. The ssrA mutant was fully invasive in phagocytic and non-phagocytic cells but failed to persist within chicken macrophages. The ability of Salmonella Enteritidis to cause disease in orally infected 1-day-old chicks was not altered when ssrA was deleted. Furthermore, caecal colonization was not affected, while spleen and liver showed reduced colonization. Following intra-peritoneal and intravenous infection of 1-day-old chicks, internal organ colonization was strongly reduced. After intravenous inoculation in adult laying hens bacterial numbers of the ssrA mutant were significantly lower in oviducts and ovaries as compared to the wild type strain. The chickens showed less reproductive tract lesions and the recovery of egg production were faster compared to the wild type strain infected chickens. These findings indicate that the SPI-2 regulator ssrA promotes reproductive tract colonization, but is not essential for intestinal colonization of chickens with the host non-specific serotype Enteritidis.  相似文献   

10.
沙门菌血清D群3个血清型FliC蛋白氨基酸序列比对分析表明肠炎沙门菌与鸡伤寒沙门菌完全相同,二者与鸡白痢沙门菌存在第91位氨基酸位点差异。本研究旨在探究肠炎沙门菌FliC蛋白第91位精氨酸突变对鞭毛形态、细菌运动性和小鼠体内定植能力的影响。运用λ-Red同源重组技术删除肠炎沙门菌CICC10467 fliC基因,构建系列反式回补突变株,通过体外生长特性试验和Western blot试验分析各菌株生长和FliC蛋白表达情况,运动性试验分析各菌株在半固体琼脂中的泳动能力,电子显微镜观察各菌株鞭毛形态,细胞感染试验分析各菌株的细胞黏附和入侵能力,动物感染试验分析各菌株的组织侵染能力。结果表明,fliC基因缺失株及点突变回补株与野生株的体外生长能力无显著差异(P ≥ 0.05)。fliC基因缺失后肠炎沙门菌不表达鞭毛蛋白,各点突变回补株与野生株鞭毛蛋白表达量无明显差异。FliC蛋白R91S突变导致肠炎沙门菌鞭毛形态由超螺旋形态转变为钝直、柔韧度减弱,运动性显著降低(P<0.000 1),对RAW264.7和HCT116细胞的黏附入侵能力显著下降(P<0.001),对BALB/c小鼠的器官侵染能力显著减弱(P<0.001)。综上表明,FliC蛋白第91位精氨酸对维持细菌运动性至关重要,第91位精氨酸突变能够显著改变肠炎沙门菌鞭毛形态,减弱肠炎沙门菌在小鼠体内定植能力。  相似文献   

11.
Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) was responsible for a worldwide pandemic during the 1980s and 1990s; however, changes in the dominant lineage before and after this event remain unknown. This study determined S. Enteritidis lineages before and after this pandemic event in Japan using multilocus sequence typing (MLST). Thirty S. Enteritidis strains were collected in Japan between 1973 and 2004, consisting of 27 human strains from individual episodes, a bovine strain, a liquid egg strain and an eggshell strain. Strains showed nine phage types and 17 pulsed-field profiles with pulsed-field gel electrophoresis. All strains had homologous type 11 sequences without any nucleotide differences in seven housekeeping genes. These MLST results suggest that S. Enteritidis with the diversities revealed by phage typing and pulsed-field profiling has a highly clonal population. Although type 11 S. Enteritidis may exhibit both pleiotropic surface structure and pulsed-field type variation, it is likely to be a stable lineage derived from an ancestor before the 1980s and/or 1990s pandemic in Japan.  相似文献   

12.
Egg contamination by Salmonella Enteritidis has remained a significant public health problem for nearly two decades, and Salmonella Heidelberg has also been recently implicated in egg-transmitted human illness. Colonization of the intestinal tract is a necessary precursor to the invasion of reproductive organs and subsequent deposition inside eggs laid by infected hens, but the relationship between the persistence of Salmonella in the intestinal tract and the likelihood of egg contamination has been uncertain. In this study, groups of laying hens were inoculated with large oral doses of strains of Salmonella Enteritidis and Salmonella Heidelberg, including variants of the original parent strains that had been reisolated from eggs laid by infected hens in a prior study. The shedding of Salmonella in voided feces was monitored for 6 wk postinoculation, and all eggs laid by infected hens between 5 and 22 days postinoculation were cultured for Salmonella in their contents. The mean duration of fecal shedding was significantly longer for the previously passaged Salmonella strains (26.7 days) than for the original parent strains (17.5 days), and the passaged strains caused a significantly higher frequency of egg contamination (6.4%) than did the parent strains (3.3%). However, the duration of fecal shedding and the frequency of egg contamination were not correlated for any of the Salmonella Enteritidis or Salmonella Heidelberg strains.  相似文献   

13.
Levels of Salmonella enterica subsp. enterica serovar Enteritidis infection and serum S. Enteritidis antibodies after experimental S. Enteritidis challenge and feed withdrawal were investigated in S. Enteritidis-vaccinated and unvaccinated hens. The results were used to determine whether formalin-inactivated S. Enteritidis vaccination can protect layer hens from S. Enteritidis challenge during feed withdrawal periods. S. Enteritidis infection rates were evaluated from cloacal swabs, eggs and organs. Serum antibody titers to deflagellated S. Enteritidis whole cells (DEWC) and S. Enteritidis FliC-specific 9-kDa polypeptide (SEp 9) were examined by commercial ELISA kits. Cloacal S. Enteritidis recovery rates were lower in the vaccinated than unvaccinated group. Recovery rates of S. Enteritidis from samples increased after feed withdrawal and decreased after re-introduction of feed. S. Enteritidis counts in cloacal swabs were lower in the vaccinated than in the unvaccinated group (P<0.05). More S. Enteritidis-positive eggs were detected from the unvaccinated group. Before S. Enteritidis challenge, the DEWC ELISA titer of the vaccinated group was higher (P<0.05) than the unvaccinated group; subsequently, the S. Enteritidis DEWC ELISA titers of both groups increased gradually. In contrast, only the vaccinated group elicited high SEp-9 antibody titer during post-challenge and feed withdrawal. Additionally, vaccinated hens yielded negative S. Enteritidis isolation rates from egg contents. There is a correlation between negative S. Enteritidis isolation rates and high SEp 9 titers in vaccinated layer hens challenged with S. Enteritidis and subjected to feed withdrawal regimens. These findings suggest the S. Enteritidis vaccination of pullets may protect against S. Enteritidis infection during forced molting and that SEp 9 titer could be a potential indicator of antibody protection against S. Enteritidis infection. The potential of the SEp 9 peptide as an antigen for S. Enteritidis vaccination in the future is worth noting.  相似文献   

14.
Japanese quails were inoculated orally with 1 x 10(10) colony-forming units of Salmonella enteritidis phage type 4 isolated from chicks to obtain information concerning S. enteritidis infection and egg transmission. The inoculation resulted in a bacteraemic infection with seeding of the liver, spleen, intestine, peritoneum, ovule, ovary and oviduct. Some infected birds showed diarrhoea, ruffled feathers, depression, loss of appetite and death. However, most infected birds remained clinically normal with normal egg production. Salmonella enteritidis was isolated from the albumen of 7, yolk of 15, shell of 13, and shell membrane of 15 of 164 eggs. The experiment suggests that S. enteritidis phage type 4 is invasive for Japanese quails and the infected eggs laid by S. enteritidis infected quails are probably the result of transovarian infection.  相似文献   

15.
ABSTRACT: Human cases of bacterial gastro-enteritis are often caused by the consumption of eggs contaminated with Salmonella species, mainly Salmonella enterica serovar Enteriditis (Salmonella Enteritidis). To reduce human exposure, in several countries worldwide surveillance programmes are implemented to detect colonized layer flocks. The sampling schemes are based on the within-flock prevalence, and, as this changes over time, knowledge of the within-flock dynamics of Salmonella Enteritidis is required. Transmission of Salmonella Enteritidis has been quantified in pairs of layers, but the question is whether the dynamics in pairs is comparable to transmission in large groups, which are more representative for commercial layer flocks. The aim of this study was to compare results of transmission experiments between pairs and groups of laying hens. Experimental groups of either 2 or 200 hens were housed at similar densities, and 1 or 4 hens were inoculated with Salmonella Enteritidis, respectively. Excretion was monitored by regularly testing of fecal samples for the presence of Salmonella Enteritidis. Using mathematical modeling, the group experiments were simulated with transmission parameter estimates from the pairwise experiments. Transmission of the bacteria did not differ significantly between pairs or groups. This finding suggests that the transmission parameter estimates from small-scale experiments might be extrapolated to the field situation.  相似文献   

16.
Salmonella enterica serovar Enteritidis is a gram-negative bacterium that negatively affects human and animal health. Many eukaryotes use antimicrobial peptides (alpha-defensins, beta-defensins, gamma-defensins, and cathelicidins) in innate immune responses to fight bacterial infections. Poultry gallinacins are the functional equivalents of mammalian beta-defensins. Two related advanced intercross lines of chickens were analyzed for association of gallinacin genotypic variation with Salmonella Enteritidis burden levels in the cecum and spleen after infection. Thirteen genes of the chicken beta-defensin cluster (GAL1-13) were sequenced from individuals of each advanced intercross line, plus the founder broiler sire and representatives of the highly inbred Leghorn and Fayoumi founder dam lines. The mean was 17 single-nucleotide polymorphisms (SNPs) per kilobase. One single-nucleotide polymorphism per gene was genotyped with SNaPshot to test for statistical associations with Salmonella Enteritidis colonization after challenge. Among the 13 gallinacin genes evaluated, the single-nucleotide polymorphisms in all genes in a cluster of three adjacent genes (GAL11, GAL12, and GAL13) were associated with bacterial load in the cecal content in the broiler x Leghorn advanced intercross line (three-gene SNP genotype effect, P < 0.008). The results strongly suggest a role of the gallinacins in defense of poultry against enteric pathogens. The use of gallinacin single-nucleotide polymorphisms as molecular markers for genetic selection for Salmonella Enteritidis resistance might result in reduced bacterial burden via development of an enhanced innate immune response.  相似文献   

17.
1. The repeatability and heritability of growth inhibition by egg albumen of two major pathogenic bacteria, a Gram-negative (Salmonella Enteritidis) and a Gram-positive (Staphyloccocus aureus) and of two antimicrobial albumen proteins, lysozyme and ovotransferrin, were estimated in commercial pedigree hens. 2. Repeatability was evaluated in 100 egg-type hens at the beginning, middle and end of the laying cycle on eggs collected for 3 weeks. Heritabilities were estimated at 36 to 40 weeks of age on 400 pedigree hens (2 eggs/hen), which were the offspring of 25 sires each mated with 4 dams. Ovotransferrin and lysozyme were quantified by ELISA. Salmonella Enteritidis (S.E.) and Staphyloccocus aureus (S.A.) were inoculated into a sample of sterilised albumen and enumerated after incubation. 3. Total protein content in albumen decreased with age of laying hens, whereas there were increases in lysozyme or ovotransferrin concentrations and in the bacteriostatic effect of albumen. 4. Repeatability for bacterial growth in albumen ranged from 0.29 to 0.39 for the number of S.E. (log cfu/ml) one day post inoculation (p.i.) but was lower and more variable at 5 d p.i. or for S.A. number. It ranged from 0.27 to 0.38 for S.E. and S.A. number at the mid period of the laying cycle. Repeatabilities were low and variable for total egg albumen protein or lysozyme and ovotranferrin concentrations (0 to 0.22). 5. Negative phenotypic correlations were observed between lysozyme concentrations and S.E. number but that between lysozyme and S.A. number was not significant. 6. Heritabilities were low (0.01 to 0.09) for protein traits. They were 0.11 for S.A. number and 0.16 for S.E. number one day p.i. 7. It appears to be more efficient to select on global bacterial growth than on specific antimicrobial proteins. The most promising trait is the number of S.E. one day p.i.  相似文献   

18.
Chicken consumption is a newly identified risk factor in Salmonella enterica serovar Enteritidis (SE) infection in humans. SE is widely distributed in commercial chicken flocks and high levels of cecal carriage and shedding may lead to broiler meat contamination. In the present study, the preventive and eliminative effect of nonimmunized freeze-dried egg yolk powder (EYP) on SE in broilers was investigated. In the prevention trial, reduced SE counts were observed in liver (P < or = 0.05), cecal contents, and fecal shedding (P < or = 0.05) in birds fed 10% or 5% EYP. Histological examination of cecal wall and cecal tonsils at 23 days postinfection indicated a lesser degree of intestinal pathology. In the elimination trial, a significantly lower (P < or = 0.05) number of SE reached the liver and spleen, and a reduction in cecal carriage and fecal shedding was observed. The histological changes in the cecal mucosa and cecal tonsils reflected an apparent inflammation and mucosal repair and also suggested that the infection had not completely resolved, confirming SE bacterial isolations in the cecal tissue. The present study indicates that supplementing the diets of broilers with 5% nonimmunized EYP, at the early stages of the growing period, reduces preharvest Salmonella load with a minimal degree of intestinal pathology.  相似文献   

19.
Salmonella enterica subspecies enterica infection remains a serious problem in a wide range of animals and in man. Poultry-derived food is the main source of human infection with the non-host-adapted serovars while fowl typhoid and pullorum disease are important diseases of poultry. We have assessed cecal colonization and immune responses of newly hatched and older chickens to Salmonella serotypes Enteritidis, Infantis, Gallinarum and Pullorum. S. Enteritidis and S. Infantis colonized the ceca more efficiently than S. Gallinarum and S. Pullorum. Salmonella infection was also associated with increased staining for B-lymphocytes and macrophages in the cecal tonsils of infected birds. S. Enteritidis infection in newly hatched birds stimulated the expression of CXCLi1 and CXCLi2 chemokines in the cecal tonsils, while S. Gallinarum up-regulated the expression of LITAF. In older chickens, S. Enteritidis infection resulted in a significantly higher expression of CXCLi2, iNOS, LITAF and IL-10 while S. Pullorum appeared to down-regulate CXCLi1 expression in the cecal tonsils. Data from spleens showed either no expression or down-regulation of the tested genes.  相似文献   

20.
本试验旨在研究芽孢杆菌制剂对"京粉一号"蛋鸡蛋品质及蛋营养成分的影响。选择健康、产蛋均匀的259日龄的"京粉一号"蛋鸡3 600只,随机分为2组,每组6个重复,每个重复300只鸡。对照组饲喂基础日粮,试验组在基础日粮中添加3%的芽孢杆菌制剂,进行为期40 d的试验。结果表明:与对照组相比,试验组蛋壳硬度、哈夫单位以及蛋黄颜色均有不同程度地提高(P>0.05);在反映蛋的物理指标上,试验组蛋黄重和蛋黄相对重显著高于对照组(P<0.05),蛋清相对重显著低于对照组(P<0.05),蛋黄蛋白比有升高的趋势(P>0.05);同时,试验组蛋黄中脂肪和水分含量显著降低(P<0.05),蛋白含量显著升高(P<0.05)。试验表明:在蛋鸡日粮中添加芽孢杆菌制剂,对蛋品质具有一定的改善作用;可显著提高蛋黄重、蛋黄相对重,降低蛋清相对重。同时,可减少蛋黄中脂肪和水分含量,增加蛋白的含量。  相似文献   

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