Sample records for salmonella enteritidis se

  1. 21 CFR 118.6 - Egg testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Egg testing for Salmonella Enteritidis (SE). 118.6... testing for Salmonella Enteritidis (SE). (a)(1) If the environmental test for pullets at 14 to 16 weeks of... requires that these eggs must be treated to achieve at least a 5-log destruction of Salmonella Enteritidis...

  2. 21 CFR 118.6 - Egg testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Egg testing for Salmonella Enteritidis (SE). 118.6... testing for Salmonella Enteritidis (SE). (a)(1) If the environmental test for pullets at 14 to 16 weeks of... requires that these eggs must be treated to achieve at least a 5-log destruction of Salmonella Enteritidis...

  3. 21 CFR 118.6 - Egg testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Egg testing for Salmonella Enteritidis (SE). 118.6... testing for Salmonella Enteritidis (SE). (a)(1) If the environmental test for pullets at 14 to 16 weeks of... requires that these eggs must be treated to achieve at least a 5-log destruction of Salmonella Enteritidis...

  4. 21 CFR 118.7 - Sampling methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Sampling methodology for Salmonella Enteritidis (SE). 118.7 Section 118.7 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....7 Sampling methodology for Salmonella Enteritidis (SE). (a) Environmental sampling. An environmental...

  5. 21 CFR 118.7 - Sampling methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Sampling methodology for Salmonella Enteritidis (SE). 118.7 Section 118.7 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....7 Sampling methodology for Salmonella Enteritidis (SE). (a) Environmental sampling. An environmental...

  6. 21 CFR 118.7 - Sampling methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Sampling methodology for Salmonella Enteritidis (SE). 118.7 Section 118.7 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....7 Sampling methodology for Salmonella Enteritidis (SE). (a) Environmental sampling. An environmental...

  7. 21 CFR 118.7 - Sampling methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Sampling methodology for Salmonella Enteritidis (SE). 118.7 Section 118.7 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....7 Sampling methodology for Salmonella Enteritidis (SE). (a) Environmental sampling. An environmental...

  8. 21 CFR 118.7 - Sampling methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Sampling methodology for Salmonella Enteritidis (SE). 118.7 Section 118.7 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....7 Sampling methodology for Salmonella Enteritidis (SE). (a) Environmental sampling. An environmental...

  9. 21 CFR 118.5 - Environmental testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Environmental testing for Salmonella Enteritidis (SE). 118.5 Section 118.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5 Environmental testing for Salmonella Enteritidis (SE). (a) Environmental testing when laying hens...

  10. 21 CFR 118.5 - Environmental testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Environmental testing for Salmonella Enteritidis (SE). 118.5 Section 118.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5 Environmental testing for Salmonella Enteritidis (SE). (a) Environmental testing when laying hens...

  11. 21 CFR 118.5 - Environmental testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Environmental testing for Salmonella Enteritidis (SE). 118.5 Section 118.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5 Environmental testing for Salmonella Enteritidis (SE). (a) Environmental testing when laying hens...

  12. 21 CFR 118.5 - Environmental testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Environmental testing for Salmonella Enteritidis (SE). 118.5 Section 118.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5 Environmental testing for Salmonella Enteritidis (SE). (a) Environmental testing when laying hens...

  13. 21 CFR 118.5 - Environmental testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Environmental testing for Salmonella Enteritidis (SE). 118.5 Section 118.5 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN....5 Environmental testing for Salmonella Enteritidis (SE). (a) Environmental testing when laying hens...

  14. 21 CFR 118.9 - Administration of the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Administration of the Salmonella Enteritidis (SE) prevention plan. 118.9 Section 118.9 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... EGGS § 118.9 Administration of the Salmonella Enteritidis (SE) prevention plan. You must have one or...

  15. 21 CFR 118.9 - Administration of the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Administration of the Salmonella Enteritidis (SE) prevention plan. 118.9 Section 118.9 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... EGGS § 118.9 Administration of the Salmonella Enteritidis (SE) prevention plan. You must have one or...

  16. 21 CFR 118.9 - Administration of the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Administration of the Salmonella Enteritidis (SE) prevention plan. 118.9 Section 118.9 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... EGGS § 118.9 Administration of the Salmonella Enteritidis (SE) prevention plan. You must have one or...

  17. 21 CFR 118.9 - Administration of the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Administration of the Salmonella Enteritidis (SE) prevention plan. 118.9 Section 118.9 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... EGGS § 118.9 Administration of the Salmonella Enteritidis (SE) prevention plan. You must have one or...

  18. 21 CFR 118.9 - Administration of the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Administration of the Salmonella Enteritidis (SE) prevention plan. 118.9 Section 118.9 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... EGGS § 118.9 Administration of the Salmonella Enteritidis (SE) prevention plan. You must have one or...

  19. 21 CFR 118.10 - Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan. 118.10 Section 118.10 Food and Drugs FOOD AND DRUG ADMINISTRATION... TRANSPORTATION OF SHELL EGGS § 118.10 Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention...

  20. 21 CFR 118.10 - Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan. 118.10 Section 118.10 Food and Drugs FOOD AND DRUG ADMINISTRATION... TRANSPORTATION OF SHELL EGGS § 118.10 Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention...

  1. 21 CFR 118.10 - Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan. 118.10 Section 118.10 Food and Drugs FOOD AND DRUG ADMINISTRATION... TRANSPORTATION OF SHELL EGGS § 118.10 Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention...

  2. 21 CFR 118.10 - Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan. 118.10 Section 118.10 Food and Drugs FOOD AND DRUG ADMINISTRATION... TRANSPORTATION OF SHELL EGGS § 118.10 Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention...

  3. 21 CFR 118.10 - Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention plan. 118.10 Section 118.10 Food and Drugs FOOD AND DRUG ADMINISTRATION... TRANSPORTATION OF SHELL EGGS § 118.10 Recordkeeping requirements for the Salmonella Enteritidis (SE) prevention...

  4. 21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 2 2012-04-01 2012-04-01 false Salmonella Enteritidis (SE) prevention measures. 118.4 Section 118.4 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... equivalent standard; (2) Environmental testing. (i) The pullet environment is tested for SE when pullets are...

  5. 21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 2 2013-04-01 2013-04-01 false Salmonella Enteritidis (SE) prevention measures. 118.4 Section 118.4 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... equivalent standard; (2) Environmental testing. (i) The pullet environment is tested for SE when pullets are...

  6. 21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 2 2014-04-01 2014-04-01 false Salmonella Enteritidis (SE) prevention measures. 118.4 Section 118.4 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... equivalent standard; (2) Environmental testing. (i) The pullet environment is tested for SE when pullets are...

  7. 21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Salmonella Enteritidis (SE) prevention measures. 118.4 Section 118.4 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... equivalent standard; (2) Environmental testing. (i) The pullet environment is tested for SE when pullets are...

  8. 21 CFR 118.4 - Salmonella Enteritidis (SE) prevention measures.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Salmonella Enteritidis (SE) prevention measures. 118.4 Section 118.4 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... equivalent standard; (2) Environmental testing. (i) The pullet environment is tested for SE when pullets are...

  9. 21 CFR 118.6 - Egg testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Egg testing for Salmonella Enteritidis (SE). 118.6... (CONTINUED) FOOD FOR HUMAN CONSUMPTION PRODUCTION, STORAGE, AND TRANSPORTATION OF SHELL EGGS § 118.6 Egg... age required by § 118.4(a) is positive, you must divert eggs to treatment (defined in § 118.3) for the...

  10. 21 CFR 118.6 - Egg testing for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 2 2011-04-01 2011-04-01 false Egg testing for Salmonella Enteritidis (SE). 118.6... (CONTINUED) FOOD FOR HUMAN CONSUMPTION PRODUCTION, STORAGE, AND TRANSPORTATION OF SHELL EGGS § 118.6 Egg... age required by § 118.4(a) is positive, you must divert eggs to treatment (defined in § 118.3) for the...

  11. Rapid real-time PCR methods to distinguish Salmonella Enteritidis wildtype field isolates from vaccine strains Salmovac SE/Gallivac SE and AviPro SALMONELLA VAC E.

    PubMed

    Maurischat, Sven; Szabo, Istvan; Baumann, Beatrice; Malorny, Burkhard

    2015-05-01

    Salmonella enterica serovar Enteritidis is a major non-typhoid Salmonella serovar causing human salmonellosis mainly associated with the consumption of poultry and products thereof. To reduce infections in poultry, S. Enteritidis live vaccine strains AviPro SALMONELLA VAC E and Salmovac SE/Gallivac SE have been licensed and used in several countries worldwide. To definitively diagnose a S. Enteritidis contamination in vaccinated herds a reliable and fast method for the differentiation between vaccine and wildtype field isolates is required. In this study, we developed and validated real-time PCR (qPCR) assays to distinguish those variants genetically. Suitable target sequences were identified by whole genome sequencing (WGS) using the Illumina MiSeq system. SNP regions in kdpA and nhaA proved to be most useful for differentiation of AviPro SALMONELLA VAC E and Salmovac SE/Gallivac SE, respectively, from wildtype strains. For each vaccine strain one TaqMan-qPCR assay and one alternative approach using High Resolution Melting (HRM) analysis was designed. All 30 Salmovac SE and 7 AviPro SALMONELLA VAC E vaccine strain reisolates tested were correctly identified by both approaches (100% inclusivity). Furthermore, all 137 (TaqMan) and 97 (HRM) Salmonella non-vaccine and related Enterobacteriaceae strains tested were excluded (100% exclusivity). The analytical detection limits were determined to be approx. 10(2) genome copies/reaction for the TaqMan and 10(4) genome copies/reaction for the HRM approach. The real-time PCR assays proved to be a reliable and fast alternative to the cultural vaccine strain identification tests helping decision makers in control measurements to take action within a shorter period of time. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Detection of Salmonella enterica serovar Enteritidis (SE) Antibodies in Serum Using A Polystyrene Bead/SE Flagella Agglutination Assay

    USDA-ARS?s Scientific Manuscript database

    Serologic screening of flocks can be an important method to detect Salmonella enteritidis (SE) infections but can be labor intensive or lack specificity. Our goal was to develop a rapid agglutination assay using SE flagella adsorbed to polystyrene beads as a simple, relatively specific test to dete...

  13. Control of Salmonella enterica serovar Enteritidis in laying hens by inactivated Salmonella Enteritidis vaccines

    PubMed Central

    de Freitas Neto, Oliveiro Caetano; Mesquita, Aline Lopes; de Paiva, Jaqueline Boldrin; Zotesso, Fábio; Berchieri Júnior, Angelo

    2008-01-01

    Salmonella Enteritidis is one of the agents that is responsible for outbreaks of human foodborne salmonellosis caused by Salmonella Enteritidis and is generally associated with the consumption of poultry products. Inactivated Salmonella Enteritidis cell vaccine is one of the available methods to control Salmonella Enteritidis in breeders and laying hens, however results in terms of efficacy vary. This vaccine has never been tested in Brazil, therefore, the present work was carried out to assess three commercial inactivated Salmonella Enteritidis vaccines allowed in Brazil. Four hundred white light variety commercial laying hens were obtained at one-day-of age. At eight weeks old, the birds were divided into four groups with one hundred animals each. Birds from three groups (V1, V2 and V3) received different intramuscular vaccines, followed by a booster dose at 16 weeks of age. Birds from another group (CG) were not vaccinated. When the laying hens were 20, 25 and 31 weeks old, 13 from each group were transferred to another room and were challenged by inoculating 2 mL neat culture of Salmonella Enteritidis. On the second day after each challenge, the caecal contents, spleen, liver and ovary of three birds from each group were analyzed for the presence of Salmonella Enteritidis. Twice a week a cloacal swab of each bird was taken and all eggs laid were examined for the presence of Salmonella Enteritidis. After four consecutive negative cloacal swabs in all the groups, the birds were sacrificed so as to examine the liver, caecal contents and ovaries. Overall, the inactivated vaccine used in group V3 reduced Salmonella Enteritidis in the feces and eggs. A very small amount of Salmonella was found in the spleen, liver, ovary and caeca of the birds in the four groups during the whole experiment. In general, inactivated Salmonella Enteritidis vaccines was able to decrease the presence of Salmonella Enteritidis in the birds and in the eggs as well. Nevertheless, they must

  14. 75 FR 18849 - Small Entity Compliance Guide: Prevention of Salmonella Enteritidis in Shell Eggs During...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-13

    ...] Small Entity Compliance Guide: Prevention of Salmonella Enteritidis in Shell Eggs During Production... ``Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation--Small... requiring shell egg producers to implement measures to prevent Salmonella Enteritidis (SE) from...

  15. Biosurfactant Produced by Salmonella Enteritidis SE86 Can Increase Adherence and Resistance to Sanitizers on Lettuce Leaves (Lactuca sativa L., cichoraceae)

    PubMed Central

    Rossi, Eliandra M.; Beilke, Luniele; Kochhann, Marília; Sarzi, Diana H.; Tondo, Eduardo C.

    2016-01-01

    Salmonella Enteritidis SE86 is an important foodborne pathogen in Southern Brazil and it is able to produce a biosurfactant. However, the importance of this compound for the microorganism is still unknown. This study aimed to investigate the influence of the biosurfactant produced by S. Enteritidis SE86 on adherence to slices of lettuce leaves and on resistance to sanitizers. First, lettuce leaves were inoculated with S. Enteritidis SE86 in order to determine the amount of biosurfactant produced. Subsequently, lettuce leaves were inoculated with S. Enteritidis SE86 with and without the biosurfactant, and the adherence and bacterial resistance to different sanitization methods were evaluated. S. Enteritidis SE86 produced biosurfactant after 16 h (emulsification index of 11 to 52.15 percent, P < 0.05) and showed greater adherence capability and resistance to sanitization methods when the compound was present. The scanning electron microscopy demonstrated that S. Enteritidis was able to adhere, form lumps, and invade the lettuce leaves’ stomata in the presence of the biosurfactant. Results indicated that the biosurfactant produced by S. Enteritidis SE86 contributed to adherence and increased resistance to sanitizers when the microorganism was present on lettuce leaves. PMID:26834727

  16. Isolation of Salmonella enterica serovar Enteritidis from houseflies (Musca domestica) found in rooms containing Salmonella serovar Enteritidis-challenged hens.

    PubMed

    Holt, Peter S; Geden, Christopher J; Moore, Randle W; Gast, Richard K

    2007-10-01

    Houseflies (Musca domestica) released into rooms containing hens challenged with Salmonella enterica serovar Enteritidis (Salmonella serovar Enteritidis) rapidly became contaminated with Salmonella serovar Enteritidis. Forty to 50% of the flies were contaminated at 48 h, and the percentage increased to 50 to 70% at 4 and 7 days postexposure and then decreased to 30% at day 15. Initial attempts at recovering surface organisms for culture using an aqueous rinse were largely unsuccessful, while cultures of internal contents readily recovered Salmonella serovar Enteritidis. However, when 0.5% detergent was incorporated into the rinse, high recovery levels of bacteria were observed from both external and internal culture regimens, indicating equal distribution of the organism on and in the fly and a tighter interaction of the organism with the host than previously thought. Salmonella serovar Enteritidis was isolated routinely from the fly gut, on rare occasions from the crop, and never from the salivary gland. Feeding contaminated flies to hens resulted in gut colonization of a third of the birds, but release of contaminated flies in a room containing previously unchallenged hens failed to result in colonization of any of the subject birds. These results indicate that flies exposed to an environment containing Salmonella serovar Enteritidis can become colonized with the organism and might serve as a source for transmission of Salmonella serovar Enteritidis within a flock situation.

  17. Isolation of Salmonella enterica Serovar Enteritidis from Houseflies (Musca domestica) Found in Rooms Containing Salmonella Serovar Enteritidis-Challenged Hens▿

    PubMed Central

    Holt, Peter S.; Geden, Christopher J.; Moore, Randle W.; Gast, Richard K.

    2007-01-01

    Houseflies (Musca domestica) released into rooms containing hens challenged with Salmonella enterica serovar Enteritidis (Salmonella serovar Enteritidis) rapidly became contaminated with Salmonella serovar Enteritidis. Forty to 50% of the flies were contaminated at 48 h, and the percentage increased to 50 to 70% at 4 and 7 days postexposure and then decreased to 30% at day 15. Initial attempts at recovering surface organisms for culture using an aqueous rinse were largely unsuccessful, while cultures of internal contents readily recovered Salmonella serovar Enteritidis. However, when 0.5% detergent was incorporated into the rinse, high recovery levels of bacteria were observed from both external and internal culture regimens, indicating equal distribution of the organism on and in the fly and a tighter interaction of the organism with the host than previously thought. Salmonella serovar Enteritidis was isolated routinely from the fly gut, on rare occasions from the crop, and never from the salivary gland. Feeding contaminated flies to hens resulted in gut colonization of a third of the birds, but release of contaminated flies in a room containing previously unchallenged hens failed to result in colonization of any of the subject birds. These results indicate that flies exposed to an environment containing Salmonella serovar Enteritidis can become colonized with the organism and might serve as a source for transmission of Salmonella serovar Enteritidis within a flock situation. PMID:17675422

  18. Electron-beam-inactivated vaccine against Salmonella enteritidis colonization in molting hens

    USDA-ARS?s Scientific Manuscript database

    Electron Beam (eBeam) ionization technology has a variety of applications in modern society. The underlying hypothesis was that electron beam (eBeam) inactivated Salmonella enterica serovar Enteritidis (SE) cells can serve as a vaccine to control Salmonella colonization and Salmonella shedding in c...

  19. Saccharomyces boulardii prevention of the hepatic injury induced by Salmonella Enteritidis infection.

    PubMed

    Wu, Daichao; Teng, Da; Wang, Xiumin; Dai, Changsong; Wang, Jianhua

    2014-10-01

    Salmonella enterica subsp. enterica serovar Enteritidis (Salmonella Enteritidis) is the predominant cause of serovar-associated food-borne outbreaks in many countries and causes significant clinical symptoms of liver injury, enteritis, and diarrheal diseases. Saccharomyces boulardii is used in clinical application for prophylaxis and the treatment of a variety of diseases caused by bacterial infection. We used a mouse model of Salmonella Enteritidis infection, which included pretreatment with S. boulardii, to reveal the protection mechanisms of S. boulardii against Salmonella Enteritidis infection, including the translocation of Salmonella Enteritidis to the liver 10 days after Salmonella Enteritidis challenge, and the colonisation of Salmonella Enteritidis and the formation of hepatic tissue lesions in mice after Salmonella Enteritidis challenge on the 10th day. Compared with Salmonella Enteritidis infection in mice, S. boulardii decreased Salmonella Enteritidis translocation to the liver by 96%, and 99% of Salmonella Enteritidis colonised the cecum on the 10th day. Saccharomyces boulardii also abated hepatic tissue injury caused by the infiltration of neutrophilic granulocytes, lymphocytes, and plasmocytes by decreasing the translocation of Salmonella to the liver. These findings demonstrated that S. boulardii is an effective agent in the prevention of the hepatic injury induced by Salmonella Enteritidis infection in a mouse model.

  20. The major sources of Salmonella enteritidis in Thailand.

    PubMed

    Sakai, T; Chalermchaikit, T

    1996-08-01

    The data of Salmonella serotypes during 1989-1993 from the World Health Organisation (WHO) National Salmonella and Shigella Center, Division of Clinical Pathology, Department of Medical Science, Ministry of Health, Thailand was analysed and found that the prevalence of Salmonella enteritidis had been dramatically increased since 1990. The average S. enteritidis isolates from human patient samples was 0.70% +/- 0.41% of the total reported Salmonella isolates during 1972-1989 and increased to 1.33%, 2.98%, 9.54%, and 16.98% in 1990, 1991, 1992, and 1993, respectively. The similar trend of S. enteritidis isolates from chicken meat samples were also observed. However, the conclusive epidemiological relationship between human and chicken S. enteritidis isolates needs to be proved by phage typing or other Salmonella typing methods.

  1. Effect of infectious bursal disease (IBD) vaccine on Salmonella Enteritidis infected chickens.

    PubMed

    Arafat, Nagah; Eladl, Abdelfattah H; Mahgoub, Hebatallah; El-Shafei, Reham A

    2017-06-22

    Chickens infected with both infectious bursal disease virus (IBDV) and Salmonella had higher mortality. In this work, we investigated the effect of IBDV vaccine (modified live-virus bursal disease vaccine, Nobilis strain 228E®) on experimentally infected chickens with Salmonella Enteritidis (SE). Four experimental groups were included in this study, negative control group, 228E®group, 228E®+SE infected group, and SE infected group. Chickens were ocularly administrated 228E® at 12days of age and orally infected with S. Enteritidis at 13days of age. Sera, intestinal fluid, blood, cloacal swabs and tissue samples were collected at 1, 2 and 3weeks post vaccination (PV). The recorded mortalities were higher in the 228E®+SE infected group, compared to the SE infected group. The anti-S. Enteritidis serum antibody titer and the intestinal mucosal IgA level were higher in the SE infected group at 2 and 3weeks PV, compared to 228E®+SE infected group. S. Enteritidis fecal shedding and organ colonization were significantly higher in the 228E®+SE infected group than the SE infected group at 2 and 3weeks PV. The 228E®+SE group had significantly lower bursa to body weight ratios at 2 and 3weeks PV, as well as had higher bursal lesion scores than the SE infected group. IBDV vaccine depressed the specific-SE systemic and mucosal antibody responses, but did not affect the specific-SE cellular immune responses. Chickens administrated IBDV vaccine, followed by S. Enteritidis infection, could cause a significant effect on the bursa of Fabricius, resulting in failure of systemic and mucosal antibody responses to the S. Enteritidis and reduce the elimination and the clearance of S. Enteritidis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Integrated farm management to prevent Salmonella Enteritidis contamination of eggs

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis (SE) in contaminated eggs is a public health hazard which may cause hospitalization or death in the elderly, infants, and individuals with impaired immune systems. Prevention of SE infection of laying hens is an essential first step in reducing SE outbreaks in humans. Multiple...

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

    PubMed

    Barbosa, Fernanda de Oliveira; Freitas Neto, Oliveiro Caetano de; Batista, Diego Felipe Alves; Almeida, Adriana Maria de; Rubio, Marcela da Silva; Alves, Lucas Bocchini Rodrigues; Vasconcelos, Rosemeire de Oliveira; Barrow, Paul Andrew; Berchieri Junior, Angelo

    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 5dpi). 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. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  4. Dynamic analysis of growth of Salmonella Enteritidis in liquid egg whites

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis (SE) is a common foodborne pathogen associated with eggs and egg products. This research was conducted to study the kinetics of growth and survival of SE in liquid egg whites (LEW). A dynamic temperature profile that exposed SE to suboptimal temperatures and below the minimu...

  5. Production of recombinant flagellin to develop ELISA-based detection of Salmonella Enteritidis.

    PubMed

    Mirhosseini, Seyed Ali; Fooladi, Abbas Ali Imani; Amani, Jafar; Sedighian, Hamid

    Food-borne diseases, caused by the pathogenic bacteria, are highly prevalent in the world. Salmonella is one of the most important bacterial genera responsible for this. Salmonella Enteritidis (SE) is one of the non-typhoid Salmonellae that can be transmitted to human from poultry products, water, and contaminated food. In recent years, new and rapid detection methods such as enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) have been developed. In this study, recombinant FliC (rFliC) was produced to be used as an antigen. The immunization was conducted in mice with the purified recombinant FliC (rFliC). The mice were subcutaneously immunized with rFliC and elicited significant rFliC specific serum IgG antibodies. An indirect ELISA system was established for the detection of Salmonella Enteritidis. Our results confirmed that the recombinant flagellin can be one of the excellent indicators for the detection of Salmonella Enteritidis. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  6. The enhanced immune responses induced by Salmonella enteritidis ghosts loaded with Neisseria gonorrhoeae porB against Salmonella in mice.

    PubMed

    Jiao, Hongmei; Yang, Hui; Zhao, Dan; He, Li; Chen, Jin; Li, Guocai

    2016-11-01

    Human health has been seriously endangered by highly prevalent salmonellosis and multidrug-resistant Salmonella strains. Current vaccines suffer from variable immune-protective effects, so more effective ones are needed to control Salmonella infection : Bacterial ghosts have been produced by the expression of lysis gene E from bacteriophage PhiX174 and can be filled with considerable exogenous substances such as DNA or drugs as a novel platform. In this study, Salmonella enteritidis (SE) ghosts were developed and loaded with Neisseria gonorrhoeae porin B (porB) to construct a novel inactive vaccine. Our new studies show that SE ghosts loaded with porB displayed increased production of pro-inflammatory cytokines (IL-1β, IL-6, IL-10 and IL-12p70) in bone marrow-derived dendritic cells (BMDCs), and elicited significantly higher specific systemic and mucosal immune responses to Salmonella than SE ghosts alone. In addition, the novel porB-loaded ghosts conferred higher protective effects on virulent Salmonella challenge. For the first time, we demonstrate that N. gonorrhoeae porB, as a novel adjuvant, can increase the immunogenicity of SE ghosts. Our studies suggested that Salmonella enteritidis ghosts loaded with Neisseria gonorrhoeae porin B might be a useful mucosal Salmonella vaccine candidate for practical use in the future. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Administration of a Salmonella Enteritidis ΔhilAssrAfliG strain by coarse spray to newly hatched broilers reduces colonization and shedding of a Salmonella Enteritidis challenge strain.

    PubMed

    De Cort, W; Haesebrouck, F; Ducatelle, R; van Immerseel, F

    2015-01-01

    Consumption of contaminated poultry meat is still an important cause of Salmonella infections in humans. Colonization inhibition (CI) occurs when a live Salmonella strain is administered to chickens and subsequently protects against challenge with another Salmonella strain belonging to the same serotype. A Salmonella Enteritidis hilAssrAfliG deletion mutant has previously been proven to reduce colonization and shedding of a wild-type Salmonella Enteritidis strain in newly hatched broilers after experimental infection. In this study, we compared two administration routes for this strain. Administering the Salmonella Enteritidis ΔhilAssrAfliG strain through drinking water on the first day of life resulted in decreased fecal shedding and cecal colonization of a wild-type Salmonella Enteritidis challenge strain administered 24 h later using a seeder-bird model. When administering the CI strain by coarse spray on newly hatched broiler chicks, an even more pronounced reduction of cecal colonization was observed, and fecal shedding of the Salmonella Enteritidis challenge strain ceased during the course of the experiment. These data suggest that administering a Salmonella Enteritidis ΔhilAssrAfliG strain to newly hatched chicks using a coarse spray is a useful and effective method that reduces colonization and shedding of a wild-type Salmonella Enteritidis strain after early challenge. © 2014 Poultry Science Association Inc.

  8. Genomic and phenotypic variation in epidemic-spanning Salmonella enterica serovar Enteritidis isolates

    PubMed Central

    2009-01-01

    Background Salmonella enterica serovar Enteritidis (S. Enteritidis) has caused major epidemics of gastrointestinal infection in many different countries. In this study we investigate genome divergence and pathogenic potential in S. Enteritidis isolated before, during and after an epidemic in Uruguay. Results 266 S. Enteritidis isolates were genotyped using RAPD-PCR and a selection were subjected to PFGE analysis. From these, 29 isolates spanning different periods, genetic profiles and sources of isolation were assayed for their ability to infect human epithelial cells and subjected to comparative genomic hybridization using a Salmonella pan-array and the sequenced strain S. Enteritidis PT4 P125109 as reference. Six other isolates from distant countries were included as external comparators. Two hundred and thirty three chromosomal genes as well as the virulence plasmid were found as variable among S. Enteritidis isolates. Ten out of the 16 chromosomal regions that varied between different isolates correspond to phage-like regions. The 2 oldest pre-epidemic isolates lack phage SE20 and harbour other phage encoded genes that are absent in the sequenced strain. Besides variation in prophage, we found variation in genes involved in metabolism and bacterial fitness. Five epidemic strains lack the complete Salmonella virulence plasmid. Significantly, strains with indistinguishable genetic patterns still showed major differences in their ability to infect epithelial cells, indicating that the approach used was insufficient to detect the genetic basis of this differential behaviour. Conclusion The recent epidemic of S. Enteritidis infection in Uruguay has been driven by the introduction of closely related strains of phage type 4 lineage. Our results confirm previous reports demonstrating a high degree of genetic homogeneity among S. Enteritidis isolates. However, 10 of the regions of variability described here are for the first time reported as being variable in S

  9. Assessing the impact of egg sweating on Salmonella Enteritidis penetration into shell eggs

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis (SE) prevalence in eggs is a major concern to the egg industry. Some research has shown that egg sweating can increase Salmonella penetration into egg contents when refrigerated eggs are moved to a warmer temperature. This occurs when eggs are tempered before wash, to minimize...

  10. In vitro studies of chicken egg yolk antibody (IgY) against Salmonella enteritidis and Salmonella typhimurium.

    PubMed

    Lee, E N; Sunwoo, H H; Menninen, K; Sim, J S

    2002-05-01

    Chicken egg yolk antibody (IgY) raised against Salmonella enteritidis or Salmonella typhimurium was found in highly specific activity levels by ELISA. S. enteritidis- and S. typhimurium-specific IgY powder, prepared by freeze-drying the egg yolk water-soluble fraction, contained 15.5 and 10.0% of specific IgY, respectively. Anti-S. enteritidis IgY cross-reacted 55.3% with S. typhimurium. The cross-reactivity of anti-S. typhimurium IgY with S. enteritidis was 42.4%. Salmonella-specific IgY was demonstrated to inhibit Salmonella growth in liquid medium. The growth rate of S. enteritidis incubated with S. enteritidis-specific IgY was fourfold less than that of the control group during a 4-to-6-h incubation. Cell counts of S. typhimurium incubated with S. typhimurium-specific IgY were reduced by 1.6 log cfu/mL in comparison to that of the control group after 6 h of incubation. The specific binding activity of IgY was further evaluated by using immunofluorescence and immunoelectron microscopy. It was found that Salmonella-specific IgY could bind to the antigens expressed on the Salmonella surface, resulting in structural alterations of the bacterial surface.

  11. Iron restriction and the growth of Salmonella enteritidis.

    PubMed Central

    Chart, H.; Rowe, B.

    1993-01-01

    Strains of Salmonella enteritidis were examined for their ability to remove ferric-ions from the iron chelating agents ovotransferrin, Desferal and EDDA. Growth of S. enteritidis phage type (PT) 4 (SE4) in trypticase soy broth containing ovotransferrin resulted in the expression of iron regulated outer membrane proteins (OMPs) of 74, 78 and 81 kDa, and unexpectedly the repression of expression of OMP C. The 38 MDa 'mouse virulence' plasmid was not required for the expression of the iron-regulated OMPs (IROMPs). SE4 was able to obtain iron bound to the iron chelator Desferal and EDDA without expressing a high-affinity iron uptake system. Strains of S. enteritidis belonging to PTs 7, 8, 13a, 23, 24 and 30 were also able to remove ferric ions from Desferal and EDDA without expressing a high-affinity iron uptake system. We conclude that strains of SE4 possess a high-affinity iron sequestering mechanism that can readily remove iron from ovotransferrin. It is likely that iron limitation, and not iron restriction, is responsible for the bacteriostatic properties of fresh egg whites. Images Fig. 2 PMID:8432322

  12. Assessing the growth and recovery of Salmonella Enteritidis SE86 after sodium dichloroisocyanurate exposure

    PubMed Central

    Ferreira, Fernanda Stoduto; Horvath, Mariana Bandeira; Tondo, Eduardo Cesar

    2013-01-01

    The objective of the present study was to assess the growth and the recovery of Salmonella (S.) Enteritidis SE86 in different diluents, culture media and using different plating methods after the exposure to 200 mg/kg sodium dichloroisocyanurate (NaDCC). Before and after NaDCC exposure, SE86 was cultured at 30 °C and 7 °C in the following diluents: Peptone water (P), Saline solution (SaS), Peptone water+Saline solution (P+SaS), Peptone water+Tween 80+Lecithin+Sodium thiosulfate (P+N) and Saline solution+Tween 80+Lecithin+Sodium thiosulfate (SaS+N). The SaS diluent was chosen because it was able to maintain cells viable without growth and was further used for plating SE86 on non selective medium (Tryptic Soy Agar-TSA) and on selective media (Mannitol Lysine Crystal Violet Brilliant Green Agar-MLCB; Brilliant Green Agar-BGA; Salmonella Shigella Agar-SS and Xylose Lysine Dextrose–XLD). The Thin Agar Layer method (TAL) i.e., selective media overlayed with non selective TSA was also evaluated. Results indicated that SE86 not exposed to NaDCC was able to grow in P, P+N, SaS+N and P+SaS, but not in SaS, that was able to maintain cells viable. SE86 exposed to NaDCC demonstrated similar counts after dilution in SaS and the plating on non selective TSA, selective media MLCB, BGA, SS and XLD and on TAL media. SE86, S. Typhimurium and S. Bredeney, exposed or not exposed to NaDCC, showed no significant differences in counts on TSA, XLD and XLD overlayed with TSA, suggesting that all those media may be used to quantify NaDCC-exposed Salmonella by plating method. PMID:24516446

  13. Breast abscess in a man due to Salmonella enterica serotype Enteritidis.

    PubMed

    Brncic, Nada; Gorup, Lari; Strcic, Miroslav; Abram, Maja; Mustac, Elvira

    2012-01-01

    Nontyphoidal salmonellae can cause breast infection only exceptionally. A case of breast abscess in a 70-year-old man due to Salmonella enterica serotype Enteritidis (Salmonella Enteritidis) is reported. The infection was successfully treated with a combination of surgical and antibiotic treatment.

  14. Breast Abscess in a Man Due to Salmonella enterica Serotype Enteritidis

    PubMed Central

    Brnčić, Nada; Strčić, Miroslav; Abram, Maja; Mustač, Elvira

    2012-01-01

    Nontyphoidal salmonellae can cause breast infection only exceptionally. A case of breast abscess in a 70-year-old man due to Salmonella enterica serotype Enteritidis (Salmonella Enteritidis) is reported. The infection was successfully treated with a combination of surgical and antibiotic treatment. PMID:22031702

  15. Genetically Similar Isolates of Salmonella enterica Serotype Enteritidis Persistent in China for a Long-Term Period.

    PubMed

    Song, Qifa; Shen, Xuanyi; Yang, Yuanbin; Zhang, Danyang; Gao, Hong

    2016-07-01

    Salmonella enterica serotype Enteritidis (S. Enteritidis) is an important causative agent of nontyphoidal salmonellosis in human populations. In this study, we collected 72 S. Enteritidis strains from 2004 to 2014 in Ningbo, mid-east China. Of the 72 strains, we identified a dominant clone of 58 strains recovered from patient's feces (n = 48), blood (n = 1), pleural effusion (n = 1), chickens (n = 3), and dessert cakes (n = 5) by pulsed-field gel electrophoresis (PFGE) and variable-number of tandem repeat analysis (MLVA). The profile arrangements of MLVA were SE1-SE2-SE3-SE5-SE6-SE8-SE9: 4-4-3-11-10-1-3. These dominant strains were susceptible to ampicillin, chloramphenicol, tetracycline, ciprofloxacin, gentamicin, cefotaxime and trimethoprim-sulfamethoxazole, and resistant to nalidixic acid. Additionally, all isolates harboured virulence genes invA, sipA, sopE, and spvB when tested by PCR. Our results reveal that genetically similar S. Enteritidis strains which accounted for several outbreaks as well as blood infection and pleural cavity infection are prevalent in China for a long-term period. This situation calls for further attention in the prevention and control of foodborne disease caused by Salmonella species. © 2016 Institute of Food Technologists®

  16. Potential use of caprylic acid in broiler chickens: effect on Salmonella enteritidis.

    PubMed

    Skřivanová, Eva; Hovorková, Petra; Čermák, Ladislav; Marounek, Milan

    2015-01-01

    The effect of dietary caprylic acid (CA) on Salmonella Enteritidis, as well as the surface treatment of chicken skin contaminated with Salmonella Enteritidis was evaluated. To evaluate the dietary effect of CA on Salmonella Enteritidis, the individually housed broiler chickens (n=48) were divided into 4 groups (positive control, negative control, 2.5 g/kg of CA in the feed, and 5 g/kg of CA in the feed). The feed of all groups, except the negative control, was artificially contaminated with Salmonella Enteritidis ATCC 13076 (10(7) colony-forming units/100 g of feed). Both concentrations of dietary CA significantly decreased counts of Salmonella Enteritidis in the crop and cecum of experimental chickens (p<0.05). The effect of CA in the crop contents was more pronounced than in the cecum. Surface treatment of chilled chicken halves with CA at 1.25 and 2.5 mg/mL significantly decreased Salmonella Enteritidis contamination of chicken skin (p<0.05). The sensory evaluation of the skin and meat showed that treatment of the skin with 1.25 mg/mL of CA worsened odor and appearance of the chicken skin, while sensory traits of chicken meat were not significantly affected. Taste and overall acceptability was not influenced by CA in both meat and skin. Treatment of the skin with 2.5 mg/mL of CA resulted in more pronounced changes of the skin odor and appearance. In conclusion, dietary CA reduced carriage of Salmonella Enteritidis in chickens, whereas surface-treatment reduced or eliminated Salmonella Enteritidis contamination in the processed bird.

  17. Loop-Mediated Isothermal Amplification of the sefA Gene for Rapid Detection of Salmonella Enteritidis and Salmonella Gallinarum in Chickens.

    PubMed

    Gong, Jiansen; Zhuang, Linlin; Zhu, Chunhong; Shi, Shourong; Zhang, Di; Zhang, Linji; Yu, Yan; Dou, Xinhong; Xu, Bu; Wang, Chengming

    2016-04-01

    Salmonella spp. pose a threat to both human and animal health, with more than 2600 serovars having been reported to date. Salmonella serovars are usually identified by slide agglutination tests, which are labor intensive and time consuming. In an attempt to develop a more rapid screening method for the major poultry Salmonella serovars, we developed a loop-mediated isothermal amplification (LAMP) assay, which directly detected the sefA gene, a fimbrial operon gene existing in several specific serovars of Salmonella enterica including the major poultry serovars, namely Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) and Salmonella enterica serovar Gallinarum (Salmonella Gallinarum). With the 177 bacterial strains we tested, positive reactions were only observed with 85 strains of serovar Salmonella Enteritidis and Salmonella Gallinarum. The detection limit of the LAMP assay was 4 CFU/reaction with genomic DNAs of Salmonella Enteritidis (ATCC 13076) from pure culture and 400 CFU/ reaction with DNA extracted from spiked chicken feces. The LAMP assay was more sensitive than conventional culture, especially without enrichment, in detecting Salmonella Enteritidis (CMCC 50041) in the spiked fecal samples. The results show the sefA LAMP method is a rapid, sensitive, specific, and practical method for directly detection of Salmonella Enteritidis and Salmonella Gallinarum in chickens. The sefA LAMP assay can potentially serve as new on-site diagnostics in the poultry industry.

  18. Effect of dietary supplementation of nitrocompounds on salmonella colonization and ileal immune gene expression in laying hens challenged with salmonella enteritidis

    USDA-ARS?s Scientific Manuscript database

    Foodborne disease caused by Salmonella Enteritidis (SE) is one of the important public health and economic concerns. A study was conducted to determine the effect of supplementation with 2-nitroethanol (NE) and 2-nitropropanol (NP) on Salmonella recovery of internal organs as well as on the immune g...

  19. Genomic epidemiology of Salmonella enterica serotype Enteritidis based on population structure of prevalent lineages

    USDA-ARS?s Scientific Manuscript database

    Salmonella enterica serotype Enteritidis (SE) is one of the most commonly reported causes of human salmonellosis. The low genetic diversity of SE measured by fingerprinting methods has made subtyping a challenge. In this study, we used whole genome sequencing to characterize a total of 125 SE and Sa...

  20. Salmonella Enteritidis deposition in eggs after experimental infection of laying hens with different oral doses

    USDA-ARS?s Scientific Manuscript database

    The continuing attribution of human Salmonella Enteritidis (SE) infections to internally contaminated eggs has necessitated the commitment of substantial public and private resources to SE testing and control programs in commercial laying flocks. Cost-effective risk reduction requires a detailed and...

  1. Colonization of mature laying hens with salmonella enteritidis by oral or intracloacal inoculation

    USDA-ARS?s Scientific Manuscript database

    A major route of Salmonella Enteritidis (SE) infection is the fecal-oral route. Evidence of SE in internal organs of laying hens once they are inoculated via the oral (OR) or intracloacal (IC) route has not been reliably demonstrated. The current study evaluated OR or IC route of inoculation of a na...

  2. Food poisoning due to Salmonella Enteritidis--a case report.

    PubMed

    Ogata, Mamoru; Ago, Kazutoshi; Ago, Mihoko; Nakashima, Hiroshi; Hayashi, Takahito

    2009-04-01

    A male in his early seventies complained of abdominal pain and diarrhea at 7h after ingesting a small piece of gratin from a box lunch prepared by a caterer. He was admitted to a hospital, but died 37 h later. Dozens of people who had eaten the same box lunch also complained of diarrhea. All of them recovered after medical treatment. A later investigation demonstrated Salmonella Enteritidis (SE) in the gratin from the box lunch. An autopsy revealed very severe typhloenteritis with edema and submucosal hemorrhage. The digestive tract contained fluid contents without foodstuffs. Bacteriological examination revealed SE in the contents of the lower ileum and large intestine. Based on these findings, we concluded that the cause of death was food poisoning due to SE. In this case, ingesting only a small piece of contaminated food caused fatal food poisoning due to SE. These results emphasize the importance of prevention against food poisoning due to Salmonella, particularly SE.

  3. Evaluation of a Multiplex PCR Assay for the Identification of Salmonella Serovars Enteritidis and Typhimurium Using Retail and Abattoir Samples.

    PubMed

    Ogunremi, Dele; Nadin-Davis, Susan; Dupras, Andrée Ann; Márquez, Imelda Gálvan; Omidi, Katayoun; Pope, Louise; Devenish, John; Burke, Teresa; Allain, Ray; Leclair, Daniel

    2017-02-01

    A multiplex PCR was developed to identify the two most common serovars of Salmonella causing foodborne illness in Canada, namely, serovars Enteritidis and Typhimurium. The PCR was designed to amplify DNA fragments from four Salmonella genes, namely, invA gene (211-bp fragment), iroB gene (309-bp fragment), Typhimurium STM 4497 (523-bp fragment), and Enteritidis SE147228 (612-bp fragment). In addition, a 1,026-bp ribosomal DNA (rDNA) fragment universally present in bacterial species was included in the assay as an internal control fragment. The detection rate of the PCR was 100% among Salmonella Enteritidis (n = 92) and Salmonella Typhimurium (n = 33) isolates. All tested Salmonella isolates (n = 194) were successfully identified based on the amplification of at least one Salmonella -specific DNA fragment. None of the four Salmonella DNA amplicons were detected in any of the non- Salmonella isolates (n = 126), indicating an exclusivity rate of 100%. When applied to crude extracts of 2,001 field isolates of Salmonella obtained during the course of a national microbiological baseline study in broiler chickens and chicken products sampled from abattoir and retail outlets, 163 isolates, or 8.1%, tested positive for Salmonella Enteritidis and another 80 isolates, or 4.0%, tested as Salmonella Typhimurium. All isolates identified by serological testing as Salmonella Enteritidis in the microbiological study were also identified by using the multiplex PCR. The new test can be used to identify or confirm pure isolates of the two serovars and is also amenable for integration into existing culture procedures for accurate detection of Salmonella colonies.

  4. Prevention of egg contamination by Salmonella Enteritidis after oral vaccination of laying hens with Salmonella Enteritidis ΔtolC and ΔacrABacrEFmdtABC mutants.

    PubMed

    Kilroy, Sofie; Raspoet, Ruth; Haesebrouck, Freddy; Ducatelle, Richard; Van Immerseel, Filip

    2016-08-12

    Vaccination of laying hens has been successfully used to reduce egg contamination by Salmonella Enteritidis, decreasing human salmonellosis cases worldwide. Currently used vaccines for layers are either inactivated vaccines or live attenuated strains produced by mutagenesis. Targeted gene deletion mutants hold promise for future vaccines, because specific bacterial functions can be removed that may improve safety and allow differentiation from field strains. In this study, the efficacy of Salmonella Enteritidis ΔtolC and ΔacrABacrEFmdtABC strains in laying hens as live vaccines was evaluated. The mutants are deficient in either the membrane channel TolC (ΔtolC) or the multi-drug efflux systems acrAB, acrEF and mdtABC (ΔacrABacrEFmdtABC). These strains have a decreased ability for gut and tissue colonization and are unable to survive in egg white, the latter preventing transmission of the vaccine strains to humans. Two groups of 30 laying hens were orally inoculated at day 1, 6 weeks and 16 weeks of age with 10(8) cfu of either vaccine strain, while a third group was left unvaccinated. At 24 weeks of age, the birds were intravenously challenged with 5 × 10(7) cfu Salmonella Enteritidis PT4 S1400/94. The vaccine strains were not shed or detected in the gut, internal organs or eggs, 2 weeks after the third vaccination. The strains significantly protected against gut and internal organ colonization, and completely prevented egg contamination by Salmonella Enteritidis under the conditions of this study. This indicates that Salmonella Enteritidis ΔtolC and ΔacrABacrEFmdtABC strains might be valuable strains for vaccination of layers against Salmonella Enteritidis.

  5. Prevalence and antibiotic resistance of Salmonella Enteritidis and Salmonella Typhimurium in raw chicken meat at retail markets in Malaysia.

    PubMed

    Thung, T Y; Mahyudin, N A; Basri, D F; Wan Mohamed Radzi, C W J; Nakaguchi, Y; Nishibuchi, M; Radu, S

    2016-08-01

    Salmonellosis is one of the major food-borne diseases in many countries. This study was carried out to determine the occurrence of Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in raw chicken meat from wet markets and hypermarkets in Selangor, as well as to determine the antibiotic susceptibility profile of S. Enteritidis and S. Typhimurium. The most probable number (MPN) in combination with multiplex polymerase chain reaction (mPCR) method was used to quantify the Salmonella spp., S. Enteritidis, and S. Typhimurium in the samples. The occurrence of Salmonella spp., S. Enteritidis, and S. Typhimurium in 120 chicken meat samples were 20.80%, 6.70%, and 2.50%, respectively with estimated quantity varying from <3 to 15 MPN/g. The antibiogram testing revealed differential multi-drug resistance among S. Enteritidis and S. Typhimurium isolates. All the isolates were resistance to erythromycin, penicillin, and vancomycin whereas sensitivity was recorded for Amoxicillin/Clavulanic acid, Gentamicin, Tetracycline, and Trimethoprim. Our findings demonstrated that the retail chicken meat could be a source of multiple antimicrobial-resistance Salmonella and may constitute a public health concern in Malaysia. © 2016 Poultry Science Association Inc.

  6. Improving Food Safety by Understanding the Evolution of Egg-contaminating Salmonella Enteritidis

    USDA-ARS?s Scientific Manuscript database

    Improving Food Safety by Understanding the Evolution of Egg-contaminating Salmonella Enteritidis Jean Guard, Veterinary Medical Officer U. S. Department of Agriculture, Athens, GA USA (jean.guard@ars.usda.gov) The curious case of egg contamination by Salmonella enterica serovar Enteritidis S. ...

  7. Visualization of gold and platinum nanoparticles interacting with Salmonella Enteritidis and Listeria monocytogenes

    PubMed Central

    Sawosz, Ewa; Chwalibog, André; Szeliga, Jacek; Sawosz, Filip; Grodzik, Marta; Rupiewicz, Marlena; Niemiec, Tomasz; Kacprzyk, Katarzyna

    2010-01-01

    Purpose Rapid development of nanotechnology has recently brought significant attention to the extraordinary biological features of nanomaterials. The objective of the present investigation was to evaluate morphological characteristics of the assembles of gold and platinum nanoparticles (nano-Au and nano-Pt respectively), with Salmonella Enteritidis (Gram-negative) and Listeria monocytogenes (Gram-positive), to reveal possibilities of constructing bacteria-nanoparticle vehicles. Methods Hydrocolloids of nano-Au or nano-Pt were added to two bacteria suspensions in the following order: nano-Au + Salmonella Enteritidis; nano-Au + Listeria monocytogenes; nano-Pt + Salmonella Enteritidis; nano-Pt + Listeria monocytogenes. Samples were inspected by transmission electron microscope. Results Visualization of morphological interaction between nano-Au and Salmonella Enteritidis and Listeria monocytogenes, showed that nano-Au were aggregated within flagella or biofilm network and did not penetrate the bacterial cell. The analysis of morphological effects of interaction of nano-Pt with bacteria revealed that nano-Pt entered cells of Listeria monocytogenes and were removed from the cells. In the case of Salmonella Enteritidis, nano-Pt were seen inside bacteria cells, probably bound to DNA and partly left bacterial cells. After washing and centrifugation, some of the nano-Pt-DNA complexes were observed within Salmonella Enteritidis. Conclusion The results indicate that the bacteria could be used as a vehicle to deliver nano-Pt to specific points in the body. PMID:20856838

  8. SALMONELLA ENTERICA SEROVAR ENTERITIDIS INFECTION MODULATES DIVERSE FUNCTIONAL PROCESSES OF CHICKEN MACROPHAGE AT THE TRANSCRIPTIONAL LEVEL

    USDA-ARS?s Scientific Manuscript database

    Salmonella enterica serovar Enteritidis (SE) is a major etiologic agent of non-typhoid salmonellosis. The organisms colonize adult chicken hosts without causing overt clinical signs. The immunological mechanisms underlying the silent and persistent infection of chickens by SE are not clearly underst...

  9. Atmospheric pressure plasma jet treatment of Salmonella Enteritidis inoculated eggshells.

    PubMed

    Moritz, Maike; Wiacek, Claudia; Koethe, Martin; Braun, Peggy G

    2017-03-20

    Contamination of eggshells with Salmonella Enteritidis remains a food safety concern. In many cases human salmonellosis within the EU can be traced back to raw or undercooked eggs and egg products. Atmospheric pressure plasma is a novel decontamination method that can reduce a wide range of pathogens. The aim of this work was to evaluate the possibility of using an effective short time cold plasma treatment to inactivate Salmonella Enteritidis on the eggshell. Therefore, artificially contaminated eggshells were treated with an atmospheric pressure plasma jet under different experimental settings with various exposure times (15-300s), distances from the plasma jet nozzle to the eggshell surface (5, 8 or 12mm), feed gas compositions (Ar, Ar with 0.2, 0.5 or 1.0% O 2 ), gas flow rates (5 and 7slm) and different inoculations of Salmonella Enteritidis (10 1 -10 6 CFU/cm 2 ). Atmospheric pressure plasma could reduce Salmonella Enteritidis on eggshells significantly. Reduction factors ranged between 0.22 and 2.27 log CFU (colony-forming units). Exposure time and, particularly at 10 4 CFU/cm 2 inoculation, feed gas had a major impact on Salmonella reduction. Precisely, longer exposure times led to higher reductions and Ar as feed gas was more effective than ArO 2 mixtures. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Egg-related Salmonella enteritidis, Italy, 1991

    PubMed Central

    Binkin, N.; Scuderi, G.; Novaco, F.; Giovanardi, G. L.; Paganelli, G.; Ferrari, G.; Cappelli, O.; Ravaglia, L.; Zilioli, F.; Amadei, V.; Magliani, W.; Viani, I.; Riccò, D.; Borrini, B.; Magri, M.; Alessandrini, A.; Bursi, G.; Barigazzi, G.; Fantasia, M.; Filetici, E.; Salmaso, S.

    1993-01-01

    In recent years, Salmonella enteritidis has become an increasingly important public health problem in Italy. In some parts of the country, the fraction of total human salmonella isolates accounted for by S. enteritidis has risen from 3-4% in the mid-1980s to more than 30% in 1990. Between 1990 and 1991, the number of reported S. enteritidis outbreaks increased more than sixfold. The 33 outbreaks reported in 1991 occurred in seven contiguous regions in northern and central Italy and were clustered in time between June and October; in the majority, products containing raw or undercooked shell eggs were implicated. Five of the egg-related outbreaks that occurred within a 30 kilometre radius over a 7-week period were investigated in detail. A phage type 1 strain containing a 38·9 MDa plasmid appeared responsible for three of the outbreaks, while in the remaining two a phage type 4 strain, also with a 38·9 MDa plasmid was isolated. Efforts are being made to enhance epidemiological surveillance and laboratory evaluation, and the use of pasteurized eggs has been recommended for high-risk populations. PMID:8472765

  11. Gene expression profiling in chicken heterophils with Salmonella enteritidis stimulation using a chicken 44 K Agilent microarray

    USDA-ARS?s Scientific Manuscript database

    Salmonella enterica serovar Enteritidis (SE) is one of the most common food-borne pathogens that cause human salmonellosis and usually results from the consumption of contaminated poultry products. The mechanism of SE resistance in chickens remains largely unknown. Previously, heterophils isolated...

  12. Oral vaccination with a live Salmonella Enteritidis/Typhimurium bivalent vaccine in layers induces cross-protection against caecal and internal organ colonization by a Salmonella Infantis strain.

    PubMed

    Eeckhaut, Venessa; Haesebrouck, Freddy; Ducatelle, Richard; Van Immerseel, Filip

    2018-05-01

    Salmonella is an important zoonotic agent, and poultry products remain one of the main sources of infection for humans. Salmonella Infantis is an emerging serotype in poultry worldwide, reflected by an increased prevalence in poultry flocks, on broiler meat and in human foodborne illness cases. In the current study, the efficacy of oral administration of a live monovalent Salmonella Enteritidis and a live bivalent Salmonella Enteritidis/Typhimurium vaccine, against a Salmonella Enteritidis and Infantis infection, was determined. Oral administration of the live vaccines to day-old chickens caused a decrease in caecal colonization by Salmonella Enteritidis, but not Infantis, at day 7, when challenged at day 2. Vaccination with the bivalent vaccine at day 1 resulted in a decreased spleen colonization by both Salmonella Infantis and Enteritidis. Twice (at day 1 and week 6) and thrice vaccination (at day 1, week 6 and 16) of laying hens with the bivalent vaccine resulted in a decreased caecal colonization by Salmonella Enteritidis and Infantis, and significantly lower oviduct colonization levels by Salmonella Enteritidis. These data show cross-protection against Salmonella Infantis by oral administration of live vaccine strains belonging to other serogroups. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Quinolone-resistant Salmonella enterica serotype Enteritidis infections associated with international travel.

    PubMed

    O'Donnell, Allison T; Vieira, Antonio R; Huang, Jennifer Y; Whichard, Jean; Cole, Dana; Karp, Beth E

    2014-11-01

    We found a strong association between nalidixic acid-resistant Salmonella enterica serotype Enteritidis infections in the United States and recent international travel by linking Salmonella Enteritidis data from the National Antimicrobial Resistance Monitoring System and the Foodborne Diseases Active Surveillance Network. Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US.

  14. Continuing multiplication of Salmonella Enteritidis strains in egg yolk during refrigeration at 7.2° C

    USDA-ARS?s Scientific Manuscript database

    The continuing attribution of human illness caused by Salmonella Enteritidis (SE) to the consumption of contaminated eggs has led to widespread implementation of risk reduction programs for commercial egg production. Prompt refrigeration of eggs to prevent bacterial multiplication to dangerously hig...

  15. Contribution of Salmonella Enteritidis virulence factors to intestinal colonization and systemic dissemination in 1-day-old chickens.

    PubMed

    Addwebi, Tarek M; Call, Douglas R; Shah, Devendra H

    2014-04-01

    Salmonella enterica serovar Enteritidis is one of the most common serovars associated with poultry and poultry product contamination in the United States. We previously identified 14 mutant strains of Salmonella Enteritidis phage type 4 (PT4) with significantly reduced invasiveness in human intestinal epithelial cells (Caco-2), chicken macrophages (HD-11), and chicken hepatocellular epithelial cells (LMH). These included Salmonella Enteritidis mutants with transposon insertions in 6 newly identified Salmonella Enteritidis-specific genes (pegD and SEN1393), and genes or genomic islands common to most other Salmonella serovars (SEN0803, SEN0034, SEN2278, and SEN3503) along with 8 genes previously known to contribute to enteric infection (hilA, pipA, fliH, fljB, csgB, spvR, and rfbMN). We hypothesized that Salmonella Enteritidis employs both common Salmonella enterica colonization factors and Salmonella Enteritidis-specific traits to establish infection in chickens. Four Salmonella Enteritidis mutants (SEN0034::Tn5, fliH::Tn5, SEN1393::Tn5, and spvR::Tn5) were indistinguishable from the isogenic wild-type strain when orally inoculated in 1-d-old chickens, whereas 2 mutants (CsgB::Tn5 and PegD::Tn5) were defective for intestinal colonization (P < 0.05) and 8 mutants (hilA::Tn5, SEN3503::Tn5, SEN0803::Tn5, SEN2278::Tn5, fljB::Tn5, rfbM::Tn5, rfbN::Tn5, and pipA::Tn5) showed significant in vivo attenuation in more than one organ (P < 0.05). Complementation studies confirmed the role of rfbN and SEN3503 during infection. This study should contribute to a better understanding of the mechanisms involved in Salmonella Enteritidis pathogenesis, and the target genes identified here could potentially serve as targets for the development of live-attenuated or subunit vaccine.

  16. In vitro evaluation of anti-infective activity of a Lactobacillus plantarum strain against Salmonella enterica serovar Enteritidis

    PubMed Central

    2013-01-01

    Background Salmonella enterica serovar Enteritidis infections are known to exhibit worldwide prevalence with increased morbidity and mortality. The conventional strategies like antibiotic therapy and vaccination have not only proved to be of sub-optimal efficacy but also led to the development of multidrug resistant strains of Salmonella. Antimicrobial activities of probiotics against various enteropathogens and other health promoting effects have assumed greater significance in recent years. The present study aims to evaluate the efficacy of a Lactobacillus plantarum strain (KSBT 56, isolated from a traditional food product of India), in preventing Salmonella enterica serovar Enteritidis growth and pathogenicity in vitro. Methods and results The cell free culture supernatant (CFCS) of KSBT 56 strain notably inhibited the growth of Salmonella Enteritidis without affecting the growth of other gram-positive lactic acid bacteria. The isolated KSBT 56 strain produces lactic acid similar to other standard probiotic strains like Lactobacillus plantarum MTCC 1407. The free radical production by KSBT 56 strain was studied by using sodC mutant of S. Enteritidis, which exhibited reduced growth in the presence of CFCS of the KSBT 56 strain, indicating the inhibitory activity of free radicals on the growth of S. Enteritidis. Our results also showed a significant reduction in the biofilm forming ability of Salmonella Enteritidis in the presence of the KSBT 56 strain (2 log cfu/ml, p = 0.01). Further, the anti-infective characteristics of KSBT 56 strain was validated by gentamicin protection assay which revealed 80% reduction in the invasion of Salmonella Enteritidis to HCT-116 cell line (Salmonella Enteritidis and KSBT 56 in a 1:1 ratio) and delayed addition of Salmonella Enteritidis by 1 h. Similarly, the reduced adhesion of Salmonella to the HCT-116 cells was observed along with the down regulation of hilA gene of Salmonella Pathogenicity Island 1 (SPI1) indicating that they

  17. The effects of polymorphisms in 7 candidate genes on resistance to Salmonella Enteritidis in native chickens.

    PubMed

    Tohidi, R; Idris, I B; Malar Panandam, J; Hair Bejo, M

    2013-04-01

    Salmonella enterica serovar Enteritidis infection is a common concern in poultry production for its negative effects on growth as well as food safety for humans. Identification of molecular markers that are linked to resistance to Salmonella Enteritidis may lead to appropriate solutions to control Salmonella infection in chickens. This study investigated the association of candidate genes with resistance to Salmonella Enteritidis in young chickens. Two native breeds of Malaysian chickens, namely, Village Chickens and Red Junglefowl, were evaluated for bacterial colonization after Salmonella Enteritidis inoculation. Seven candidate genes were selected on the basis of their physiological role in immune response, as determined by prior studies in other genetic lines: natural resistance-associated protein 1 (NRAMP1), transforming growth factor β3 (TGFβ3), transforming growth factor β4 (TGFβ4), inhibitor of apoptosis protein 1 (IAP1), caspase 1 (CASP1), lipopolysaccharide-induced tumor necrosis factor (TNF) α factor (LITAF), and TNF-related apoptosis-inducing ligand (TRAIL). Polymerase chain reaction-RFLP was used to identify polymorphisms in the candidate genes; all genes exhibited polymorphisms in at least one breed. The NRAMP1-SacI polymorphism correlated with the differences in Salmonella Enteritidis load in the cecum (P = 0.002) and spleen (P = 0.01) of Village Chickens. Polymorphisms in the restriction sites of TGFβ3-BsrI, TGFβ4-MboII, and TRAIL-StyI were associated with Salmonella Enteritidis burden in the cecum, spleen, and liver of Village Chickens and Red Junglefowl (P < 0.05). These results indicate that the NRAMP1, TGFβ3, TGFβ4, and TRAIL genes are potential candidates for use in selection programs for increasing genetic resistance against Salmonella Enteritidis in native Malaysian chickens.

  18. Predictive Modeling for the Growth of Salmonella Enteritidis in Chicken Juice by Real-Time Polymerase Chain Reaction.

    PubMed

    Noviyanti, Fia; Hosotani, Yukie; Koseki, Shigenobu; Inatsu, Yasuhiro; Kawasaki, Susumu

    2018-04-02

    The goals of this study were to monitor the growth kinetics of Salmonella Enteritidis in chicken juice using real-time polymerase chain reaction (PCR) and to evaluate its efficacy by comparing the results with an experimental database. Salmonella Enteritidis was inoculated in chicken juice samples at an initial inoculum of 10 4 CFU/mL with inoculated samples incubated at six different temperatures (10, 15, 20, 25, 30, and 35°C). Sampling was carried out for 36 h to observe the growth of Salmonella Enteritidis. The total DNA was extracted from the samples, and the copy number of the Salmonella invasion gene (invA) was quantified by real-time PCR and converted to Salmonella Enteritidis cell concentration. Growth kinetics data were analyzed by the Baranyi and Roberts model to obtain growth parameters, whereas the Ratkowsky's square-root model was used to describe the effect of the interactions between growth parameters and temperature on the growth of Salmonella Enteritidis. The growth parameters of Salmonella Enteritidis obtained from an experiment conducted at a constant temperature were validated with growth data from chicken juice samples that were incubated under fluctuating temperature conditions between 5°C and 30°C for 30-min periods. A high correlation was observed between maximum growth rate (μ max ) and storage temperature, indicating that the real-time PCR-monitoring method provides a precise estimation of Salmonella Enteritidis growth in food material with a microbial flora. Moreover, the μ max data reflected data from microbial responses viewer database and ComBase. The results of this study suggested that real-time PCR monitoring provides a precise estimation of Salmonella Enteritidis growth in food materials with a background microbial flora.

  19. Salmonella Enteritidis with double deletion in phoP fliC and a competitive exclusion culture elicit substantial additive protective effects against Salmonella exposure in newly hatched chicks.

    PubMed

    Methner, U; Berndt, A; Locke, M

    2017-10-27

    A live Salmonella Enteritidis vaccine (SE147N ΔphoP fliC), able to express both a homologous intestinal colonisation-inhibition effect and a systemic invasion-inhibition effect, was tested for its potential to generate a postulated additive protective effect in case of combined application with a competitive exclusion (CE) culture against Salmonella exposure in very young chicks. Both, SE147N ΔphoP fliC and the CE culture alone were highly protective against systemic and intestinal colonisation of the challenge strain in case of moderate Salmonella exposure, consequently, additive protective effects in combined use could not be detected. However, in case of high Salmonella Enteritidis challenge with 10 6 cfu/bird at day 3 of life the combination of the ΔphoP fliC vaccine and the CE culture resulted in a protective effect much more pronounced than either of the single preparations and most substantial compared to untreated control birds. The term additive protective effects reflects the recognition that exclusion effects by gut flora cultures and inhibition effects by Salmonella vaccines are caused by different mechanisms. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Salmonella enteritidis Effector AvrA Stabilizes Intestinal Tight Junctions via the JNK Pathway.

    PubMed

    Lin, Zhijie; Zhang, Yong-Guo; Xia, Yinglin; Xu, Xiulong; Jiao, Xinan; Sun, Jun

    2016-12-23

    Salmonella pathogenesis studies to date have focused on Salmonella typhimurium, and the pathogenesis of a second major serotype, Salmonella enteritidis, is poorly understood. Salmonella spp. possess effector proteins that display biochemical activities and modulate host functions. Here, we generated a deletion mutant of the effector AvrA, S.E-AvrA - , and a plasmid-mediated complementary strain, S.E-AvrA - /pAvrA + (S.E-AvrA + ), in S. Enteritidis. Using in vitro and in vivo infection models, we showed that AvrA stabilizes epithelial tight junction (TJ) proteins, such as ZO-1, in human intestinal epithelial cells. Transepithelial electrical resistance was significantly higher in cells infected with S.E-AvrA + than in cells infected with S.E-AvrA - Inhibition of the JNK pathway suppresses the disassembly of TJ proteins; we found that enteritidis AvrA inhibited JNK activity in cells infected with wild type or S.E-AvrA + strains. Therefore, Enteritidis AvrA-induced ZO-1 stability is achieved via suppression of the JNK pathway. Furthermore, the S.E-AvrA - strain led to enhanced bacterial invasion, both in vitro and in vivo Taken together, our data reveal a novel role for AvrA in S. Enteritidis: Enteritidis AvrA stabilizes intestinal TJs and attenuates bacterial invasion. The manipulation of JNK activity and TJs in microbial-epithelial interactions may be a novel therapeutic approach for the treatment of infectious diseases. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Salmonella enteritidis Effector AvrA Stabilizes Intestinal Tight Junctions via the JNK Pathway*

    PubMed Central

    Lin, Zhijie; Zhang, Yong-Guo; Xia, Yinglin; Xu, Xiulong; Jiao, Xinan

    2016-01-01

    Salmonella pathogenesis studies to date have focused on Salmonella typhimurium, and the pathogenesis of a second major serotype, Salmonella enteritidis, is poorly understood. Salmonella spp. possess effector proteins that display biochemical activities and modulate host functions. Here, we generated a deletion mutant of the effector AvrA, S.E-AvrA−, and a plasmid-mediated complementary strain, S.E-AvrA−/pAvrA+ (S.E-AvrA+), in S. Enteritidis. Using in vitro and in vivo infection models, we showed that AvrA stabilizes epithelial tight junction (TJ) proteins, such as ZO-1, in human intestinal epithelial cells. Transepithelial electrical resistance was significantly higher in cells infected with S.E-AvrA+ than in cells infected with S.E-AvrA−. Inhibition of the JNK pathway suppresses the disassembly of TJ proteins; we found that enteritidis AvrA inhibited JNK activity in cells infected with wild type or S.E-AvrA+ strains. Therefore, Enteritidis AvrA-induced ZO-1 stability is achieved via suppression of the JNK pathway. Furthermore, the S.E-AvrA− strain led to enhanced bacterial invasion, both in vitro and in vivo. Taken together, our data reveal a novel role for AvrA in S. Enteritidis: Enteritidis AvrA stabilizes intestinal TJs and attenuates bacterial invasion. The manipulation of JNK activity and TJs in microbial-epithelial interactions may be a novel therapeutic approach for the treatment of infectious diseases. PMID:27875307

  2. Isolation and characterization of Salmonella Enteritidis and Salmonella Typhimurium from chicken meat in Egypt.

    PubMed

    Tarabees, Reda; Elsayed, Mohamed S A; Shawish, Reyad; Basiouni, Shereen; Shehata, Awad A

    2017-04-30

    Salmonella enterica serovars Enteritidis and Typhimurium represent the major serovars associated with human salmonellosis. Contamination of meat products with these serovars is considered the main source of infection. In this study, 100 raw chicken meat samples were investigated for the presence of Salmonella spp., which were subsequently identified based on biochemical and serological tests as well as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) profile. Furthermore, the isolated serovars were examined using multiplex polymerase chain reaction (PCR) for the presence of virulence genes suspected to have a role in infection. S. Enteritidis was isolated from two samples (2%), while S. Typhimurium was isolated from three samples (3%) of chicken meat. Of the 17 examined virulence genes using multiplex PCR, the sitC, sopB, sifA, lpfC, spaN, sipB, invA, spiA, and msgA genes were detected in S. Enteritidis. However, the sitC, iroN, sopB, sifA, lpfC, spaN, sipB, invA, and tolC genes were successfully amplified in S. Typhimurium. The detection of S. Enteritidis and S. Typhimurium in meat, even at low incidence, has important implications. In addition, the data presented here is the first attempt to identify a wide range of virulence genes in Egyptian Salmonella isolates recovered from meat products. A strict public health and food safety regime is urgently needed in order to decrease the human health hazard risk associated with salmonellosis.

  3. Differential levels of cecal colonization by Salmonella Enteritidis in chickens triggers distinct immune kinome profiles

    USDA-ARS?s Scientific Manuscript database

    Salmonella enterica serovar Enteritidis are facultative intracellular bacteria that cause disease in numerous species. Salmonella-related infections originating from poultry and/or poultry products are a major cause of human foodborne illness, and S. Enteritidis is the leading cause worldwide. Des...

  4. Whole Genome Sequence Analysis of Salmonella Enteritidis Isolated from Wild Mice

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis is a foodborne pathogen of global concern because of the high frequency isolated from foods and patients. Draft genomes of 64 S. Enteritidis strains from intestines and spleens of mice were reported. The availability of these genomes provides useful information on genomic dive...

  5. Similarities between Salmonella Enteritidis isolated from humans and captive wild animals in South Africa.

    PubMed

    Smith, Anthony M; Ismail, Husna; Henton, Maryke M; Keddy, Karen H

    2014-12-15

    Salmonella is well recognized as an aetiological agent of gastrointestinal and diarrhoeal disease. Salmonella enterica serotype Enteritidis (Salmonella Enteritidis) is one of the commonest serotypes associated with foodborne illness. In South Africa, we compared Salmonella Enteritidis strains isolated from humans with gastroenteritis and strains isolated from captive wild animals, between June 2011 and July 2012. Bacteria were phenotypically characterized using standard microbiological techniques. Genotypic relatedness of isolates was investigated by pulsed-field gel electrophoresis (PFGE) analysis. a diversity of 27 PFGE patterns amongst 196 human non-invasive isolates was shown; two PFGE patterns predominated and accounted for 74% of all human isolates. Human isolates showed a 12% prevalence rate for nalidixic acid resistance. Animal isolates from 5 different sources were investigated. With the exception of an isolate from a ground hornbill, all animal isolates (jaguar, crocodile, lion and poultry) showed PFGE pattern matches to a human isolate. Animal isolates showed susceptibility to all antimicrobial agents tested, with the exception of nalidixic acid resistance in isolates from the lion and poultry source. Our data showed similarities between Salmonella Enteritidis strains isolated from humans and captive wild animals, suggesting a probable common source for strains from humans and animals.

  6. Composition of Gut Microbiota Influences Resistance of Newly Hatched Chickens to Salmonella Enteritidis Infection

    PubMed Central

    Varmuzova, Karolina; Kubasova, Tereza; Davidova-Gerzova, Lenka; Sisak, Frantisek; Havlickova, Hana; Sebkova, Alena; Faldynova, Marcela; Rychlik, Ivan

    2016-01-01

    Since poultry is a very common source of non-typhoid Salmonella for humans, different interventions aimed at decreasing the prevalence of Salmonella in chickens are understood as an effective measure for decreasing the incidence of human salmonellosis. One such intervention is the use of probiotic or competitive exclusion products. In this study we tested whether microbiota from donor hens of different age will equally protect chickens against Salmonella Enteritidis infection. Newly hatched chickens were therefore orally inoculated with cecal extracts from 1-, 3-, 16-, 28-, and 42-week-old donors and 7 days later, the chickens were infected with S. Enteritidis. The experiment was terminated 4 days later. In the second experiment, groups of newly hatched chickens were inoculated with cecal extracts of 35-week-old hens either on day 1 of life followed by S. Enteritidis infection on day 2 or were infected with S. Enteritidis infection on day 1 followed by therapeutic administration of the cecal extract on day 2 or were inoculated on day 1 of life with a mixture of the cecal extract and S. Enteritidis. This experiment was terminated when the chickens were 5 days old. Both Salmonella culture and chicken gene expression confirmed that inoculation of newly hatched chickens with microbiota from 3-week-old or older chickens protected them against S. Enteritidis challenge. On the other hand, microbiota from 1-week-old donors failed to protect chickens against S. Enteritidis challenge. Microbiota from 35-week-old hens protected chickens even 24 h after administration. However, simultaneous or therapeutic microbiota administration failed to protect chickens against S. Enteritidis infection. Gut microbiota can be used as a preventive measure against S. Enteritidis infection but its composition and early administration is critical for its efficacy. PMID:27379083

  7. Multiplication of Salmonella Enteritidis in egg yolks after inoculation outside, on, and inside vitelline membranes and storage at different temperatures

    USDA-ARS?s Scientific Manuscript database

    Prompt refrigeration to restrict bacterial growth is important for reducing egg-borne transmission of Salmonella enterica serovar Enteritidis (SE). The nutrient-rich yolk interior is a relatively infrequent location for initial SE deposition in eggs, but migration across the vitelline membrane can ...

  8. Public health investigations of Salmonella Enteritidis in catering raw shell eggs, 2002-2004.

    PubMed

    Little, C L; Surman-Lee, S; Greenwood, M; Bolton, F J; Elson, R; Mitchell, R T; Nichols, G N; Sagoo, S K; Threlfall, E J; Ward, L R; Gillespie, I A; O'Brien, S

    2007-06-01

    In response to a dramatic change in the epidemiology of Salmonella Enteritidis in England and Wales thought to be associated with raw shell eggs, the Health Protection Agency initiated public health investigations to establish the incidence of Salmonella contamination and origin of eggs used by catering premises implicated in outbreaks of Salm. Enteritidis. Between October 2002 and November 2004, 16 971 eggs were sampled and Salmonella were recovered from 3.4%. Salmonella was isolated from 5.5% and 6.3% of Spanish and eggs of unknown origin, respectively, used in catering premises linked to outbreaks, a level significantly higher than that (1.1%) found in nonLion Quality UK eggs sampled. The small sample of UK Lion Quality eggs tested (reflecting their lack of use in premises visited) did not contain Salmonella. Several phage types of Salm. Enteritidis other than phage type 4 (PT 4) were identified with nonUK eggs. Eggs from Spain were implicated as a major source of infection. Eggs were contaminated more frequently with Salmonella when shells were dirty and/or cracked, and stored at above 8 degrees C. The use of Spanish eggs by the catering sector has been identified as a consistent significant factor in many of the outbreaks caused by Salm. Enteritidis nonPT4 in England and Wales during 2002-2004. Advice to caterers and hospitals that raw shell eggs should not be used in food that will either not be cooked or only lightly cooked should be reinforced.

  9. Direct construction of predictive models for describing growth Salmonella enteritidis in liquid eggs – a one-step approach

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to develop a new approach using a one-step approach to directly construct predictive models for describing the growth of Salmonella Enteritidis (SE) in liquid egg white (LEW) and egg yolk (LEY). A five-strain cocktail of SE, induced to resist rifampicin at 100 mg/L, ...

  10. Horizontal transmission of Salmonella Enteritidis in experimentally infected laying hens housed in conventional or enriched cages.

    PubMed

    Gast, Richard K; Guraya, Rupa; Jones, Deana R; Anderson, Kenneth E

    2014-12-01

    The majority of human illnesses caused by Salmonella Enteritidis are attributed to contaminated eggs, and the prevalence of this pathogen in commercial laying flocks has been identified as a leading epidemiologic risk factor. Flock housing and management systems can affect opportunities for the introduction, transmission, and persistence of foodborne pathogens in poultry. The animal welfare implications of different types of housing for laying hens have been widely discussed in recent years, but the food safety consequences of these production systems remain incompletely understood. The present study assessed the effects of 2 different housing systems (conventional cages and colony cages enriched with perching and nesting areas) on the horizontal transmission of experimentally introduced Salmonella Enteritidis infection within groups of laying hens. In each of 2 trials, 136 hens were distributed among cages of both housing systems and approximately one-third of the hens in each cage were orally inoculated with doses of 10(8) cfu of Salmonella Enteritidis (phage type 13a in one trial and phage type 4 in the other). At regular intervals through 23 d postinoculation, cloacal swabs were collected from all hens (inoculated and uninoculated) and cultured for Salmonella Enteritidis. Horizontal contact transmission of infection was observed for both Salmonella Enteritidis strains, reaching peak prevalence values of 27.1% of uninoculated hens in conventional cages and 22.7% in enriched cages. However, no significant differences (P > 0.05) in the overall frequencies of horizontal Salmonella Enteritidis transmission were evident between the 2 types of housing. These results suggest that opportunities for Salmonella Enteritidis infection to spread horizontally throughout laying flocks may be similar in conventional and enriched cage-based production systems. ©2014 Poultry Science Association Inc.

  11. National surveillance of Salmonella Enteritidis in commercial eggs in Japan.

    PubMed

    Esaki, H; Shimura, K; Yamazaki, Y; Eguchi, M; Nakamura, M

    2013-05-01

    A total of 105 033 eggs were collected across Japan from June 2010 to January 2011 and tested for Salmonella Enteritidis to provide data for the risk profiling of S. Enteritidis in eggs by the Food Safety Commission of Japan. S. Enteritidis isolates were recovered from three samples (20 eggs/sample) and these samples were different in regard to sampling period, grading and packaging centre and farm. The prevalence of S. Enteritidis in commercial eggs in Japan is estimated at ~0.003% which was a tenfold decrease in prevalence compared to similar surveillance in the mid 1990s. The decrease in the contamination in commercial eggs is considered a contributory factor in the decrease of foodborne diseases associated with S. Enteritidis in this period.

  12. Microarray-based detection of Salmonella enterica serovar Enteritidis genes involved in chicken reproductive tract colonization.

    PubMed

    Raspoet, R; Appia-Ayme, C; Shearer, N; Martel, A; Pasmans, F; Haesebrouck, F; Ducatelle, R; Thompson, A; Van Immerseel, F

    2014-12-01

    Salmonella enterica serovar Enteritidis has developed the potential to contaminate table eggs internally, by colonization of the chicken reproductive tract and internalization in the forming egg. The serotype Enteritidis has developed mechanisms to colonize the chicken oviduct more successfully than other serotypes. Until now, the strategies exploited by Salmonella Enteritidis to do so have remained largely unknown. For that reason, a microarray-based transposon library screen was used to identify genes that are essential for the persistence of Salmonella Enteritidis inside primary chicken oviduct gland cells in vitro and inside the reproductive tract in vivo. A total of 81 genes with a potential role in persistence in both the oviduct cells and the oviduct tissue were identified. Major groups of importance include the Salmonella pathogenicity islands 1 and 2, genes involved in stress responses, cell wall, and lipopolysaccharide structure, and the region-of-difference genomic islands 9, 21, and 40. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  13. Salmonella Enteritidis organ invasion and egg contamination in experimentally infected laying hens housed in conventional or enriched cages.

    USDA-ARS?s Scientific Manuscript database

    Both disease surveillance and epidemiologic analyses have confirmed a strong association between human salmonellosis and the prevalence of Salmonella Enteritidis (SE) in commercial egg flocks. The majority of human illnesses caused by this pathogen are attributed to contaminated eggs. Animal welfare...

  14. Colonization of internal organs by Salmonella Enteritidis in experimentally infected laying hens housed in enriched colony cages at different stocking densities

    USDA-ARS?s Scientific Manuscript database

    The frequency of human infections with Salmonella Enteritidis (SE) has been linked to contaminated eggs and thus to SE prevalence in commercial egg-laying flocks. Contamination of the edible contents of eggs is a consequence of SE colonization of reproductive tissues in systemically infected hens. T...

  15. [Genetic mechanisms of Salmonella enteritidis biodiversity and clinical features of salmonellosis].

    PubMed

    Mavziutov, A R; Murzabaeva, R T; Nazmutdinova, R G; Mirsaiapova, I A

    2010-01-01

    To assess prevalence of fragments of Escherichia coli pathogenicity islands in Salmonella enteritidis strains as well as to study clinical signs of disease caused by these strains in adults. Ninety-six patients with salmonellosis were studied. Ninety strains of S. enteritidis were isolated and tested by PCR for the presence of genes associated with pathogenicity islands of E. coli: hlyA, hlyB, sfaG, and sfaA. It was determined that DNA fragments homologous to pathogenicity islands of E. coli were present in 87 (96.7%) of S. enteritidis clinical isolates. Disease caused by Salmonella strains which possess only sfaG was mostly mild--7 (33.3%), whereas strains which had sfaG with fragments of hlyA and/or hlyB caused severe disease--7 (50%). sfaA fragments were found mostly in combination with other genes. In such cases the disease was mostly severe--6 (42.8%). Correlation between presence of E. coli pathogenicity islands in Salmonella spp., their antibiotic resistance and severity of infection was established.

  16. Chicken innate immune response to oral infection with Salmonella enterica serovar Enteritidis

    PubMed Central

    2013-01-01

    The characterization of the immune response of chickens to Salmonella infection is usually limited to the quantification of expression of genes coding for cytokines, chemokines or antimicrobial peptides. However, processes occurring in the cecum of infected chickens are likely to be much more diverse. In this study we have therefore characterized the transcriptome and proteome in the chicken cecum after infection with Salmonella Enteritidis. Using a combination of 454 pyrosequencing, protein mass spectrometry and quantitative real-time PCR, we identified 48 down- and 56 up-regulated chicken genes after Salmonella Enteritidis infection. The most inducible gene was that coding for MMP7, exhibiting a 5952 fold induction 9 days post-infection. An induction of greater than 100 fold was observed for IgG, IRG1, SAA, ExFABP, IL-22, TRAP6, MRP126, IFNγ, iNOS, ES1, IL-1β, LYG2, IFIT5, IL-17, AVD, AH221 and SERPIN B. Since prostaglandin D2 synthase was upregulated and degrading hydroxyprostaglandin dehydrogenase was downregulated after the infection, prostaglandin must accumulate in the cecum of chickens infected with Salmonella Enteritidis. Finally, above mentioned signaling was dependent on the presence of a SPI1-encoded type III secretion system in Salmonella Enteritidis. The inflammation lasted for 2 weeks after which time the expression of the “inflammatory” genes returned back to basal levels and, instead, the expression of IgA and IgG increased. This points to an important role for immunoglobulins in the restoration of homeostasis in the cecum after infection. PMID:23687968

  17. Investigation of Salmonella Enteritidis outbreaks in South Africa using multi-locus variable-number tandem-repeats analysis, 2013-2015.

    PubMed

    Muvhali, Munyadziwa; Smith, Anthony Marius; Rakgantso, Andronica Moipone; Keddy, Karen Helena

    2017-10-02

    Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) has become a significant pathogen in South Africa, and the need for improved molecular surveillance of this pathogen has become important. Over the years, multi-locus variable-number tandem-repeats analysis (MLVA) has become a valuable molecular subtyping technique for Salmonella, particularly for highly homogenic serotypes such as Salmonella Enteritidis. This study describes the use of MLVA in the molecular epidemiological investigation of outbreak isolates in South Africa. Between the years 2013 and 2015, the Centre for Enteric Diseases (CED) received 39 Salmonella Enteritidis isolates from seven foodborne illness outbreaks, which occurred in six provinces. MLVA was performed on all isolates. Three MLVA profiles (MLVA profiles 21, 22 and 28) were identified among the 39 isolates. MLVA profile 28 accounted for 77% (30/39) of the isolates. Isolates from a single outbreak were grouped into a single MLVA profile. A minimum spanning tree (MST) created from the MLVA data showed a close relationship between MLVA profiles 21, 22 and 28, with a single VNTR locus difference between them. MLVA has proven to be a reliable method for the molecular epidemiological investigation of Salmonella Enteritidis outbreaks in South Africa. These foodborne outbreaks emphasize the importance of the One Health approach as an essential component for combating the spread of zoonotic pathogens such as Salmonella Enteritidis.

  18. [Use of bacteriphages against Salmonella Enteritidis: a prevention tool].

    PubMed

    García, Cristina; Marín, Clara; Catalá-Gregori, Pablo; Soriano, Jose Miguel

    2015-06-01

    Salmonellosis is a highly prevalent disease still searching for preventive tools to avoid contamination level priority public health. The in vitro effect of bacteriophages against Salmonella enteritidis was evaluated as a prevention tool. Two tests with three concentrations of bacteriophages were conducted against two strains of Salmonella Enteritidis inoculated in fresh faeces of laying hens. Each test had a positive control. Thus, four groups in each test were evaluated. Each experimental group included two replicates, and three plates were incubated per replicate. The concentrations tested were three: commercial solution (5 × 10(7) pfu/mL), and two dilutions (1/10 and 1/30). One of the strains tested was CECT 4300, a certified strain of Colección Española de Cultivo Tipo and the other a field isolated strain in a sacrificed hen farm. Both strains were inoculated at 1.3 × 10(5) cfu/g of faeces in each of the four groups. Isolation and identification of bacteria by ISO6579 was done at various times after inoculation: 1 minute, 24 hours and 7 days. In the first test, with certified strain, Salmonella was isolated in all groups at time 1 minute. After 24 hours, Salmonella was isolated in all groups except in one of the replicas treated with 1/10 dilution of bacteriophages, one of the other replica plate treated with 1/10 dilution, and two plates of the two replicas treated with the commercial solution. After 7 days, the bacteria were not isolated from any of the experimental groups. In the second test, with the field strain, Salmonella was isolated in all groups at time 1 minute. After 24 hours, Salmonella was isolated in all groups except in one of the replicas treated with 1/10 dilution of bacteriophages and the two replicas treated with the commercial solution. Salmonella was not isolated in any of the experimental groups at 7 days. The use of bacteriophages reduced Salmonella enteritidis isolates in faeces at 24 hours after the application, so it could be

  19. Characteristics of invasion-reduced hilA gene mutant of Salmonella Enteritidis in vitro and in vivo.

    PubMed

    Lv, Shuang; Si, Wei; Yu, Shenye; Li, Zhaoli; Wang, Xiumei; Chen, Liping; Zhang, Wanjiang; Liu, Siguo

    2015-08-01

    Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) is a facultative intracellular pathogen that causes huge losses in poultry industry and also food poisoning in humans due to its being a food-borne pathogen. Functions of Invasion-related genes need to be explored, as invasion is a key step for Salmonella infection. In this study, a transposon mutant library of Salmonella Enteritidis isolate SM6 was constructed and screened for the invasion-related genes via incubation with Caco-2 cells. Three stably attenuated mutants were identified for significantly reduced invasion with insertions all in hilA (hyperinvasive locus A) gene. We constructed and evaluated the hilA deletion mutant in vivo and in vitro. SM6△hilA showed significantly reduced ability to invade Caco-2 cells and decreased pathogenicity in chicks. However, the bacterial load and pathological damage in the cecum were significantly higher than those in the SM6 in vivo. Present results provide new evidences for pathogenicity research on Salmonella Enteritidis. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. A comparison of transmission characteristics of Salmonella enterica serovar Enteritidis between pair-housed and group-housed laying hens.

    PubMed

    Thomas, Ekelijn; Bouma, Annemarie; Klinkenberg, Don

    2011-02-23

    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.

  1. Molecular epidemiological characteristics of Salmonella enterica serovars Enteritidis, Typhimurium and Livingstone strains isolated in a Tunisian university hospital.

    PubMed

    Ktari, Sonia; Ksibi, Boutheina; Gharsallah, Houda; Mnif, Basma; Maalej, Sonda; Rhimi, Fouzia; Hammami, Adnene

    2016-03-01

    Enteritidis, Typhimurium and Livingstone are the main Salmonella enterica serovars recovered in Tunisia. Here, we aimed to assess the genetic diversity of fifty-seven Salmonella enterica strains from different sampling periods, origins and settings using pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST) and multi-locus variable-number tandem repeat analysis (MLVA). Salmonella Enteritidis, isolated from human and food sources from two regions in Sfax in 2007, were grouped into one cluster using PFGE. However, using MLVA these strains were divided into two clusters. Salmonella Typhimurium strains, recovered in 2012 and represent sporadic cases of human clinical isolates, were included in one PFGE cluster. Nevertheless, the MLVA technique, divided Salmonella Typhimurium isolates into six clusters with diversity index reaching (DI = 0.757). For Salmonella Livingstone which was responsible of two nosocomial outbreaks during 2000-2003, the PFGE and MLVA methods showed that these strains were genetically closely related. Salmonella Enteritidis and Salmonella Livingstone populations showed a single ST lineage ST11 and ST543 respectively. For Salmonella Typhimurium, two MLST sequence types ST19 and ST328 were defined. Salmonella Enteritidis and Salmonella Typhimurium strains were clearly differentiated by MLVA which was not the case using PFGE. © 2015 APMIS. Published by John Wiley & Sons Ltd.

  2. Biofilm formation by Salmonella Enteritidis and Salmonella Typhimurium isolated from avian sources is partially related with their in vivo pathogenicity.

    PubMed

    Borges, Karen Apellanis; Furian, Thales Quedi; de Souza, Sara Neves; Menezes, Rafaela; de Lima, Diane Alves; Fortes, Flávia Bornancini Borges; Salle, Carlos Tadeu Pippi; Moraes, Hamilton Luiz Souza; Nascimento, Vladimir Pinheiro

    2018-03-22

    Salmonella Enteritidis and Salmonella Typhimurium are among the most prevalent serotypes isolated from salmonellosis outbreaks and poultry. Salmonella spp. have the capacity to form biofilms on several surfaces, which can favour survival in hostile environments, such as slaughterhouses. Salmonella strains present differences in pathogenicity. However, there is little information regarding the pathogenicity of S. Enteritidis and S. Typhimurium isolated from avian sources and their relationship to biofilm production. The aim of this study was to use a novel pathogenicity index and a biofilm production assay to evaluate their relationships within these serotypes. In addition, we detected the presence of the spiA and agfA genes in these strains. Biofilm formation was investigated at two temperatures (37 °C and 28 °C) using microtiter plate assay, and the results were compared with the individual pathogenicity index of each strain. PCR was used to detect spiA and agfA, virulence genes associated with biofilm production. S. Enteritidis and S. Typhimurium strains were capable of producing biofilm at 37 °C and 28 °C. Sixty-two percent and 59.5% of S. Enteritidis and 73.8% and 46.2% of S. Typhimurium produced biofilm at 37 °C and 28 °C, respectively. Biofilm production at 37 °C was significantly higher in both serotypes. Only S. Enteritidis was capable of adhering strongly at both temperatures. Biofilm production was related to pathogenicity index only at 28 °C for S. Enteritidis. spiA and agfA were found in almost all strains and were not statistically associated with biofilm production. Copyright © 2018 Elsevier Ltd. All rights reserved.

  3. Validation of cooking methods using shell eggs inoculated with Salmonella serotypes Enteritidis and Heidelberg.

    PubMed

    Davis, A L; Curtis, P A; Conner, D E; McKee, S R; Kerth, L K

    2008-08-01

    Salmonella enterica serotype Enteritidis has long been associated with eggs, and more recently, Salmonella enterica serotype Heidelberg has also become associated with eggs. This study was undertaken to determine whether Salmonella Enteritidis and Salmonella Heidelberg are effectively eliminated from eggs by various cooking methods. Seven cooking methods were chosen--hard and soft cooked, scrambled, over easy, sunny-side up, poached, and free poached--and a pan insert and the free-flowing method were used. Shell eggs, purchased from a grocery store, were inoculated with Salmonella and cooked. The cooked eggs were analyzed by USDA-approved methods for Salmonella recovery. Findings indicated that existing cooking methods for the hard-cooked, soft-cooked, and poaching methods were safe. However, the same was not true for the current sunny-side-up, over-easy, and scrambled egg cooking methods.

  4. Prevention of Salmonella enteritidis in shell eggs during production, storage, and transportation. Final rule.

    PubMed

    2009-07-09

    The Food and Drug Administration (FDA) is issuing a final rule that requires shell egg producers to implement measures to prevent Salmonella Enteritidis (SE) from contaminating eggs on the farm and from further growth during storage and transportation, and requires these producers to maintain records concerning their compliance with the rule and to register with FDA. FDA is taking this action because SE is among the leading bacterial causes of foodborne illness in the United States, and shell eggs are a primary source of human SE infections. The final rule will reduce SE-associated illnesses and deaths by reducing the risk that shell eggs are contaminated with SE.

  5. The relationship between the numbers of Salmonella Enteritidis, Salmonella Heidelberg, or Salmonella Hadar colonizing reproductive tissues of experimentally infected laying hens and deposition inside eggs.

    PubMed

    Gast, Richard K; Guraya, Rupa; Guard, Jean; Holt, Peter S

    2011-06-01

    Contamination of eggs by Salmonella Enteritidis has been a prominent cause of human illness for several decades and is the focus of a recently implemented national regulatory plan for egg-producing flocks in the United States. Salmonella Heidelberg has also been identified as an egg-transmitted pathogen. The deposition of Salmonella strains inside eggs is a consequence of reproductive tract colonization in infected laying hens, but prior research has not determined the relationship between the numbers of Salmonella that colonize reproductive organs and the associated frequency of egg contamination. In the present study, groups of laying hens in two trials were experimentally infected with large oral doses of strains of Salmonella Enteritidis (phage type 13a), Salmonella Heidelberg, or Salmonella Hadar. Reproductive tissues of selected hens were cultured to detect and enumerate Salmonella at 5 days postinoculation, and the interior contents of eggs laid between 6 and 25 days postinoculation were tested for contamination. Significantly more internally contaminated eggs were laid by hens infected with Salmonella Enteritidis (3.58%) than with strains of either Salmonella Heidelberg (0.47%) or Salmonella Hadar (0%). However, no significant differences were observed between Salmonella strains in either isolation frequency or the number of colony-forming units (CFU) isolated from ovaries or oviducts. Salmonella isolation frequencies ranged from 20.8% to 41.7% for ovaries and from 8.3% to 33.3% for oviducts. Mean Salmonella colonization levels ranged from 0.10 to 0.51 log CFU/g for ovaries and from 0.25 to 0.46 log CFU/g for oviducts. Although parallel rank-orders were observed for Salmonella enumeration (in both ovaries and oviducts) and egg contamination frequency, a statistically significant relationship could not be established between these two parameters of infection.

  6. Effective characterization of Salmonella Enteritidis by most probable number (MPN) followed by multiplex polymerase chain reaction (PCR) methods.

    PubMed

    Zappelini, Lincohn; Martone-Rocha, Solange; Dropa, Milena; Matté, Maria Helena; Tiba, Monique Ribeiro; Breternitz, Bruna Suellen; Razzolini, Maria Tereza Pepe

    2017-02-01

    Nontyphoidal Salmonella (NTS) is a relevant pathogen involved in gastroenteritis outbreaks worldwide. In this study, we determined the capacity to combine the most probable number (MPN) and multiplex polymerase chain reaction (PCR) methods to characterize the most important Salmonella serotypes in raw sewage. A total of 499 isolates were recovered from 27 raw sewage samples and screened using two previously described multiplex PCR methods. From those, 123 isolates were selected based on PCR banding pattern-identical or similar to Salmonella Enteritidis and Salmonella Typhimurium-and submitted to conventional serotyping. Results showed that both PCR assays correctly serotyped Salmonella Enteritidis, however, they presented ambiguous results for Salmonella Typhimurium identification. These data highlight that MPN and multiplex PCR can be useful methods to describe microbial quality in raw sewage and suggest two new PCR patterns for Salmonella Enteritidis identification.

  7. Modeling the effect of temperature on survival rate of Salmonella Enteritidis in yogurt.

    PubMed

    Szczawiński, J; Szczawińska, M E; Łobacz, A; Jackowska-Tracz, A

    2014-01-01

    The aim of the study was to determine the inactivation rates of Salmonella Enteritidis in commercially produced yogurt and to generate primary and secondary mathematical models to predict the behaviour of these bacteria during storage at different temperatures. The samples were inoculated with the mixture of three S. Enteritidis strains and stored at 5 degrees C, 10 degrees C, 15 degrees C, 20 degrees C and 25 degrees C for 24 h. The number of salmonellae was determined every two hours. It was found that the number of bacteria decreased linearly with storage time in all samples. Storage temperature and pH of yogurt significantly influenced survival rate of S. Enteritidis (p < 0.05). In samples kept at 5 degrees C the number of salmonellae decreased at the lowest rate, whereas at 25 degrees C the reduction in number of bacteria was the most dynamic. The natural logarithm of mean inactivation rates of Salmonella calculated from primary model was fitted to two secondary models: linear and polynomial. Equations obtained from both secondary models can be applied as a tool for prediction of inactivation rate of Salmonella in yogurt stored under temperature range from 5 to 25 degrees C; however, polynomial model gave the better fit to the experimental data.

  8. Reduction of Salmonella Enteritidis in the spleens of hens by bacterins that vary in fimbrial protein SefD.

    PubMed

    Sanchez-Ingunza, Roxana; Guard, Jean; Morales, Cesar A; Icard, Alan H

    2015-10-01

    The objective of this research was to determine whether variation in the presence of fimbrial protein SefD would impact efficacy of bacterins as measured by recovery of Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) from the spleens of hens. Two bacterins were prepared that varied in SefD content. Also, two adjuvants were tested, namely, water-in-oil and aluminum hydroxide gel (alum). Control groups for both adjuvant preparations included infected nonvaccinated hens and uninfected nonvaccinated hens. At 21 days postinfection, Salmonella Enteritidis was recovered from 69.7%, 53.1%, and 86.0% from the spleens of all hens vaccinated with bacterins lacking SefD, bacterins that included SefD, and infected nonvaccinated control hens, respectively. No Salmonella was recovered from uninfected nonvaccinates. Results from individual trials showed that both bacterins reduced positive spleens, but that the one with SefD was more efficacious. Alum adjuvant had fewer side effects on hens and egg production as compared to water-in-oil. However, adjuvant did not change the relative recovery of Salmonella Enteritidis from spleens. These results suggest that SefD is a promising target antigen for improving the efficacy of immunotherapy in hens, and is intended to reduce Salmonella Enteritidis in the food supply.

  9. Spread of a Major Clone of Salmonella enterica Serotype Enteritidis in Poultry and in Salmonellosis Outbreaks in Southern Brazil.

    PubMed

    Borges, Karen Apellanis; Furian, Thales Quedi; de Souza, Sara Neves; Tondo, Eduardo César; Streck, André Felipe; Salle, Carlos Tadeu Pippi; de Souza Moraes, Hamilton Luiz; do Nascimento, Vladimir Pinheiro

    2017-01-01

    Salmonella spp. are among the most important agents of foodborne diseases all over the world. Human Salmonella outbreaks are often associated with the consumption of poultry products (meat and eggs), and one of the most prevalent serotypes associated with these products is Salmonella Enteritidis. Brazil is one of the most important poultry exporters in the world. In southern Brazil, three closely related clones of Salmonella Enteritidis have been responsible for the majority of foodborne Salmonella outbreaks over the past decade. However, until now, there has been little information regarding the clonal relationship among the Brazilian Salmonella strains of avian origin and those involved in foodborne outbreaks. Therefore, the aim of the present study was to complete the molecular characterization of Salmonella Enteritidis strains isolated from poultry and food sources involved in Salmonella outbreaks. PCR ribotyping was performed to discriminate the strains into different ribotype profiles according to the banding pattern amplification. This technique was able to differentiate the Salmonella Enteritidis strains into two banding patterns: R2 and R4. R2 accounted for 98.7% of the strains. DNA sequencing of the 600-bp fragment, present in all ribotypes, was applied to confirm this result. The sequences generated showed high levels of similarity, ranging from 99.7 to 100%, and were grouped into a single cluster. These results suggest that there is a clonal relationship among the Salmonella Enteritidis strains responsible for several salmonellosis outbreaks and the strains collected from poultry sources.

  10. Immunogenicity of Salmonella enterica serovar Enteritidis virulence protein, InvH, and cross-reactivity of its antisera with Salmonella strains.

    PubMed

    Dehghani, Behzad; Rasooli, Iraj; Gargari, Seyed Latif Mousavi; Nadooshan, Mohammad Reza Jalali; Owlia, Parviz; Nazarian, Shahram

    2013-02-22

    Acellular vaccines containing bacterial immunodominant components such as surface proteins may be potent alternatives to live attenuated vaccines in order to reduce salmonellosis risk to human health. invH gene, an important part of needle complex in type three secretion system (TTSS) plays important role in efficient bacterial adherence and entry into epithelial cells. In this work we hypothesize that use of a 15 kDa recombinant InvH as Salmonella enterica serovar Enteritidis surface protein could provoke antibody production in mouse and would help us study feasibility of its potential for diagnosis and/or a recombinant vaccine. The purified InvH provoked significant rise of IgG in mice. Active protection induced by immunization with InvH against variable doses of S. enterica serovar Enteritidis, indicated that the immunized mice were completely protected against challenge with 10(4) LD(50). The immunoreaction of sera from immunized mice with other Salmonella strains or cross reaction with sera of Salmonella strains inoculated mice is indicative of possessing by Salmonella strains of the surface protein, InvH, that can be employed in both prophylactic and diagnostic measures against S. enterica. Bacteria free spleen and ileum of the immunized mice in this study indicate that the invH gene affects bacterial invasion. Efficacy of the virulence protein, InvH, in shuttling into host cells in injectisome of S. enterica serovar Enteritidis and inhibition of this phenomenon by active immunization was shown in this study. In conclusion immunization with InvH protein can develop protection against S. enterica serovar Enteritidis infections. InvH in Salmonella strains can be exploited in protective measures as well as a diagnostic tool in Salmonella infections. Copyright © 2012 Elsevier GmbH. All rights reserved.

  11. Multiplication in Egg Yolk and Survival in Egg Albumen of Genetically and Phenotypically Characterized Salmonella Enteritidis Strains.

    PubMed

    Gast, Richard K; Guard, Jean; Guraya, Rupa; Locatelli, Aude

    2018-06-01

    Prompt refrigeration of eggs to prevent the multiplication of Salmonella Enteritidis to high levels during storage is an important practice for reducing the risk of egg-transmitted human illness. The efficacy of egg refrigeration for achieving this goal depends on the interaction among the location of contamination, the ability of contaminant strains to survive or multiply, and the rate at which growth-restricting temperatures are attained. The present study assessed the significance of several characterized genetic and phenotypic properties for the capabilities of 10 Salmonella Enteritidis isolates to multiply rapidly in egg yolk and survive for several days in egg albumen during unrefrigerated (25°C) storage. The growth of small numbers of each Salmonella Enteritidis strain (approximately 10 1 CFU/mL) inoculated into egg yolk samples was determined after 6 and 24 h of incubation. The survival of larger numbers of Salmonella Enteritidis (approximately 10 5 CFU/mL) inoculated into albumen samples was determined at 24 and 96 h of incubation. In yolk, the inoculated Salmonella Enteritidis strains multiplied to mean levels of approximately 10 2.6 CFU/mL after 6 h of incubation and 10 8.3 CFU/mL after 24 h. In albumen, mean levels of approximately 10 4.6 CFU/mL Salmonella Enteritidis were maintained through 96 h. The concentrations of the various Salmonella strains after incubation in either yolk or albumen were distributed over relatively narrow ranges of values. Significant ( P < 0.01) differences observed among individual strains suggested that maintenance of the fimbrial gene sefD may have positive genetic selection value by improving fitness to grow inside egg yolk, whereas the antibiotic resistance gene bla TEM-1 tet(A) appeared to have negative genetic selection value by decreasing fitness to survive in egg albumen.

  12. Effect of in-feed supplementation of trans-cinnamaldehyde and caprylic acid on chicken cecal microbiome in response to Salmonella Enteritidis

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis (SE) is a major foodborne pathogen causing enteric illnesses in humans, with undercooked eggs and poultry meat as the primary sources of infection. Our previous research revealed that in-feed supplementation of two GRAS (generally recognized as safe)-status, natural compounds,...

  13. Migration of Salmonella serotypes Heidelberg and Enteritidis in previously frozen chicken breast meat.

    PubMed

    Tozzo, Kamila; Neto, Américo F G; Spercoski, Katherinne M; Ronnau, Milton; Soares, Vanessa M; Bersot, Luciano S

    2018-02-01

    Salmonella spp. have been shown to migrate to the internal regions of meat cuts. Storage conditions and the presence of proteolytic microbiota can influence this process. Our study assessed the impact of storage time, temperature, and the presence of proteolytic psychrotrophic bacteria on migration. Samples of previously frozen chicken breast with skin and bone were then sterilized using gamma ray irradiation and a cobalt-60 source (11 KGy) and them were inoculated with cultures of S. Enteritidis, S. Enteritidis and psychrotrophs, S. Heidelberg, or S. Heidelberg and psychrotrophs. Inoculated samples were stored for 6, 12, 24, 48, or 168 h at 2, 7, or -30 °C. After treatment, samples were divided into similar-sized segments and bacterial counts were determined in different regions (A - superface, B - intermediate region, and C - internal region). S. Heidelberg and S. Enteritidis both demonstrated successful internal migration for each time, temperature, and bacterial combination (p < 0.05). Our data revealed that Salmonella migration proceeded for 24 h, but slowed at 48 h (p < 0.05). S. Enteritidis with psychrotrophs showed a low amount of internal migration (p < 0.05). We therefore conclude that Salmonella spp. are able to migrate into the internal regions of meat cuts in a short period of time, even at low temperatures. The presence of proteolytic psychrotrophs inhibits the migration of S. Enteritidis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Ethanol adaptation induces direct protection and cross-protection against freezing stress in Salmonella enterica serovar Enteritidis.

    PubMed

    He, S; Zhou, X; Shi, C; Shi, X

    2016-03-01

    Salmonella enterica serovar Enteritidis (Salm. Enteritidis) encounters mild ethanol stress during its life cycle. However, adaptation to a stressful condition may affect bacterial resistance to subsequent stresses. Hence, this work was undertaken to investigate the influences of ethanol adaptation on stress tolerance of Salm. Enteritidis. Salmonella Enteritidis was subjected to different ethanol adaptation treatments (2·5-10% ethanol for 1 h). Cellular morphology and tolerance to subsequent environmental stresses (15% ethanol, -20°C, 4°C, 50°C and 10% NaCl) were evaluated. It was found that 10% was the maximum ethanol concentration that allowed growth of the target bacteria. Ethanol adaptation did not cause cell-surface damage in Salm. Enteritidis as revealed by membrane permeability measurements and electron micrograph analysis. Salmonella Enteritidis adapted with 2·5-10% ethanol displayed an enhanced resistance to a 15%-ethanol challenge compared with an unchallenged control. The maximum ethanol resistance was observed when ethanol concentration used for ethanol adaptation was increased to 5·0%. Additionally, pre-adaptation to 5·0% ethanol cross-protected Salm. Enteritidis against -20°C, but not against 4°C, 50°C or 10% NaCl. Ethanol adaptation provided Salm. Enteritidis direct protection from a high level ethanol challenge and cross-protection from freezing, but not other stresses tested (low temperature, high salinity or high temperature). The results are valuable in developing adequate and efficient control measures for Salm. Enteritidis in foods. © 2016 The Society for Applied Microbiology.

  15. Salmonella enteritidis and Arizona hinshawii isolated from wild sandhill cranes

    USGS Publications Warehouse

    Windingstad, R.M.; Trainer, D.O.; Duncan, R.M.

    1977-01-01

    Salmonella enteritidis serotype Rubislaw and Arizona hinshawii were isolated from cloacal swabs of "healthy" live-trapped sandhill cranes (Grus canadensis tabida) in Indiana and Wisconsin. These respective isolations were the first reported from wild sandhill cranes.

  16. Salmonella Enteritidis flagellar mutants have a colonization benefit in the chicken oviduct.

    PubMed

    Kilroy, Sofie; Raspoet, Ruth; Martel, An; Bosseler, Leslie; Appia-Ayme, Corinne; Thompson, Arthur; Haesebrouck, Freddy; Ducatelle, Richard; Van Immerseel, Filip

    2017-02-01

    Egg borne Salmonella Enteritidis is still a major cause of human food poisoning. Eggs can become internally contaminated following colonization of the hen's oviduct. In this paper we aimed to analyze the role of flagella of Salmonella Enteritidis in colonization of the hen's oviduct. Using a transposon library screen we showed that mutants lacking functional flagella are significantly more efficient in colonizing the hen's oviduct in vivo. A micro-array analysis proved that transcription of a number of flagellar genes is down-regulated inside chicken oviduct cells. Flagella contain flagellin, a pathogen associated molecular pattern known to bind to Toll-like receptor 5, activating a pro-inflammatory cascade. In vitro tests using primary oviduct cells showed that flagellin is not involved in invasion. Using a ligated loop model, a diminished inflammatory reaction was seen in the oviduct resulting from injection of an aflagellated mutant compared to the wild-type. It is hypothesized that Salmonella Enteritidis downregulates flagellar gene expression in the oviduct and consequently prevents a flagellin-induced inflammatory response, thereby increasing its oviduct colonization efficiency. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Genetic diversity of human isolates of Salmonella enterica serovar Enteritidis in Malaysia.

    PubMed

    Bakeri, S A; Yasin, R M; Koh, Y T; Puthucheary, S D; Thong, K L

    2003-01-01

    The study was undertaken to determine clonal relationship and genetic diversity of the human strains of Salmonella enterica serovar Enteritidis isolated from 1995 to 2002 from different parts of Malaysia. Antimicrobial susceptibility test, plasmid profiling and pulsed-field gel electrophoresis were applied to analyse 65 human isolates of S. Enteritidis obtained over an eight year period from different parts of Malaysia. Four nonhuman isolates were included for comparison. A total of 14 distinct XbaI-pulsed-field profiles (PFPs) were observed, although a single PFP X1 was predominant and this particular clone was found to be endemic in Malaysia. The incidence of drug resistant S. Enteritidis remained relatively low with only 37% of the strains analysed being resistant to one or more antimicrobial agents. All except one resistant strain carried at least one plasmid ranging in size from 3.7 to 62 MDa giving nine plasmid profiles. The three isolates from raw milk and one from well-water had similar PFPs to that of the human isolates. Salmonella Enteritidis strains were more diverse than was previously thought. Fourteen subtypes were noted although one predominant clone persisted in Malaysia. The combination of pulsed-field gel electrophoresis, plasmid profiling and antibiograms provided additional discrimination to the highly clonal strains of S. Enteritidis. This is the first report to assess the genotypes of the predominant clinical S. Enteritidis in different parts of the country. As S. Enteritidis is highly endemic in Malaysia, the data generated would be useful for tracing the source during outbreaks of gastroenteritis in the study area.

  18. Mathematical modeling and validation of growth of Salmonella Enteritidis and background microorganisms in potato salad – one-step kinetic analysis and model development

    USDA-ARS?s Scientific Manuscript database

    This study was conducted to examine the growth of Salmonella Enteritidis (SE) in potato salad caused by cross-contamination and temperature abuse, and develop mathematical models to predict its growth. The growth of SE was investigated under constant temperature conditions (8, 10, 15, 20, 25, 30, a...

  19. The SPR detection of Salmonella enteritidis in food using aptamers as recongnition elements

    NASA Astrophysics Data System (ADS)

    Di, W. T.; Du, X. W.; Pan, M. F.; Wang, J. P.

    2017-09-01

    In this experiment, a fast, accurate, non-destructive, unmarked and simple-operation detection method for Salmonella enteritidis in food was established by the BI-3000 plasma resonance biosensor (SPR). This article establishes a method of using nucleic acid aptamer as immune recognition element in SPR which can be employed to detect Salmonella enteritidis in food for the first time. The experimental conditions were screened and the experimental scheme was validated and applied. The best flow rate was 5μL/min, the best concentration of the aptamers was 180mM, and the best regenerating solution was the 20mM NaOH. This method had almost no cross-reactivity. Besides, we established a standard curve of Salmonella enteritidis and SPR signal, with the detection limit of 2 cfu/mL. Finally, we tested the samples of chicken, pork, shrimp and fish purchased from supermarkets. The method has the advantages of short time, low detection limit and easy operation, which can be used for a large number of food samples.

  20. Glycerol supplementation enhances the protective effect of dietary FloraMax-B11 against Salmonella Enteritidis colonization in neonate broiler chickens.

    PubMed

    Delgado, R; Latorre, J D; Vicuña, E; Hernandez-Velasco, X; Vicente, J L; Menconi, A; Kallapura, G; Layton, S; Hargis, B M; Téllez, G

    2014-09-01

    Two independent trials were conducted in the present study to evaluate the effect of 5% glycerol supplementation combined with dietary FloraMax-B11 (FM) against Salmonella Enteritidis colonization in neonate broiler chickens. In each trial, 60 chicks were randomly assigned into 4 groups. Group 1 received a control diet. Group 2 received a control diet supplemented with 5% glycerol. Group 3 received a control diet supplemented with FM, and group 4 received a control diet supplemented with 5% glycerol and FM. At placement, chickens were challenged with Salmonella Enteritidis at 10(4) cfu/bird. In each trial, 12 chicks were humanely killed 72 h postchallenge, respectively, for Salmonella Enteritidis colonization. Supplementation of 5% glycerol or FM by themselves, showed no significant effect on Salmonella Enteritidis recovery or incidence when compared with control nontreated chickens in both trials. However, no detectable Salmonella Enteritidis was observed in the chickens that received the supplementation of 5% glycerol combined with FM in both trials. Further studies are in progress in older birds to substantiate these findings. © 2014 Poultry Science Association Inc.

  1. Refined Live Attenuated Salmonella enterica Serovar Typhimurium and Enteritidis Vaccines Mediate Homologous and Heterologous Serogroup Protection in Mice

    PubMed Central

    Schmidlein, Patrick; Simon, Raphael; Pasetti, Marcela F.; Galen, James E.; Levine, Myron M.

    2015-01-01

    Invasive nontyphoidal Salmonella (NTS) infections constitute a major health problem among infants and toddlers in sub-Saharan Africa; these infections also occur in infants and the elderly in developed countries. We genetically engineered a Salmonella enterica serovar Typhimurium strain of multilocus sequence type 313, the predominant genotype circulating in sub-Saharan Africa. We evaluated the capacities of S. Typhimurium and Salmonella enterica serovar Enteritidis ΔguaBA ΔclpX live oral vaccines to protect mice against a highly lethal challenge dose of the homologous serovar and determined protection against other group B and D serovars circulating in sub-Saharan Africa. The vaccines S. Typhimurium CVD 1931 and S. Enteritidis CVD 1944 were immunogenic and protected BALB/c mice against 10,000 50% lethal doses (LD50) of S. Typhimurium or S. Enteritidis, respectively. S. Typhimurium CVD 1931 protected mice against the group B serovar Salmonella enterica serovar Stanleyville (91% vaccine efficacy), and S. Enteritidis CVD 1944 protected mice against the group D serovar Salmonella enterica serovar Dublin (85% vaccine efficacy). High rates of survival were observed when mice were infected 12 weeks postimmunization, indicating that the vaccines elicited long-lived protective immunity. Whereas CVD 1931 did not protect against S. Enteritidis R11, CVD 1944 did mediate protection against S. Typhimurium D65 (81% efficacy). These findings suggest that a bivalent (S. Typhimurium and S. Enteritidis) vaccine would provide broad protection against the majority of invasive NTS infections in sub-Saharan Africa. PMID:26351285

  2. Validation of Single and Pooled Manure Drag Swabs for the Detection of Salmonella Serovar Enteritidis in Commercial Poultry Houses.

    PubMed

    Kinde, Hailu; Goodluck, Helen A; Pitesky, Maurice; Friend, Tom D; Campbell, James A; Hill, Ashley E

    2015-12-01

    Single swabs (cultured individually) are currently used in the Food and Drug Administration (FDA) official method for sampling the environment of commercial laying hens for the detection of Salmonella enterica ssp. serovar Enteritidis (Salmonella Enteritidis). The FDA has also granted provisional acceptance of the National Poultry Improvement Plan's (NPIP) Salmonella isolation and identification methodology for samples taken from table-egg layer flock environments. The NPIP method, as with the FDA method, requires single-swab culturing for the environmental sampling of laying houses for Salmonella Enteritidis. The FDA culture protocol requires a multistep culture enrichment broth, and it is more labor intensive than the NPIP culture protocol, which requires a single enrichment broth. The main objective of this study was to compare the FDA single-swab culturing protocol with that of the NPIP culturing protocol but using a four-swab pool scheme. Single and multi-laboratory testing of replicate manure drag swab sets (n  =  525 and 672, respectively) collected from a Salmonella Enteritidis-free commercial poultry flock was performed by artificially contaminating swabs with either Salmonella Enteritidis phage type 4, 8, or 13a at one of two inoculation levels: low, x¯  = 2.5 CFU (range 2.5-2.7), or medium, x¯  = 10.0 CFU (range 7.5-12). For each replicate, a single swab (inoculated), sets of two swabs (one inoculated and one uninoculated), and sets of four swabs (one inoculated and three uninoculated), testing was conducted using the FDA or NPIP culture method. For swabs inoculated with phage type 8, the NPIP method was more efficient (P < 0.05) for all swab sets at both inoculation levels than the reference method. The single swabs in the NPIP method were significantly (P < 0.05) better than four-pool swabs in detecting Salmonella Enteritidis at the lower inoculation level. In the collaborative study (n  =  13 labs) using Salmonella Enteritidis phage

  3. Optical immunosensors for detection of Listeria monocytogenes and Salmonella enteritidis from food

    NASA Astrophysics Data System (ADS)

    Bhunia, Arun K.; Geng, Tao; Lathrop, Amanda; Valadez, Angela; Morgan, Mark T.

    2004-03-01

    Listeria monocytogenes and Salmonella are two major foodborne pathogens of significant concern. Two optical evanescent wave immunosensors were evaluated for detection: Antibody-coupled fiber-optic biosensor and a surface plasmon resonant (SPR) immunosensor. In the fiber-optic sensor, polyclonal antibodies for the test organisms were immobilized on polystyrene fiber wave -guides using streptavidin - biotin chemistry. Cyanine 5 -labeled monoclonal antibodies C11E9 (for L. monocytogenes) and SF-11 (for Salmonella Enteritidis) were used to generate a specific fluorescent signal. Signal acquisition was performed by launching a laser-light (635 nm) from an Analyte-2000. This immunosensor was able to detect 103 - 109 cfu/ml of L. monocytogenes or 106-109 cfu/ml of Salmonella Enteritidis and the assays were conducted at near real-time with results obtained within one hour of sampling. The assays were specific and showed signal even in the presence of other microorganisms such as E. coli, Enterococcus faecalis or Salmonella Typhimurium. In the SPR system, IAsys instrument (resonant mirror sensor) was used. Monoclonal antibody-C11E9 was directly immobilized onto a carboxylate cuvette. Whole Listeria cells at various concentrations did not yield any signal while surface protein extracts did. Crude protein extracts from L. monocytogenes and L. innocua had average binding responses of around 150 arc sec (0.25 ng/mm2), which was significantly different from L. grayi, L. ivanovii, or L. welshimeri with average responses of <48 arc sec. Both fiber-optic and SPR sensors show promise in near real-time detection of foodborne L. monocytogenes and Salmonella Enteritidis.

  4. Salmonella Enteritidis infections associated with a contaminated immersion blender at a cAMP.

    PubMed

    Daly, Elizabeth R; Smith, Colleen M; Wikoff, Peter; Seiferth, John; Finnigan, Jayne; Nadeau, Alisha M; Welch, Joyce J

    2010-09-01

    More than 100 foodborne salmonellosis outbreaks occur each year in the United States. Contaminated food preparation equipment is implicated in approximately 32% of Salmonella outbreaks with a known source. In April 2009, we investigated reported Salmonella infections at a camp in New Hampshire. Camp attendees were contacted to complete a standard questionnaire. The questionnaire asked about foods eaten while at the camp, symptoms of gastrointestinal illness, visits to healthcare providers, and specimen submission for pathogen testing. Laboratory and environmental investigations were conducted, including testing of foods and food preparation equipment. A total of 133 ill persons, including 47 laboratory-confirmed Salmonella Enteritidis infections, were identified during this investigation. A total of 142 (80%) of 178 camp attendees completed a standard questionnaire and 109 cases of gastrointestinal illness and 33 healthy individuals were identified. Statistical analysis of survey data indicated that people who ate pudding were 15 times more likely to become ill with salmonellosis than those who did not eat pudding (risk ratio, 15.2; 95% confidence interval, 2.3-102.3). Salmonella Enteritidis was identified in leftover pudding and in the internal mixing components of the blender used to mix the pudding. All patient, food, and blender isolates exhibited the same pulsed-field gel electrophoresis pattern. This outbreak of Salmonella Enteritidis was caused by a Salmonella-contaminated hand-held immersion blender used to prepare pudding at a camp. A malfunctioning blender shaft seal is suspected to have resulted in contamination of the blender and subsequently pudding prepared using the blender.

  5. Antimicrobial activity of basil (Ocimum basilicum) oil against Salmonella enteritidis in vitro and in food.

    PubMed

    Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat

    2010-01-01

    Nine essential oils were examined for antimicrobial activity against reference and clinical strains of Salmonella Enteritidis. Based on the size of the inhibition zone and the minimal inhibitory concentration, basil oil had the strongest antimicrobial activity against all the tested bacteria, and S. Enteritidis SE3 was the most sensitive strain to all the tested oils. Gas chromatography/mass spectrometry analysis revealed that the major constituents of the oil were linalool (64.35%), 1,8-cineole (12.28%), eugenol (3.21%), germacrene D (2.07%), alpha-terpineol (1.64%), and rho-cymene (1.03%). When applied in nham, a fermented pork sausage, experimentally inoculated with S. Enteritidis SE3 and stored at 4 degrees C, basil oil inhibited the bacterium in a dose-dependent fashion. Basil oil at a concentration of 50 ppm reduced the number of bacteria in the food from 5 to 2log cfu/g after storage for 3 d. An unmeasurable level of the bacterium in the food was observed at days 2 and 3 of storage when 100 and 150 ppm of basil oil was used, respectively. Sensory evaluation suggested that the addition of 100 but not of 150 ppm to nham would be acceptable to consumers. The results from this study confirm the potential use of basil oil as an antimicrobial agent to control S. Enteritidis in food.

  6. Effect of dietary bacteriophage supplementation in internal organs, fecal excretion and ileal immune response in laying hens challenged with salmonella enteritidis

    USDA-ARS?s Scientific Manuscript database

    A study was conducted to evaluate the role of bacteriophage (BP) against Salmonella enterica serovar Enteritidis (SE) internal organs colonization and ileum immune response in laying hens. Hens were challenged both orally and intracloacally with 108 cfu/mL cells of nalidixic acid resistant Salmonell...

  7. DNA aptamer-based colorimetric detection platform for Salmonella Enteritidis.

    PubMed

    Bayraç, Ceren; Eyidoğan, Füsun; Avni Öktem, Hüseyin

    2017-12-15

    Food safety is a major issue to protect public health and a key challenge is to find detection methods for identification of hazards in food. Food borne infections affects millions of people each year and among pathogens, Salmonella Enteritidis is most widely found bacteria causing food borne diseases. Therefore, simple, rapid, and specific detection methods are needed for food safety. In this study, we demonstrated the selection of DNA aptamers with high affinity and specificity against S. Enteritidis via Cell Systematic Evolution of Ligands by Exponential Enrichment (Cell-SELEX) and development of sandwich type aptamer-based colorimetric platforms for its detection. Two highly specific aptamers, crn-1 and crn-2, were developed through 12 rounds of selection with K d of 0.971µM and 0.309µM, respectively. Both aptamers were used to construct sandwich type capillary detection platforms. With the detection limit of 10 3 CFU/mL, crn-1 and crn-2 based platforms detected target bacteria specifically based on color change. This platform is also suitable for detection of S. Enteritidis in complex food matrix. Thus, this is the first to demonstrate use of Salmonella aptamers for development of the colorimetric aptamer-based detection platform in its identification and detection with naked eye in point-of-care. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. FUNCTIONS EXERTED BY THE VIRULENCE ASSOCIATED TYPE THREE SECRETION SYSTEMS DURING SALMONELLA ENTERICA SEROVAR ENTERITIDIS INFECTION OF CHICKEN OVIDUCT EPITHELIAL CELLS AND MACROPHAGES

    USDA-ARS?s Scientific Manuscript database

    Salmonella enterica serovar, Enteritidis (SE) infection of chicken is a major contributing factor to non-typhoidal salmonellosis. The roles of the type three secretion systems (T3SS-1 and T3SS-2) in the pathogenesis of SE infection of chickens are poorly understood. In this study, the functions exer...

  9. Refined live attenuated Salmonella enterica serovar Typhimurium and Enteritidis vaccines mediate homologous and heterologous serogroup protection in mice.

    PubMed

    Tennant, Sharon M; Schmidlein, Patrick; Simon, Raphael; Pasetti, Marcela F; Galen, James E; Levine, Myron M

    2015-12-01

    Invasive nontyphoidal Salmonella (NTS) infections constitute a major health problem among infants and toddlers in sub-Saharan Africa; these infections also occur in infants and the elderly in developed countries. We genetically engineered a Salmonella enterica serovar Typhimurium strain of multilocus sequence type 313, the predominant genotype circulating in sub-Saharan Africa. We evaluated the capacities of S. Typhimurium and Salmonella enterica serovar Enteritidis ΔguaBA ΔclpX live oral vaccines to protect mice against a highly lethal challenge dose of the homologous serovar and determined protection against other group B and D serovars circulating in sub-Saharan Africa. The vaccines S. Typhimurium CVD 1931 and S. Enteritidis CVD 1944 were immunogenic and protected BALB/c mice against 10,000 50% lethal doses (LD50) of S. Typhimurium or S. Enteritidis, respectively. S. Typhimurium CVD 1931 protected mice against the group B serovar Salmonella enterica serovar Stanleyville (91% vaccine efficacy), and S. Enteritidis CVD 1944 protected mice against the group D serovar Salmonella enterica serovar Dublin (85% vaccine efficacy). High rates of survival were observed when mice were infected 12 weeks postimmunization, indicating that the vaccines elicited long-lived protective immunity. Whereas CVD 1931 did not protect against S. Enteritidis R11, CVD 1944 did mediate protection against S. Typhimurium D65 (81% efficacy). These findings suggest that a bivalent (S. Typhimurium and S. Enteritidis) vaccine would provide broad protection against the majority of invasive NTS infections in sub-Saharan Africa. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  10. Reduction of Salmonella Enteritidis in the spleens of hens by bacterins that vary in fimbrial protein SefD

    USDA-ARS?s Scientific Manuscript database

    Gene sefD is part of operon sefABCD, and it is required for production of the SEF14 fimbria by Salmonella Enteritidis. We compared strains that varied in SefD content for their ability to reduce recovery of Salmonella Enteritidis from the spleens of hens infected by parenteral challenge. The two bac...

  11. Role of the sseK1 gene in the pathogenicity of Salmonella enterica serovar enteritidis in vitro and in vivo.

    PubMed

    Yang, Yadong; Yu, Chuan; Ding, Ke; Zhang, Chunjie; Liao, Chengshui; Jia, Yanyan; Li, Jing; Cheng, Xiangchao

    2018-04-01

    Salmonella enteritidis is a common food-borne pathogen associated with consumption of contaminated poultry meat and eggs, which frequently causes gastroenteritis in humans. Salmonella secreted effector K1 (SseK1), as a translocated and secreted protein has been identified to be essential for the virulence of Salmonella typhimurium in host cells. However, the role of the sseK1 gene in the pathogenicity of S. enteritidis remain unclear. In this study, a sseK1 deletion mutant of S. enteritidis was constructed and its biological characteristics were examined. It was found that the sseK1 deletion mutant did not affect the growth, adherence and invasion of Salmonella enteritidis when compared to the wild-type S. enteritidis. However, the mutant showed decreased formation of biofilm and significantly reduced intracellular survival of bacteria in activated mouse peritoneal macrophages, as well as showed reduced pathogenicity to a murine model by increasing the lethal dose 50% (LD 50 ) value and decreasing the proliferation ratio of bacteria in vivo. Taken together, this study determined an important role for SseK1 in the pathogenicity of S. enteritidis in vitro and in vivo. Copyright © 2018 Elsevier Ltd. All rights reserved.

  12. An evaluation of the effect of sodium bisulfate as a feed additive on Salmonella enterica serotype Enteritidis in experimentally infected broilers.

    PubMed

    Kassem, I I; Sanad, Y M; Stonerock, R; Rajashekara, G

    2012-04-01

    The colonization of broiler chickens with Salmonella can pose serious health and economic risks for both consumers and the poultry industry. Because colonization with Salmonella can lead to subsequent contamination of chicken carcasses during processing, preemptive control measures should include the reduction of this pathogen in chickens before slaughter. In this study, we evaluated the effect of sodium bisulfate, a potential antimicrobial feed additive, on Salmonella colonization of experimentally infected broiler chickens. Two hundred and forty 1-d-old chickens were infected orally with Salmonella enterica serotype Enteritidis and divided into 4 groups (each comprised of 60 chickens). Three groups received different concentrations of sodium bisulfate integrated into their feed, while the feed of the fourth group (positive control) was not treated. At time points before the broilers' slaughter age, different organs/tissues (liver, spleen, cecum, and bone marrow) and feces were aseptically collected and tested for the occurrence and density of Salmonella Enteritidis. Our results show that at 3 d postinfection, high colonization with Salmonella Enteritidis was detected and affected all tested tissues and fecal samples. Although colonization decreased across time, Salmonella Enteritidis persisted in the cecum, feces, spleen, and bone marrow, but not in the liver, until slaughter age. Furthermore, the addition of sodium bisulfate to the feed did not significantly reduce Salmonella Enteritidis numbers in infected chickens or affect the shedding of the pathogen.

  13. Colonisation of poultry by Salmonella Enteritidis S1400 is reduced by combined administration of Lactobacillus salivarius 59 and Enterococcus faecium PXN-33.

    PubMed

    Carter, Alun; Adams, Martin; La Ragione, Roberto M; Woodward, Martin J

    2017-02-01

    Salmonella Enteritidis remains a significant issue within the poultry industry and one potential solution is to use probiotic bacteria to prevent Salmonella colonisation through competitive exclusion (CE). We demonstrate that combined administration of Lactobacillus salivarius 59 and Enterococcus faecium PXN33 were effective competitive excluders of Salmonella Enteritidis S1400 in poultry. Two models were developed to evaluate the efficacy of probiotic where birds received Salmonella Enteritidis S1400 by a) oral gavage and b) sentinel bird to bird transmission. A statistically significant (p<0.001) 2 log reduction of Salmonella Enteritidis S1400 colonisation was observed in the ileum, caecum and colon at day 43 using combined administration of the two probiotic bacteria. However, no Salmonella Enteritidis S1400 colonisation reduction was observed when either probiotic was administered individually. In the sentinel bird model the combined probiotic administered at days 12 and 20 was more effective than one-off or double administrations at age 1 and 12days. In vitro cell free culture supernatant studies suggest the mechanism of Salmonella Enteritidis S1400 inhibition was due to a reduction in pH by the probiotic bacteria. Our current study provides further evidence that probiotics can significantly reduce pathogenic bacterial colonisation in poultry and that mixed preparation of probiotics provide superior performance when compared to individual bacterial preparations. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Cross-protection against Salmonella enteritidis infection in mice. III. Delayed hypersensitivity reaction and clearance of the challenge organism.

    PubMed

    Padmanaban, V D; Mittal, K R

    1979-01-01

    Mice were immunized with live vaccines and with live vaccines with complete adjuvant incorporating Salmonella enteritidis, Salmonella typhi-murium, Salmonella gallinarum or Salmonella pullorum. On the 21st day after vacination, the hypersensitivity reactions elicited by the mice to extracts of the challenge organism (S. enteritidis 5694 SMR) were assessed. The degree of delayed hypersensitivity reaction was compared with the level of protection induced by the vaccine. The role in protection of delayed hypersensitivity is discussed. Clearance of the challenge organism from the liver of previously vaccinated and unvaccinated mice was assessed quantitatively.

  15. Development of a Novel, Rapid Multiplex Polymerase Chain Reaction Assay for the Detection and Differentiation of Salmonella enterica Serovars Enteritidis and Typhimurium Using Ultra-Fast Convection Polymerase Chain Reaction.

    PubMed

    Kim, Tae-Hoon; Hwang, Hyun Jin; Kim, Jeong Hee

    2017-10-01

    Salmonella enterica serovars Enteritidis and Typhimurium are the most common causative agents of human nontyphoidal salmonellosis. The rapid detection and timely treatment of salmonellosis are important to increase the curative ratio and prevent spreading of the disease. In this study, we developed a rapid multiplex convection polymerase chain reaction (PCR) method to detect Salmonella spp. and differentiate Salmonella Enteritidis and Salmonella Typhimurium. We used the invA gene for Salmonella spp. detection. Salmonella Enteritidis-specific primers and Salmonella Typhimurium-specific primers were designed using the insertion element (IE) and spy genes, respectively. The primer set for Salmonella spp. detection clearly detected both Salmonella Enteritidis and Salmonella Typhimurium after a 21-min amplification reaction. Serovar-specific primer sets for Salmonella Enteritidis and Salmonella Typhimurium specifically detected each target species in a 21-min amplification reaction. We were able to detect Salmonella spp. at a single copy level in the singleplex mode. The limits of detection for Salmonella Enteritidis and Salmonella Typhimurium were 30 copies in both the singleplex and multiplex modes. The PCR run time could be reduced to 10.5 min/15 cycles. The multiplex convection PCR method developed in this study could detect the Salmonella spp. Salmonella Enteritidis and Salmonella Typhimurium in artificially contaminated milk with as few as 10 0 colony-forming unit/mL after 4-h enrichment. The PCR assay developed in this study provides a rapid, specific, and sensitive method for the detection of Salmonella spp. and the differentiation of Salmonella Enteritidis and Salmonella Typhimurium.

  16. Activation of chicken bone marrow-derived dendritic cells induced by a Salmonella Enteritidis ghost vaccine candidate.

    PubMed

    Kamble, N M; Jawale, C V; Lee, J H

    2016-10-01

    Bacterial Ghost-based vaccine development has been applied to a variety of gram-negative bacteria. Developed Salmonella Enteritidis (S. Enteritidis) ghost are promising vaccine candidates because of their immunogenic and enhanced biosafety potential. In this study, we aimed to evaluate the immunostimulatory effect of a S. Enteritidis ghost vaccine on the maturation of chicken bone marrow-derived dendritic cells (chBM-DCs) in vitro The immature chBM-DCs were stimulated with S. Enteritidis ghost vaccine candidate. The vaccine efficiently stimulated maturation events in chBM-DCs, indicated by up-regulated expression of CD40, CD80, and MHC-II molecules. Immature BM-DCs responded to stimulation with S. Enteritidis ghost by increased expression of IL-6 and IL-12p40 cytokines. Also, S. Enteritidis ghost stimulated chBM-DCs induced the significant expression of IFN-γ and IL-2 in co-cultured autologous CD4+ T cells. In conclusion, our data suggest that S. Enteritidis ghost vaccine candidate is capable of activating and interacting with chBM-DCs. The results from current study may help for rational designing of Salmonella ghost based heterologous antigen delivery platforms to dendritic cells. © 2016 Poultry Science Association Inc.

  17. Method for the detection of Salmonella enterica serovar Enteritidis

    DOEpatents

    Agron, Peter G.; Andersen, Gary L.; Walker, Richard L.

    2008-10-28

    Described herein is the identification of a novel Salmonella enterica serovar Enteritidis locus that serves as a marker for DNA-based identification of this bacterium. In addition, three primer pairs derived from this locus that may be used in a nucleotide detection method to detect the presence of the bacterium are also disclosed herein.

  18. Distinct Salmonella Enteritidis lineages associated with enterocolitis in high-income settings and invasive disease in low-income settings

    PubMed Central

    Feasey, Nicholas A.; Hadfield, James; Keddy, Karen H.; Dallman, Timothy J; Jacobs, Jan; Deng, Xiangyu; Wigley, Paul; Barquist, Lars; Langridge, Gemma C.; Feltwell, Theresa; Harris, Simon R.; Mather, Alison E.; Fookes, Maria; Aslett, Martin; Msefula, Chisomo; Kariuki, Samuel; Maclennan, Calman A.; Onsare, Robert S.; Weill, François-Xavier; Le Hello, Simon; Smith, Anthony M.; McClelland, Michael; Desai, Prerak; Parry, Christopher M.; Cheesbrough, John; French, Neil; Campos, Josefina; Chabalgoity, Jose A.; Betancor, Laura; Hopkins, Katie L.; Nair, Satheesh; Humphrey, Tom J.; Lunguya, Octavie; Cogan, Tristan A.; Tapia, Milagritos D.; Sow, Samba O.; Tennant, Sharon M.; Bornstein, Kristin; Levine, Myron M.; Lacharme-Lora, Lizeth; Everett, Dean B.; Kingsley, Robert A.; Parkhill, Julian; Heyderman, Robert S.; Dougan, Gordon

    2016-01-01

    An epidemiological paradox surrounds Salmonella enterica serovar Enteritidis. In high-income settings, it has been responsible for an epidemic of poultry-associated, self-limiting enterocolitis, whilst in sub-Saharan Africa it is a major cause of invasive nontyphoidal Salmonella disease, associated with high case-fatality. Whole-genome sequence analysis of 675 isolates of S. Enteritidis from 45 countries reveals the existence of a global epidemic clade and two novel clades of S. Enteritidis that are each geographically restricted to distinct regions of Africa. The African isolates display genomic degradation, a novel prophage repertoire and have an expanded, multidrug resistance plasmid. S. Enteritidis is a further example of a Salmonella serotype that displays niche plasticity, with distinct clades that enable it to become a prominent cause of gastroenteritis in association with the industrial production of eggs, and of multidrug resistant, bloodstream invasive infection in Africa. PMID:27548315

  19. Distinct Salmonella Enteritidis lineages associated with enterocolitis in high-income settings and invasive disease in low-income settings.

    PubMed

    Feasey, Nicholas A; Hadfield, James; Keddy, Karen H; Dallman, Timothy J; Jacobs, Jan; Deng, Xiangyu; Wigley, Paul; Barquist, Lars; Langridge, Gemma C; Feltwell, Theresa; Harris, Simon R; Mather, Alison E; Fookes, Maria; Aslett, Martin; Msefula, Chisomo; Kariuki, Samuel; Maclennan, Calman A; Onsare, Robert S; Weill, François-Xavier; Le Hello, Simon; Smith, Anthony M; McClelland, Michael; Desai, Prerak; Parry, Christopher M; Cheesbrough, John; French, Neil; Campos, Josefina; Chabalgoity, Jose A; Betancor, Laura; Hopkins, Katie L; Nair, Satheesh; Humphrey, Tom J; Lunguya, Octavie; Cogan, Tristan A; Tapia, Milagritos D; Sow, Samba O; Tennant, Sharon M; Bornstein, Kristin; Levine, Myron M; Lacharme-Lora, Lizeth; Everett, Dean B; Kingsley, Robert A; Parkhill, Julian; Heyderman, Robert S; Dougan, Gordon; Gordon, Melita A; Thomson, Nicholas R

    2016-10-01

    An epidemiological paradox surrounds Salmonella enterica serovar Enteritidis. In high-income settings, it has been responsible for an epidemic of poultry-associated, self-limiting enterocolitis, whereas in sub-Saharan Africa it is a major cause of invasive nontyphoidal Salmonella disease, associated with high case fatality. By whole-genome sequence analysis of 675 isolates of S. Enteritidis from 45 countries, we show the existence of a global epidemic clade and two new clades of S. Enteritidis that are geographically restricted to distinct regions of Africa. The African isolates display genomic degradation, a novel prophage repertoire, and an expanded multidrug resistance plasmid. S. Enteritidis is a further example of a Salmonella serotype that displays niche plasticity, with distinct clades that enable it to become a prominent cause of gastroenteritis in association with the industrial production of eggs and of multidrug-resistant, bloodstream-invasive infection in Africa.

  20. Persistence of fecal shedding of Salmonella Enteritidis by experimentally infected laying hens housed in conventional or enriched cages.

    PubMed

    Gast, Richard K; Guraya, Rupa; Jones, Deana R; Anderson, Kenneth E

    2015-07-01

    Salmonella Enteritidis can be deposited inside eggs laid by infected hens, so the prevalence of this pathogen in commercial egg-producing flocks is an important risk factor for human illness. Opportunities for the introduction, transmission, and persistence of salmonellae in poultry are potentially influenced by flock housing and management systems. Animal welfare concerns have spurred the development of alternatives to traditional cage-based housing. However, the consequences of poultry housing systems for food safety have not been fully resolved by prior research. The present study assessed the effects of two different housing systems (conventional cages and colony cages enriched with perching and nesting areas) on the persistence of fecal shedding of Salmonella Enteritidis by groups of experimentally infected laying hens. In each of two trials, 136 hens were distributed among cages of both housing systems and orally inoculated with doses of 10(8) cfu of Salmonella Enteritidis (phage type 13a in one trial and phage type 4 in the other). At weekly intervals, samples of voided feces were collected from beneath each cage and cultured to detect Salmonella Enteritidis. Fecal shedding of Salmonella Enteritidis was detected for up to 8 wk post-inoculation by hens housed in enriched colony cages and 10 wk by hens housed in conventional cages. For both trials combined, the frequency of positive fecal cultures was significantly (P < 0.05) greater for conventional cages than for enriched colony cages at 1 wk (84.7 vs. 71.5%), 2 wk (54.2 vs. 31.3%), 3 wk (21.5 vs. 7.6%), and 4 wk (9.7 vs. 2.8%) post-inoculation. These results demonstrate that the susceptibility of hens to intestinal colonization by Salmonella Enteritidis can differ between conventional and enriched cage-based production systems, although this effect does not necessarily translate into a corresponding difference in the longer-term persistence of fecal shedding. © 2015 Poultry Science Association Inc.

  1. Cross-Contamination and Biofilm Formation by Salmonella enterica Serovar Enteritidis on Various Cutting Boards.

    PubMed

    Dantas, Stéfani T A; Rossi, Bruna F; Bonsaglia, Erika C R; Castilho, Ivana G; Hernandes, Rodrigo T; Fernandes, Ary; Rall, Vera L M

    2018-02-01

    Cross-contamination is one of the main factors related to foodborne outbreaks. This study aimed to analyze the cross-contamination process of Salmonella enterica serovar Enteritidis from poultry to cucumbers, on various cutting board surfaces (plastic, wood, and glass) before and after washing and in the presence and absence of biofilm. Thus, 10 strains of Salmonella Enteritidis were used to test cross-contamination from poultry to the cutting boards and from thereon to cucumbers. Moreover, these strains were evaluated as to their capacity to form biofilm on hydrophobic (wood and plastic) and hydrophilic materials (glass). We recovered the 10 isolates from all unwashed boards and from all cucumbers that had contacted them. After washing, the recovery ranged from 10% to 100%, depending on the board material. In the presence of biofilm, the recovery of salmonellae was 100%, even after washing. Biofilm formation occurred more on wood (60%) and plastic (40%) than glass (10%) boards, demonstrating that bacteria adhered more to a hydrophobic material. It was concluded that the cutting boards represent a critical point in cross-contamination, particularly in the presence of biofilm. Salmonella Enteritidis was able to form a biofilm on these three types of cutting boards but glass showed the least formation.

  2. Genomic comparison of the closely-related Salmonella enterica serovars enteritidis, dublin and gallinarum

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Matthews, T. David; Schmieder, Robert; Silva, Genivaldo G. Z.

    The Salmonella enterica serovars Enteritidis, Dublin, and Gallinarum are closely related but differ in virulence and host range. To identify the genetic elements responsible for these differences and to better understand how these serovars are evolving, we sequenced the genomes of Enteritidis strain LK5 and Dublin strain SARB12 and compared these genomes to the publicly available Enteritidis P125109, Dublin CT 02021853 and Dublin SD3246 genome sequences. We also compared the publicly available Gallinarum genome sequences from biotype Gallinarum 287/91 and Pullorum RKS5078. Using bioinformatic approaches, we identified single nucleotide polymorphisms, insertions, deletions, and differences in prophage and pseudogene content betweenmore » strains belonging to the same serovar. Through our analysis we also identified several prophage cargo genes and pseudogenes that affect virulence and may contribute to a host-specific, systemic lifestyle. These results strongly argue that the Enteritidis, Dublin and Gallinarum serovars of Salmonella enterica evolve by acquiring new genes through horizontal gene transfer, followed by the formation of pseudogenes. As a result, the loss of genes necessary for a gastrointestinal lifestyle ultimately leads to a systemic lifestyle and niche exclusion in the host-specific serovars.« less

  3. Genomic comparison of the closely-related Salmonella enterica serovars enteritidis, dublin and gallinarum

    DOE PAGES

    Matthews, T. David; Schmieder, Robert; Silva, Genivaldo G. Z.; ...

    2015-06-03

    The Salmonella enterica serovars Enteritidis, Dublin, and Gallinarum are closely related but differ in virulence and host range. To identify the genetic elements responsible for these differences and to better understand how these serovars are evolving, we sequenced the genomes of Enteritidis strain LK5 and Dublin strain SARB12 and compared these genomes to the publicly available Enteritidis P125109, Dublin CT 02021853 and Dublin SD3246 genome sequences. We also compared the publicly available Gallinarum genome sequences from biotype Gallinarum 287/91 and Pullorum RKS5078. Using bioinformatic approaches, we identified single nucleotide polymorphisms, insertions, deletions, and differences in prophage and pseudogene content betweenmore » strains belonging to the same serovar. Through our analysis we also identified several prophage cargo genes and pseudogenes that affect virulence and may contribute to a host-specific, systemic lifestyle. These results strongly argue that the Enteritidis, Dublin and Gallinarum serovars of Salmonella enterica evolve by acquiring new genes through horizontal gene transfer, followed by the formation of pseudogenes. As a result, the loss of genes necessary for a gastrointestinal lifestyle ultimately leads to a systemic lifestyle and niche exclusion in the host-specific serovars.« less

  4. Genomic Comparison of the Closely-Related Salmonella enterica Serovars Enteritidis, Dublin and Gallinarum

    PubMed Central

    Matthews, T. David; Schmieder, Robert; Silva, Genivaldo G. Z.; Busch, Julia; Cassman, Noriko; Dutilh, Bas E.; Green, Dawn; Matlock, Brian; Heffernan, Brian; Olsen, Gary J.; Farris Hanna, Leigh; Schifferli, Dieter M.; Maloy, Stanley; Dinsdale, Elizabeth A.; Edwards, Robert A.

    2015-01-01

    The Salmonella enterica serovars Enteritidis, Dublin, and Gallinarum are closely related but differ in virulence and host range. To identify the genetic elements responsible for these differences and to better understand how these serovars are evolving, we sequenced the genomes of Enteritidis strain LK5 and Dublin strain SARB12 and compared these genomes to the publicly available Enteritidis P125109, Dublin CT 02021853 and Dublin SD3246 genome sequences. We also compared the publicly available Gallinarum genome sequences from biotype Gallinarum 287/91 and Pullorum RKS5078. Using bioinformatic approaches, we identified single nucleotide polymorphisms, insertions, deletions, and differences in prophage and pseudogene content between strains belonging to the same serovar. Through our analysis we also identified several prophage cargo genes and pseudogenes that affect virulence and may contribute to a host-specific, systemic lifestyle. These results strongly argue that the Enteritidis, Dublin and Gallinarum serovars of Salmonella enterica evolve by acquiring new genes through horizontal gene transfer, followed by the formation of pseudogenes. The loss of genes necessary for a gastrointestinal lifestyle ultimately leads to a systemic lifestyle and niche exclusion in the host-specific serovars. PMID:26039056

  5. Experimental infection of Salmonella Enteritidis by the poultry red mite, Dermanyssus gallinae.

    PubMed

    Valiente Moro, C; Chauve, C; Zenner, L

    2007-05-31

    Dermanyssus gallinae is an important ectoparasite of laying hens in Europe and it is suspected of being a vector of pathogens. We carried out an in vitro study to evaluate the role of D. gallinae as a vector of Salmonella enterica subsp. enterica serotype Enteritidis. Two means of infecting the mite were tested: through the blood meal and after cuticular contact. Mites became carriers of Salmonella immediately after the infection with 29% and 53%, respectively, for oral route and cuticular contact. This percentage increased over time until it reached 95% (D7) and 80% (D14). The numerical identification of bacteria on the selective medium SM ID demonstrated the multiplication of Salmonella inside previously infected mites. In addition, transovarial passage as well as transstadial passage (from N1 to N2 stages) were demonstrated. Moreover, the observation of a negative effect of Salmonella on Dermanyssus oviposition was also observed. Finally, previously infected mites were able to contaminate the blood during the blood meal. Therefore, it appears that D. gallinae may act as a biological vector of S. Enteritidis under experimental conditions. It may represent a suitable environment for the development of Salmonella and could be an additional factor for the persistence of salmonellosis infection between successive flocks.

  6. Comparison of Salmonella enteritidis phage types isolated from layers and humans in Belgium in 2005.

    PubMed

    Welby, Sarah; Imberechts, Hein; Riocreux, Flavien; Bertrand, Sophie; Dierick, Katelijne; Wildemauwe, Christa; Hooyberghs, Jozef; Van der Stede, Yves

    2011-08-01

    The aim of this study was to investigate the available results for Belgium of the European Union coordinated monitoring program (2004/665 EC) on Salmonella in layers in 2005, as well as the results of the monthly outbreak reports of Salmonella Enteritidis in humans in 2005 to identify a possible statistical significant trend in both populations. Separate descriptive statistics and univariate analysis were carried out and the parametric and/or non-parametric hypothesis tests were conducted. A time cluster analysis was performed for all Salmonella Enteritidis phage types (PTs) isolated. The proportions of each Salmonella Enteritidis PT in layers and in humans were compared and the monthly distribution of the most common PT, isolated in both populations, was evaluated. The time cluster analysis revealed significant clusters during the months May and June for layers and May, July, August, and September for humans. PT21, the most frequently isolated PT in both populations in 2005, seemed to be responsible of these significant clusters. PT4 was the second most frequently isolated PT. No significant difference was found for the monthly trend evolution of both PT in both populations based on parametric and non-parametric methods. A similar monthly trend of PT distribution in humans and layers during the year 2005 was observed. The time cluster analysis and the statistical significance testing confirmed these results. Moreover, the time cluster analysis showed significant clusters during the summer time and slightly delayed in time (humans after layers). These results suggest a common link between the prevalence of Salmonella Enteritidis in layers and the occurrence of the pathogen in humans. Phage typing was confirmed to be a useful tool for identifying temporal trends.

  7. Modeling the survival of Salmonella Enteritidis and Salmonella Typhimurium during the fermentation of yogurt.

    PubMed

    Savran, Derya; Pérez-Rodríguez, Fernando; Halkman, A Kadir

    2018-03-01

    The objective of this study was to evaluate the behavior of Salmonella Enteritidis and Salmonella Typhimurium, the two most important serovars of salmonellosis , during the fermentation of yogurt. The microorganisms were enumerated in milk throughout the fermentation process at three initial inoculum levels (3, 5 and 7 log CFU/mL). DMFit software was used in the fitting procedure of the data (IFR, Norwich, UK, Version 3.5). The data provided sigmoidal curves that were successfully displayed with the Baranyi model. The results showed that the initial inoculum level did not affect the growth for both pathogens; thus, the µ max values (maximum specific growth rate) did not significantly differ across all the contamination levels, ranging from 0.26 to 0.38 for S. Enteritidis and from 0.50 to 0.56 log CFU/g/h for S. Typhimurium ( P > 0.05). However, the µ max values significantly differed between the two serovars ( P < 0.05). The λ values (lag time) did not have a clear trend in either of the pathogens. The present study showed that Salmonella can survive the fermentation process of milk even at a low contamination level. In addition, the models presented in this study can be used in quantitative risk assessment studies to estimate the threat to consumers.

  8. Transposon mutagenesis of Salmonella Enteritidis identifies genes that contribute to invasiveness in human and chicken cells and survival in egg albumen.

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis is the world’s leading cause of food borne salmonellosis and illness in people is linked strongly to its contamination of eggs produced by otherwise healthy appearing hens. Salmonella Enteritidis is noted for generating exceptional strain heterogeneity despite having a clonal ...

  9. Increased lymphocyte subpopulations and macrophages in the ovaries and oviducts of laying hens infected with Salmonella enterica serovar Enteritidis.

    PubMed

    Withanage, G S K; Sasai, K; Fukata, T; Miyamoto, T; Lillehoj, H S; Baba, E

    2003-12-01

    Salmonella enterica serovar Enteritidis (SE) is a causative agent for human food poisoning cases throughout the world. The ovaries and the oviducts of the laying hens are the major sites of SE colonization from which vertical transmission to eggs occurs. In this study, Salmonella-induced changes in T lymphocytes, B lymphocytes and macrophages in the ovaries and oviducts were assessed after primary and secondary experimental inoculations of laying hen with SE. Statistically significant increases in the numbers of T cells (both CD4+ and CD8+) and macrophages were observed 7 to 14 days after primary inoculation, followed by a peak in B-cell numbers from the 14th day post-primary inoculation onwards in the secretory areas of the oviducts. The peak in lymphocyte numbers immediately preceded a decline in the rate of SE recovery from the reproductive tract beginning at day 14. The correlation of decreased Salmonella recovery with elevated lymphocyte and macrophage numbers strongly suggests that local cell-mediated immunity is involved in controlling SE injection in the ovaries and oviducts.

  10. High Incubation Temperature and Threonine Dietary Level Improve Ileum Response Against Post-Hatch Salmonella Enteritidis Inoculation in Broiler Chicks

    PubMed Central

    de Oliveira, Heraldo Bezerra; Campos, Danila Barreiro; Guerra, Ricardo Romão; Costa, Fernando Guilherme Perazzo

    2015-01-01

    This study assessed the effect of both embryonic thermal manipulation and dietary threonine level on the response of broilers inoculated with Salmonella Enteritidis, considering bacterial counts in the cecal contents, intestinal morphology, mucin and heat shock protein 70 gene expression, body weight and weight gain. Thermal manipulation was used from 11 days of incubation until hatch, defining three treatments: standard (37.7°C), continuous high temperature (38.7°C) and continuous low temperature (36.7°C). After hatch, chicks were distributed according to a 3x2+1 factorial arrangement (three temperatures and two threonine levels and one sham-inoculated control). At two days of age, all chicks were inoculated with Salmonella Enteritidis, except for the sham-inoculated control group. There was no interaction between the factors on any analyses. High temperature during incubation was able to reduce colonization by Salmonella Enteritidis in the first days, reducing both Salmonella counts and the number of positive birds. It also increased mucin expression and decreased Hsp70 expression compared with other inoculated groups. High temperature during incubation and high threonine level act independently to reduce the negative effects associated to Salmonella Enteritidis infection on intestinal morphology and performance, with results similar to sham-inoculated birds. The findings open new perspectives for practical strategies towards the pre-harvest Salmonella control in the poultry industry. PMID:26131553

  11. Colonization of reproductive organs and internal contamination of eggs after experimental infection of laying hens with Salmonella heidelberg and Salmonella enteritidis.

    PubMed

    Gast, Richard K; Guard-Bouldin, Jean; Holt, Peter S

    2004-12-01

    Internal contamination of eggs laid by hens infected with Salmonella enteritidis has been a prominent international public health issue since the mid-1980s. Considerable resources have been committed to detecting and controlling S. enteritidis infections in commercial laying flocks. Recently, the Centers for Disease Control and Prevention also reported a significant association between eggs or egg-containing foods and S. heidelberg infections in humans. The present study sought to determine whether several S. heidelberg isolates obtained from egg-associated human disease outbreaks were able to colonize reproductive tissues and be deposited inside eggs laid by experimentally infected hens in a manner similar to the previously documented behavior of S. enteritidis. In two trials, groups of laying hens were orally inoculated with large doses of four S. heidelberg strains and an S. enteritidis strain that consistently caused egg contamination in previous studies. All five Salmonella strains (of both serotypes) colonized the intestinal tracts and invaded the livers, spleens, ovaries, and oviducts of inoculated hens, with no significant differences observed between the strains for any of these parameters. All four S. heidelberg strains were recovered from the interior liquid contents of eggs laid by infected hens, although at lower frequencies (between 1.1% and 4.5%) than the S. enteritidis strain (7.0%).

  12. Comparison of 7 culture methods for Salmonella serovar Enteritidis and Salmonella serovar Typhimurium isolation in poultry feces.

    PubMed

    Rodríguez, Francisco I; Procura, Francisco; Bueno, Dante J

    2018-06-26

    The present work compared 7 different culture methods and 3 selective-differential plating media for Salmonella ser. Enteritidis (SE) and S. ser. Typhimurium (ST) isolation using artificially contaminated poultry feces. The sensitivity (Se) and accuracy (AC) values increased when Modified Semisolid Rappaport Vassiliadis (MSRV) methods were used in place of the Tetrathionate (TT) or Tetrathionate Hajna broth (TTH) method in the enrichment step. However, there was no significant difference between the pre-enrichment incubation at 4 to 6 and 18 to 24 h for MSRV5 and MSRV24 methods, respectively. All Salmonella strains were recovered in the lowest dilutions tested for MSRV24 and 3 out of 4 for MSRV5 methods (2 to 10 cfu/25 g). The TT and TTH methods showed a detection limit between 2.2 × 101 and 1.0 × 106 cfu/25 g of fecal sample. The agreement was variable between the methods. However, there was a very good agreement between the MSRV5 and MSRV24 methods, and between tetrathionate direct (TTD, no pre-enrichment media used) and buffered peptone water 18 to 24 h Tetrathionate broth combination (TT24 method) for Salmonella strains. The 3 selective-differential plating media showed an agreement between fair and excellent. They performed a high Se and AC in the MSRV methods for Salmonella strains. There was a significant difference between center and periphery for MSRV methods, and there was a fair agreement between them for all strains. The MSRV methods are better than TT/TTH methods for the isolation of different strains of SE and ST in poultry fecal samples. The MSRV5 method can be used to reduce the time for the detection of SE and ST in these samples. Furthermore, a loopful of the periphery of the growth should be streaked onto differential-selective plating media, even in the absence of halo, to decrease the number of false negative results.

  13. Multistate outbreak of Salmonella enterica serotype enteritidis infection associated with pet guinea pigs.

    PubMed

    Bartholomew, Michael L; Heffernan, Richard T; Wright, Jennifer G; Klos, Rachel F; Monson, Timothy; Khan, Sofiya; Trees, Eija; Sabol, Ashley; Willems, Robert A; Flynn, Raymond; Deasy, Marshall P; Jones, Benjamen; Davis, Jeffrey P

    2014-06-01

    Salmonella causes about one million illnesses annually in the United States. Although most infections result from foodborne exposures, animal contact is an important mode of transmission. We investigated a case of Salmonella enterica serotype Enteritidis (SE) sternal osteomyelitis in a previously healthy child who cared for two recently deceased guinea pigs (GPs). A case was defined as SE pulsed-field gel electrophoresis (PFGE) XbaI pattern JEGX01.0021, BlnI pattern JEGA26.0002 (outbreak strain) infection occurring during 2010 in a patient who reported GP exposure. To locate outbreak strain isolates, PulseNet and the US Department of Agriculture National Veterinary Service Laboratories (NVSL) databases were queried. Outbreak strain isolates underwent multilocus variable-number tandem repeat analysis (MLVA). Traceback and environmental investigations were conducted at homes, stores, and breeder or broker facilities. We detected 10 cases among residents of eight states and four NVSL GP outbreak strain isolates. One patient was hospitalized; none died. The median patient age was 9.5 (range, 1-61) years. Among 10 patients, two purchased GPs at independent stores, and three purchased GPs at different national retail chain (chain A) store locations; three were chain A employees and two reported GP exposures of unknown characterization. MLVA revealed four related patterns. Tracebacks identified four distributors and 92 sources supplying GPs to chain A, including one breeder potentially supplying GPs to all case-associated chain A stores. All environmental samples were Salmonella culture-negative. A definitive SE-contaminated environmental source was not identified. Because GPs can harbor Salmonella, consumers and pet industry personnel should be educated regarding risks.

  14. Guar meal diets as an alternative approach to inducing molt and improving Salmonella enteritidis resistance in late-phase laying hens.

    PubMed

    Gutierrez, O; Zhang, C; Caldwell, D J; Carey, J B; Cartwright, A L; Bailey, C A

    2008-03-01

    Induced molting of laying hens is a practice used by commercial egg producers to increase the productive lifetime of their flock. However, the conventional method of inducing molt, which involves removal of feed, water, or both as well as a reduction in photoperiod to less than a natural day has drawn criticism due to animal welfare and food safety concerns. The objective of this study was to explore the efficacy of diets containing high levels of guar meal (GM) in inducing molt and reducing susceptibility to Salmonella Enteritidis colonization in late-phase laying hens. Late-phase (68 wk old) Lohmann laying hens were either full-fed standard laying hen diets (nonmolted control), induced to molt by feed withdrawal, or full-fed standard laying hen diets containing 20% GM with or without 250 units/kg of mannanase Hemicell supplementation. On the fourth day of treatment, all hens were orally challenged with SE (1.65 x 10(7) cfu). Hens were killed and evaluated for Salmonella colonization and differences in organ weights 5 d postinoculation. Salmonella Enteritidis present in crop, liver, ovary, and cecal contents were significantly reduced by feeding GM with enzyme supplementation compared with feed withdrawal hens. No significant differences were observed in reproductive tract weights of molted groups, although a difference in liver weight was detected. Results indicate that feeding diets containing 20% GM are as effective as complete feed withdrawal with respect to inducing molt with the added benefit of improved resistance to Salmonella Enteritidis colonization and translocation.

  15. A genome-wide screen identifies Salmonella Enteritidis lipopolysaccharide biosynthesis and the HtrA heat shock protein as crucial factors involved in egg white persistence at chicken body temperature.

    PubMed

    Raspoet, R; Shearer, N; Appia-Ayme, C; Haesebrouck, F; Ducatelle, R; Thompson, A; Van Immerseel, F

    2014-05-01

    Eggs contaminated with Salmonella Enteritidis are an important source of human foodborne Salmonella infections. Salmonella Enteritidis is able to contaminate egg white during formation of the egg within the chicken oviduct, and it has developed strategies to withstand the antimicrobial properties of egg white to survive in this hostile environment. The mechanisms involved in the persistence of Salmonella Enteritidis in egg white are likely to be complex. To address this issue, a microarray-based transposon library screen was performed to identify genes necessary for survival of Salmonella Enteritidis in egg white at chicken body temperature. The majority of identified genes belonged to the lipopolysaccharide biosynthesis pathway. Additionally, we provide evidence that the serine protease/heat shock protein (HtrA) appears essential for the survival of Salmonella Enteritidis in egg white at chicken body temperature.

  16. Comparison of four molecular methods to type Salmonella Enteritidis strains.

    PubMed

    Campioni, Fábio; Pitondo-Silva, André; Bergamini, Alzira M M; Falcão, Juliana P

    2015-05-01

    This study compared the pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), multilocus variable-number of tanden-repeat analysis (MLVA), and multilocus sequence typing (MLST) methods for typing 188 Salmonella Enteritidis strains from different sources isolated over a 24-year period in Brazil. PFGE and ERIC-PCR were more efficient than MLVA for subtyping the strains. However, MLVA provided additional epidemiological information for those strains. In addition, MLST showed the Brazilian strains as belonging to the main clonal complex of S. Enteritidis, CC11, and provided the first report of two new STs in the S. enterica database but could not properly subtype the strains. Our results showed that the use of PFGE or ERIC-PCR together with MLVA is suitable to efficiently subtype S. Enteritidis strains and provide important epidemiological information. © 2015 APMIS. Published by John Wiley & Sons Ltd.

  17. Colonization of internal organs by Salmonella Enteritidis in experimentally infected laying hens housed in enriched colony cages at different stocking densities.

    PubMed

    Gast, Richard K; Guraya, Rupa; Jones, Deana R; Anderson, Kenneth E; Karcher, Darrin M

    2016-06-01

    Epidemiologic analyses have linked the frequency of human infections with Salmonella enterica subspecies enterica serovar Enteritidis to the consumption of contaminated eggs and thus to the prevalence of this pathogen in commercial egg-laying flocks. Contamination of the edible contents of eggs by Salmonella Enteritidis is a consequence of the colonization of reproductive tissues in systemically infected hens. The animal welfare implications of laying hen housing systems have been widely debated, but no definitive consensus has yet emerged about the food safety significance of poultry housing options. The present study sought to determine the effects of two different bird stocking densities on the invasion of internal organs by Salmonella Enteritidis in groups of experimentally infected laying hens housed in colony cages enriched with perching and nesting areas. In two trials, groups of laying hens were distributed at two different stocking densities into colony cages and (along with a group housed in conventional cages) orally inoculated with doses of 1.0 × 10(7) cfu of Salmonella Enteritidis. At 5 to 6 d post-inoculation, hens were euthanized and samples of internal organs were removed for bacteriologic culturing. For both trials combined, Salmonella Enteritidis was recovered at a significantly (P < 0.05) greater frequency from hens in enriched colony cages at the higher stocking density than at the lower density from livers (75.0% vs. 51.4%) and ovaries (51.4% vs. 30.6%). However, spleens from hens in enriched colony cages at the higher stocking density were significantly less often positive for Salmonella Enteritidis than from hens in conventional cages at that same density (90.3% vs. 68.1%). These results suggest that stocking density can influence the susceptibility of hens to Salmonella Enteritidis, but other housing systems parameters may also contribute to the outcome of infections. Published by Oxford University Press on behalf of Poultry Science

  18. Feed Supplementation with Red Seaweeds, Chondrus crispus and Sarcodiotheca gaudichaudii, Reduce Salmonella Enteritidis in Laying Hens

    PubMed Central

    Kulshreshtha, Garima; Rathgeber, Bruce; MacIsaac, Janice; Boulianne, Martine; Brigitte, Lehoux; Stratton, Glenn; Thomas, Nikhil A.; Critchley, Alan T.; Hafting, Jeff; Prithiviraj, Balakrishnan

    2017-01-01

    Salmonella Enteritidis is vertically transmitted to eggs from laying hens through infected ovaries and oviducts. S. Enteritidis can also penetrate the eggshell from contaminated feces. Reducing S. Enteritidis in laying hens is vital to provide safer eggs and minimize the spread of salmonellosis to humans. Antibiotics have been widely used to control bacterial diseases in broilers and laying hens. However, there is a major concern that the use of antibiotics leads to the development of antibiotic resistance and adverse effects on microbiota of the treated birds. Thus, there is an interest in developing alternatives to antibiotics, such as dietary prebiotics. In the present study, feed supplemented with the red seaweeds: Chondrus crispus (CC) or Sarcodiotheca gaudichaudii (SG), was offered to laying hens late in production to control S. Enteritidis. Diets contained one of the following; 2% or 4% Chondrus crispus (CC2, and CC4, respectively) or Sarcodiotheca gaudichaudii (SG2 and SG4, respectively). Chlortetracycline was used in the positive control diet. During week-4, 48 birds were orally challenged with 2 × 109 CFU/mL of S. Enteritidis. Eggs and fecal samples were collected 1, 3, 5, and 7 days’ post inoculation. Birds were euthanized and organs (ceca, ovary, liver, and spleen) were sampled and analyzed for the presence of S. Enteritidis, 7 days’ post inoculation. Results showed that seaweed reduced the negative effect on body weight and egg production in S. Enteritidis-challenged laying hens. Analysis of fecal samples showed that the antibiotic (CTC) reduced S. Enteritidis in the intestinal tract and fecal samples, 3 days’ post inoculation. Fecal samples from Chlortetracycline and CC4 supplemented birds tested negative for S. Enteritidis on days 5 and 7 post inoculation (lowest detection limit = 10-1). S. Enteritidis colonization in the ceca was also significantly reduced in birds fed CC (4%) and Chlortetracycline. Blood serum profiles revealed that there

  19. Effect of coffee filtrate, methylglyoxal, glyoxal, and caffeine on Salmonella typhimurium and S. enteritidis survival in ground chicken breasts.

    PubMed

    Maletta, Anne B; Were, Lilian M

    2012-02-01

    The antimicrobial effect of roasted coffee filtrate (CF) and dicarbonyls on Salmonella Typhimurium and Salmonella Enteritidis in raw ground chicken breast meat (GCB) was investigated. Coffee was brewed and filtered before addition to GCB. Coffee filtrate with and without added caffeine, methylglyoxal, and/or glyoxal was added to GCB and then inoculated with Salmonella Typhimurium and Salmonella Enteritidis. Ground chicken samples were stomached with peptone water at days 1, 3, 5, and 7, plated on XLD agar with a TSA overlay, and Salmonella survivors were enumerated. CF alone gave less than a 1 Log reduction in all runs compared to control GCB with no treatment. Methylglyoxal (2.28 mg/g GCB) had the greatest antimicrobial effect against Salmonella Typhimurium and Salmonella Enteritidis in GCB with average Log reductions of 2.27 to 3.23, respectively, over the 7 d duration of the experiment compared to control GCB with no treatment. A 1 Log reduction was observed in GCB with CF, 0.93 mg glyoxal, and 1 mg caffeine/g chicken compared to the control and GCB with only CF. Heat-produced coffee compounds could potentially reduce Salmonella in retail ground chicken and chicken products. © 2011 Institute of Food Technologists®

  20. Epigenetic modification: possible approach to reduce Salmonella enterica serovar enteritidis susceptibility under stress conditions.

    PubMed

    Soleimani, A F; Zulkifli, I; Hair-Bejo, M; Ebrahimi, M; Jazayeri, S D; Hashemi, S R; Meimandipour, A; Goh, Y M

    2012-01-01

    Stressors may influence chicken susceptibility to pathogens such as Salmonella enterica. Feed withdrawal stress can cause changes in normal intestinal epithelial structure and may lead to increased attachment and colonization of Salmonella. This study aimed to investigate modulatory effects of epigenetic modification by feed restriction on S. enterica serovar Enteritidis colonization in broiler chickens subjected to feed withdrawal stress. Chicks were divided into four groups: ad libitum feeding; ad libitum feeding with 24-h feed withdrawal on day 42; 60% feed restriction on days 4, 5, and 6; and 60% feed restriction on days 4, 5, and 6 with 24-h feed withdrawal on day 42. Attachment of S. Enteritidis to ileal tissue was determined using an ex vivo ileal loop assay, and heat shock protein 70 (Hsp70) expression was evaluated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blotting. Feed withdrawal stress increased S. Enteritidis attachment to ileal tissue. However, following feed withdrawal the epigenetically modified chickens had significantly lower attachment of S. Enteritidis than their control counterparts. A similar trend with a very positive correlation was observed for Hsp70 expression. It appears that epigenetic modification can enhance resistance to S. Enteritidis colonization later in life in chickens under stress conditions. The underlying mechanism could be associated with the lower Hsp70 expression in the epigenetically modified chickens.

  1. Genetic diversity and antimicrobial resistance pattern of Salmonella enterica serovar Enteritidis clinical isolates in Thailand.

    PubMed

    Utrarachkij, Fuangfa; Nakajima, Chie; Siripanichgon, Kanokrat; Changkaew, Kanjana; Thongpanich, Yuwanda; Pornraungwong, Srirat; Suthienkul, Orasa; Suzuki, Yasuhiko

    2016-04-01

    To trace the history of antimicrobial resistance in Salmonella enterica serovar Enteritidis (S. Enteritidis, SE) circulating in Thailand, we characterised clinical isolates obtained during 2004-2007. Antimicrobial resistance profiles, multi-locus variable number tandem repeat analysis (MLVA) types and 3 representative virulence determinants (spvA, sodCI and sopE) were established from SE isolates (n = 192) collected from stool and blood of patients throughout Thailand during the period 2004-2007. Resistance was found in SE against 10 out of 11 antimicrobials studied. The highest resistance ratios were observed for nalidixic acid (83.2%), ciprofloxacin (51.1%) and ampicillin (50.5%), and 25.5% were multidrug resistant. Based on five polymorphic tandem repeat loci analysis, MLVA identified 20 distinct types with three closely related predominant types. A significant increase of AMP resistance from 2004 to 2006 was strongly correlated with that of a MLVA type, 5-5-11-7-3. The usage of antimicrobials in human medicine or farm settings might act as selective pressures and cause the spread of resistant strains. Hence, a strict policy on antimicrobial usage needs to be implemented to achieve the control of resistant SE in Thailand. Copyright © 2015 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  2. Protective action of Lactobacillus kefir carrying S-layer protein against Salmonella enterica serovar Enteritidis.

    PubMed

    Golowczyc, M A; Mobili, P; Garrote, G L; Abraham, A G; De Antoni, G L

    2007-09-30

    Eight Lactobacillus kefir strains isolated from different kefir grains were tested for their ability to antagonize Salmonella enterica serovar Enteritidis (Salmonella enteritidis) interaction with epithelial cells. L. kefir surface properties such as autoaggregation and coaggregation with Salmonella and adhesion to Caco-2/TC-7 cells were evaluated. L. kefir strains showed significantly different adhesion capacities, six strains were able to autoaggregate and four strains coaggregated with Salmonella. Coincubation of Salmonella with coaggregating L. kefir strains significantly decreased its capacity to adhere to and to invade Caco-2/TC-7 cells. This was not observed with non coaggregating L. kefir strains. Spent culture supernatants of L. kefir contain significant amounts of S-layer proteins. Salmonella pretreated with spent culture supernatants (pH 4.5-4.7) from all tested L. kefir strains showed a significant decrease in association and invasion to Caco-2/TC-7 cells. Artificially acidified MRS containing lactic acid to a final concentration and pH equivalent to lactobacilli spent culture supernatants did not show any protective action. Pretreatment of this pathogen with spent culture supernatants reduced microvilli disorganization produced by Salmonella. In addition, Salmonella pretreated with S-layer proteins extracted from coaggregating and non coaggregating L. kefir strains were unable to invade Caco-2/TC-7 cells. After treatment, L. kefir S-layer protein was detected associated with Salmonella, suggesting a protective role of this protein on association and invasion.

  3. Molecular characterization of Salmonella enterica serotype Enteritidis isolates from food and human samples by serotyping, antimicrobial resistance, plasmid profiling, (GTG)5-PCR and ERIC-PCR.

    PubMed

    Fardsanei, F; Nikkhahi, F; Bakhshi, B; Salehi, T Z; Tamai, I A; Soltan Dallal, M M

    2016-11-01

    In recent years, Salmonella enterica serovar Enteritidis has been a primary cause of human salmonellosis in many countries. The major objective of this study was to investigate genetic diversity among Salmonella Enteritidis strains from different origins (food and human) by Enterobacterial Repetitive Intergenic Consensus (ERIC) -PCR, as well as to assess their plasmid profiling and antimicrobial resistance. A total of 30 Salmonella Enteritidis isolates, 15 from food samples (chicken, lamb, beef and duck meats) and 15 from clinical samples were collected in Tehran. Identification of isolates as Salmonella was confirmed by using conventional standard biochemical and serological tests. Multiplex-PCR was used for serotyping of isolates to identify Salmonella Enteritidis. Antimicrobial susceptibility testing to 16 agents founds drug resistance patterns among Salmonella Enteritidis isolates. No resistance was observed to cephalexin, ceftriaxone, ceftazidime and cefotaxime, ciprofloxacin, imipenem or meropenem, chloramphenicol and gentamicin. The highest resistance (96.7%) was observed to nitrofurantoin. Seven plasmid profiles (P1-P7) were detected, and a 68-kb plasmid was found in all isolates. Two different primers; ERIC and (GTG)5 were used for genotyping, which each produced four profiles. The majority of clinical and food isolates fell into two separate common types (CTs) with a similar percentage of 95% by ERIC-PCR. Using primer (GTG)5, 29 isolates incorporated in three CTs with 70% of isolates showing a single banding pattern. Limited genetic diversity among human and food isolates of Salmonella Enteritidis may indicate that contaminated foods were possibly the source of human salmonellosis. These results confirmed that ERIC-PCR genotyping has limited discriminatory power for Salmonella Enteritidis of different origin.

  4. Development of multiplex PCR assay for simultaneous detection of Salmonella genus, Salmonella subspecies I, Salm. Enteritidis, Salm. Heidelberg and Salm. Typhimurium.

    PubMed

    Park, S H; Ricke, S C

    2015-01-01

    The aim of this research was to develop multiplex PCR assay that could simultaneously detect Salmonella genus, Salmonella subsp. I, Salm. Enteritidis, Heidelberg and Typhimurium because these Salmonella serovars are the most common isolates associated with poultry products. Five primers were utilized to establish multiplex PCR and applied to Salmonella isolates from chickens and farm environments. These isolates were identified as Salmonella subsp. I and 16 of 66 isolates were classified as Salm. Enteritidis, while Heidelberg or Typhimurium was not detected. We also spiked three Salmonella strains on chicken breast meat to evaluate the specificity and sensitivity of multiplex PCR as well as qPCR to optimize quantification of Salmonella in these samples. The optimized multiplex PCR and qPCR could detect approx. 2·2 CFU of Salmonella per gram after 18 h enrichment. The multiplex PCR and qPCR would provide rapid and consistent results. Also, these techniques would be useful for the detection and quantification of Salmonella in contaminated poultry, foods and environmental samples. The strategy for the rapid detection of Salmonella serovars in poultry is needed to further reduce the incidence of salmonellosis in humans. The optimized multiplex PCR will be useful to detect prevalent Salmonella serovars in poultry products. © 2014 The Society for Applied Microbiology.

  5. [Epidemiologic pattern of Salmonella enteritidis and Salmonella typhimurium in man--analysis of Salmonella cases in the Brandenburg federal territory].

    PubMed

    Käsbohrer, A; Talaska, T; Bögel, K; Stöhr, K; Lehmacher, W

    1993-01-01

    It is the aim of this study (a study that is part of several investigations suggested and coordinated by the WHO) to highlight the characteristics of transmission of Salmonella in humans. To achieve this, the data collected on the basis of the Federal German Law governing Epidemic Diseases as well as additionally available data (sex, serovar, differentiation between diseased and symptomatic, allocation to the respective place of residence) have been evaluated. In this manner it became possible to determine different patterns of incidence of the presently most frequently occurring salmonella serovars S. enteritidis and S. typhimurium in respect of distribution according to age and sex, as well as the dynamics of distribution and incidence in terms of time. The incidence is particularly remarkable among children, where S. enteritidis is not so frequent as S. typhimurium. There is a distinct trend towards the male sex among children in respect of the infestation. The fact that women are particularly often asymptomatic carriers points to the special nature of the sources and paths of infection. In respect of seasonal dynamics S. typhimurium does not have a specific summer peak, contrary to S. enteritidis, and this too indicates that the mechanisms of transmission or multiplication differ from one another. The study presented here is supplemented by the evaluation of further parameters to support and promote on-target epidemiological investigation into the various problems raised by this and other studies.

  6. Efficacy of jatropha curcas plant extract against the survival of salmonella enteritidis

    USDA-ARS?s Scientific Manuscript database

    Introduction: The use of plant-derived antimicrobials has shown to be effective at inhibiting microbial growth. Although Jatropha curcas is known to possess antimicrobial properties, its efficacy against Salmonella Enteritidis has not yet been investigated. Purpose: The purpose of this study was...

  7. Risks Involved in the Use of Enrofloxacin for Salmonella Enteritidis or Salmonella Heidelberg in Commercial Poultry

    PubMed Central

    Morales-Barrera, Eduardo; Calhoun, Nicole; Lobato-Tapia, Jose L.; Lucca, Vivian; Prado-Rebolledo, Omar; Hernandez-Velasco, Xochitl; Merino-Guzman, Ruben; Petrone-García, Victor M.; Latorre, Juan D.; Mahaffey, Brittany D.; Teague, Kyle D.; Graham, Lucas E.; Wolfenden, Amanda D.; Baxter, Mikayla F. A.; Hargis, Billy M.; Tellez, Guillermo

    2016-01-01

    The objectives of the present study were to evaluate the risks involved in the use of Enrofloxacin for Salmonella Enteritidis (SE) or Salmonella Heidelberg (SH) in commercial poultry and determine the effects of a probiotic as an antibiotic alternative. Two experiments were conducted to evaluate the risks involved in the use of Enrofloxacin for SE or SH in commercial poultry. Experiment 1 consisted of two trials. In each trial, chickens were assigned to one of three groups; control + SE challenged; Enrofloxacin 25 mg/kg + SE; and Enrofloxacin 50 mg/kg + SE. Chickens received Enrofloxacin in the drinking water from days 1 to 5 of age. On day 6, all groups received fresh water without any treatment. All chickens were orally gavaged with 107 cfu/chick of SE at 7 days of age and euthanized on 8 days of age. In Experiment 2, turkey poults were assigned to one of the three groups; control + SH; probiotic + SH; and Enrofloxacin 50 mg/kg + SH. Poults received probiotic or Enrofloxacin in the drinking water from days 1 to 5 of age. On day 6, poults received fresh water without any treatment. Poults were orally gavaged with 107 cfu/poult of SH at 7 days of age. Poults were weighed and humanely killed 24 h post-SH challenge to evaluate serum concentration of fluorescein isothiocyanate-dextran to evaluate intestinal permeability, metagenomics, and SH infection. In both trials of Experiment 1, chickens treated with Enrofloxacin were more susceptible to SE organ invasion and intestinal colonization when compared with control non-treated chickens (P < 0.05). In Experiment 2, poults treated with 50 mg/kg of Enrofloxacin showed an increase in body weight, however, this group also showed an increase in SH susceptibility, intestinal permeability, and lower proportion of Firmicutes and Bacteroidetes, but with control group had the highest proportion of Proteobacteria. By contrast, poults that received the probiotic had the highest

  8. Risks Involved in the Use of Enrofloxacin for Salmonella Enteritidis or Salmonella Heidelberg in Commercial Poultry.

    PubMed

    Morales-Barrera, Eduardo; Calhoun, Nicole; Lobato-Tapia, Jose L; Lucca, Vivian; Prado-Rebolledo, Omar; Hernandez-Velasco, Xochitl; Merino-Guzman, Ruben; Petrone-García, Victor M; Latorre, Juan D; Mahaffey, Brittany D; Teague, Kyle D; Graham, Lucas E; Wolfenden, Amanda D; Baxter, Mikayla F A; Hargis, Billy M; Tellez, Guillermo

    2016-01-01

    The objectives of the present study were to evaluate the risks involved in the use of Enrofloxacin for Salmonella Enteritidis (SE) or Salmonella Heidelberg (SH) in commercial poultry and determine the effects of a probiotic as an antibiotic alternative. Two experiments were conducted to evaluate the risks involved in the use of Enrofloxacin for SE or SH in commercial poultry. Experiment 1 consisted of two trials. In each trial, chickens were assigned to one of three groups; control + SE challenged; Enrofloxacin 25 mg/kg + SE; and Enrofloxacin 50 mg/kg + SE. Chickens received Enrofloxacin in the drinking water from days 1 to 5 of age. On day 6, all groups received fresh water without any treatment. All chickens were orally gavaged with 10(7) cfu/chick of SE at 7 days of age and euthanized on 8 days of age. In Experiment 2, turkey poults were assigned to one of the three groups; control + SH; probiotic + SH; and Enrofloxacin 50 mg/kg + SH. Poults received probiotic or Enrofloxacin in the drinking water from days 1 to 5 of age. On day 6, poults received fresh water without any treatment. Poults were orally gavaged with 10(7) cfu/poult of SH at 7 days of age. Poults were weighed and humanely killed 24 h post-SH challenge to evaluate serum concentration of fluorescein isothiocyanate-dextran to evaluate intestinal permeability, metagenomics, and SH infection. In both trials of Experiment 1, chickens treated with Enrofloxacin were more susceptible to SE organ invasion and intestinal colonization when compared with control non-treated chickens (P < 0.05). In Experiment 2, poults treated with 50 mg/kg of Enrofloxacin showed an increase in body weight, however, this group also showed an increase in SH susceptibility, intestinal permeability, and lower proportion of Firmicutes and Bacteroidetes, but with control group had the highest proportion of Proteobacteria. By contrast, poults that received the probiotic had the highest

  9. Effects of thermosonication on the fate of Escherichia coli O157:H7 and Salmonella Enteritidis in mango juice.

    PubMed

    Kiang, W-S; Bhat, R; Rosma, A; Cheng, L-H

    2013-04-01

    In this study, the effects of thermosonication and thermal treatment on Escherichia coli O157:H7 and Salmonella Enteritidis in mango juice were investigated at 50 and 60°C. Besides, nonlethal injury of Salm. Enteritidis after both treatments was also examined. The highest inactivation was attained with thermosonication at 60°C. The inactivation rate was different for both pathogens, and Salm. Enteritidis was found to be more sensitive to thermosonication than E. coli O157:H7. Salmonella Enteritidis was recovered in all treated samples, except those subjected to more than 5-min thermosonication at 60°C. It was found that the introduction of high-intensity ultrasound enhanced the inactivation of pathogens compared to thermal treatment alone. On the other hand, Salm. Enteritidis was detected in a number of samples following incubation in universal pre-enrichment broth, but no growth was detected after incubation in mango juice. Fruit juices are commonly heat treated to inactivate micro-organisms and enzymes. However, excessive heat treatments may result in undesirable changes in juice quality. Treatment by power ultrasound, a nonthermal technology, may be an alternative processing technique to pasteurize fruit juices. This study highlights the effectiveness of thermosonication in inactivating Escherichia coli O157:H7 and Salmonella Enteritidis in mango juice. © 2012 The Society for Applied Microbiology.

  10. Integrated surveillance and potential sources of Salmonella enteritidis in human cases in Canada from 2003 to 2009.

    PubMed

    Nesbitt, A; Ravel, A; Murray, R; McCormick, R; Savelli, C; Finley, R; Parmley, J; Agunos, A; Majowicz, S E; Gilmour, M

    2012-10-01

    Salmonella enteritidis has emerged as the most prevalent cause of human salmonellosis in Canada. Recent trends of S. enteritidis subtypes and their potential sources were described by integrating Salmonella data from several Canadian surveillance and monitoring programmes. A threefold increase in S. enteritidis cases from 2003 to 2009 was identified to be primarily associated with phage types 13, 8 and 13a. Other common phage types (4, 1, 6a) showed winter seasonality and were more likely to be associated with cases linked to international travel. Conversely, phage types 13, 8 and 13a had summer seasonal peaks and were associated with cases of domestically acquired infections. During agri-food surveillance, S. enteritidis was detected in various commodities, most frequently in chicken (with PT13, PT8 and PT13a predominating). Antimicrobial resistance was low in human and non-human isolates. Continued integrated surveillance and collaborative prevention and control efforts are required to mitigate future illness.

  11. Integrated surveillance and potential sources of Salmonella Enteritidis in human cases in Canada from 2003 to 2009

    PubMed Central

    NESBITT, A.; RAVEL, A.; MURRAY, R.; McCORMICK, R.; SAVELLI, C.; FINLEY, R.; PARMLEY, J.; AGUNOS, A.; MAJOWICZ, S. E.; GILMOUR, M.

    2012-01-01

    SUMMARY Salmonella Enteritidis has emerged as the most prevalent cause of human salmonellosis in Canada. Recent trends of S. Enteritidis subtypes and their potential sources were described by integrating Salmonella data from several Canadian surveillance and monitoring programmes. A threefold increase in S. Enteritidis cases from 2003 to 2009 was identified to be primarily associated with phage types 13, 8 and 13a. Other common phage types (4, 1, 6a) showed winter seasonality and were more likely to be associated with cases linked to international travel. Conversely, phage types 13, 8 and 13a had summer seasonal peaks and were associated with cases of domestically acquired infections. During agri-food surveillance, S. Enteritidis was detected in various commodities, most frequently in chicken (with PT13, PT8 and PT13a predominating). Antimicrobial resistance was low in human and non-human isolates. Continued integrated surveillance and collaborative prevention and control efforts are required to mitigate future illness. PMID:22166269

  12. Epigenetic Modification of TLRs in Leukocytes Is Associated with Increased Susceptibility to Salmonella enteritidis in Chickens

    PubMed Central

    Zhao, Guiping; Zheng, Maiqing; Li, Peng; Wang, Huihua; Zhu, Yun; Chen, Jilan; Wen, Jie

    2012-01-01

    Toll-like receptors (TLRs) signaling pathways are the first lines in defense against Salmonella enteritidis (S. enteritidis) infection but the molecular mechanism underlying susceptibility to S. enteritidis infection in chicken remains unclear. SPF chickens injected with S. enteritidis were partitioned into two groups, one consisted of those from Salmonella-susceptible chickens (died within 5 d after injection, n = 6), the other consisted of six Salmonella-resistant chickens that survived for 15 d after injection. The present study shows that the bacterial load in susceptible chickens was significantly higher than that in resistant chickens and TLR4, TLR2-1 and TLR21 expression was strongly diminished in the leukocytes of susceptible chickens compared with those of resistant chickens. The induction of expression of pro-inflammatory cytokine genes, IL-6 and IFN-β, was greatly enhanced in the resistant but not in susceptible chickens. Contrasting with the reduced expression of TLR genes, those of the zinc finger protein 493 (ZNF493) gene and Toll-interacting protein (TOLLIP) gene were enhanced in the susceptible chickens. Finally, the expression of TLR4 in peripheral blood mononuclear cells (PBMCs) infected in vitro with S. enteritidis increased significantly as a result of treatment with 5-Aza-2-deoxycytidine (5-Aza-dc) while either 5-Aza-dc or trichostatin A was effective in up-regulating the expression of TLR21 and TLR2-1. DNA methylation, in the predicted promoter region of TLR4 and TLR21 genes, and an exonic CpG island of the TLR2-1 gene was significantly higher in the susceptible chickens than in resistant chickens. Taken together, the results demonstrate that ZNF493-related epigenetic modification in leukocytes probably accounts for increased susceptibility to S. enteritidis in chickens by diminishing the expression and response of TLR4, TLR21 and TLR2-1. PMID:22438967

  13. Conjugal Transfer of the Pathogenicity Island ROD21 in Salmonella enterica serovar Enteritidis Depends on Environmental Conditions

    PubMed Central

    Salazar-Echegarai, Francisco J.; Tobar, Hugo E.; Nieto, Pamela A.; Riedel, Claudia A.; Bueno, Susan M.

    2014-01-01

    Unstable pathogenicity islands are chromosomal elements that can be transferred from one bacterium to another. Salmonella enterica serovar Enteritidis (S. Enteritidis) is a pathogenic bacterium containing such unstable pathogenicity islands. One of them, denominated ROD21, is 26.5 kb in size and capable of excising from the chromosome in certain culture conditions, as well as during bacterial infection of phagocytic cells. In this study we have evaluated whether ROD21 can be effectively transferred from one bacterium to another. We generated a donor and several recipient strains of S. Enteritidis to carry out transfer assays in liquid LB medium. These assays showed that ROD21 is effectively transferred from donor to recipient strains of S. Enteritidis and S. Typhimurium. When Escherichia coli was used as the recipient strain, ROD21 transfer failed to be observed. Subsequently, we showed that a conjugative process was required for the transfer of the island and that changes in temperature and pH increased the transfer frequency between Salmonella strains. Our data indicate that ROD21 is an unstable pathogenicity island that can be transferred by conjugation in a species-specific manner between Salmonellae. Further, ROD21 transfer frequency increases in response to environmental changes, such as pH and temperature. PMID:24705125

  14. Interdigitated microelectrode based impedance biosensor for detection of salmonella enteritidis in food samples

    NASA Astrophysics Data System (ADS)

    Kim, G.; Morgan, M.; Hahm, B. K.; Bhunia, A.; Mun, J. H.; Om, A. S.

    2008-03-01

    Salmonella enteritidis outbreaks continue to occur, and S. enteritidis-related outbreaks from various food sources have increased public awareness of this pathogen. Conventional methods for pathogens detection and identification are labor-intensive and take days to complete. Some immunological rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown great potential for the rapid detection of foodborne pathogens. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on avidin-biotin binding on the surface of the IME to form an active sensing layer. To increase the sensitivity of the sensor, three types of sensors that have different electrode gap sizes (2 μm, 5 μm, 10 μm) were fabricated and tested. The impedimetric biosensor could detect 103 CFU/mL of Salmonella in pork meat extract with an incubation time of 5 minutes. This method may provide a simple, rapid and sensitive method to detect foodborne pathogens.

  15. Antimicrobial Resistance and Molecular Characterization of Salmonella enterica Serovar Enteritidis from Retail Chicken Products in Shanghai, China.

    PubMed

    Zhou, Xiujuan; Xu, Li; Xu, Xuebin; Zhu, Yuding; Suo, Yujuan; Shi, Chunlei; Shi, Xianming

    2018-05-30

    Salmonella enterica serovar Enteritidis is the leading global cause of salmonellosis. A total of 146 Salmonella Enteritidis isolates obtained from retail chicken products in Shanghai, China were characterized for their antimicrobial susceptibilities, virulence and antibiotic resistance gene profiles, and molecular subtypes using pulsed-field gel electrophoresis (PFGE). Approximately 42% (61/146) of the isolates were susceptible to all 13 antimicrobials tested. More than half of the isolates (50.70%) were resistant to ampicillin, 49.32% to sulfisoxazole, 17.12% to tetracycline, and 15.75% to doxycycline. Thirty (20.55%) isolates were resistant to three or more antimicrobials. The avrA, mgtC, and sopE virulence genes were identified in all isolates, while 97.2% and 92.4% were positive for bcfC and spvC genes, respectively. Genes associated with resistance to streptomycin (aadA), β-lactams (blaTEM, blaCMY, blaSHV, and blaCTX), tetracycline (tetA and tetB), and sulfonamides (sulI, sulII, and sulIII) were detected among corresponding resistant isolates. A total of 41 PFGE patterns were identified from 77 antimicrobial resistance (AMR) isolates and were primarily grouped into seven clusters (A-G), each with 90% similarity. The majority of Salmonella Enteritidis isolates (63.63%, 49/77) shared the same PFGE cluster, indicating potential cross contamination during processing and cutting or working during retailing and marketing. A significantly (p < 0.05) lower percentage (<25%) of isolates belonging to clusters D and E were resistant to sulfisoxazole compared with those belonging to clusters A, B, C, F, and G (>80%), indicating that sulfisoxazole resistance might be associated with genetic content (PFGE profiles) of Salmonella Enteritidis. This study provides important and updated information about the baseline antimicrobial-resistant data for food safety risk assessment of Salmonella Enteritidis from retailed chicken in Shanghai, which is the first step for the

  16. Vectorial competence of larvae and adults of Alphitobius diaperinus in the transmission of Salmonella enteritidis in poultry.

    PubMed

    Leffer, Andreia M; Kuttel, Javier; Martins, Lidiane M; Pedroso, Antonio Carlos; Astolfi-Ferreira, Claudete S; Ferreira, Fernando; Ferreira, Antonio J Piantino

    2010-06-01

    The ingestion of food products originating from poultry infected with Salmonella spp. is one of the major causes of food poisoning in humans. The control of poultry salmonellosis is particularly difficult since birds are asymptomatic and numerous factors may expedite the maintenance of bacteria in poultry production facilities. The aim of the study was to determine the vectorial capacity of adults and larvae of Alphitobius diaperinus (Coleoptera: Tenebrionidae) in the experimental transmission of Salmonella Enteritidis phage type 4 to 1-day-old specific pathogen-free White Leghorn chicks. Adult insects and larvae were starved for 1 day, fed for 24 h or 7 days on sterile ration that had been treated with Salmonella Enteritidis phage type 4, and the levels of bacterial infection were determined. Infected adult insects and larvae were fed to groups of day-old chicks, after which bacteria were recovered from cecum, liver, and spleen samples over a 7-day period. Infected larvae were more efficient than adult insects in transmitting Salmonella Enteritidis to chicks. Higher concentrations of bacteria could be reisolated from the cecum, liver, and spleen of chicks that had ingested infected larvae compared with those that had ingested infected adults. The control of A. diaperinus, and particularly of the larvae, represents a critical factor in the reduction of Salmonella spp. in poultry farms.

  17. Yerba mate enhances probiotic bacteria growth in vitro but as a feed additive does not reduce Salmonella Enteritidis colonization in vivo.

    PubMed

    Gonzalez-Gil, Francisco; Diaz-Sanchez, Sandra; Pendleton, Sean; Andino, Ana; Zhang, Nan; Yard, Carrie; Crilly, Nate; Harte, Federico; Hanning, Irene

    2014-02-01

    Yerba mate (Ilex paraguariensis) is a tea known to have beneficial effects on human health and antimicrobial activity against some foodborne pathogens. Thus, the application of yerba mate as a feed additive for broiler chickens to reduce Salmonella colonization was evaluated. The first in vitro evaluation was conducted by suspending Salmonella Enteritidis and lactic acid bacteria (LAB) in yerba mate extract. The in vivo evaluations were conducted using preventative and horizontal transmission experiments. In all experiments, day-of-hatch chicks were treated with one of the following 1) no treatment (control); 2) ground yerba mate in feed; 3) probiotic treatment (Lactobacillus acidophilus and Pediococcus; 9:1 administered once on day of hatch by gavage); or 4) both yerba mate and probiotic treatments. At d 3, all chicks were challenged with Salmonella Enteritidis (preventative experiment) or 5 of 20 chicks (horizontal transmission experiment). At d 10, all birds were euthanized, weighed, and cecal contents enumerated for Salmonella. For the in vitro evaluation, antimicrobial activity was observed against Salmonella and the same treatment enhanced growth of LAB. For in vivo evaluations, none of the yerba mate treatments significantly reduced Salmonella Enteritidis colonization, whereas the probiotic treatment significantly reduced Salmonella colonization in the horizontal transmission experiment. Yerba mate decreased chicken BW and decreased the performance of the probiotic treatment when used in combination. In conclusion, yerba mate had antimicrobial activity against foodborne pathogens and enhanced the growth of LAB in vitro, but in vivo yerba mate did not decrease Salmonella Enteritidis colonization.

  18. Salmonella enterica serotype enteritidis in French Polynesia, South Pacific, 2008-2013.

    PubMed

    Le Hello, Simon; Maillard, Fiona; Mallet, Henri-Pierre; Daudens, Elise; Levy, Marc; Roy, Valérie; Branaa, Philippe; Bertrand, Sophie; Fabre, Laetitia; Weill, François-Xavier

    2015-06-01

    Outbreaks of Salmonella enterica serotype Enteritidis infections associated with eggs occurred in French Polynesia during 2008-2013. Molecular analysis of isolates by using clustered regularly interspaced short palindromic repeat polymorphisms and multilocus variable-number tandem-repeat analysis was performed. This subtyping made defining the epidemic strain, finding the source, and decontaminating affected poultry flocks possible.

  19. Changes in the Salmonella enterica Enteritidis phenotypes in presence of acyl homoserine lactone quorum sensing signals.

    PubMed

    Campos-Galvão, Maria Emilene Martino; Ribon, Andrea Oliveira Barros; Araújo, Elza Fernandes; Vanetti, Maria Cristina Dantas

    2016-05-01

    Quorum sensing is used by bacteria to coordinate gene expression in response to population density and involves the production, detection and response to extracellular signaling molecules known as autoinducers (AIs). Salmonella does not synthesize the AI-1, acyl homoserine lactone (AHL) common to gram-negative bacteria; however, it has a receptor for AI-1, the SdiA protein. The effect of SdiA in modulating phenotypes of Salmonella has not been elucidated. In this report, we provide evidence that the AIs-1 affect Salmonella enterica serovar Enteritidis behavior by enhancing the biofilm formation and expression of virulence genes under anaerobic conditions. Biofilm formation by Salmonella was detected by the crystal violet method and by scanning electron microscopy. The presence of AHLs, particularly C12-HSL, increased biofilm formation and promoted expression of biofilm formation genes (lpfA, fimF, fliF, glgC) and virulence genes (hilA, invA, invF). Our results demonstrated that AHLs produced by other organisms played an important role in virulence phenotypes of Salmonella Enteritidis. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Oral immunisation of laying hens with the live vaccine strains of TAD Salmonella vac E and TAD Salmonella vac T reduces internal egg contamination with Salmonella Enteritidis.

    PubMed

    Gantois, Inne; Ducatelle, Richard; Timbermont, Leen; Boyen, Filip; Bohez, Lotte; Haesebrouck, Freddy; Pasmans, Frank; van Immerseel, Filip

    2006-09-11

    Eggs are a major source of human infections with Salmonella. Therefore controlling egg contamination in laying hen flocks is one of the main targets for control programmes. A study was carried out to assess the effect of oral vaccination with TAD Salmonella vac E, TAD Salmonella vac T and with both vaccines TAD Salmonella vac E and TAD Salmonella vac T, on colonization of the reproductive tract and internal egg contamination of laying hens with Salmonella Enteritidis. Three groups of 30 laying hens were vaccinated at 1 day, 6 weeks and 16 weeks of age with either one of the vaccine strains, or a combination of both vaccine strains, while a fourth group was left unvaccinated. At 24 weeks of age, the birds were intravenously challenged with 0.5 ml containing 5 x 10(7)cfu Salmonella Enteritidis PT4 S1400/94. The number of oviducts from which Salmonella was isolated, was significantly lower in the vaccinated than in the non-vaccinated hens at 3 weeks post-challenge. Significantly less egg contents were Salmonella positive in the birds vaccinated with TAD Salmonella vac E or TAD Salmonella vac T (12/105 batches of eggs in both groups) than in the unvaccinated birds (28/105 batches of eggs). Internal egg contamination in the hens vaccinated with both TAD Salmonella vac E and TAD Salmonella vac T was even more reduced, as over the whole experiment, only one batch of eggs was positive. In conclusion, these data indicate that vaccination of laying hens with these live vaccines could be considered as a valuable tool in controlling internal egg contamination.

  1. Purulent pericarditis with Salmonella enteritidis in a patient with CD4/CD8 depression.

    PubMed

    Takamiya, Yosuke; Shirai, Kazuyuki; Fujino, Masahiro; Miller, Nathan; Tsuchiya, Yoshihiro; Okabe, Masanori; Saku, Keijiro

    2008-06-01

    A 65-year-old man was admitted for high-grade fever with a shaking chill and general fatigue. Chest X-ray showed cardiomegaly, and echocardiography revealed a large amount of pericardial effusion. Emergency pericardiocentesis was performed, and Salmonella enteritidis was found in pericardial fluids. We diagnosed purulent pericarditis with S. enteritidis, and administered antibiotics. While high-grade fever resolved 10 days after beginning of treatment, effusive-constrictive pericarditis (ECP) without definite symptoms persisted for 2 months. Because of the improvement of his hemodynamic states on cardiac catheterization after 1 year, an operative procedure was not required. He was diagnosed as having CD4/CD8 depression without apparent diseases. There are few reports of pericarditis with S. enteritidis, and we believe this case might be only the second recorded case of ECP with S. enteritidis.

  2. The relationship between the immune response and susceptibility to Salmonella enterica serovar Enteritidis infection in the laying hen

    USDA-ARS?s Scientific Manuscript database

    Chicken eggs are one of the main sources of human salmonellosis, with Salmonella enterica serovar Enteritidis the most frequent cause of human non-typhoid salmonellosis. Laying hens colonized with S. Enteritidis generally do not show clinical signs. The bacteria colonize and invade the intestinal ...

  3. Red Seaweeds Sarcodiotheca gaudichaudii and Chondrus crispus down Regulate Virulence Factors of Salmonella Enteritidis and Induce Immune Responses in Caenorhabditis elegans.

    PubMed

    Kulshreshtha, Garima; Borza, Tudor; Rathgeber, Bruce; Stratton, Glenn S; Thomas, Nikhil A; Critchley, Alan; Hafting, Jeff; Prithiviraj, Balakrishnan

    2016-01-01

    Red seaweeds are a rich source of unique bioactive compounds and secondary metabolites that are known to improve human and animal health. S. Enteritidis is a broad range host pathogen, which contaminates chicken and poultry products that end into the human food chain. Worldwide, Salmonella outbreaks have become an important economic and public health concern. Moreover, the development of resistance in Salmonella serovars toward multiple drugs highlights the need for alternative control strategies. This study evaluated the antimicrobial property of red seaweeds extracts against Salmonella Enteritidis using the Caenorhabditis elegans infection model. Six red seaweed species were tested for their antimicrobial activity against S. Enteritidis and two, Sarcodiotheca gaudichaudii (SG) and Chondrus crispus (CC), were found to exhibit such properties. Spread plate assay revealed that SG and CC (1%, w/v) significantly reduced the growth of S. Enteritidis. Seaweed water extracts (SWE) of SG and CC, at concentrations from 0.4 to 2 mg/ml, significantly reduced the growth of S. Enteritidis (log CFU 4.5-5.3 and log 5.7-6.0, respectively). However, methanolic extracts of CC and SG did not affect the growth of S. Enteritidis. Addition of SWE (0.2 mg/ml, CC and SG) significantly decreased biofilm formation and reduced the motility of S. Enteritidis. Quantitative real-time PCR analyses showed that SWE (CC and SG) suppressed the expression of quorum sensing gene sdiA and of Salmonella Pathogenesis Island-1 (SPI-1) associated genes sipA and invF, indicating that SWE might reduce the invasion of S. Enteritidis in the host by attenuating virulence factors. Furthermore, CC and SG water extracts significantly improved the survival of infected C. elegans by impairing the ability of S. Enteritidis to colonize the digestive tract of the nematode and by enhancing the expression of C. elegans immune responsive genes. As the innate immune response pathways of C. elegans and mammals show a high

  4. Red Seaweeds Sarcodiotheca gaudichaudii and Chondrus crispus down Regulate Virulence Factors of Salmonella Enteritidis and Induce Immune Responses in Caenorhabditis elegans

    PubMed Central

    Kulshreshtha, Garima; Borza, Tudor; Rathgeber, Bruce; Stratton, Glenn S.; Thomas, Nikhil A.; Critchley, Alan; Hafting, Jeff; Prithiviraj, Balakrishnan

    2016-01-01

    Red seaweeds are a rich source of unique bioactive compounds and secondary metabolites that are known to improve human and animal health. S. Enteritidis is a broad range host pathogen, which contaminates chicken and poultry products that end into the human food chain. Worldwide, Salmonella outbreaks have become an important economic and public health concern. Moreover, the development of resistance in Salmonella serovars toward multiple drugs highlights the need for alternative control strategies. This study evaluated the antimicrobial property of red seaweeds extracts against Salmonella Enteritidis using the Caenorhabditis elegans infection model. Six red seaweed species were tested for their antimicrobial activity against S. Enteritidis and two, Sarcodiotheca gaudichaudii (SG) and Chondrus crispus (CC), were found to exhibit such properties. Spread plate assay revealed that SG and CC (1%, w/v) significantly reduced the growth of S. Enteritidis. Seaweed water extracts (SWE) of SG and CC, at concentrations from 0.4 to 2 mg/ml, significantly reduced the growth of S. Enteritidis (log CFU 4.5–5.3 and log 5.7–6.0, respectively). However, methanolic extracts of CC and SG did not affect the growth of S. Enteritidis. Addition of SWE (0.2 mg/ml, CC and SG) significantly decreased biofilm formation and reduced the motility of S. Enteritidis. Quantitative real-time PCR analyses showed that SWE (CC and SG) suppressed the expression of quorum sensing gene sdiA and of Salmonella Pathogenesis Island-1 (SPI-1) associated genes sipA and invF, indicating that SWE might reduce the invasion of S. Enteritidis in the host by attenuating virulence factors. Furthermore, CC and SG water extracts significantly improved the survival of infected C. elegans by impairing the ability of S. Enteritidis to colonize the digestive tract of the nematode and by enhancing the expression of C. elegans immune responsive genes. As the innate immune response pathways of C. elegans and mammals show a

  5. Studies on the effects of phosphine on Salmonella enterica serotype Enteritidis in culture medium and in black pepper (Piper nigrum).

    PubMed

    Castro, M F P M; Rezende, A C B; Benato, E A; Valentini, S R T; Furlani, R P Z; Tfouni, S A V

    2011-04-01

    The effect of phosphine on Salmonella enterica serotype Enteritidis inoculated in culture medium and in black pepper grains (Piper nigrum), as well as on the reduction of the microbial load of the dried and moisturized product, was verified. The postfumigation effect was verified in inoculated samples with 0.92 and 0.97 water activity (a(w)) exposed to 6 g/m(3) phosphine for 72 h, dried to 0.67 a(w), and stored for 24, 48, and 72 h. No decreases were observed in Salmonella Enteritidis populations in culture medium when fumigant concentrations up to 6 g/m(3) were applied for 48 h at 35°C. However, the colonies showed reductions in size and atypical coloration as the phosphine concentration increased. No reduction in Salmonella counts occurred on the inoculated dried samples after fumigation. On the other hand, when phosphine at concentrations of 6 g/m(3) was applied on moisturized black pepper for 72 h, decreases in Salmonella counts of around 80% were observed. The counts of total aerobic mesophilic bacterium populations of the dried and moisturized black pepper were not affected by the fumigant treatment. The results of the postfumigation studies indicated that Salmonella Enteritidis was absent in the fumigated grains after drying and storage for 72 h, indicating a promising application for this technique. It was concluded that for Salmonella Enteritidis control, phosphine fumigation could be applied to black pepper grains before drying and the producers should rigidly follow good agricultural practices, mainly during the drying process, in order to avoid product recontamination. Additional work is needed to confirm the findings with more Salmonella serotypes and strains.

  6. Allelic variation in TLR4 is linked to resistance to Salmonella Enteritidis infection in chickens.

    PubMed

    Li, Peng; Wang, Huihua; Zhao, Xingwang; Gou, Zhongyong; Liu, Ranran; Song, Yongmei; Li, Qinghe; Zheng, Maiqing; Cui, Huanxian; Everaert, Nadia; Zhao, Guiping; Wen, Jie

    2017-07-01

    Salmonella Enteritidis (SE) is a foodborne pathogen that negatively affects both animal and human health. Polymorphisms of the TLR4 gene may affect recognition by Toll-like receptor 4 (TLR4) of bacterial lipopolysaccharide (LPS), leading to differences in host resistance to pathogenic infections. The present study has investigated polymorphic loci of chicken TLR4 (ChTLR4) in ten chicken breeds, electrostatic potentials of mutant structures of TLR4, and a linkage analysis between allelic variation and survival ratio to infection with SE in specific-pathogen-free (SPF) White Leghorns. A total of 19 Single Nucleotide Polymorphisms (SNPs), of which 10 were novel, were found in chicken breeds. Seven newly identified amino acid variants (C68G, G674A, G782A, A896T, T959G, T986A, and A1104C) and previously reported important mutations (G247A, G1028A, C1147T, and A1832G) were demonstrated in the extracellular domain of the ChTLR4 gene. Significant changes in surface electrostatic potential of the ectodomain of TLR4, built by homology modeling, were observed at the Glu83Lys (G247A), Arg298Ser (A896T), Ser368Arg (A1104C), and Gln611Arg (A1832G) substitutions. Linkage analysis showed that one polymorphic locus G247A of TLR4 gene, common in all breeds examined, was significantly associated with increased resistance to SE in SPF White Leghorns chicks (log-rank P-value = 0.04). The genotypes from A1832G SNPs did not show statistically significant survival differences. This study has provided the first direct evidence that G247A substitution in ChTLR4 is associated with increased resistance to Salmonella Enteritidis. © 2017 Poultry Science Association Inc.

  7. Difference in cellular damage and cell death in thermal death time disks and high hydrostatic pressure treated Salmonella Enteritidis (ATCC13076) in liquid whole egg

    USDA-ARS?s Scientific Manuscript database

    Differences in membrane damage including leakage of intracellular UV-materials and loss of viability of Salmonella Enteritidis (ATCC13076) in liquid whole egg (LWE) following thermal-death-time (TDT) disk and high hydrostatic pressure treatments were examined. Salmonella enteritidis was inoculated ...

  8. Integration host factor is important for biofilm formation by Salmonella enterica Enteritidis.

    PubMed

    Leite, Bruna; Werle, Catierine Hirsch; Carmo, Camila Pinheiro do; Nóbrega, Diego Borin; Milanez, Guilherme Paier; Culler, Hebert Fabricio; Sircili, Marcelo Palma; Alvarez-Martinez, Cristina E; Brocchi, Marcelo

    2017-08-31

    Salmonella enterica Enteritidis forms biofilms and survives in agricultural environments, infecting poultry and eggs. Bacteria in biofilms are difficult to eradicate compared to planktonic cells, causing serious problems in industry and public health. In this study, we evaluated the role of ihfA and ihfB in biofilm formation by S. enterica Enteritidis by employing different microbiology techniques. Our data indicate that ihf mutant strains are impaired in biofilm formation, showing a reduction in matrix formation and a decrease in viability and metabolic activity. Phenotypic analysis also showed that deletion of ihf causes a deficiency in curli fimbriae expression, cellulose production and pellicle formation. These results show that integration host factor has an important regulatory role in biofilm formation by S. enterica Enteritidis. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. wksl3, a New Biocontrol Agent for Salmonella enterica Serovars Enteritidis and Typhimurium in Foods: Characterization, Application, Sequence Analysis, and Oral Acute Toxicity Study

    PubMed Central

    Kang, Hyun-Wol; Kim, Jae-Won; Jung, Tae-Sung

    2013-01-01

    Of the Salmonella enterica serovars, S. Enteritidis and S. Typhimurium are responsible for most of the Salmonella outbreaks implicated in the consumption of contaminated foods in the Republic of Korea. Because of the widespread occurrence of antimicrobial-resistant Salmonella in foods and food processing environments, bacteriophages have recently surfaced as an alternative biocontrol tool. In this study, we isolated a virulent bacteriophage (wksl3) that could specifically infect S. Enteritidis, S. Typhimurium, and several additional serovars. Transmission electron microscopy revealed that phage wksl3 belongs to the family Siphoviridae. Complete genome sequence analysis and bioinformatic analysis revealed that the DNA of phage wksl3 is composed of 42,766 bp with 64 open reading frames. Since it does not encode any phage lysogeny factors, toxins, pathogen-related genes, or food-borne allergens, phage wksl3 may be considered a virulent phage with no side effects. Analysis of genetic similarities between phage wksl3 and four of its relatives (SS3e, vB_SenS-Ent1, SE2, and SETP3) allowed wksl3 to be categorized as a SETP3-like phage. A single-dose test of oral toxicity with BALB/c mice resulted in no abnormal clinical observations. Moreover, phage application to chicken skin at 8°C resulted in an about 2.5-log reduction in the number of Salmonella bacteria during the test period. The strong, stable lytic activity, the significant reduction of the number of S. Enteritidis bacteria after application to food, and the lack of clinical symptoms of this phage suggest that wksl3 may be a useful agent for the protection of foods against S. Enteritidis and S. Typhimurium contamination. PMID:23335772

  10. Tolerance of Salmonella Enteritidis and Staphylococcus aureus to surface cleaning and household bleach.

    PubMed

    Kusumaningrum, H D; Paltinaite, R; Koomen, A J; Hazeleger, W C; Rombouts, F M; Beumer, R R

    2003-12-01

    Effective cleaning and sanitizing of food preparation sites is important because pathogens are readily spread to food contact surfaces after preparation of contaminated raw products. Tolerance of Salmonella Enteritidis and Staphylococcus aureus to surface cleaning by wiping with regular, microfiber, and antibacterial-treated cloths was investigated. Wiping with cleaning cloths resulted in a considerable reduction of microorganisms from surfaces, despite the greater difficulty in removing S. aureus than Salmonella Enteritidis. Depending on the cloth type, S. aureus were reduced on surfaces from initial numbers of approximately 10(5) CFU/100 cm2 to numbers from less than 4 CFU/100 cm2 (below the detection limit) to 100 CFU/100 cm2. Directly after the cloths were used to clean the contaminated surfaces, they contained high numbers of bacteria (10(4) to 10(5) CFU/100 cm2), except for the disposable antibacterial-treated cloths, in which no bacteria could be detected. The tolerance of these pathogens to sodium hypochlorite was studied in the suspension test and in cloths. S. aureus showed a better tolerance for sodium hypochlorite than Salmonella Enteritidis. Inactivation of microorganisms in cloths required a higher concentration of sodium hypochlorite than was needed in the suspension test. Repeated exposure to sodium hypochlorite, however, resulted in an increase in susceptibility to this compound. This study provides essential information about the transfer of bacteria when wiping surfaces and highlights the need for a hygiene procedure with cleaning cloths that sufficiently avoids cross-contamination in the household environment.

  11. Transposon Mutagenesis of Salmonella enterica Serovar Enteritidis Identifies Genes That Contribute to Invasiveness in Human and Chicken Cells and Survival in Egg Albumen

    PubMed Central

    Zhou, Xiaohui; Kim, Hye-Young; Call, Douglas R.; Guard, Jean

    2012-01-01

    Salmonella enterica serovar Enteritidis is an important food-borne pathogen, and chickens are a primary reservoir of human infection. While most knowledge about Salmonella pathogenesis is based on research conducted on Salmonella enterica serovar Typhimurium, S. Enteritidis is known to have pathobiology specific to chickens that impacts epidemiology in humans. Therefore, more information is needed about S. Enteritidis pathobiology in comparison to that of S. Typhimurium. We used transposon mutagenesis to identify S. Enteritidis virulence genes by assay of invasiveness in human intestinal epithelial (Caco-2) cells and chicken liver (LMH) cells and survival within chicken (HD-11) macrophages as a surrogate marker for virulence. A total of 4,330 transposon insertion mutants of an invasive G1 Nalr strain were screened using Caco-2 cells. This led to the identification of attenuating mutations in a total of 33 different loci, many of which include genes previously known to contribute to enteric infection (e.g., Salmonella pathogenicity island 1 [SPI-1], SPI-4, SPI-5, CS54, fliH, fljB, csgB, spvR, and rfbMN) in S. Enteritidis and other Salmonella serovars. Several genes or genomic islands that have not been reported previously (e.g., SPI-14, ksgA, SEN0034, SEN2278, and SEN3503) or that are absent in S. Typhimurium or in most other Salmonella serovars (e.g., pegD, SEN1152, SEN1393, and SEN1966) were also identified. Most mutants with reduced Caco-2 cell invasiveness also showed significantly reduced invasiveness in chicken liver cells and impaired survival in chicken macrophages and in egg albumen. Consequently, these genes may play an important role during infection of the chicken host and also contribute to successful egg contamination by S. Enteritidis. PMID:22988017

  12. Salmonella enteritidis outbreak in a restaurant chain: the continuing challenges of prevention.

    PubMed Central

    Vugia, D. J.; Mishu, B.; Smith, M.; Tavris, D. R.; Hickman-Brenner, F. W.; Tauxe, R. V.

    1993-01-01

    In 1990, a Salmonella enteritidis (SE) outbreak occurred in a restaurant chain in Pennsylvania. To determine its cause(s), we conducted a case-control study and a cohort study at one restaurant, and a survey of restaurants. Egg dishes were associated with illness (P = 0.03). Guests from one hotel eating at the restaurant had a diarrhoeal attack rate of 14%, 4.7-fold higher than among those not eating there (P = 0.04). There were no differences in egg handling between affected and unaffected restaurants. Eggs supplied to affected restaurants were medium grade AA eggs from a single farm, and were reportedly refrigerated during distribution. Human and hen SE isolates were phage type 8 and had similar plasmid profiles and antibiograms. We estimate the prevalence of infected eggs during the outbreak to be as high as 1 in 12. Typical restaurant egg-handling practices and refrigeration during distribution appear to be insufficient by themselves to prevent similar outbreaks. PMID:8432323

  13. APTES Functionalized Iron Oxide-Silver Magnetic Hetero-Nanocomposites for Selective Capture and Rapid Removal of Salmonella enteritidis from Aqueous Solution

    NASA Astrophysics Data System (ADS)

    Trang, Vu Thi; Dinh, Ngo Xuan; Lan, Hoang; Tam, Le Thi; Huy, Tran Quang; Tuan, Pham Anh; Phan, Vu Ngoc; Le, Anh-Tuan

    2018-02-01

    Magnetic nanomaterials, as a promising platform for the fast and sensitive detection of bacterial pathogens, have attracted increasing interest from researchers in recent years. In this work, by utilizing a two-step synthetic technique consisting of co-precipitation and subsequent hydrothermal reaction, followed by functionalization steps with (3-aminopropyl)triethoxysilane (APTES) and the antibody against Salmonella enteritidis, antibody-conjugated Fe3O4-Ag@APTES hetero-nanocomposites were successfully prepared. Due to the specific antibody, the developed Fe3O4-Ag@APTES@SE-Ab conjugates are capable of selectively capturing S. enteritidis at a low concentration of about 101 CFU/mL. Moreover, the prepared magnetic conjugates also revealed that the S. enteritidis could be rapidly removed from water solution in 20 min by using an external magnetic field with a removal efficiency obtained of ˜ 91.36%. These results indicated that the Fe3O4-Ag@APTES@SE-Ab conjugates are promising for the rapid selective capture and removal of bacterial pathogens from aqueous environments, and can be used for improving the detection quality of pathogens in water samples using immunosensor-based diagnostic tests.

  14. Differential roles for pathogenicity islands SPI-13 and SPI-8 in the interaction of Salmonella Enteritidis and Salmonella Typhi with murine and human macrophages.

    PubMed

    Espinoza, Rodrigo A; Silva-Valenzuela, Cecilia A; Amaya, Fernando A; Urrutia, Ítalo M; Contreras, Inés; Santiviago, Carlos A

    2017-02-15

    Salmonella pathogenicity island (SPI)-13 is conserved in many serovars of S. enterica, including S. Enteritidis, S. Typhimurium and S. Gallinarum. However, it is absent in typhoid serovars such as S. Typhi and Paratyphi A, which carry SPI-8 at the same genomic location. Because the interaction with macrophages is a critical step in Salmonella pathogenicity, in this study we investigated the role played by SPI-13 and SPI-8 in the interaction of S. Enteritidis and S. Typhi with cultured murine (RAW264.7) and human (THP-1) macrophages. Our results showed that SPI-13 was required for internalization of S. Enteritidis in murine but not human macrophages. On the other hand, SPI-8 was not required for the interaction of S. Typhi with human or murine macrophages. Of note, the presence of an intact copy of SPI-13 in a S. Typhi mutant carrying a deletion of SPI-8 did not improve its ability to be internalized by, or survive in human or murine macrophages. Altogether, our results point out to different roles for SPI-13 and SPI-8 during Salmonella infection. While SPI-13 contributes to the interaction of S. Enteritidis with murine macrophages, SPI-8 is not required in the interaction of S. Typhi with murine or human macrophages. We hypothesized that typhoid serovars have lost SPI-13 and maintained SPI-8 to improve their fitness during another phase of human infection.

  15. Nuggets of Wisdom: Salmonella Enteritidis Outbreaks and the Case for New Rules on Uncooked Frozen Processed Chicken.

    PubMed

    Hobbs, J Leigh; Warshawsky, Bryna; Maki, Anne; Zittermann, Sandra; Murphy, Allana; Majury, Anna; Middleton, Dean

    2017-04-01

    In 2014 and 2015, three Canadian Salmonella serotype Enteritidis outbreak investigations implicated uncooked, frozen, processed chicken products produced at the same establishment, namely establishment A. In November 2014, a sustained increase in the number of reported domestically acquired Salmonella Enteritidis cases in Ontario led to the first outbreak investigation, which implicated uncooked, frozen, processed chicken products produced at establishment A. In June 2015, the identification of pulsed-field gel electrophoresis patterns that had not been previously reported in Canada led to a national Salmonella Enteritidis investigation. Of 51 cases reported nationally, 35 were from Ontario. Uncooked, frozen, processed chicken products produced at establishment A were identified as the source of the outbreak, and public health action was taken as a result of this second investigation. In September 2015, a sustained increase in the number of domestically acquired Salmonella Enteritidis PT13a cases in Ontario led to a third outbreak investigation, which identified a total of 36 PT13a cases. Uncooked, frozen, processed chicken products produced at establishment A were again identified as the source of the outbreak. Outbreaks have been linked to uncooked, frozen, processed chicken products since the late 1990s. Information collected during the three outbreak investigations, and from other jurisdictions, suggests that the breaded and prebrowned appearance of the product, as well as factors related to product packaging and marketing, result in consumer misperception that this raw product is cooked. This misperception may result in mishandling and improper cooking. The three outbreaks described in this article highlight the potential ongoing risks to consumers from these products and support interventions to prevent contamination at the source level and infection at the consumer level.

  16. Type 1 fimbriae are important factors limiting the dissemination and colonization of mice by Salmonella Enteritidis and contribute to the induction of intestinal inflammation during Salmonella invasion

    PubMed Central

    Kuźmińska-Bajor, Marta; Grzymajło, Krzysztof; Ugorski, Maciej

    2015-01-01

    We have recently shown that Salmonella Gallinarum type 1 fimbriae with endogenous mannose-resistant (MR) variant of the FimH protein increase systemic dissemination of S. Gallinarum and colonization of internal organs in comparison to the S. Gallinarum fimH knockout strain or the mutant expressing mannose-sensitive (MS) FimH variant from S. Enteritidis. Elaborating from these studies, we proposed that MS variants of FimH are advantageous in gastrointestinal infections, in contrast to MR FimH variants which decrease intestinal colonization and promote their systemic spreading. To support our hypothesis, we carried out in vivo studies using mice infected with wild-type S. Enteritidis and its fimH knockout strain (S. Enteritidis), which was characterized by significantly lower adhesion and invasiveness of murine ICE-1 intestinal cells. Using bioluminescence imaging, we observed that the loss of MS FimH adhesin correlates well with the highly increased colonization of mice by these bacteria. The appearance of the mutant strain was observed much earlier than wild-type Salmonella, and mice infected with 104–107 S. Enteritidis fimH::kan CFUs had significantly (P < 0.05) shorter infection-free time than animals inoculated with wild-type S. Enteritidis. Infections caused by non-typhoid Salmonella, such as S. Enteritidis, are associated with massive inflammation of the lamina propria and lymph nodes in the intestinal tract. Therefore, we evaluated the role of MS type 1 fimbriae in the induction of cytokine expression and secretion, using murine ICE-1 intestinal cells. We showed that the expression, as well as secretion, of Il-1b, Il-6, Il-10, and Il-12b was significantly higher in cells infected with wild-type S. Enteritidis compared to cells infected with the mutant strain. Based on our results, we propose that type 1 fimbriae may play an important role in the pathogenicity of S. Enteritidis and may contribute to an intestinal inflammatory response. PMID:25914682

  17. Influence of ethanol adaptation on Salmonella enterica serovar Enteritidis survival in acidic environments and expression of acid tolerance-related genes

    USDA-ARS?s Scientific Manuscript database

    Aims: Salmonella enterica serovar Enteritidis (S. Enteritidis) can encounter mild ethanol stress during its life cycle. However, adaptation to a stressful condition may affect bacterial resistance to subsequent stresses. Hence, this work was undertaken to investigate the influences of ethanol adapta...

  18. Persistence of fecal shedding of Salmonella Enteritidis by experimentally infected laying hens housed in conventional or enriched cages

    USDA-ARS?s Scientific Manuscript database

    : Because Salmonella Enteritidis can be deposited inside eggs laid by infected hens, the prevalence of this pathogen in commercial egg-producing flocks is an important risk factor for human illness. Opportunities for the introduction, transmission, and persistence of salmonellae in poultry are poten...

  19. Persistence of fecal shedding of Salmonella Enteritidis by experimentally infected laying hens housed in conventional or enriched cages.

    USDA-ARS?s Scientific Manuscript database

    Because Salmonella Enteritidis can be deposited inside eggs laid by infected hens, the prevalence of this pathogen in commercial egg-producing flocks is an important risk factor for human illness. Opportunities for the introduction, transmission, and persistence of salmonellae in poultry are potenti...

  20. Investigation of an outbreak of Salmonella enteritidis gastroenteritis associated with consumption of eggs in a restaurant chain in Maryland.

    PubMed

    Lin, F Y; Morris, J G; Trump, D; Tilghman, D; Wood, P K; Jackman, N; Israel, E; Libonati, J P

    1988-10-01

    Salmonella enteritidis ser. enteritidis was isolated from patrons and employees of three restaurants in a restaurant chain in Maryland during August and September 1985. Isolates from all three restaurants had identical plasmid profiles; this profile was present in 13 of 40 randomly selected S. enteritidis isolates received by the Maryland state health department laboratory during a comparable time period. The outbreak in one restaurant resulted in at least 71 illnesses, with 17 persons known to have been hospitalized. Scrambled eggs served on a "breakfast bar" were implicated as the vehicle of transmission in this restaurant, with eggs a possible vehicle in another of the three restaurants. The data point out the risks associated with improper handling of eggs in food service establishments, provide further evidence for the observed association between S. enteritidis and eggs in the northeastern United States, and demonstrate the utility of plasmid analysis in investigation of outbreaks involving common Salmonella serotypes.

  1. Genetic and Phenotypic Characterization of a Salmonella enterica serovar Enteritidis Emerging Strain with Superior Intra-macrophage Replication Phenotype

    PubMed Central

    Shomer, Inna; Avisar, Alon; Desai, Prerak; Azriel, Shalhevet; Smollan, Gill; Belausov, Natasha; Keller, Nathan; Glikman, Daniel; Maor, Yasmin; Peretz, Avi; McClelland, Michael; Rahav, Galia; Gal-Mor, Ohad

    2016-01-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the ubiquitous Salmonella serovars worldwide and a major cause of food-born outbreaks, which are often associated with poultry and poultry derivatives. Here we report a nation-wide S. Enteritidis clonal outbreak that occurred in Israel during the last third of 2015. Pulsed field gel electrophoresis and whole genome sequencing identified genetically related strains that were circulating in Israel as early as 2008. Global comparison linked this outbreak strain to several clinical and marine environmental isolates that were previously isolated in California and Canada, indicating that similar strains are prevalent outside of Israel. Phenotypic comparison between the 2015 outbreak strain and other clinical and reference S. Enteritidis strains showed only limited intra-serovar phenotypic variation in growth in rich medium, invasion into Caco-2 cells, uptake by J774.1A macrophages, and host cell cytotoxicity. In contrast, significant phenotypic variation was shown among different S. Enteritidis isolates when biofilm-formation, motility, invasion into HeLa cells and uptake by THP-1 human macrophages were studied. Interestingly, the 2015 outbreak clone was found to possess superior intra-macrophage replication ability within both murine and human macrophages in comparison to the other S. Enteritidis strains studied. This phenotype is likely to play a role in the virulence and host-pathogen interactions of this emerging clone. PMID:27695450

  2. Salmonella enterica Serotype Enteritidis in French Polynesia, South Pacific, 2008–2013

    PubMed Central

    Maillard, Fiona; Mallet, Henri-Pierre; Daudens, Elise; Levy, Marc; Roy, Valérie; Branaa, Philippe; Bertrand, Sophie; Fabre, Laetitia; Weill, François-Xavier

    2015-01-01

    Outbreaks of Salmonella enterica serotype Enteritidis infections associated with eggs occurred in French Polynesia during 2008–2013. Molecular analysis of isolates by using clustered regularly interspaced short palindromic repeat polymorphisms and multilocus variable-number tandem-repeat analysis was performed. This subtyping made defining the epidemic strain, finding the source, and decontaminating affected poultry flocks possible. PMID:25988406

  3. Effect of dietary Bacillus coagulans supplementation on growth performance and immune responses of broiler chickens challenged by Salmonella enteritidis.

    PubMed

    Zhen, Wenrui; Shao, Yujing; Gong, Xiuyan; Wu, Yuanyuan; Geng, Yanqiang; Wang, Zhong; Guo, Yuming

    2018-04-11

    This study was conducted to evaluate the protective efficacy of dietary Bacillus coagulans (B. coagulans) supplementation in birds receiving Salmonella enteritidis (SE). Two hundred and forty 1-day-old Cobb broilers were randomly assigned to 2 × 2 factorial arrangements of treatments with 2 levels of dietary B. coagulans (0 or 400 mg/kg) and 2 levels of SE challenge (0 or 1 × 109 SE between d 9 to 11). Results showed that SE infection did not affect growth performance, but caused intestinal inflammation and barrier function impairment by reducing intestinal goblet cells and beneficial bacteria numbers, increasing cecal Salmonella colonization and liver Salmonella invasion, downregulating jejunal mucin-2 (at 7 and 17 d post-infection, DPI), TLR2 (at 7 and 17 DPI), TLR4 (at 17 DPI), TNFSF15 (at 7 and 17 DPI) gene mRNA levels, and upregulating jejunal IFN-γ mRNA levels (at 17 DPI) compared to uninfected birds. Moreover, SE infection also elevated the concentration of jejunal anti-Salmonella IgA and sera anti-Salmonella IgG compared to uninfected birds. However, chickens received B. coagulans diets showed significant increase in body weight gain and weight gain to feed intake ratio from d 15 to 21, alkaline phosphatase activity (at 7 DPI), cecal Lactobacilli and Bifidobacterium numbers (at 7 DPI; at 17 DPI), villous height: crypt ratio (at 17 DPI), and goblet cell numbers (at 7 and 17 DPI), whereas exhibiting reduced jejunal crypt depth (at 17 DPI), cecal Escherichia coli (at 7, 17, and 31 DPI), and Salmonella (at 7 and 17 DPI) levels compared with the non-supplemented birds, regardless of SE infection. In addition, B. coagulans supplement upregulated lysozyme mRNA levels (at 17 DPI), downregulated IFN-γ mRNA levels (at 7 and 17 DPI), showed an increased trend in Fowlicidin-2 mRNA levels (at 7 DPI) and a reduced trend in liver Salmonella load compared to the non-supplemented control. These data indicated that B. coagulans has a protective effect in SE infected

  4. Multi-locus variable-number tandem repeat analysis for outbreak studies of Salmonella enterica serotype Enteritidis

    PubMed Central

    Malorny, Burkhard; Junker, Ernst; Helmuth, Reiner

    2008-01-01

    Background Salmonella enterica subsp. enterica serotype Enteritidis is known as an important and pathogenic clonal group which continues to cause worldwide sporadic cases and outbreaks in humans. Here a new multiple-locus variable-number tandem repeat analysis (MLVA) method is reported for highly-discriminative subtyping of Salmonella Enteritidis. Emphasis was given on the most predominant phage types PT4 and PT8. The method comprises multiplex PCR specifically amplifying repeated sequences from nine different loci followed by an automatic fragment size analysis using a multicolor capillary electrophoresis instrument. A total of 240 human, animal, food and environmental isolates of S. Enteritidis including 23 definite phage types were used for development and validation. Furthermore, the MLVA types were compared to the phage types of several isolates from two recent outbreaks to determine the concordance between both methods and to estimate their in vivo stability. The in vitro stability of the two MLVA types specifically for PT4 and PT8 strains were determined by multiple freeze-thaw cycles. Results Seventy-nine different MLVA types were identified in 240 S. Enteritidis strains. The Simpson's diversity index for the MLVA method was 0.919 and Nei diversity values for the nine VNTR loci ranged from 0.07 to 0.65. Twenty-four MLVA types could be assigned to 62 PT4 strains and 21 types to 81 PT8 strains. All outbreak isolates had an indistinguishable outbreak specific MLVA type. The in vitro stability experiments showed no changes of the MLVA type compared to the original isolate. Conclusion This MLVA method is useful to discriminate S. Enteritidis strains even within a single phage type. It is easy in use, fast, and cheap compared to other high-resolution molecular methods and therefore an important tool for surveillance and outbreak studies for S. Enteritidis. PMID:18513386

  5. Multiplication of Salmonella Enteritidis in egg yolks after inoculation outside, on, and inside vitelline membranes and storage at different temperatures

    USDA-ARS?s Scientific Manuscript database

    Prompt refrigeration to restrict bacterial growth is a widely acknowledged practice for reducing the risk of egg-borne transmission of Salmonella Enteritidis to consumers. A recently published federal regulation for S. Enteritidis control requires eggs to be refrigerated within 36 after they are la...

  6. Longitudinal Monitoring of Successive Commercial Layer Flocks for Salmonella enterica Serovar Enteritidis.

    PubMed

    Denagamage, Thomas N; Patterson, Paul; Wallner-Pendleton, Eva; Trampel, Darrell; Shariat, Nikki; Dudley, Edward G; Jayarao, Bhushan M; Kariyawasam, Subhashinie

    2016-11-01

    The Pennsylvania Egg Quality Assurance Program (EQAP) provided the framework for Salmonella Enteritidis (SE) control programs, including the Food and Drug Administration (FDA) mandated Final Egg Rule, for commercial layer facilities throughout the United States. Although flocks with ≥3000 birds must comply with the FDA Final Egg Rule, smaller flocks are exempted from the rule. As a result, eggs produced by small layer flocks may pose a greater public health risk than those from larger flocks. It is also unknown if the EQAPs developed with large flocks in mind are suitable for small- and medium-sized flocks. Therefore, a study was performed to evaluate the effectiveness of best management practices included in EQAPs in reducing SE contamination of small- and medium-sized flocks by longitudinal monitoring of their environment and eggs. A total of 59 medium-sized (3000 to 50,000 birds) and small-sized (<3000 birds) flocks from two major layer production states of the United States were enrolled and monitored for SE by culturing different types of environmental samples and shell eggs for two consecutive flock cycles. Isolated SE was characterized by phage typing, pulsed-field gel electrophoresis (PFGE), and clustered regularly interspaced short palindromic repeats-multi-virulence-locus sequence typing (CRISPR-MVLST). Fifty-four Salmonella isolates belonging to 17 serovars, 22 of which were SE, were isolated from multiple sample types. Typing revealed that SE isolates belonged to three phage types (PTs), three PFGE fingerprint patterns, and three CRISPR-MVLST SE Sequence Types (ESTs). The PT8 and JEGX01.0004 PFGE pattern, the most predominant SE types associated with foodborne illness in the United States, were represented by a majority (91%) of SE. Of the three ESTs observed, 85% SE were typed as EST4. The proportion of SE-positive hen house environment during flock cycle 2 was significantly less than the flock cycle 1, demonstrating that current EQAP practices were

  7. Survival and Filamentation of Salmonella enterica Serovar Enteritidis PT4 and Salmonella enterica Serovar Typhimurium DT104 at Low Water Activity

    PubMed Central

    Mattick, K. L.; Jørgensen, F.; Legan, J. D.; Cole, M. B.; Porter, J.; Lappin-Scott, H. M.; Humphrey, T. J.

    2000-01-01

    In this study we investigated the long-term survival of and morphological changes in Salmonella strains at low water activity (aw). Salmonella enterica serovar Enteritidis PT4 and Salmonella enterica serovar Typhimurium DT104 survived at low aw for long periods, but minimum humectant concentrations of 8% NaCl (aw, 0.95), 96% sucrose (aw, 0.94), and 32% glycerol (aw, 0.92) were bactericidal under most conditions. Salmonella rpoS mutants were usually more sensitive to bactericidal levels of NaCl, sucrose, and glycerol. At a lethal aw, incubation at 37°C resulted in more rapid loss of viability than incubation at 21°C. At aw values of 0.93 to 0.98, strains of S. enterica serovar Enteritidis and S. enterica serovar Typhimurium formed filaments, some of which were at least 200 μm long. Filamentation was independent of rpoS expression. When the preparations were returned to high-aw conditions, the filaments formed septa, and division was complete within approximately 2 to 3 h. The variable survival of Salmonella strains at low aw highlights the importance of strain choice when researchers produce modelling data to simulate worst-case scenarios or conduct risk assessments based on laboratory data. The continued increase in Salmonella biomass at low aw (without a concomitant increase in microbial count) would not have been detected by traditional microbiological enumeration tests if the tests had been performed immediately after low-aw storage. If Salmonella strains form filaments in food products that have low aw values (0.92 to 0.98), there are significant implications for public health and for designing methods for microbiological monitoring. PMID:10742199

  8. Inactivation of salmonella in shell eggs by hot water immersion and its effect on quality

    USDA-ARS?s Scientific Manuscript database

    Thermal inactivation kinetics of heat resistant strains of Salmonella Enteritidis in shell eggs processed by hot water immersion were determined, and the effects of the processing on egg quality were evaluated. Shell eggs were inoculated with a composite of heat resistant Salmonella Enteritidis (SE)...

  9. Salmonella enterica Serovar Enteritidis, England and Wales, 1945–2011

    PubMed Central

    Lane, Christopher R.; LeBaigue, Susan; Esan, Oluwaseun B.; Awofisyo, Adedoyin A.; Adams, Natalie L.; Fisher, Ian S.T.; Grant, Kathie A.; Peters, Tansy M.; Larkin, Lesley; Davies, Robert H.

    2014-01-01

    In England and Wales, the emergence of Salmonella enterica serovar Enteritidis resulted in the largest and most persistent epidemic of foodborne infection attributable to a single subtype of any pathogen since systematic national microbiological surveillance was established. We reviewed 67 years of surveillance data to examine the features, underlying causes, and overall effects of S. enterica ser. Enteritidis. The epidemic was associated with the consumption of contaminated chicken meat and eggs, and a decline in the number of infections began after the adoption of vaccination and other measures in production and distribution of chicken meat and eggs. We estimate that >525,000 persons became ill during the course of the epidemic, which caused a total of 6,750,000 days of illness, 27,000 hospitalizations, and 2,000 deaths. Measures undertaken to control the epidemic have resulted in a major reduction in foodborne disease in England and Wales. PMID:24960614

  10. In ovo evaluation of FloraMax®-B11 on Marek´s disease HVT vaccine protective efficacy, hatchability, microbiota composition, morphometric analysis, and Salmonella Enteritidis infection in broiler chickens

    USDA-ARS?s Scientific Manuscript database

    Four experiments were conducted to evaluate the effect of in ovo administration of FloraMax®-B11 (FM) on Marek´s disease (MD) herpesvirus of turkeys (HVT) vaccine protective efficacy, hatchability, microbiota composition, morphometric analysis and Salmonella enteritidis (SE) infection in chickens. I...

  11. "Omics" of Food-Borne Gastroenteritis: Global Proteomic and Mutagenic Analysis of Salmonella enterica Serovar Enteritidis.

    PubMed

    Arunima, Aryashree; Yelamanchi, Soujanya D; Padhi, Chandrashekhar; Jaiswal, Sangeeta; Ryan, Daniel; Gupta, Bhawna; Sathe, Gajanan; Advani, Jayshree; Gowda, Harsha; Prasad, T S Keshava; Suar, Mrutyunjay

    2017-10-01

    Salmonella Enteritidis causes food-borne gastroenteritis by the two type three secretion systems (TTSS). TTSS-1 mediates invasion through intestinal lining, and TTSS-2 facilitates phagocytic survival. The pathogens' ability to infect effectively under TTSS-1-deficient background in host's phagocytes is poorly understood. Therefore, pathobiological understanding of TTSS-1-defective nontyphoidal Salmonellosis is highly important. We performed a comparative global proteomic analysis of the isogenic TTSS-1 mutant of Salmonella Enteritidis (M1511) and its wild-type isolate P125109. Our results showed 43 proteins were differentially expressed. Functional annotation further revealed that differentially expressed proteins belong to pathogenesis, tRNA and ncRNA metabolic processes. Three proteins, tryptophan subunit alpha chain, citrate lyase subunit alpha, and hypothetical protein 3202, were selected for in vitro analysis based on their functional annotations. Deletion mutants generated for the above proteins in the M1511 strain showed reduced intracellular survival inside macrophages in vitro. In sum, this study provides mass spectrometry-based evidence for seven hypothetical proteins, which will be subject of future investigations. Our study identifies proteins influencing virulence of Salmonella in the host. The study complements and further strengthens previously published research on proteins involved in enteropathogenesis of Salmonella and extends their role in noninvasive Salmonellosis.

  12. Salmonella enteritidis outbreak in Thailand: study by random amplified polymorphic DNA (RAPD) analysis.

    PubMed

    Kantama, L; Jayanetra, P

    1996-03-01

    An outbreak of Salmonella enteritidis in Thailand was reported in 1990. The majority of isolates were found in chicken and human throughout the country. The continuation of a high rate of spreading which is presently continuing prompted us to investigate possible clonal involvement in the outbreak. One hundred and twenty five isolates of S. enteritidis which were isolated between 1990-1993 were clonally identified by the technique of Random Amplified Polymorphic DNA (RAPD) analysis. Eight profiles were found indicating the presence of 8 clones, designated no. 1-8. The predominant clone was profile no. 4 which was encountered in 93.6% of tested isolates while the rest of the profile comprised only 0.8-1.6%. The predominant clone was distributed mainly in isolates from chickens and humans which is suggestive that the profile no. 4 is the major clone involved in this outbreak and that chickens were the source of S. enteritidis infection. The information from the Microbiology Laboratory at Ramathibodi Hospital revealed that nearly 40% of S. enteritidis were isolated from blood specimens. This may reflect the invasiveness of S. enteritidis in Thailand. We concluded that the outbreak involved the single clone, RAPD profile no. 4 which may disperse dominantly during the epidemic.

  13. Curcuma and Scutellaria plant extracts protect chickens against inflammation and Salmonella Enteritidis infection.

    PubMed

    Varmuzova, Karolina; Matulova, Marta Elsheimer; Gerzova, Lenka; Cejkova, Darina; Gardan-Salmon, Delphine; Panhéleux, Marina; Robert, Fabrice; Sisak, Frantisek; Havlickova, Hana; Rychlik, Ivan

    2015-09-01

    After a ban on the use of antibiotics as growth promoters in farm animals in the European Union in 2006, an interest in alternative products with antibacterial or anti-inflammatory properties has increased. In this study, we therefore tested the effects of extracts from Curcuma longa and Scutellaria baicalensis used as feed additives against cecal inflammation induced by heat stress or Salmonella Enteritidis (S. Enteritidis) infection in chickens. Curcuma extract alone was not enough to decrease gut inflammation induced by heat stress. However, a mixture of Curcuma and Scutellaria extracts used as feed additives decreased gut inflammation induced by heat or S. Enteritidis, decreased S. Enteritidis counts in the cecum but was of no negative effect on BW or humoral immune response. Using next-generation sequencing of 16S rRNA we found out that supplementation of feed with the 2 plant extracts had no effect on microbiota diversity. However, if the plant extract supplementation was provided to the chickens infected with S. Enteritidis, Faecalibacterium, and Lactobacillus, both bacterial genera with known positive effects on gut health were positively selected. The supplementation of chicken feed with extracts from Curcuma and Scutelleria thus may be used in poultry production to effectively decrease gut inflammation and increase chicken performance. © 2015 Poultry Science Association Inc.

  14. Salmonella Enteritidis Deposition inside Eggs after Experimental Infection of Laying Hens with Different Oral Doses

    USDA-ARS?s Scientific Manuscript database

    The continuing attribution of human Salmonella Enteritidis infections to internally contaminated eggs has necessitated the commitment of substantial public and private resources to risk reduction and testing programs for commercial laying flocks. Cost-effective risk reduction requires a detailed und...

  15. Detection of Salmonella enteritidis Using a Miniature Optical Surface Plasmon Resonance Biosensor

    NASA Astrophysics Data System (ADS)

    Son, J. R.; Kim, G.; Kothapalli, A.; Morgan, M. T.; Ess, D.

    2007-04-01

    The frequent outbreaks of foodborne illness demand rapid detection of foodborne pathogens. Unfortunately, conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Biosensors have shown great potential for the rapid detection of foodborne pathogens. Surface plasmon resonance (SPR) sensors have been widely adapted as an analysis tool for the study of various biological binding reactions. SPR biosensors could detect antibody-antigen bindings on the sensor surface by measuring either a resonance angle or refractive index value. In this study, the feasibility of a miniature SPR sensor (Spreeta, TI, USA) for detection of Salmonella enteritidis has been evaluated. Anti-Salmonella antibodies were immobilized on the gold sensor surface by using neutravidin. Salmonella could be detected by the Spreeta biosensor at concentrations down to 105 cfu/ml.

  16. Molecular detection and antimicrobial resistance profile of zoonotic Salmonella enteritidis isolated from broiler chickens in Kohat, Pakistan.

    PubMed

    Asif, Muhammad; Rahman, Hazir; Qasim, Muhammad; Khan, Taj Ali; Ullah, Waheed; Jie, Yan

    2017-05-01

    Salmonella enteritidis infection is a frequently encountered zoonotic problem, occurring with concerning regularity in recent years on a worldwide basis. The study that we undertook for the first time detected S. enteritidis and associated antimicrobial resistance pattern in broiler chickens. A total of 150 different poultry samples were first enriched and grown on selective media, and then processed for molecular detection of S. enteritidis by amplification of the spvb gene. The overall detection rate of S. enteritidis was 23.3% (n=35), while an increased detection rate of S. enteritidis was found in the chicken breast tissue (n=9; 30%). When antibiogram was tested for S. enteritidis against common antibiotics, increased resistance to ampicillin (n=29; 82.2%), tetracycline (n=28; 80%), augmentin (n=27; 77.14%), and chloramphenicol (n=19; 54.2%) was observed. Multidrug resistance was reported in 54.8% (n=19) of the S. enteritidis isolates, while 20% (n=07) of isolates were extensively drug resistant. The present study for the first time reports S. enteritidis on the basis of spvb gene detection. The increased drug resistance in S. enteritidis is an emerging problem that could negatively impact efforts to prevent and treat broiler-transmitted S. enteritidis. Copyright © 2017. Published by Elsevier Taiwan LLC.

  17. Detection of egg yolk antibodies reflecting Salmonella enteritidis infections using a surface plasmon resonance biosensor.

    PubMed

    Thomas, Ekelijn; Bouma, Annemarie; van Eerden, Ellen; Landman, Wil J M; van Knapen, Frans; Stegeman, Arjan; Bergwerff, Aldert A

    2006-08-31

    A surface plasmon resonance (SPR) biosensor assay was developed on the basis of a lipopolysaccharide antigen of Salmonella enterica serovar enteritidis (S. enterica serovar enteritidis) to detect egg yolk antibodies against S. enterica serovar enteritidis. This biosensor assay was compared to two commercial ELISA kits based on LPS antigen and flagellar antigen. A number of 163 egg yolk and combined egg white and yolk samples from chickens experimentally infected with S. enterica serovar enteritidis and 90 egg yolk and combined egg white and yolk samples from uninfected chickens were analyzed. Receiver operating characteristic analysis of the data calculated a diagnostic sensitivity of 82% and a diagnostic specificity of 100%. The within-day coefficient of variation of a positive internal-control egg yolk was 1%. The SPR biosensor assay was able to detect antibodies in a significantly higher percentage of known positive samples than the commercial ELISA's. The anticipated use of the SPR biosensor assay is to determine the S. enterica serovar enteritidis serostatus of non-vaccinated layer hens.

  18. Salmonella enteritidis infections associated with foods purchased from mobile lunch trucks--Alberta, Canada, October 2010-February 2011.

    PubMed

    2013-07-19

    During October 2010-February 2011, an outbreak of 91 Salmonella Enteritidis (SE) infections in Alberta, Canada, was investigated by a local public health department (Alberta Health Services, Calgary Zone). Index cases initially were linked through a common history of consumption of food purchased from mobile food-vending vehicles (lunch trucks) operating at worksites in Alberta. Further investigation implicated one catering company that supplied items for the lunch trucks and other vendors. In 85 cases, patients reported consumption of food prepared by the catering company in the 7 days before illness. Six patients were employees of the catering company, and two food samples collected from the catering company were positive for SE. Foods likely were contaminated directly or indirectly through the use of illegally sourced, SE-contaminated eggs at the implicated catering facility and by catering employees who were infected with SE. Public health interventions put into place to control the outbreak included screening employees for Salmonella, excluding those infected from food-handling duties, and training employees in safe food-handling procedures. No further outbreak cases were identified after full implementation of the interventions. This investigation highlights the potential for lunch trucks to be a source of foodborne illness and the need for robust regulatory compliance monitoring of lunch trucks and their food suppliers.

  19. The evaluation and application of multilocus variable number tandem repeat analysis (MLVA) for the molecular epidemiological study of Salmonella enterica subsp. enterica serovar Enteritidis infection.

    PubMed

    Liu, Yao; Shi, Xiaolu; Li, Yinghui; Chen, Qiongcheng; Jiang, Min; Li, Wanli; Qiu, Yaqun; Lin, Yiman; Jiang, Yixiang; Kan, Biao; Sun, Qun; Hu, Qinghua

    2016-01-29

    Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is one of the most prevalent Salmonella serotypes that cause gastroenteritis worldwide and the most prevalent serotype causing Salmonella infections in China. A rapid molecular typing method with high throughput and good epidemiological discrimination is urgently needed for detecting the outbreaks and finding the source for effective control of S. Enteritidis infections. In this study, 194 strains which included 47 from six outbreaks that were well-characterized epidemiologically were analyzed with pulse field gel electrophoresis (PFGE) and multilocus variable number tandem repeat analysis (MLVA). Seven VNTR loci published by the US Center for Disease Control and Prevention (CDC) were used to evaluate and develop MLVA scheme for S. Enteritidis molecular subtyping by comparing with PFGE, and then MLVA was applied to the suspected outbreaks detection. All S. Enteritidis isolates were analyzed with MLVA to establish a MLVA database in Shenzhen, Guangdong province, China to facilitate the detection of S. Enteritidis infection clusters. There were 33 MLVA types and 29 PFGE patterns among 147 sporadic isolates. These two measures had Simpson indices of 0.7701 and 0.8043, respectively, which did not differ significantly. Epidemiological concordance was evaluated by typing 47 isolates from six epidemiologically well-characterized outbreaks and it did not differ for PFGE and MLVA. We applied the well established MLVA method to detect two S. Enteritidis foodborne outbreaks and find their sources successfully in 2014. A MLVA database of 491 S. Enteritidis strains isolated from 2004 to 2014 was established for the surveillance of clusters in the future. MLVA typing of S. Enteritidis would be an effective tool for early warning and epidemiological surveillance of S. Enteritidis infections.

  20. Salmonella Enteritidis Strains from Poultry Exhibit Differential Responses to Acid Stress, Oxidative Stress, and Survival in the Egg Albumen

    PubMed Central

    Casavant, Carol; Hawley, Quincy; Addwebi, Tarek; Call, Douglas R.; Guard, Jean

    2012-01-01

    Abstract Salmonella Enteritidis is the major foodborne pathogen that is primarily transmitted by contaminated chicken meat and eggs. We recently demonstrated that Salmonella Enteritidis strains from poultry differ in their ability to invade human intestinal cells and cause disease in orally challenged mice. Here we hypothesized that the differential virulence of Salmonella Enteritidis strains is due to the differential fitness in the adverse environments that may be encountered during infection in the host. The responses of a panel of six Salmonella Enteritidis strains to acid stress, oxidative stress, survival in egg albumen, and the ability to cause infection in chickens were analyzed. This analysis allowed classification of strains into two categories, stress-sensitive and stress-resistant, with the former showing significantly (p<0.05) reduced survival in acidic (gastric phase of infection) and oxidative (intestinal and systemic phase of infection) stress. Stress-sensitive strains also showed impaired intestinal colonization and systemic dissemination in orally inoculated chickens and failed to survive/grow in egg albumen. Comparative genomic hybridization microarray analysis revealed no differences at the discriminatory level of the whole gene content between stress-sensitive and stress-resistant strains. However, sequencing of rpoS, a stress-regulatory gene, revealed that one of the three stress-sensitive strains carried an insertion mutation in the rpoS resulting in truncation of σS. Finding that one of the stress-sensitive strains carried an easily identifiable small polymorphism within a stress-response gene suggests that the other strains may also have small polymorphisms elsewhere in the genome, which likely impact regulation of stress or virulence associated genes in some manner. PMID:22304629

  1. Genomic Epidemiology of Salmonella enterica Serotype Enteritidis based on Population Structure of Prevalent Lineages

    PubMed Central

    Desai, Prerak T.; den Bakker, Henk C.; Mikoleit, Matthew; Tolar, Beth; Trees, Eija; Hendriksen, Rene S.; Frye, Jonathan G.; Porwollik, Steffen; Weimer, Bart C.; Wiedmann, Martin; Weinstock, George M.; Fields, Patricia I.; McClelland, Michael

    2014-01-01

    Salmonella enterica serotype Enteritidis is one of the most commonly reported causes of human salmonellosis. Its low genetic diversity, measured by fingerprinting methods, has made subtyping a challenge. We used whole-genome sequencing to characterize 125 S. enterica Enteritidis and 3 S. enterica serotype Nitra strains. Single-nucleotide polymorphisms were filtered to identify 4,887 reliable loci that distinguished all isolates from each other. Our whole-genome single-nucleotide polymorphism typing approach was robust for S. enterica Enteritidis subtyping with combined data for different strains from 2 different sequencing platforms. Five major genetic lineages were recognized, which revealed possible patterns of geographic and epidemiologic distribution. Analyses on the population dynamics and evolutionary history estimated that major lineages emerged during the 17th–18th centuries and diversified during the 1920s and 1950s. PMID:25147968

  2. Gene Expression Profiles of Chicken Embryo Fibroblasts in Response to Salmonella Enteritidis Infection.

    PubMed

    Szmolka, Ama; Wiener, Zoltán; Matulova, Marta Elsheimer; Varmuzova, Karolina; Rychlik, Ivan

    2015-01-01

    The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only 'non-immune' genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria.

  3. Signature-tagged mutagenesis screening revealed a novel smooth-to-rough transition determinant of Salmonella enterica serovar Enteritidis.

    PubMed

    Jiao, Yang; Guo, Rongxian; Tang, Peipei; Kang, Xilong; Yin, Junlei; Wu, Kaiyue; Geng, Shizhong; Li, Qiuchun; Sun, Jun; Xu, Xiulong; Zhou, Xiaohui; Gan, Junji; Jiao, Xinan; Liu, Xiufan; Pan, Zhiming

    2017-03-03

    Salmonella enterica serovar Enteritidis (S. Enteritidis) has emerged as one of the most important food-borne pathogens for humans. Lipopolysaccharide (LPS), as a component of the outer membrane, is responsible for the virulence and smooth-to-rough transition in S. Enteritidis. In this study, we screened S. Enteritidis signature-tagged transposon mutant library using monoclonal antibody against somatic O 9 antigen (O 9 MAb) and O 9 factor rabbit antiserum to identify novel genes that are involved in smooth-to-rough transition. A total of 480 mutants were screened and one mutant with transposon insertion in rfbG gene had smooth-to-rough transition phenotype. In order to verify the role of rfbG gene, an rfbG insertion or deletion mutant was constructed using λ-Red recombination system. Phenotypic and biological analysis revealed that rfbG insertion or deletion mutants were similar to the wild-type strain in growth rate and biochemical properties, but the swimming motility was reduced. SE Slide Agglutination test and ELISA test showed that rfbG mutants do not stimulate animals to produce agglutinating antibody. In addition, the half-lethal dose (LD 50 ) of the rfbG deletion mutant strain was 10 6.6 -fold higher than that of the parent strain in a mouse model when injected intraperitoneally. These data indicate that the rfbG gene is involved in smooth-to-rough transition, swimming motility and virulence of S. Enteritidis. Furthermore, somatic O-antigen antibody-based approach to screen signature-tagged transposon mutants is feasible to clarify LPS biosynthesis and to find suitable markers in DIVA-vaccine research.

  4. Molecular detection assay of five Salmonella serotypes of public interest: Typhimurium, Enteritidis, Newport, Heidelberg, and Hadar.

    PubMed

    Bugarel, M; Tudor, A; Loneragan, G H; Nightingale, K K

    2017-03-01

    Foodborne illnesses due to Salmonella represent an important public-health concern worldwide. In the United States, a majority of Salmonella infections are associated with a small number of serotypes. Furthermore, some serotypes that are overrepresented among human disease are also associated with multi-drug resistance phenotypes. Rapid detection of serotypes of public-health concern might help reduce the burden of salmonellosis cases and limit exposure to multi-drug resistant Salmonella. We developed a two-step real-time PCR-based rapid method for the identification and detection of five Salmonella serotypes that are either overrepresented in human disease or frequently associated with multi-drug resistance, including serotypes Enteritidis, Typhimurium, Newport, Hadar, and Heidelberg. Two sets of four markers were developed to detect and differentiate the five serotypes. The first set of markers was developed as a screening step to detect the five serotypes; whereas, the second set was used to further distinguish serotypes Heidelberg, Newport and Hadar. The utilization of these markers on a two-step investigation strategy provides a diagnostic specificity of 97% for the detection of Typhimurium, Enteritidis, Heidelberg, Infantis, Newport and Hadar. The diagnostic sensitivity of the detection makers is >96%. The availability of this two-step rapid method will facilitate specific detection of Salmonella serotypes that contribute to a significant proportion of human disease and carry antimicrobial resistance. Published by Elsevier B.V.

  5. Immunogenicity and protective efficacy of a Salmonella Enteritidis sptP mutant as a live attenuated vaccine candidate.

    PubMed

    Lin, Zhijie; Tang, Peipei; Jiao, Yang; Kang, Xilong; Li, Qiuchun; Xu, Xiulong; Sun, Jun; Pan, Zhiming; Jiao, Xinan

    2017-06-24

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a highly adaptive pathogen in both humans and animals. As a Salmonella Type III secretion system (T3SS) effector, Salmonella protein tyrosine phosphatase (SptP) is critical for virulence in this genus. To investigate the feasibility of using C50336ΔsptP as a live attenuated oral vaccine in mice, we generated the sptP gene deletion mutant C50336ΔsptP in S. Enteritidis strain C50336 by λ-Red mediated recombination and evaluated the protective ability of the S. Enteritidis sptP mutant strain C50336ΔsptP against mice salmonellosis. We found that C50336ΔsptP was a highly immunogenic, effective, and safe vaccine in mice. Compared to wild-type C50336, C50336ΔsptP showed reduced virulence as confirmed by the 50% lethal dose (LD 50 ) in orally infected mice. C50336ΔsptP also showed decreased bacterial colonization both in vivo and in vitro. Immunization with C50336ΔsptP had no significant effect on body weight and did not result in obvious clinical symptoms relative to control animals treated with phosphate-buffered saline (PBS), but induced humoral and cellular immune responses at 12 and 26 days post inoculation. Immunization with 1 × 10 8 colony-forming units (CFU) C50336ΔsptP per mouse provided 100% protection against subsequent challenge with the wild-type C50336 strain, and immunized mice showed mild and temporary clinical symptoms as compared to those of control group. These results demonstrate that C50336ΔsptP can be a live attenuated oral vaccine for salmonellosis.

  6. Feed deprivation affects crop environment and modulates Salmonella enteritidis colonization and invasion of leghorn hens.

    PubMed

    Durant, J A; Corrier, D E; Byrd, J A; Stanker, L H; Ricke, S C

    1999-05-01

    Leghorn hens over 50 weeks of age were assigned to two treatment groups designated as either unmolted controls or molted. A forced molt was induced by a 9-day feed withdrawal, and each hen was challenged orally with 10(5) Salmonella enteritidis organisms on day 4 of feed withdrawal. On days 4 and 9 of molt, the numbers of lactobacilli and the concentrations of lactate, acetate, propionate, and butyrate, and total volatile fatty acids in the crops decreased while crop pH increased significantly (P < 0.05) in the molted hens compared to the controls. S. enteritidis crop and cecal colonization, in addition to spleen and liver invasion, increased significantly (P < 0.05) in the molted hens compared to the controls. The invasive phenotype of Salmonella spp. is complex and requires several virulence genes which are regulated by the transcriptional activator HilA. Samples of the crop contents from the molted and unmolted birds were pooled separately, centrifuged, and filter sterilized. The sterile crop contents were then used to measure the expression of hilA. By using a lacZY transcriptional fusion to the hilA gene in S. enteritidis, we found that hilA expression was 1.6- to 2.1-fold higher in the crop contents from molted birds than in those from control birds in vitro. The results of the study suggest that the changes in the microenvironment of the crop caused by feed deprivation are important regulators of S. enteritidis survival and influence the susceptibility of molted hens to S. enteritidis infections. Furthermore, our in vitro results on the expression of hilA suggest that the change in crop environment during feed withdrawal has the potential to significantly affect virulence by increasing the expression of genes necessary for intestinal invasion.

  7. Feed Deprivation Affects Crop Environment and Modulates Salmonella enteritidis Colonization and Invasion of Leghorn Hens

    PubMed Central

    Durant, Juliet A.; Corrier, Donald E.; Byrd, J. Allen; Stanker, Larry H.; Ricke, Steven C.

    1999-01-01

    Leghorn hens over 50 weeks of age were assigned to two treatment groups designated as either unmolted controls or molted. A forced molt was induced by a 9-day feed withdrawal, and each hen was challenged orally with 105 Salmonella enteritidis organisms on day 4 of feed withdrawal. On days 4 and 9 of molt, the numbers of lactobacilli and the concentrations of lactate, acetate, propionate, and butyrate, and total volatile fatty acids in the crops decreased while crop pH increased significantly (P < 0.05) in the molted hens compared to the controls. S. enteritidis crop and cecal colonization, in addition to spleen and liver invasion, increased significantly (P < 0.05) in the molted hens compared to the controls. The invasive phenotype of Salmonella spp. is complex and requires several virulence genes which are regulated by the transcriptional activator HilA. Samples of the crop contents from the molted and unmolted birds were pooled separately, centrifuged, and filter sterilized. The sterile crop contents were then used to measure the expression of hilA. By using a lacZY transcriptional fusion to the hilA gene in S. enteritidis, we found that hilA expression was 1.6- to 2.1-fold higher in the crop contents from molted birds than in those from control birds in vitro. The results of the study suggest that the changes in the microenvironment of the crop caused by feed deprivation are important regulators of S. enteritidis survival and influence the susceptibility of molted hens to S. enteritidis infections. Furthermore, our in vitro results on the expression of hilA suggest that the change in crop environment during feed withdrawal has the potential to significantly affect virulence by increasing the expression of genes necessary for intestinal invasion. PMID:10223980

  8. Distribution of Salmonella Enteritidis in internal organs and variation of cecum microbiota in chicken after oral challenge.

    PubMed

    Zeng, Jinxin; Lei, Changwei; Wang, Yulong; Chen, Yanpeng; Zhang, Xiuzhong; Kang, Zhuangzhuang; Zhai, Xiwen; Ye, Xiaolan; Wang, Hongning

    2018-06-12

    The aim study was to explore the distribution of Salmonella Enteritidis (S. enteritidis) in internal organs and variation of cecum microbiota in newly hatched chicken after oral challenge during a 21-day period. The quantities of S. enteritidis DNA in different internal organs (heart, liver, spleen, stomach, pancreas, small intestine, blood and cecum contents) were determined by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR). The result showed that all of the above-mentioned samples were positive at 12 h post inoculation (PI) after oral challenge. The highest copy numbers of S. enteritidis in all tissue were heart and liver, with about 2 × 10 2 to 6 × 10 6 copies of DNA target sequences/0.5 g. The copy number of S. enteritidis in the stomach was only lower than the heart and liver. The blood at 8 d PI, the pancreas at 10 d PI, the heart at 14 d PI and the stomach at 17 d PI didn't have a positive result. However, the liver, spleen, cecum contents and small intestine were all positive during the 21-day period. The cecum contents at 0 d PI, 4 d PI and 10 d PI from the control group and experiment group were collected for bacterial 16 S rRNA sequencing targeting the V3-V4 hypervariable region. The result showed that at the 0 d PI, the main cecum microbiota ingredient of the two-day old chicken was Enterobacteriaceae (Proteobacteria) and the other microbiology species were fewer. At the 10 d PI, the microbiota ingredient of cecum became abundant and stable mainly including the families Ruminococcaceae (Firmicutes), Enterobacteriaceae (Proteobacteria), Lachnospiraceae (Firmicutes) and clostridiacaea (Firmicutes) both of the two group, suggesting Salmonella infection with 2-day old chicken might not significantly change cecum microbiota community. The study indicated the major organs, which carried numerous S. enteritidis, providing a significantly guideline for salmonella detection in poultry and revealed the main

  9. Heat stress decreases expression of the cytokines, avian β-defensins 4 and 6 and Toll-like receptor 2 in broiler chickens infected with Salmonella Enteritidis.

    PubMed

    Quinteiro-Filho, W M; Calefi, A S; Cruz, D S G; Aloia, T P A; Zager, A; Astolfi-Ferreira, C S; Piantino Ferreira, J A; Sharif, S; Palermo-Neto, J

    2017-04-01

    A high ambient temperature is a highly relevant stressor in poultry production. Heat stress (HS) has been reported to reduce animal welfare, performance indices and increase Salmonella susceptibility. Salmonella spp. are major zoonotic pathogen that cause over 1 billion of human infections worldwide annually. Therefore, the current study was designed to analyze the effect of heat stress on Salmonella infection in chickens through modulation of the immune responses. Salmonella Enteritidis was inoculated via gavage at one day of age (10 6 cfu/mL). Heat stress 31±1°C was applied from 35 to 41 days of age. Broiler chickens were divided into the following groups of 12 chickens: control (C); heat stress (HS31°C); S. Enteritidis positive control (PC); and S. Enteritidis+heat stress (PHS31°C). We observed that heat stress increased corticosterone serum levels. Concomitantly heat stress decreased (1) the IgA and IFN-γ plasmatic levels; (2) the mRNA expression of IL-6, IL-12 in spleen and IL-1β, IL-10, TGF-β in cecal tonsils; (3) the mRNA expression of AvBD4 and AvBD6 in cecal tonsils; and (4) the mRNA expression of TLR2 in spleen and cecal tonsils of chickens infected with S. Enteritidis (PHS31°C group). Heat stress also increased Salmonella colonization in the crop and caecum as well as Salmonella invasion to the spleen, liver and bone marrow, showing a deficiency in the control of S. Enteritidis induced infection. Together, the present data suggested that heat stress activated hypothalamus-pituitary-adrenal (HPA) axis, as observed by the increase in the corticosterone levels, which in turn presumably decreases the immune system activity, leading to an impairment of the intestinal mucosal barrier and increasing chicken susceptibility to the invasion of different organs by S. Enteritidis . Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Control of Salmonella Enteritidis in turkeys using organic acids and competitive exclusion product.

    PubMed

    Milbradt, E L; Zamae, J R; Araújo Júnior, J P; Mazza, P; Padovani, C R; Carvalho, V R; Sanfelice, C; Rodrigues, D M; Okamoto, A S; Andreatti Filho, R L

    2014-08-01

    To evaluate the use of organic acids (OAs) and competitive exclusion (CE) product administered continuously in the feed and transiently in drinking water on the control of Salmonella enterica subspecie enterica serotype Enteritidis (SE) prior to slaughter. The influence of treatments were evaluated on pH, population of the lactic acid bacteria (LAB) and bacteria of the family Enterobacteriaceae, concentration of volatile fatty acids and SE colonization in the crop and caecum. The birds were challenged with SE 24 h before being slaughtered, and then, the caeca and crop were removed and subjected to SE counts. Continuous administration of OAs reduced the population of bacteria from the Enterobacteriaceae family in both crop and caecum, positively influenced the butyric acid concentration and reduced SE colonization in the caecum. The diet supplemented with CE product positively influenced the quantity of LAB in the crop and caecum, elevated the butyric acid concentration and reduced both Enterobacteriaceae quantity and SE colonization in the caecum. There was no effect from administering the treatments via drinking water on the variables measured. Continuous supplementation in feed with OAs and CE product reduced SE colonization of the caeca. Supplementation of OAs and CE product in diet to turkeys can reduce the SE load, potentially leading to a lower contamination risk of meat during slaughter. © 2014 The Society for Applied Microbiology.

  11. Linear antigenic mapping of flagellin (FliC) from Salmonella enterica serovar Enteritidis with yeast surface expression system.

    PubMed

    Wang, Gaoling; Shi, Bingtian; Li, Tao; Zuo, Teng; Wang, Bin; Si, Wei; Xin, Jiuqing; Yang, Kongbin; Shi, Xuanlin; Liu, Siguo; Liu, Henggui

    2016-02-29

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major cause of food-borne illness around the world and can have significant health implications in humans, poultry and other animals. Flagellin (FliC) is the primary component of bacterial flagella. It has been shown that the FliC of S. Enteritidis is a significant antigenic structure and can elicit strong humoral responses against S. Enteritidis infection in chickens. Here, we constructed a FliC antigen library using a yeast surface expression system. Yeast cells expressing FliC peptide antigens were labeled with chicken sera against S. Enteritidis and sorted using FACS. The analyses of FliC peptides revealed that the FliC linear antigenicity in chickens resided on three domains which were able to elicit strong humoral responses in vivo. Animal experiments further revealed that the antibodies elicited by these antigenic domains were able to significantly inhibit the invasion of S. Enteritidis into the liver and spleen of chickens. These findings will facilitate our better understanding of the humoral responses elicited by FliC in chickens upon infection by S. Enteritidis. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Growth of Mycobacterium avium subsp. paratuberculosis, Escherichia coli, and Salmonella Enteritidis during Preparation and Storage of Yogurt

    PubMed Central

    Cirone, K.; Huberman, Y.; Morsella, C.; Méndez, L.; Jorge, M.; Paolicchi, F.

    2013-01-01

    The purpose of this study was to determine the viability of Mycobacterium avium subsp. paratuberculosis (MAP), Escherichia coli (E. coli), and Salmonella Enteritidis (S. Enteritidis) during preparation and refrigerated storage of yogurt. Three yogurts were prepared using pasteurized commercial milk. Each yogurt was artificially contaminated with (1) MAP, (2) E. coli + S. Enteritidis, and (3) MAP + E. coli + S. Enteritidis. Samples were taken during and after the fermentation process until day 20 after inoculation. MAP was not detected during their preparation and short-term storage but was recuperated after starting at 180 min after inoculation storage. Live bacterial counts of E. coli, and S. Enteritidis increased during the first 24 hours, followed by a slight decrease towards the end of the study. In this study it was shown how MAP, E. coli, and S. Enteritidis resisted the acidic conditions generated during the preparation of yogurt and low storage temperatures. This work contributes to current knowledge regarding survival of MAP, E. coli, and S. Enteritidis during preparation and refrigerated storage of yogurt and emphasizes the need to improve hygiene measures to ensure the absence of these pathogenic microorganisms in dairy products. PMID:24455399

  13. A pulsed field gel electrophoresis (PFGE) study that suggests a major world-wide clone of Salmonella enterica serovar Enteritidis.

    PubMed

    Pang, Jen-Chieh; Chiu, Tsai-Hsin; Helmuth, Reiner; Schroeter, Andreas; Guerra, Beatriz; Tsen, Hau-Yang

    2007-05-30

    Since human infections by Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) have been increasing world-wide over the past years and epidemiological studies have implicated the consumption of meat, poultry, eggs and egg products, elucidation of the predominant subtypes for this Salmonella spp. is important. In this study, 107 poultry and food isolates of Salmonella Enteritidis obtained from Germany were analyzed by pulsed field gel electrophoresis (PFGE), and the subtypes were compared with those of the 124 human isolates obtained in Taiwan. Results showed that for these 107 poultry and food isolates, when XbaI, SpeI and NotI were used for chromosomal DNA digestion followed by PFGE analysis, a total of 19, 20 and 19 PFGE patterns, respectively, were identified. Of them, 51 (47.7%), 52 (48.6%) and 42 (39.3%) strains belong to a single pattern of X3, S3 and N3, respectively, and 34 strains belong to a pattern combination of X3S3N3, which was the major subtype. When PFGE patterns of these 107 German isolates were compared with those of the 124 human isolates obtained in Taiwan, pattern combination of X3S3N3 was found as the most common pattern shared by isolates from both areas. PT4 is a major phage type for German and Taiwan isolates. Although most of the X3S3N3 strains are of this phage type, some strains of other PFGE patterns are also of this phage type. Since strains used in this study were unrelated, i.e., they were isolated from different origins in areas geographically far apart from each other, the PFGE study suggests a major world-wide clone of S. enterica serovar Enteritidis.

  14. Four linked outbreaks of Salmonella enteritidis phage type 4 infection--the continuing egg threat.

    PubMed

    Ejidokun, O O; Killalea, D; Cooper, M; Holmyard, S; Cross, A; Kemp, C

    2000-06-01

    Four outbreaks of Salmonella enteritidis phage type (PT) 4 occurred among guests at functions for which a single commercial caterer supplied food. Retrospective cohort studies were used to describe the epidemiology of three of these outbreaks and identify the vehicle(s) responsible. Of 172 guests at these three events, 47 fitted the clinical case definition for illness and 24 cases were confirmed to have S. enteritidis PT4 infection. Food containing raw egg was identified epidemiologically as the likely vehicle of infection in two of the three outbreaks (odds ratios (OR) and 95% confidence intervals 9.1 (2.2-39.9) and 6.9 (1.2-46.4)). Logistic regression analysis yielded OR = 10.7 (p = 0.0022) and OR = 9.3 (p = 0.015) for egg consumption in two of the outbreaks. These outbreaks highlighted the continuing need to remind the public and commercial caterers of the potential high risks of contracting salmonella from shell eggs. Education of caterers includes advice to obtain eggs and other products from reputable and identifiable suppliers.

  15. Ovine serum immunoglobulin has immunomodulatory effects in growing rats gavaged with Salmonella enteritidis.

    PubMed

    Balan, Prabhu; Han, Kyoung-Sik; Rutherfurd-Markwick, Kay; Singh, Harjinder; Moughan, Paul J

    2011-05-01

    In this study, we aimed to determine whether orally administered ovine serum Ig modulate aspects of immunity and associated gut microflora in growing rats challenged with Salmonella enteritidis. The 4 groups consisted of rats fed a casein-based control diet (BD; ungavaged) and 3 groups of rats gavaged with 1 × 10(7) viable Salmonella enteritidis and fed a BD diet, a BD diet containing freeze-dried ovine Ig (FDOI), or a BD diet containing inactivated ovine Ig (IOI). The rats were randomly allocated to 1 of the 4 diets (n = 15) and consumed it for 18 d. They were orally gavaged on d 15. Phagocytic activity of peripheral blood leukocyte and lymphocyte proliferation in the presence of the concanavalin A (ConA) were greater (P < 0.05) in the ungavaged BD- and gavaged FDOI-fed rats than in the gavaged rats fed either the BD or IOI diet. ConA-stimulated Peyer's patch cells and splenocytes from the gavaged rats fed the FDOI diet produced more IFNγ, IgA, and IgG than the gavaged rats fed either the BD or IOI diet (P < 0.05). The gavaged FDOI-fed rats had higher ileal and colonic digesta and plasma concentrations of anti-Salmonella secretory sIgA and secretory sIgG (P < 0.05). DNA analysis of a denatured gradient gel electrophoresis profile revealed that 6 of 10 bands had sequence similarity to probiotic strains of bacteria in the ileum and colon of the gavaged FDOI-fed rats. In conclusion, an ovine Ig fraction modulated various indices of immune function and associated gut microflora in growing rats inoculated with Salmonella.

  16. Detecting Salmonella Enteritidis in Laying Hens and Eggs after Experimental Infection at Different Oral Dose Levels

    USDA-ARS?s Scientific Manuscript database

    The attribution of human illness to eggs contaminated with Salmonella Enteritidis has led to substantial commitments of resources (by both government and industry) to risk reduction and testing programs in egg-laying flocks. Cost-effective application of testing requires a thorough understanding of ...

  17. Commercially laid eggs vs. discarded hatching eggs: contamination by Salmonella spp.

    PubMed

    Kottwitz, Luciana B M; Leão, Joice Aparecida; Back, Alberto; Rodrigues, Dalia dos P; Magnani, Marciane; de Oliveira, Tereza C R M

    2013-01-01

    Salmonella enterica is frequently associated with outbreaks of human salmonellosis, and products of avian origin, such as eggs and chicken meat, are the main vehicles of its transmission. The present study describes the occurrence of different serovars of Salmonella enterica and phagotypes of S. enterica serovar Enteritidis in eggs destined for human consumption. Four thousand eggs obtained from commercial egg laying farms and one thousand discarded hatching eggs from broiler farms, which were acquired at farmers' markets and informal shops, were analyzed. Salmonella spp. was isolated from 52.0% of the discarded hatching eggs, in which the predominant serovar was Enteritidis (84.6%), and the predominant Salmonella Enteritidis phagotype (PT) was PT7 (26.9%). Salmonella spp. was not isolated from eggs obtained from commercial egg laying farms. The antimicrobial resistance profile showed that 23.1% (n = 6) of the SE strains were resistant to nalidixic acid. The results suggest that the consumption of discarded hatching eggs represents an important source of Salmonella transmission to humans.

  18. Dam methylation is required for efficient biofilm production in Salmonella enterica serovar Enteritidis.

    PubMed

    Aya Castañeda, María del Rosario; Sarnacki, Sebastián Hernán; Noto Llana, Mariángeles; López Guerra, Adriana Gabriela; Giacomodonato, Mónica Nancy; Cerquetti, María Cristina

    2015-01-16

    The ecological success of Salmonella enterica to survive in different environments is due, in part, to the ability to form biofilms, something which is especially important for food industry. The aim of the current study was to evaluate the involvement of Dam methylation in biofilm production in S. Enteritidis strains. The ability to generate biofilms was analyzed in wild type and dam mutant strains. In S. Enteritidis, the absence of Dam affected the capacity to develop pellicles at the air-liquid interface and reduced the ability to form biofilm on polystyrene surfaces. Curli and cellulose production, determined by Congo red and calcofluor assays, were affected in dam mutant strains. Relative quantitative real-time PCR experiments showed that the expression of csgD and csgA genes is reduced in mutants lacking dam gene with respect to the wild type strains, whereas transcript levels of bcsA are not affected in the absence of Dam. To our knowledge, this is the first report on the participation of Dam methylation on biofilm production in Enteritidis or any other serovar of S. enterica. Results presented here suggest that changes in gene expression required for biofilm production are finely regulated by Dam methylation. Thus, Dam methylation could modulate csgD expression and upregulate the expression of factors related with biofilm production, including curli and cellulose. This study contributes to the understanding of biofilm regulation in Salmonella spp. and to the design of new strategies to prevent food contamination and humans and animals infections. Copyright © 2014. Published by Elsevier B.V.

  19. Gene Expression Profiles of Chicken Embryo Fibroblasts in Response to Salmonella Enteritidis Infection

    PubMed Central

    Szmolka, Ama; Wiener, Zoltán; Matulova, Marta Elsheimer; Varmuzova, Karolina; Rychlik, Ivan

    2015-01-01

    The response of chicken to non-typhoidal Salmonella infection is becoming well characterised but the role of particular cell types in this response is still far from being understood. Therefore, in this study we characterised the response of chicken embryo fibroblasts (CEFs) to infection with two different S. Enteritidis strains by microarray analysis. The expression of chicken genes identified as significantly up- or down-regulated (≥3-fold) by microarray analysis was verified by real-time PCR followed by functional classification of the genes and prediction of interactions between the proteins using Gene Ontology and STRING Database. Finally the expression of the newly identified genes was tested in HD11 macrophages and in vivo in chickens. Altogether 19 genes were induced in CEFs after S. Enteritidis infection. Twelve of them were also induced in HD11 macrophages and thirteen in the caecum of orally infected chickens. The majority of these genes were assigned different functions in the immune response, however five of them (LOC101750351, K123, BU460569, MOBKL2C and G0S2) have not been associated with the response of chicken to Salmonella infection so far. K123 and G0S2 were the only ’non-immune’ genes inducible by S. Enteritidis in fibroblasts, HD11 macrophages and in the caecum after oral infection. The function of K123 is unknown but G0S2 is involved in lipid metabolism and in β-oxidation of fatty acids in mitochondria. PMID:26046914

  20. Genetic Diversity and Evolution of Salmonella enterica Serovar Enteritidis Strains with Different Phage Types

    PubMed Central

    Pettengill, James; Strain, Errol; Allard, Marc W.; Ahmed, Rafiq; Zhao, Shaohua; Brown, Eric W.

    2014-01-01

    Phage typing has been used for the epidemiological surveillance of Salmonella enterica serovar Enteritidis for over 2 decades. However, knowledge of the genetic and evolutionary relationships between phage types is very limited, making differences difficult to interpret. Here, single nucleotide polymorphisms (SNPs) identified from whole-genome comparisons were used to determine the relationships between some S. Enteritidis phage types (PTs) commonly associated with food-borne outbreaks in the United States. Emphasis was placed on the predominant phage types PT8, PT13a, and PT13 in North America. With >89,400 bp surveyed across 98 S. Enteritidis isolates representing 14 distinct phage types, 55 informative SNPs were discovered within 23 chromosomally anchored loci. To maximize the discriminatory and evolutionary partitioning of these highly homogeneous strains, sequences comprising informative SNPs were concatenated into a single combined data matrix and subjected to phylogenetic analysis. The resultant phylogeny allocated most S. Enteritidis isolates into two distinct clades (clades I and II) and four subclades. Synapomorphic (shared and derived) sets of SNPs capable of distinguishing individual clades/subclades were identified. However, individual phage types appeared to be evolutionarily disjunct when mapped to this phylogeny, suggesting that phage typing may not be valid for making phylogenetic inferences. Furthermore, the set of SNPs identified here represents useful genetic markers for strain differentiation of more clonal S. Enteritidis strains and provides core genotypic markers for future development of a SNP typing scheme with S. Enteritidis. PMID:24574287

  1. A national outbreak of Salmonella enteritidis infections from ice cream. The Investigation Team.

    PubMed

    Hennessy, T W; Hedberg, C W; Slutsker, L; White, K E; Besser-Wiek, J M; Moen, M E; Feldman, J; Coleman, W W; Edmonson, L M; MacDonald, K L; Osterholm, M T

    1996-05-16

    In September 1994, the Minnesota Department of Health detected an increase in the number of reports of Salmonella enteritidis infections. After a case-control study implicated a nationally distributed brand of ice cream (Schwan's) in the outbreak, the product was recalled and further epidemiologic and microbiologic investigations were conducted. We defined an outbreak-associated case of S. enteritidis infection as one in which S. enteritidis was cultured from a person who became ill in September or October 1994. We established national surveillance and surveyed customers of the implicated manufacturer. The steps involved in the manufacture of ice cream associated with cases of S. enteritidis infection were compared with those of products not known to be associated with infection matched for the date of manufacture. Cultures for bacteria were obtained from ice cream samples, the ice cream plant, and tanker trailers that had transported the ice cream base (premix) to the plant. We estimate that S. enteritidis gastroenteritis developed in 224,000 persons in the United States after they ate Schwan's ice cream. The attack rate for consumers was 6.6 percent. Ice cream associated with infection contained a higher percentage of premix that had been transported by tanker that had carried nonpasteurized eggs immediately before (P = 0.02). S. enteritidis was isolated from 8 of 226 ice cream products (3 percent), but not from environmental samples obtained from the ice cream plant (n = 157) or tanker trailers (n = 204). This nationwide outbreak of salmonellosis was most likely the result of contamination of pasteurized ice cream premix during transport in tanker trailers that had previously carried nonpasteurized liquid eggs containing S. enteritidis. To prevent further outbreaks, food products not destined for repasteurization should be transported in dedicated containers.

  2. Inactivation of Salmonella Enteritidis on lettuces used by minimally processed vegetable industries.

    PubMed

    Silveira, Josete Bailardi; Hessel, Claudia Titze; Tondo, Eduardo Cesar

    2017-01-30

    Washing and disinfection methods used by minimally processed vegetable industries of Southern Brazil were reproduced in laboratory in order to verify their effectiveness to reduce Salmonella Enteritidis SE86 (SE86) on lettuce. Among the five industries investigated, four carried out washing with potable water followed by disinfection with 200 ppm sodium hypochlorite during different immersion times. The washing procedure alone decreased approximately 1 log CFU/g of SE86 population and immersion times of 1, 2, 5, and 15 minutes in disinfectant solution demonstrated reduction rates ranging from 2.06±0.10 log CFU/g to 3.01±0.21 log CFU/g. Rinsing alone was able to reduce counts from 0.12±0.63 log CFU/g to 1.90±1.07 log CFU/g. The most effective method was washing followed by disinfection with 200 ppm sodium hypochlorite for 15 minutes and final rinse with potable water, reaching 5.83 log CFU/g of reduction. However, no statistical differences were observed on the reduction rates after different immersion times. A time interval of 1 to 2 minutes may be an advantage to the minimally vegetable processed industries in order to optimize the process without putting at risk food safety.

  3. Influence of ethanol adaptation on Salmonella enterica serovar Enteritidis survival in acidic environments and expression of acid tolerance-related genes.

    PubMed

    He, Shoukui; Cui, Yan; Qin, Xiaojie; Zhang, Fen; Shi, Chunlei; Paoli, George C; Shi, Xianming

    2018-06-01

    Cross-protection to environmental stresses by ethanol adaptation in Salmonella poses a great threat to food safety because it can undermine food processing interventions. The ability of Salmonella enterica serovar Enteritidis (S. Enteritidis) to develop acid resistance following ethanol adaptation (5% ethanol for 1 h) was evaluated in this study. Ethanol-adapted S. Enteritidis mounted cross-tolerance to malic acid (a two-fold increase in minimum bactericidal concentration), but not to acetic, ascorbic, lactic, citric and hydrochloric acids. The population of S. Enteritidis in orange juice (pH 3.77) over a 48-h period was not significantly (p > 0.05) influenced by ethanol adaptation. However, an increased survival by 0.09-1.02 log CFU/ml was noted with ethanol-adapted cells of S. Enteritidis compared to non-adapted cells in apple juice (pH 3.57) stored at 25 °C (p < 0.05), but not at 4 °C. RT-qPCR revealed upregulation of two acid tolerance-related genes, rpoS (encoding σ S ) and SEN1564A (encoding an acid shock protein), following ethanol adaptation. The relative expression level of the acid resistance gene hdeB did not change. The resistance phenotypes and transcriptional profiles of S. Enteritidis suggest some involvement of rpoS and SEN1564A in the ethanol-induced acid tolerance mechanism. Copyright © 2017. Published by Elsevier Ltd.

  4. Quinolone Resistance Determinants of Clinical Salmonella Enteritidis in Thailand.

    PubMed

    Utrarachkij, Fuangfa; Nakajima, Chie; Changkwanyeun, Ruchirada; Siripanichgon, Kanokrat; Kongsoi, Siriporn; Pornruangwong, Srirat; Changkaew, Kanjana; Tsunoda, Risa; Tamura, Yutaka; Suthienkul, Orasa; Suzuki, Yasuhiko

    2017-10-01

    Salmonella Enteritidis has emerged as a global concern regarding quinolone resistance and invasive potential. Although quinolone-resistant S. Enteritidis has been observed with high frequency in Thailand, information on the mechanism of resistance acquisition is limited. To elucidate the mechanism, a total of 158 clinical isolates of nalidixic acid (NAL)-resistant S. Enteritidis were collected throughout Thailand, and the quinolone resistance determinants were investigated in the context of resistance levels to NAL, norfloxacin (NOR), and ciprofloxacin (CIP). The analysis of point mutations in type II topoisomerase genes and the detection of plasmid-mediated quinolone resistance genes showed that all but two harbored a gyrA mutation, the qnrS1 gene, or both. The most commonly affected codon in mutant gyrA was 87, followed by 83. Double codon mutation in gyrA was found in an isolate with high-level resistance to NAL, NOR, and CIP. A new mutation causing serine to isoleucine substitution at codon 83 was identified in eight isolates. In addition to eighteen qnrS1-carrying isolates showing nontypical quinolone resistance, one carrying both the qnrS1 gene and a gyrA mutation also showed a high level of resistance. Genotyping by multilocus variable number of tandem repeat analysis suggested a possible clonal expansion of NAL-resistant strains nationwide. Our data suggested that NAL-resistant isolates with single quinolone resistance determinant may potentially become fluoroquinolone resistant by acquiring secondary determinants. Restricted therapeutic and farming usage of quinolones is strongly recommended to prevent the emergence of fluoroquinolone-resistant isolates.

  5. Assessing the impact of egg sweating on Salmonella Enteritidis penetration into shell eggs.

    PubMed

    Gradl, Janet A; Curtis, Patricia A; Jones, Deana R; Anderson, Kenneth E

    2017-07-01

    Salmonella Enteritidis (SE) prevalence in eggs is a major concern to the egg industry. Some research has shown that egg sweating can increase Salmonella penetration into egg contents when refrigerated eggs are moved to a warmer temperature. This occurs when eggs are tempered before wash, to minimize thermal cracks. The effect of egg sweating on SE penetration into shell eggs over a 6 week storage period at 4°C was assessed. A 2 × 2 factorial of SE inoculation and egg sweating was utilized. Treatments included (SES) nalidixic acid (NA)-resistant SE inoculated and sweated, (SENS) NA-resistant SE inoculated and not sweated, (NSES) buffered peptone water (BPW) inoculated and sweated, and (NSENS) BPW inoculated and not sweated. Eggs were inoculated with 108 SE. Eggs formed condensation for approximately 17 min in a 32°C incubator. Shell rinse, shell emulsion, and egg contents were sampled then enumerated and assessed for prevalence of SE over a 6 wk storage period at 4°C. After wk 1, the SENS shell rinse had higher SE counts (0.32 log10 CFU/mL) than the other 3 treatments, where no SE was enumerated. A significant week by treatment interaction was found for the shell rinse SE detection (P < 0.05). In subsequent weeks, no SE counts were obtained from the egg shell rinse, shell emulsion, or egg contents. The SENS shell rinses had significantly higher SE prevalence than the SES rinses in weeks 1 (100% vs. 34.3%), 2 (57.6% vs. 22.2%), and 3 (38.2% vs. 11.1%) (P < 0.05). In samples from weeks 4, 5, and 6, there was no difference in SE prevalence between SES and SENS. Egg sweating did not increase SE penetration into the shell emulsion across treatment or week (P < 0.05). The decreasing trend of SE prevalence obtained over the study period indicate that refrigeration is effective at inhibiting SE growth. These results indicate that egg sweating occurring under common US egg handling practices is not harmful to egg safety. © 2017 Poultry Science Association Inc.

  6. Heat transfer models for predicting Salmonella enteritidis in shell eggs through supply chain distribution.

    PubMed

    Almonacid, S; Simpson, R; Teixeira, A

    2007-11-01

    Egg and egg preparations are important vehicles for Salmonella enteritidis infections. The influence of time-temperature becomes important when the presence of this organism is found in commercial shell eggs. A computer-aided mathematical model was validated to estimate surface and interior temperature of shell eggs under variable ambient and refrigerated storage temperature. A risk assessment of S. enteritidis based on the use of this model, coupled with S. enteritidis kinetics, has already been reported in a companion paper published earlier in JFS. The model considered the actual geometry and composition of shell eggs and was solved by numerical techniques (finite differences and finite elements). Parameters of interest such as local (h) and global (U) heat transfer coefficient, thermal conductivity, and apparent volumetric specific heat were estimated by an inverse procedure from experimental temperature measurement. In order to assess the error in predicting microbial population growth, theoretical and experimental temperatures were applied to a S. enteritidis growth model taken from the literature. Errors between values of microbial population growth calculated from model predicted compared with experimentally measured temperatures were satisfactorily low: 1.1% and 0.8% for the finite difference and finite element model, respectively.

  7. Horizontal transmission of Salmonella Enteritidis in experimentally infected laying hens housed in conventional or enriched cages

    USDA-ARS?s Scientific Manuscript database

    The majority of human illnesses caused by Salmonella Enteritidis are attributed to contaminated eggs, and the prevalence of this pathogen in commercial laying flocks has been identified as a leading epidemiologic risk factor. Flock housing and management systems can affect opportunities for the intr...

  8. [Characteristics of drug resistance and molecular typing research for Salmonella Enteritidis isolated in Henan province from 2011 to 2013].

    PubMed

    Zhao, Jiayong; Zhang, Yukai; Xie, Zhiqiang; Pan, Jingjing; Su, Jia; Mu, Yujiao; Huang, Xueyong; Zhang, Baifan; Xia, Shengli

    2016-03-01

    To investigate the antimicrobial resistance status and pulsed field gel electrophoresis (PFGE) patterns of Salmonella Enteritidis (S.Enteritidis) strains in Henan province. S. Enteritidis strains were isolated from seven sentinel hospitals from March 2011 to December 2013. According to molecular typing and Salmonella (Kirby-Bauer, K-B) drug susceptibility testing method published by the international PulseNet bacterial infectious disease monitoring network and USA Clinical and Laboratory Standards Institute (CLSI), we analyzed drug sensitivity of 8 kinds antibiotics and PFGE molecule characteristics of 120 S. Enteritidis isolates from seven sentinel hospitals. Among 120 strains of S. Enteritidis, 77 were isolated from male patients, 43 from female patients. A total of 78 strains S. Enteritidis were isolated from young children ranged from 0 to 5 years old (65.0%), including 57 strains isolated from 6 months to 2 years old (47.5%). The isolated time mainly centralized on May to October of the year, 11 strains isolated in March-April (9.2%), 48 were in May-July (40.0%),54 in August-October (45.0%), 7 in other months, with a typical summer seasonal characteristics. The resistance rate of 120 strains S. Enteritidis to ampicillin was 50.0% (n=60); to ceftazidime was 14.2% (n=17), to cefotaxime was 18.3% (n=22); to cefepime was 5.8% (n=7); to nalidixic acid was 61.7% (n=74); to ciprofloxacin was 8.3% (n=10), to norfloxacin was 5.8% (n=7); to gentamicin was 42.5% (n=51); to streptomycin was 21.7% (n=26); to chloramphenicol was 30.0% (n=36); resistance to methicillin benzyl ammonium was 11.7% (n=14), compound sulfamethoxazole resistance rate was 71.7% (n=86); the tetracycline resistant rate was 47.5% (n=57). All 120 strains of S. Enteritidis had different levels of resistance to 8 kinds of antibiotics, all strains were multidrug resistant strains, 28 isolates were resistant to 3-4 kinds of antibiotics (23.3%), 38 isolates were resistant to 5-6 kinds of antibiotics (31

  9. [Properties of Salmonella isolates in the Czech Republic].

    PubMed

    Srámová, H; Karpísková, R; Dĕdicová, D; Sisák, F; Rychlík, I

    1999-08-01

    Based on a grant project "Use and importance of epidemiological markers in Salmonella enteritidis and Salmonella typhimurium in the spread of salmonelloses in children under two years of age" implemented in 1995 to 1997, the authors investigated epidemiological markers in 1,186 salmonella isolates; the strains were isolated from faeces of 838 sick children, from 266 faeces of their contacts, from 49 specimens of incriminated foods and from 33 smears from the children's environment. Of 1,186 Salmonella isolates 999 were strains of S. enteritidis, 39 strains of S. typhimurium and 148 strains were not identified. The markers of Salmonella isolates were investigated from the aspect of biotyping--98% S. enteritidis were formed by the biovar Jena. 2% by biovar Essen; sensitivity to antibiotics--94.5% Salmonella strains were sensitive to 12 selected antibiotics, 2.9% were resistant and in 2.6% the resistance was in the intermediate zone; phagotyping--in 808 strains of S. enteritidis PT 8--88% predominated, in S. typhimurium DT 104 and DT 141; assessment of plasmid profiles--in strains of S. enteritidis plasmid 55 kb predominated, in three strains of S. typhimurium a plasmid size 95 kb; virulence--was compared in 43 strains isolated from hospitalized children with a severe clinical course with 39 strains from children treated at home. In vitro tests revealed that hospitalization of affected children was associated with virulence of the strains (SE phagotype 8) and not with age. The presented results are discussed with regard to the epidemiological situation in the Czech Republic and in the world.

  10. Evaluation of suspension array analysis for detection of egg yolk antibodies against Salmonella Enteritidis.

    PubMed

    Thomas, M E; Klinkenberg, D; Bergwerff, A A; van Eerden, E; Stegeman, J A; Bouma, A

    2010-06-01

    Salmonella enterica serovar Enteritidis (SE) is an important source of food-related diarrhoea in humans, and table eggs are considered the primordial source of contamination of the human food chain. Using eggs collected at egg-packing stations as samples could be a convenient strategy to detect colonization of layer flocks. The aim of this study was to evaluate egg yolk anti-Salmonella antibody detection using suspension array analysis. An egg yolk panel from contact-infected and non-colonized laying hens was used for the evaluation. Receiver Operating Characteristic (ROC) curves were generated to define a cut-off value and to assess the overall accuracy of the assay. The diagnostic sensitivity and specificity were estimated by maximum likelihood. Sensitivity was quantified on hen level and on sample level, and also quantified as a function of time since colonization. The area under the ROC curve was estimated at 0.984 (se 0.006, P<0.001). Of all colonized contact-infected hens, 67.6% [95% CI: 46.8, 100] developed an antibody response, which was detectable 17.4 days [14.3, 26.9] after colonization. In total, 98% [95.4, 99.4] of the 'immunopositive' hens had test positive eggs. The overall sensitivity of the immunological test was 66.7% [45.9, 98.7] and the specificity was 98.5% [97.8, 99.1]. This study provided essential parameters for optimizing surveillance programs based on detection of antibodies, and indicates that immunology based on examination of egg yolk gives important information about the Salmonella status of the flock. (c) 2010 Elsevier B.V. All rights reserved.

  11. Genetic diversity and virulence genes of Salmonella enterica subspecies enterica serotype Enteritidis isolated from meats and eggs.

    PubMed

    Fardsanei, Fatemeh; Soltan Dallal, Mohammad Mehdi; Douraghi, Masoumeh; Zahraei Salehi, Taghi; Mahmoodi, Mahmood; Memariani, Hamed; Nikkhahi, Farhad

    2017-06-01

    Salmonella enterica subspecies enterica serotype Enteritidis (S. Enteritidis) is one of the leading causes of food-borne gastroenteritis associated with the consumption of contaminated food products of animal origin. Little is known about the genetic diversity and virulence content of S. Enteritidis isolated from poultry meats and eggs in Iran. A total of 34 S. Enteritidis strains were collected from different food sources of animal origin in Tehran from May 2015 to July 2016. All of the S. Enteritidis strains were serotyped, antimicrobial susceptibility tested, and characterized for virulence genes. Pulsed-field gel electrophoresis (PFGE) was also applied for comparison of genetic relatedness. All of the strains harbored invA, hilA, ssrA, sefA, spvC, and sipA genes. A high prevalence of resistance against certain antibiotics such as cefuroxime (79.4%), nalidixic acid (47%), and ciprofloxacin (44.2%) was also observed. Regarding PFGE, S. Enteritidis strains from different sources showed considerable overlap, suggesting the lack of diversity among these isolates. Moreover, no correlation between virulence profiles or antibiotypes and PFGE clusters was observed. In conclusion, our study provided valuable information on virulence gene content, antibiotic resistance, and genetic diversity of S. Enteritidis isolated from food sources. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Contamination of eggs by Salmonella Enteritidis in experimentally infected laying hens housed in conventional or enriched cages

    USDA-ARS?s Scientific Manuscript database

    Both epidemiologic analyses and active disease surveillance confirm an ongoing strong association between human salmonellosis and the prevalence of Salmonella Enteritidis in commercial egg flocks. The majority of human illnesses caused by this pathogen are attributed to the consumption of contaminat...

  13. Antimicrobial effect of blueberry (Vaccinium corymbosum L.) extracts against the growth of Listeria monocytogenes and Salmonella Enteritidis

    USDA-ARS?s Scientific Manuscript database

    We studied the antimicrobial effects of berry extracts obtained from four cultivars (Elliott, Darrow, Bluecrop and Duke) of blueberry (Vaccinium corymbosum L.) on the growth of Listeria monocytogenes and Salmonella Enteritidis. The minimal inhibitory concentration (MIC) and minimal bactericidal conc...

  14. Development of classification models to detect Salmonella Enteritidis and Salmonella Typhimurium found in poultry carcass rinses by visible-near infrared hyperspectral imaging

    NASA Astrophysics Data System (ADS)

    Seo, Young Wook; Yoon, Seung Chul; Park, Bosoon; Hinton, Arthur; Windham, William R.; Lawrence, Kurt C.

    2013-05-01

    Salmonella is a major cause of foodborne disease outbreaks resulting from the consumption of contaminated food products in the United States. This paper reports the development of a hyperspectral imaging technique for detecting and differentiating two of the most common Salmonella serotypes, Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST), from background microflora that are often found in poultry carcass rinse. Presumptive positive screening of colonies with a traditional direct plating method is a labor intensive and time consuming task. Thus, this paper is concerned with the detection of differences in spectral characteristics among the pure SE, ST, and background microflora grown on brilliant green sulfa (BGS) and xylose lysine tergitol 4 (XLT4) agar media with a spread plating technique. Visible near-infrared hyperspectral imaging, providing the spectral and spatial information unique to each microorganism, was utilized to differentiate SE and ST from the background microflora. A total of 10 classification models, including five machine learning algorithms, each without and with principal component analysis (PCA), were validated and compared to find the best model in classification accuracy. The five machine learning (classification) algorithms used in this study were Mahalanobis distance (MD), k-nearest neighbor (kNN), linear discriminant analysis (LDA), quadratic discriminant analysis (QDA), and support vector machine (SVM). The average classification accuracy of all 10 models on a calibration (or training) set of the pure cultures on BGS agar plates was 98% (Kappa coefficient = 0.95) in determining the presence of SE and/or ST although it was difficult to differentiate between SE and ST. The average classification accuracy of all 10 models on a training set for ST detection on XLT4 agar was over 99% (Kappa coefficient = 0.99) although SE colonies on XLT4 agar were difficult to differentiate from background microflora. The average classification

  15. Risk factors for the occurrence of sporadic Salmonella enterica serotype enteritidis infections in children in France: a national case-control study.

    PubMed Central

    Delarocque-Astagneau, E.; Desenclos, J. C.; Bouvet, P.; Grimont, P. A.

    1998-01-01

    To determine risk factors associated with the occurrence of sporadic cases of Salmonella enteritidis infections among children in France, we conducted a matched case-control study. Cases were identified between 1 March and 30 September 1995. One hundred and five pairs of cases and controls matched for age and place of residence were interviewed. In the 1-5 years age group, illness was associated with the consumption of raw eggs or undercooked egg-containing foods (OR 2.4, 95% CI 1.2-4.8). Storing eggs more than 2 weeks after purchase was associated with Salmonella enteritidis infection (OR 3.8, 95% CI 1.4-10.2), particularly during the summer period (OR 6.0, 95% CI 1.3-26.8). Cases were more likely to report a case of diarrhoea in the household 10-3 days before the onset of symptoms, particularly in the age group < or = 1 year (P = 0.01). This study confirms the link between eggs and the occurrence of sporadic cases of Salmonella enteritidis among children, highlights the potential role of prolonged egg storage and underlines the role of person-to-person transmission in infants. PMID:10030705

  16. Dose-response effects in an outbreak of Salmonella enteritidis.

    PubMed Central

    Mintz, E. D.; Cartter, M. L.; Hadler, J. L.; Wassell, J. T.; Zingeser, J. A.; Tauxe, R. V.

    1994-01-01

    The effects of ingested Salmonella enteritidis (SE) dose on incubation period and on the severity and duration of illness were estimated in a cohort of 169 persons who developed gastroenteritis after eating hollandaise sauce made from grade-A shell eggs. The cohort was divided into three groups based on self-reported dose of sauce ingested. As dose increased, median incubation period decreased (37 h in the low exposure group v. 21 h in the medium exposure group v. 17.5 h in the high exposure group, P = 0.006) and greater proportions reported body aches (71 v. 85 v. 94%, P = 0.0009) and vomiting (21 v. 56 v. 57%, P = 0.002). Among 118 case-persons who completed a follow-up questionnaire, increased dose was associated with increases in median weight loss in kilograms (3.2 v. 4.5 v. 5.0, P = 0.0001), maximum daily number of stools (12.5 v. 15.0 v. 20.0, P = 0.02), subjective rating of illness severity (P = 0.0007), and the number of days of confinement to bed (3.0 v. 6.5 v. 6.5, P = 0.04). In this outbreak, ingested dose was an important determinant of the incubation period, symptoms and severity of acute salmonellosis. PMID:8119352

  17. Dose-response effects in an outbreak of Salmonella enteritidis.

    PubMed

    Mintz, E D; Cartter, M L; Hadler, J L; Wassell, J T; Zingeser, J A; Tauxe, R V

    1994-02-01

    The effects of ingested Salmonella enteritidis (SE) dose on incubation period and on the severity and duration of illness were estimated in a cohort of 169 persons who developed gastroenteritis after eating hollandaise sauce made from grade-A shell eggs. The cohort was divided into three groups based on self-reported dose of sauce ingested. As dose increased, median incubation period decreased (37 h in the low exposure group v. 21 h in the medium exposure group v. 17.5 h in the high exposure group, P = 0.006) and greater proportions reported body aches (71 v. 85 v. 94%, P = 0.0009) and vomiting (21 v. 56 v. 57%, P = 0.002). Among 118 case-persons who completed a follow-up questionnaire, increased dose was associated with increases in median weight loss in kilograms (3.2 v. 4.5 v. 5.0, P = 0.0001), maximum daily number of stools (12.5 v. 15.0 v. 20.0, P = 0.02), subjective rating of illness severity (P = 0.0007), and the number of days of confinement to bed (3.0 v. 6.5 v. 6.5, P = 0.04). In this outbreak, ingested dose was an important determinant of the incubation period, symptoms and severity of acute salmonellosis.

  18. Characterization of Chicken Spleen Transcriptome after Infection with Salmonella enterica Serovar Enteritidis

    PubMed Central

    Matulova, Marta; Rajova, Jana; Vlasatikova, Lenka; Volf, Jiri; Stepanova, Hana; Havlickova, Hana; Sisak, Frantisek; Rychlik, Ivan

    2012-01-01

    In this study we were interested in identification of new markers of chicken response to Salmonella Enteritidis infection. To reach this aim, gene expression in the spleens of naive chickens and those intravenously infected with S. Enteritidis with or without previous oral vaccination was determined by 454 pyrosequencing of splenic mRNA/cDNA. Forty genes with increased expression at the level of transcription were identified. The most inducible genes encoded avidin (AVD), extracellular fatty acid binding protein (EXFABP), immune responsive gene 1 (IRG1), chemokine ah221 (AH221), trappin-6-like protein (TRAP6) and serum amyloid A (SAA). Using cDNA from sorted splenic B-lymphocytes, macrophages, CD4, CD8 and γδ T-lymphocytes, we found that the above mentioned genes were preferentially expressed in macrophages. AVD, EXFABP, IRG1, AH221, TRAP6 and SAA were induced also in the cecum of chickens orally infected with S. Enteritidis on day 1 of life or day 42 of life. Unusual results were obtained for the immunoglobulin encoding transcripts. Prior to the infection, transcripts coding for the constant parts of IgM, IgY, IgA and Ig light chain were detected in B-lymphocytes. However, after the infection, immunoglobulin encoding transcripts were expressed also by T-lymphocytes and macrophages. Expression of AVD, EXFABP, IRG1, AH221, TRAP6, SAA and all immunoglobulin genes can be therefore used for the characterization of the course of S. Enteritidis infection in chickens. PMID:23094107

  19. Sensitivity to disinfection of bacterial indicator organisms for monitoring the Salmonella Enteritidis status of layer farms after cleaning and disinfection.

    PubMed

    Dewaele, I; Ducatelle, R; Herman, L; Heyndrickx, M; De Reu, K

    2011-06-01

    The present study evaluated Escherichia coli, Enterococcus faecalis, and Enterococcus hirae as potential indicator organisms for the possible Salmonella Enteritidis (SE) presence in layer farms after cleaning and disinfection by comparing their susceptibility to disinfection. A quantitative suspension disinfection test according to European Standard EN1656 was performed using disinfection products CID20 and Virocid (both from CID Lines, Ieper, Belgium). In a preliminary test, the sensitivity to both disinfection products was compared between ATCC strains of SE, E. coli, En. faecalis, and En. hirae. The sensitivity of SE to disinfection was most comparable to that of E. coli. A second disinfection test compared the elimination of E. coli to SE ATCC strains as well as field strains. Results showed no significant effect regarding the strain (P > 0.05 for CID20 and Virocid), meaning that no difference was detected in sensitivity toward disinfection. When comparing the sensitivity in general at species level for all concentrations of disinfectant used, no significant difference was found between E. coli and SE in sensitivity to Virocid (P > 0.05). In conclusion, because of its similar response to disinfection in a suspension disinfection test, E. coli could be used as an indicator for possible Salmonella presence after cleaning and disinfection.

  20. Emergence of host-adapted Salmonella Enteritidis through rapid evolution in an immunocompromised host.

    PubMed

    Klemm, Elizabeth J; Gkrania-Klotsas, Effrossyni; Hadfield, James; Forbester, Jessica L; Harris, Simon R; Hale, Christine; Heath, Jennifer N; Wileman, Thomas; Clare, Simon; Kane, Leanne; Goulding, David; Otto, Thomas D; Kay, Sally; Doffinger, Rainer; Cooke, Fiona J; Carmichael, Andrew; Lever, Andrew Ml; Parkhill, Julian; MacLennan, Calman A; Kumararatne, Dinakantha; Dougan, Gordon; Kingsley, Robert A

    2016-03-01

    Host adaptation is a key factor contributing to the emergence of new bacterial, viral and parasitic pathogens. Many pathogens are considered promiscuous because they cause disease across a range of host species, while others are host-adapted, infecting particular hosts 1 . Host adaptation can potentially progress to host restriction where the pathogen is strictly limited to a single host species and is frequently associated with more severe symptoms. Host-adapted and host-restricted bacterial clades evolve from within a broader host-promiscuous species and sometimes target different niches within their specialist hosts, such as adapting from a mucosal to a systemic lifestyle. Genome degradation, marked by gene inactivation and deletion, is a key feature of host adaptation, although the triggers initiating genome degradation are not well understood. Here, we show that a chronic systemic non-typhoidal Salmonella infection in an immunocompromised human patient resulted in genome degradation targeting genes that are expendable for a systemic lifestyle. We present a genome-based investigation of a recurrent blood-borne Salmonella enterica serotype Enteritidis ( S . Enteritidis) infection covering 15 years in an interleukin (IL)-12 β-1 receptor-deficient individual that developed into an asymptomatic chronic infection. The infecting S. Enteritidis harbored a mutation in the mismatch repair gene mutS that accelerated the genomic mutation rate. Phylogenetic analysis and phenotyping of multiple patient isolates provides evidence for a remarkable level of within-host evolution that parallels genome changes present in successful host-restricted bacterial pathogens but never before observed on this timescale. Our analysis identifies common pathways of host adaptation and demonstrates the role that immunocompromised individuals can play in this process.

  1. 9 CFR 147.11 - Laboratory procedure recommended for the bacteriological examination of salmonella.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... procedure recommended for the bacteriological examination of salmonella. (a) For egg- and meat-type chickens... the bacteriological examination of salmonella. 147.11 Section 147.11 Animals and Animal Products... 25 birds, and birds from Salmonella enteritidis (SE) positive environments should be cultured in...

  2. 9 CFR 147.11 - Laboratory procedure recommended for the bacteriological examination of salmonella.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... procedure recommended for the bacteriological examination of salmonella. (a) For egg- and meat-type chickens... the bacteriological examination of salmonella. 147.11 Section 147.11 Animals and Animal Products... 25 birds, and birds from Salmonella enteritidis (SE) positive environments should be cultured in...

  3. 9 CFR 147.11 - Laboratory procedure recommended for the bacteriological examination of salmonella.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... procedure recommended for the bacteriological examination of salmonella. (a) For egg- and meat-type chickens... the bacteriological examination of salmonella. 147.11 Section 147.11 Animals and Animal Products... 25 birds, and birds from Salmonella enteritidis (SE) positive environments should be cultured in...

  4. 9 CFR 147.11 - Laboratory procedure recommended for the bacteriological examination of salmonella.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... procedure recommended for the bacteriological examination of salmonella. (a) For egg- and meat-type chickens... the bacteriological examination of salmonella. 147.11 Section 147.11 Animals and Animal Products... 25 birds, and birds from Salmonella enteritidis (SE) positive environments should be cultured in...

  5. 9 CFR 147.11 - Laboratory procedure recommended for the bacteriological examination of salmonella.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... procedure recommended for the bacteriological examination of salmonella. (a) For egg- and meat-type chickens... the bacteriological examination of salmonella. 147.11 Section 147.11 Animals and Animal Products... 25 birds, and birds from Salmonella enteritidis (SE) positive environments should be cultured in...

  6. On the Evolutionary History, Population Genetics and Diversity among Isolates of Salmonella Enteritidis PFGE Pattern JEGX01.0004

    PubMed Central

    Allard, Marc W.; Luo, Yan; Strain, Errol; Pettengill, James; Timme, Ruth; Wang, Charles; Li, Cong; Keys, Christine E.; Zheng, Jie; Stones, Robert; Wilson, Mark R.; Musser, Steven M.; Brown, Eric W.

    2013-01-01

    Facile laboratory tools are needed to augment identification in contamination events to trace the contamination back to the source (traceback) of Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis). Understanding the evolution and diversity within and among outbreak strains is the first step towards this goal. To this end, we collected 106 new S. Enteriditis isolates within S. Enteriditis Pulsed-Field Gel Electrophoresis (PFGE) pattern JEGX01.0004 and close relatives, and determined their genome sequences. Sources for these isolates spanned food, clinical and environmental farm sources collected during the 2010 S. Enteritidis shell egg outbreak in the United States along with closely related serovars, S. Dublin, S. Gallinarum biovar Pullorum and S. Gallinarum. Despite the highly homogeneous structure of this population, S. Enteritidis isolates examined in this study revealed thousands of SNP differences and numerous variable genes (n = 366). Twenty-one of these genes from the lineages leading to outbreak-associated samples had nonsynonymous (causing amino acid changes) changes and five genes are putatively involved in known Salmonella virulence pathways. While chromosome synteny and genome organization appeared to be stable among these isolates, genome size differences were observed due to variation in the presence or absence of several phages and plasmids, including phage RE-2010, phage P125109, plasmid pSEEE3072_19 (similar to pSENV), plasmid pOU1114 and two newly observed mobile plasmid elements pSEEE1729_15 and pSEEE0956_35. These differences produced modifications to the assembled bases for these draft genomes in the size range of approximately 4.6 to 4.8 mbp, with S. Dublin being larger (∼4.9 mbp) and S. Gallinarum smaller (4.55 mbp) when compared to S. Enteritidis. Finally, we identified variable S. Enteritidis genes associated with virulence pathways that may be useful markers for the development of rapid surveillance and typing

  7. A large outbreak of Salmonella enteritidis phage type 4 associated with eggs from overseas.

    PubMed Central

    Stevens, A.; Joseph, C.; Bruce, J.; Fenton, D.; O'Mahony, M.; Cunningham, D.; O'Connor, B.; Rowe, B.

    1989-01-01

    In February 1989 the largest reported outbreak to date in the United Kingdom of Salmonella enteritidis phage type 4 (PT4) infection occurred following a wedding reception at a hotel. One hundred and seventy-three people met the case definition of illness of whom 118 had the organism isolated from their stools. A further 17 were found to be S. enteritidis PT4 positive, but were asymptomatic. Lightly-cooked, egg-based sauces were the epidemiologically proven vehicles of infection. Investigations showed this outbreak to be the first to implicate imported European eggs as the source of infection. An unusual feature of this outbreak was a reported incubation period of less than 3 h for some of the confirmed cases of salmonellosis. PMID:2691263

  8. Infection of Mice by Salmonella enterica Serovar Enteritidis Involves Additional Genes That Are Absent in the Genome of Serovar Typhimurium

    PubMed Central

    Silva, Cecilia A.; Blondel, Carlos J.; Quezada, Carolina P.; Porwollik, Steffen; Andrews-Polymenis, Helene L.; Toro, Cecilia S.; Zaldívar, Mercedes; Contreras, Inés

    2012-01-01

    Salmonella enterica serovar Enteritidis causes a systemic, typhoid-like infection in newly hatched poultry and mice. In the present study, a library of 54,000 transposon mutants of S. Enteritidis phage type 4 (PT4) strain P125109 was screened for mutants deficient in the in vivo colonization of the BALB/c mouse model using a microarray-based negative-selection screening. Mutants in genes known to contribute to systemic infection (e.g., Salmonella pathogenicity island 2 [SPI-2], aro, rfa, rfb, phoP, and phoQ) and enteric infection (e.g., SPI-1 and SPI-5) in this and other Salmonella serovars displayed colonization defects in our assay. In addition, a strong attenuation was observed for mutants in genes and genomic islands that are not present in S. Typhimurium or in most other Salmonella serovars. These genes include a type I restriction/modification system (SEN4290 to SEN4292), the peg fimbrial operon (SEN2144A to SEN2145B), a putative pathogenicity island (SEN1970 to SEN1999), and a type VI secretion system remnant SEN1001, encoding a hypothetical protein containing a lysin motif (LysM) domain associated with peptidoglycan binding. Proliferation defects for mutants in these individual genes and in exemplar genes for each of these clusters were confirmed in competitive infections with wild-type S. Enteritidis. A ΔSEN1001 mutant was defective for survival within RAW264.7 murine macrophages in vitro. Complementation assays directly linked the SEN1001 gene to phenotypes observed in vivo and in vitro. The genes identified here may perform novel virulence functions not characterized in previous Salmonella models. PMID:22083712

  9. Multiplication in egg yolk and survival in egg albumen of genetically and phenotypically characterized Salmonella Enteritidis strains

    USDA-ARS?s Scientific Manuscript database

    Prompt refrigeration of eggs to prevent the multiplication of Salmonella Enteritidis to high levels during storage is an important practice for reducing the risk of egg-transmitted human illness. The efficacy of egg refrigeration for achieving this goal depends on the location of contamination, the ...

  10. Drastic decrease of Salmonella Enteritidis isolated from humans in Belgium in 2005, shift in phage types and influence on foodborne outbreaks

    PubMed Central

    COLLARD, J. M.; BERTRAND, S.; DIERICK, K.; GODARD, C.; WILDEMAUWE, C.; VERMEERSCH, K.; DUCULOT, J.; VAN IMMERSEEL, F.; PASMANS, F.; IMBERECHTS, H.; QUINET, C.

    2008-01-01

    SUMMARY In Belgium, non-typhoidal salmonellosis and campylobacteriosis are the two most frequently reported foodborne illnesses. During 2005, a 71% decrease of Salmonella Enteritidis infections compared with the average annual number cases in the period 2000–2004 was recorded by the Belgian National Reference Centre for Salmonella and Shigella. After the peak of 1999, the total number of salmonellosis cases decreased gradually, with the exception of 2003 when an increase was again recorded due to the rise of isolates belonging to the serotype Enteritidis. PT4, the predominant phage type of serotype Enteriditis over recent years (except in 2003), became the second most prevalent phage type in 2005 after PT21. We present in this paper the epidemiology (incidence and trends) of human salmonellosis in Belgium and assess the role of the vaccination programme in layer flocks on the decline of the incidence of human salmonellosis and foodborne outbreaks due to S. Enteritidis. PMID:17645812

  11. Frequency and persistence of fecal shedding of Salmonella Enteritidis by experimentally infected laying hens housed in enriched colony cages at different stocking densities

    USDA-ARS?s Scientific Manuscript database

    Human infections with Salmonella Enteritidis are often attributed to the consumption of contaminated eggs, so the prevalence of this pathogen in egg-laying poultry is an important public health risk factor. Numerous and complex environmental influences on Salmonella persistence and transmission are ...

  12. Frequency and persistence of fecal shedding of Salmonella Enteritidis by experimentally infected laying hens housed in enriched colony cages at different stocking densities

    USDA-ARS?s Scientific Manuscript database

    Human Salmonella Enteritidis infections are often linked with consuming contaminated eggs, so the prevalence of this pathogen in egg-laying poultry is an important risk factor for public health. Salmonella persistence and transmission in commercial egg producing flocks are influenced by the complex ...

  13. Outer Membrane Vesicles Derived from Salmonella Enteritidis Protect against the Virulent Wild-Type Strain Infection in a Mouse Model.

    PubMed

    Liu, Qiong; Yi, Jie; Liang, Kang; Zhang, Xiangmin; Liu, Qing

    2017-08-28

    Foodborne contamination and salmonellosis caused by Salmonella Enteritidis ( S . Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S . Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S . Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold LD 50 ) of the wild-type S . Enteritidis infection. These results indicated that S . Enteritidis OMVs might be an ideal vaccine strategy for preventing S . Enteritidis diseases.

  14. Chicken-Specific Kinome Array Reveals that Salmonella enterica Serovar Enteritidis Modulates Host Immune Signaling Pathways in the Cecum to Establish a Persistence Infection

    PubMed Central

    Kogut, Michael H.; Swaggerty, Christina L.; Byrd, James Allen; Selvaraj, Ramesh; Arsenault, Ryan J.

    2016-01-01

    Non-typhoidal Salmonella enterica induces an early, short-lived pro-inflammatory response in chickens that is asymptomatic of clinical disease and results in a persistent colonization of the gastrointestinal (GI) tract that transmits infections to naïve hosts via fecal shedding of bacteria. The underlying mechanisms that control this persistent colonization of the ceca of chickens by Salmonella are only beginning to be elucidated. We hypothesize that alteration of host signaling pathways mediate the induction of a tolerance response. Using chicken-specific kinomic immune peptide arrays and quantitative RT-PCR of infected cecal tissue, we have previously evaluated the development of disease tolerance in chickens infected with Salmonella enterica serovar Enteritidis (S. Enteritidis) in a persistent infection model (4–14 days post infection). Here, we have further outlined the induction of an tolerance defense strategy in the cecum of chickens infected with S. Enteritidis beginning around four days post-primary infection. The response is characterized by alterations in the activation of T cell signaling mediated by the dephosphorylation of phospholipase c-γ1 (PLCG1) that inhibits NF-κB signaling and activates nuclear factor of activated T-cells (NFAT) signaling and blockage of interferon-γ (IFN-γ) production through the disruption of the JAK-STAT signaling pathway (dephosphorylation of JAK2, JAK3, and STAT4). Further, we measured a significant down-regulation reduction in IFN-γ mRNA expression. These studies, combined with our previous findings, describe global phenotypic changes in the avian cecum of Salmonella Enteritidis-infected chickens that decreases the host responsiveness resulting in the establishment of persistent colonization. The identified tissue protein kinases also represent potential targets for future antimicrobial compounds for decreasing Salmonella loads in the intestines of food animals before going to market. PMID:27472318

  15. Chicken-Specific Kinome Array Reveals that Salmonella enterica Serovar Enteritidis Modulates Host Immune Signaling Pathways in the Cecum to Establish a Persistence Infection.

    PubMed

    Kogut, Michael H; Swaggerty, Christina L; Byrd, James Allen; Selvaraj, Ramesh; Arsenault, Ryan J

    2016-07-27

    Non-typhoidal Salmonella enterica induces an early, short-lived pro-inflammatory response in chickens that is asymptomatic of clinical disease and results in a persistent colonization of the gastrointestinal (GI) tract that transmits infections to naïve hosts via fecal shedding of bacteria. The underlying mechanisms that control this persistent colonization of the ceca of chickens by Salmonella are only beginning to be elucidated. We hypothesize that alteration of host signaling pathways mediate the induction of a tolerance response. Using chicken-specific kinomic immune peptide arrays and quantitative RT-PCR of infected cecal tissue, we have previously evaluated the development of disease tolerance in chickens infected with Salmonella enterica serovar Enteritidis (S. Enteritidis) in a persistent infection model (4-14 days post infection). Here, we have further outlined the induction of an tolerance defense strategy in the cecum of chickens infected with S. Enteritidis beginning around four days post-primary infection. The response is characterized by alterations in the activation of T cell signaling mediated by the dephosphorylation of phospholipase c-γ1 (PLCG1) that inhibits NF-κB signaling and activates nuclear factor of activated T-cells (NFAT) signaling and blockage of interferon-γ (IFN-γ) production through the disruption of the JAK-STAT signaling pathway (dephosphorylation of JAK2, JAK3, and STAT4). Further, we measured a significant down-regulation reduction in IFN-γ mRNA expression. These studies, combined with our previous findings, describe global phenotypic changes in the avian cecum of Salmonella Enteritidis-infected chickens that decreases the host responsiveness resulting in the establishment of persistent colonization. The identified tissue protein kinases also represent potential targets for future antimicrobial compounds for decreasing Salmonella loads in the intestines of food animals before going to market.

  16. Postharvest transfer and survival of Salmonella enterica serotype enteritidis on living lettuce.

    PubMed

    Waitt, J A; Kuhn, D D; Welbaum, G E; Ponder, M A

    2014-02-01

    The potential for postharvest transfer of Salmonella to 'living lettuce' is not well understood. In this study, the transfer of Salmonella enterica Enteritidis (6 log CFU g(-1) ) from worker hands or contaminated roots to leaves of living lettuce was quantified. Transfer rates of Salmonella from contaminated gloves to sequentially handled lettuce heads ranged from 94% to head 1, 82% to head 2 and 69% to head 3. On average, 2.9 ± 0.1 log CFU g(-1) (64%) Salmonella was transferred from inoculated roots to leaves resulting from typical postharvest handling activities for living lettuce. Salmonella persisted on leaves stored at recommended storage temperatures (4°C) and increased 0.5 log CFU g(-1) when stored at temperature abuse conditions (12°C). Salmonella increased 1.6 log CFU g(-1) on roots after 18-day storage at 12°C, emphasizing the need to maintain temperature control to reduce the risk of human illness. Hydroponically grown lettuce packaged in plastic clamshells with intact roots, marketed as 'living lettuce', is increasing in popularity due to its extended shelf life. This study demonstrates the transfer of Salmonella from contaminated worker hands and contaminated roots to leaves where it persisted at 4°C for 18 day. Temperature abuse (12°C) increased Salmonella on roots and leaves. These findings suggest that failure to maintain temperatures below 12°C can pose a risk for consumers purchasing living lettuce at markets where recommended storage temperatures are not maintained. © 2013 The Society for Applied Microbiology.

  17. Lipopolysaccharide-Specific but Not Anti-Flagellar Immunoglobulin A Monoclonal Antibodies Prevent Salmonella enterica Serotype Enteritidis Invasion and Replication within HEp-2 Cell Monolayers

    PubMed Central

    Iankov, Ianko D.; Petrov, Dragomir P.; Mladenov, Ivan V.; Haralambieva, Iana H.; Mitov, Ivan G.

    2002-01-01

    The protective potential of immunoglobulin A (IgA) monoclonal antibodies (MAbs) directed against O and H antigens of Salmonella enterica serotype Enteritidis to prevent bacterial adhesion to and invasion of HEp-2 cells was evaluated. Although anti-flagellar IgA MAbs showed strong agglutinating capacities, they did not protect cell monolayers. In contrast, IgA MAbs specific for the O:9 epitope of Salmonella lipopolysaccharide antigen alone prevented S. enterica serotype Enteritidis entry and replication within HEp-2 cells, and the protection was not mediated by direct binding of antibodies to bacterial adhesins or by agglutination of microorganisms. PMID:11854252

  18. Combining Lactic Acid Spray with Near-Infrared Radiation Heating To Inactivate Salmonella enterica Serovar Enteritidis on Almond and Pine Nut Kernels

    PubMed Central

    Ha, Jae-Won

    2015-01-01

    The aim of this study was to investigate the efficacy of near-infrared radiation (NIR) heating combined with lactic acid (LA) sprays for inactivating Salmonella enterica serovar Enteritidis on almond and pine nut kernels and to elucidate the mechanisms of the lethal effect of the NIR-LA combined treatment. Also, the effect of the combination treatment on product quality was determined. Separately prepared S. Enteritidis phage type (PT) 30 and non-PT 30 S. Enteritidis cocktails were inoculated onto almond and pine nut kernels, respectively, followed by treatments with NIR or 2% LA spray alone, NIR with distilled water spray (NIR-DW), and NIR with 2% LA spray (NIR-LA). Although surface temperatures of nuts treated with NIR were higher than those subjected to NIR-DW or NIR-LA treatment, more S. Enteritidis survived after NIR treatment alone. The effectiveness of NIR-DW and NIR-LA was similar, but significantly more sublethally injured cells were recovered from NIR-DW-treated samples. We confirmed that the enhanced bactericidal effect of the NIR-LA combination may not be attributable to cell membrane damage per se. NIR heat treatment might allow S. Enteritidis cells to become permeable to applied LA solution. The NIR-LA treatment (5 min) did not significantly (P > 0.05) cause changes in the lipid peroxidation parameters, total phenolic contents, color values, moisture contents, and sensory attributes of nut kernels. Given the results of the present study, NIR-LA treatment may be a potential intervention for controlling food-borne pathogens on nut kernel products. PMID:25911473

  19. In-Feed Supplementation of trans-Cinnamaldehyde Reduces Layer-Chicken Egg-Borne Transmission of Salmonella enterica Serovar Enteritidis

    PubMed Central

    Upadhyaya, Indu; Upadhyay, Abhinav; Kollanoor-Johny, Anup; Mooyottu, Shankumar; Baskaran, Sangeetha A.; Yin, Hsin-Bai; Schreiber, David T.; Khan, Mazhar I.; Darre, Michael J.; Curtis, Patricia A.

    2015-01-01

    Salmonella enterica serovar Enteritidis is a major foodborne pathogen in the United States, causing gastroenteritis in humans, primarily through consumption of contaminated eggs. Chickens are the reservoir host of S. Enteritidis. In layer hens, S. Enteritidis colonizes the intestine and migrates to various organs, including the oviduct, leading to egg contamination. This study investigated the efficacy of in-feed supplementation with trans-cinnamaldehyde (TC), a generally recognized as safe (GRAS) plant compound obtained from cinnamon, in reducing S. Enteritidis cecal colonization and systemic spread in layers. Additionally, the effect of TC on S. Enteritidis virulence factors critical for macrophage survival and oviduct colonization was investigated in vitro. The consumer acceptability of eggs was also determined by a triangle test. Supplementation of TC in feed for 66 days at 1 or 1.5% (vol/wt) for 40- or 25-week-old layer chickens decreased the amounts of S. Enteritidis on eggshell and in yolk (P < 0.001). Additionally, S. Enteritidis persistence in the cecum, liver, and oviduct in TC-supplemented birds was decreased compared to that in controls (P < 0.001). No significant differences in feed intake, body weight, or egg production in birds or in consumer acceptability of eggs were observed (P > 0.05). In vitro cell culture assays revealed that TC reduced S. Enteritidis adhesion to and invasion of primary chicken oviduct epithelial cells and reduced S. Enteritidis survival in chicken macrophages (P < 0.001). Follow-up gene expression analysis using real-time quantitative PCR (qPCR) showed that TC downregulated the expression of S. Enteritidis virulence genes critical for chicken oviduct colonization (P < 0.001). The results suggest that TC may potentially be used as a feed additive to reduce egg-borne transmission of S. Enteritidis. PMID:25710365

  20. RNA Sequencing Reveals Differences between the Global Transcriptomes of Salmonella enterica Serovar Enteritidis Strains with High and Low Pathogenicities

    PubMed Central

    2014-01-01

    Salmonella enterica serovar Enteritidis is one of the important causes of bacterial food-borne gastroenteritis worldwide. Field strains of S. Enteritidis are relatively genetically homogeneous; however, they show extensive phenotypic diversity and differences in virulence potential. RNA sequencing (RNA-Seq) was used to characterize differences in the global transcriptome between several genetically similar but phenotypically diverse poultry-associated field strains of S. Enteritidis grown in laboratory medium at avian body temperature (42°C). These S. Enteritidis strains were previously characterized as high-pathogenicity (HP; n = 3) and low-pathogenicity (LP; n = 3) strains based on both in vitro and in vivo virulence assays. Using the negative binomial distribution-based statistical tools edgeR and DESeq, 252 genes were identified as differentially expressed in LP strains compared with their expression in the HP strains (P < 0.05). A majority of genes (235, or 93.2%) showed significantly reduced expression, whereas a few genes (17, or 6.8%) showed increased expression in all LP strains compared with HP strains. LP strains showed a unique transcriptional profile that is characterized by significantly reduced expression of several transcriptional regulators and reduced expression of genes involved in virulence (e.g., Salmonella pathogenicity island 1 [SPI-1], SPI-5, and fimbrial and motility genes) and protection against osmotic, oxidative, and other stresses, such as iron-limiting conditions commonly encountered within the host. Several functionally uncharacterized genes also showed reduced expression. This study provides a first concise view of the global transcriptional differences between field strains of S. Enteritidis with various levels of pathogenicity, providing the basis for future functional characterization of several genes with potential roles in virulence or stress regulation of S. Enteritidis. PMID:24271167

  1. Studies of contamination of three broiler breeder houses with Salmonella enteritidis before and after cleansing and disinfection.

    PubMed

    Davies, R H; Wray, C

    1996-01-01

    Three broiler breeder houses on three different sites were sampled before and after cleansing and disinfection. None of the farms achieved total elimination of Salmonella enteritidis from the poultry house environment but substantial improvements were seen when errors in the cleansing and disinfection protocol in the first house had been corrected. Fundamental errors such as over-dilution and inconsistent application of disinfectants were observed despite supervision of the process by technical advisors. In each of the three poultry units failure to eliminate a mouse population that was infected with S. enteritidis was likely to be the most important hazard for the next flock.

  2. Toll-Like Receptor Activation by Generalized Modules for Membrane Antigens from Lipid A Mutants of Salmonella enterica Serovars Typhimurium and Enteritidis.

    PubMed

    Rossi, Omar; Caboni, Mariaelena; Negrea, Aurel; Necchi, Francesca; Alfini, Renzo; Micoli, Francesca; Saul, Allan; MacLennan, Calman A; Rondini, Simona; Gerke, Christiane

    2016-04-01

    Invasive nontyphoidal Salmonella (iNTS) disease is a neglected disease with high mortality in children and HIV-positive individuals in sub-Saharan Africa, caused primarily by Africa-specific strains of Salmonella enterica serovars Typhimurium and Enteritidis. A vaccine using GMMA (generalized modules for membrane antigens) fromS.Typhimurium andS.Enteritidis containing lipid A modifications to reduce potential in vivo reactogenicity is under development. GMMA with penta-acylated lipid A showed the greatest reduction in the level of cytokine release from human peripheral blood monocytes from that for GMMA with wild-type lipid A. Deletion of the lipid A modification genes msbB and pagP was required to achieve pure penta-acylation. Interestingly, ΔmsbBΔ pagP GMMA from S. Enteritidis had a slightly higher stimulatory potential than those from S. Typhimurium, a finding consistent with the higher lipopolysaccharide (LPS) content and Toll-like receptor 2 (TLR2) stimulatory potential of the former. Also, TLR5 ligand flagellin was found in Salmonella GMMA. No relevant contribution to the stimulatory potential of GMMA was detected even when the flagellin protein FliC from S. Typhimurium was added at a concentration as high as 10% of total protein, suggesting that flagellin impurities are not a major factor for GMMA-mediated immune stimulation. Overall, the stimulatory potential of S. Typhimurium and S. Enteritidis ΔmsbB ΔpagP GMMA was close to that of Shigella sonnei GMMA, which are currently in phase I clinical trials. Copyright © 2016 Rossi et al.

  3. Application of Fourier transform infrared spectroscopy and chemometrics for differentiation of Salmonella enterica serovar Enteritidis phage types.

    PubMed

    Preisner, Ornella; Guiomar, Raquel; Machado, Jorge; Menezes, José Cardoso; Lopes, João Almeida

    2010-06-01

    Fourier transform infrared (FT-IR) spectroscopy and chemometric techniques were used to discriminate five closely related Salmonella enterica serotype Enteritidis phage types, phage type 1 (PT1), PT1b, PT4b, PT6, and PT6a. Intact cells and outer membrane protein (OMP) extracts from bacterial cell membranes were subjected to FT-IR analysis in transmittance mode. Spectra were collected over a wavenumber range from 4,000 to 600 cm(-1). Partial least-squares discriminant analysis (PLS-DA) was used to develop calibration models based on preprocessed FT-IR spectra. The analysis based on OMP extracts provided greater separation between the Salmonella Enteritidis PT1-PT1b, PT4b, and PT6-PT6a groups than the intact cell analysis. When these three phage type groups were considered, the method based on OMP extract FT-IR spectra was 100% accurate. Moreover, complementary local models that considered only the PT1-PT1b and PT6-PT6a groups were developed, and the level of discrimination increased. PT1 and PT1b isolates were differentiated successfully with the local model using the entire OMP extract spectrum (98.3% correct predictions), whereas the accuracy of discrimination between PT6 and PT6a isolates was 86.0%. Isolates belonging to different phage types (PT19, PT20, and PT21) were used with the model to test its robustness. For the first time it was demonstrated that FT-IR analysis of OMP extracts can be used for construction of robust models that allow fast and accurate discrimination of different Salmonella Enteritidis phage types.

  4. Evaluation of a selected lactic acid bacteria-based probiotic on Salmonella enterica serovar Enteritidis colonization and intestinal permeability in broiler chickens.

    PubMed

    Prado-Rebolledo, Omar F; Delgado-Machuca, Jaime de Jesus; Macedo-Barragan, Rafael J; Garcia-Márquez, Luis J; Morales-Barrera, Jesus E; Latorre, Juan D; Hernandez-Velasco, Xochitl; Tellez, Guillermo

    2017-02-01

    Two experiments were conducted to evaluate the effect of a lactic acid bacteria-based probiotic (FloraMax-B11 ® ) against Salmonella enterica serovar Enteritidis intestinal colonization and intestinal permeability in broiler chickens. Experiment 1 consisted of two independent trials. In each trial, day-old broiler chicks were assigned to one of two groups: control + S. Enteritidis or probiotic + S. Enteritidis. At 72 h post-S. Enteritidis challenge, haematology and caecal content were evaluated for S. Enteritidis colonization. In Experiment 2, day-old broiler chicks were assigned to one of four groups: negative control; probiotic; control + S. Enteritidis; or probiotic + S. Enteritidis. At 72 h post-S. Enteritidis challenge, chickens in all groups were given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). In both trials of Experiment 1, a significant reduction (P < 0.05) in colony-forming units/gram of S. Enteritidis in caecal content and a reduction in the incidence of S. Enteritidis enriched caecal samples were observed in probiotic + S. Enteritidis chickens. In addition, significant heterophilia and lymphopaenia were observed in control + S. Enteritidis chickens. In Experiment 2, a decrease in numbers of S. Enteritidis in caeca were observed in probiotic + S. Enteritidis chickens when compared to control + S. Enteritidis. Also, an increase in serum FITC-d concentration was detected in control + S. Enteritidis. These results suggest that early infection with S. Enteritidis can increase intestinal permeability, but the adverse effects can be prevented by the administration of the probiotic tested.

  5. Numbers of Salmonella enteritidis in the contents of naturally contaminated hens' eggs.

    PubMed Central

    Humphrey, T. J.; Whitehead, A.; Gawler, A. H.; Henley, A.; Rowe, B.

    1991-01-01

    Over 5700 hens eggs from 15 flocks naturally infected with Salmonella enteritidis were examined individually for the presence of the organism in either egg contents or on shells. Thirty-two eggs (0.6%) were positive in the contents. In the majority, levels of contamination were low. Three eggs, however, were found to contain many thousands of cells. In eggs where it was possible to identify the site of contamination, the albumen was more frequently positive than the yolk. Storage at room temperature had no significant effect on the prevalence of salmonella-positive eggs but those held for more than 21 days were more likely (P less than 0.01) to be heavily contaminated. In batches of eggs where both shells and contents were examined, 1.1% were positive on the former site and 0.9% in the latter. PMID:2050203

  6. Whole-Genome Sequencing Analysis of Salmonella enterica Serovar Enteritidis Isolates in Chile Provides Insights into Possible Transmission between Gulls, Poultry, and Humans.

    PubMed

    Toro, Magaly; Retamal, Patricio; Ayers, Sherry; Barreto, Marlen; Allard, Marc; Brown, Eric W; Gonzalez-Escalona, Narjol

    2016-10-15

    Salmonella enterica subsp. enterica serotype Enteritidis is a major cause of human salmonellosis worldwide; however, little is known about the genetic relationships between S Enteritidis clinical strains and S Enteritidis strains from other sources in Chile. We compared the whole genomes of 30 S Enteritidis strains isolated from gulls, domestic chicken eggs, and humans in Chile, to investigate their phylogenetic relationships and to establish their relatedness to international strains. Core genome multilocus sequence typing (cgMLST) analysis showed that only 246/4,065 shared loci differed among these Chilean strains, separating them into two clusters (I and II), with cluster II being further divided into five subclusters. One subcluster (subcluster 2) contained strains from all surveyed sources that differed at 1 to 18 loci (of 4,065 loci) with 1 to 18 single-nucleotide polymorphisms (SNPs), suggesting interspecies transmission of S Enteritidis in Chile. Moreover, clusters were formed by strains that were distant geographically, which could imply that gulls might be spreading the pathogen throughout the country. Our cgMLST analysis, using other S Enteritidis genomes available in the National Center for Biotechnology Information (NCBI) database, showed that S Enteritidis strains from Chile and the United States belonged to different lineages, which suggests that S Enteritidis regional markers might exist and could be used for trace-back investigations. This study highlights the importance of gulls in the spread of Salmonella Enteritidis in Chile. We revealed a close genetic relationship between some human and gull S Enteritidis strains (with as few as 2 of 4,065 genes being different), and we also found that gull strains were present in clusters formed by strains isolated from other sources or distant locations. Together with previously published evidence, this suggests that gulls might be spreading this pathogen between different regions in Chile and that some of

  7. Whole-Genome Sequencing Analysis of Salmonella enterica Serovar Enteritidis Isolates in Chile Provides Insights into Possible Transmission between Gulls, Poultry, and Humans

    PubMed Central

    Ayers, Sherry; Barreto, Marlen; Allard, Marc; Brown, Eric W.

    2016-01-01

    ABSTRACT Salmonella enterica subsp. enterica serotype Enteritidis is a major cause of human salmonellosis worldwide; however, little is known about the genetic relationships between S. Enteritidis clinical strains and S. Enteritidis strains from other sources in Chile. We compared the whole genomes of 30 S. Enteritidis strains isolated from gulls, domestic chicken eggs, and humans in Chile, to investigate their phylogenetic relationships and to establish their relatedness to international strains. Core genome multilocus sequence typing (cgMLST) analysis showed that only 246/4,065 shared loci differed among these Chilean strains, separating them into two clusters (I and II), with cluster II being further divided into five subclusters. One subcluster (subcluster 2) contained strains from all surveyed sources that differed at 1 to 18 loci (of 4,065 loci) with 1 to 18 single-nucleotide polymorphisms (SNPs), suggesting interspecies transmission of S. Enteritidis in Chile. Moreover, clusters were formed by strains that were distant geographically, which could imply that gulls might be spreading the pathogen throughout the country. Our cgMLST analysis, using other S. Enteritidis genomes available in the National Center for Biotechnology Information (NCBI) database, showed that S. Enteritidis strains from Chile and the United States belonged to different lineages, which suggests that S. Enteritidis regional markers might exist and could be used for trace-back investigations. IMPORTANCE This study highlights the importance of gulls in the spread of Salmonella Enteritidis in Chile. We revealed a close genetic relationship between some human and gull S. Enteritidis strains (with as few as 2 of 4,065 genes being different), and we also found that gull strains were present in clusters formed by strains isolated from other sources or distant locations. Together with previously published evidence, this suggests that gulls might be spreading this pathogen between different regions

  8. Gene expression response of Salmonella enterica serotype Enteritidis phage type 8 to the subinhibitory concentrations of the plant-derived compounds,trans-cinnamaldehyde,and eugenol

    USDA-ARS?s Scientific Manuscript database

    Background: Salmonella Enteritidis phage type 8 (PT8) is a major poultry-associated Salmonella strain implicated in foodborne outbreaks in the United States. We previously reported that two GRAS-status, plant-derived compounds, trans-cinnamaldehyde (TC) and eugenol (EG) significantly reduced S. Ent...

  9. Phylogenetic structure of European Salmonella Enteritidis outbreak correlates with national and international egg distribution network

    PubMed Central

    Inns, Thomas; Jombart, Thibaut; Ashton, Philip; Loman, Nicolas; Chatt, Carol; Messelhaeusser, Ute; Rabsch, Wolfgang; Simon, Sandra; Nikisins, Sergejs; Bernard, Helen; le Hello, Simon; Jourdan da-Silva, Nathalie; Kornschober, Christian; Mossong, Joel; Hawkey, Peter; de Pinna, Elizabeth; Grant, Kathie; Cleary, Paul

    2016-01-01

    Outbreaks of Salmonella Enteritidis have long been associated with contaminated poultry and eggs. In the summer of 2014 a large multi-national outbreak of Salmonella Enteritidis phage type 14b occurred with over 350 cases reported in the United Kingdom, Germany, Austria, France and Luxembourg. Egg supply network investigation and microbiological sampling identified the source to be a Bavarian egg producer. As part of the international investigation into the outbreak, over 400 isolates were sequenced including isolates from cases, implicated UK premises and eggs from the suspected source producer. We were able to show a clear statistical correlation between the topology of the UK egg distribution network and the phylogenetic network of outbreak isolates. This correlation can most plausibly be explained by different parts of the egg distribution network being supplied by eggs solely from independent premises of the Bavarian egg producer (Company X). Microbiological sampling from the source premises, traceback information and information on the interventions carried out at the egg production premises all supported this conclusion. The level of insight into the outbreak epidemiology provided by whole-genome sequencing (WGS) would not have been possible using traditional microbial typing methods. PMID:28348865

  10. Phylogenetic structure of European Salmonella Enteritidis outbreak correlates with national and international egg distribution network.

    PubMed

    Dallman, Tim; Inns, Thomas; Jombart, Thibaut; Ashton, Philip; Loman, Nicolas; Chatt, Carol; Messelhaeusser, Ute; Rabsch, Wolfgang; Simon, Sandra; Nikisins, Sergejs; Bernard, Helen; le Hello, Simon; Jourdan da-Silva, Nathalie; Kornschober, Christian; Mossong, Joel; Hawkey, Peter; de Pinna, Elizabeth; Grant, Kathie; Cleary, Paul

    2016-08-01

    Outbreaks of Salmonella Enteritidis have long been associated with contaminated poultry and eggs. In the summer of 2014 a large multi-national outbreak of Salmonella Enteritidis phage type 14b occurred with over 350 cases reported in the United Kingdom, Germany, Austria, France and Luxembourg. Egg supply network investigation and microbiological sampling identified the source to be a Bavarian egg producer. As part of the international investigation into the outbreak, over 400 isolates were sequenced including isolates from cases, implicated UK premises and eggs from the suspected source producer. We were able to show a clear statistical correlation between the topology of the UK egg distribution network and the phylogenetic network of outbreak isolates. This correlation can most plausibly be explained by different parts of the egg distribution network being supplied by eggs solely from independent premises of the Bavarian egg producer (Company X). Microbiological sampling from the source premises, traceback information and information on the interventions carried out at the egg production premises all supported this conclusion. The level of insight into the outbreak epidemiology provided by whole-genome sequencing (WGS) would not have been possible using traditional microbial typing methods.

  11. Recovery of salmonella serovar enteritidis from inoculated broiler hatching eggs using shell rinse and shell crush sampling methods

    USDA-ARS?s Scientific Manuscript database

    This study compared the recovery of Salmonella from hatching eggs using three sampling methods (eggshell rinsing, eggshell crush following a previous rinse, and eggshell crush without previous rinse). Eggshells were drop-inoculated with approximately 10, 100, or 1,000 cfu/eggshell of S. Enteritidis...

  12. [Bacteremia associated with mycotic aneurysm of the transversal aortic arch and myocarditis caused by Salmonella enteritidis].

    PubMed

    Martínez-Martínez, L; Mesa, E; Rodríguez, J E; Sánchez, M P; Ugarte, J; Algora Weber, A; Dámaso, D; Daza, R M; Mendaza, P

    1989-02-01

    A 60-year-old male with diabetes mellitus had Salmonella enteritidis bacteremia associated with mycotic aneurysm of the transverse aortic arc and myocarditis. Antibiotic therapy with ampicillin and chloramphenicol was ineffective despite the fact that the microorganism was sensitive in vitro to those antimicrobials, and the patient had a progressive clinical deterioration which culminated in death.

  13. Adhesion of Salmonella Enteritidis and Listeria monocytogenes on stainless steel welds.

    PubMed

    Casarin, Letícia Sopeña; Brandelli, Adriano; de Oliveira Casarin, Fabrício; Soave, Paulo Azevedo; Wanke, Cesar Henrique; Tondo, Eduardo Cesar

    2014-11-17

    Pathogenic microorganisms are able to adhere on equipment surfaces, being possible to contaminate food during processing. Salmonella spp. and Listeria monocytogenes are important pathogens that can be transmitted by food, causing severe foodborne diseases. Most surfaces of food processing industry are made of stainless steel joined by welds. However currently, there are few studies evaluating the influence of welds in the microorganism's adhesion. Therefore the purpose of the present study was to investigate the adhesion of Salmonella Enteritidis and L. monocytogenes on surface of metal inert gas (MIG), and tungsten inert gas (TIG) welding, as well as to evaluate the cell and surface hydrophobicities. Results demonstrated that both bacteria adhered to the surface of welds and stainless steel at same levels. Despite this, bacteria and surfaces demonstrated different levels of hydrophobicity/hydrophilicity, results indicated that there was no correlation between adhesion to welds and stainless steel and the hydrophobicity. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Multi-laboratory validation study of multilocus variable-number tandem repeat analysis (MLVA) for Salmonella enterica serovar Enteritidis, 2015

    PubMed Central

    Peters, Tansy; Bertrand, Sophie; Björkman, Jonas T; Brandal, Lin T; Brown, Derek J; Erdõsi, Tímea; Heck, Max; Ibrahem, Salha; Johansson, Karin; Kornschober, Christian; Kotila, Saara M; Le Hello, Simon; Lienemann, Taru; Mattheus, Wesley; Nielsen, Eva Møller; Ragimbeau, Catherine; Rumore, Jillian; Sabol, Ashley; Torpdahl, Mia; Trees, Eija; Tuohy, Alma; de Pinna, Elizabeth

    2017-01-01

    Multilocus variable-number tandem repeat analysis (MLVA) is a rapid and reproducible typing method that is an important tool for investigation, as well as detection, of national and multinational outbreaks of a range of food-borne pathogens. Salmonella enterica serovar Enteritidis is the most common Salmonella serovar associated with human salmonellosis in the European Union/European Economic Area and North America. Fourteen laboratories from 13 countries in Europe and North America participated in a validation study for MLVA of S. Enteritidis targeting five loci. Following normalisation of fragment sizes using a set of reference strains, a blinded set of 24 strains with known allele sizes was analysed by each participant. The S. Enteritidis 5-loci MLVA protocol was shown to produce internationally comparable results as more than 90% of the participants reported less than 5% discrepant MLVA profiles. All 14 participating laboratories performed well, even those where experience with this typing method was limited. The raw fragment length data were consistent throughout, and the inter-laboratory validation helped to standardise the conversion of raw data to repeat numbers with at least two countries updating their internal procedures. However, differences in assigned MLVA profiles remain between well-established protocols and should be taken into account when exchanging data. PMID:28277220

  15. Multi-laboratory validation study of multilocus variable-number tandem repeat analysis (MLVA) for Salmonella enterica serovar Enteritidis, 2015.

    PubMed

    Peters, Tansy; Bertrand, Sophie; Björkman, Jonas T; Brandal, Lin T; Brown, Derek J; Erdõsi, Tímea; Heck, Max; Ibrahem, Salha; Johansson, Karin; Kornschober, Christian; Kotila, Saara M; Le Hello, Simon; Lienemann, Taru; Mattheus, Wesley; Nielsen, Eva Møller; Ragimbeau, Catherine; Rumore, Jillian; Sabol, Ashley; Torpdahl, Mia; Trees, Eija; Tuohy, Alma; de Pinna, Elizabeth

    2017-03-02

    Multilocus variable-number tandem repeat analysis (MLVA) is a rapid and reproducible typing method that is an important tool for investigation, as well as detection, of national and multinational outbreaks of a range of food-borne pathogens. Salmonella enterica serovar Enteritidis is the most common Salmonella serovar associated with human salmonellosis in the European Union/European Economic Area and North America. Fourteen laboratories from 13 countries in Europe and North America participated in a validation study for MLVA of S. Enteritidis targeting five loci. Following normalisation of fragment sizes using a set of reference strains, a blinded set of 24 strains with known allele sizes was analysed by each participant. The S. Enteritidis 5-loci MLVA protocol was shown to produce internationally comparable results as more than 90% of the participants reported less than 5% discrepant MLVA profiles. All 14 participating laboratories performed well, even those where experience with this typing method was limited. The raw fragment length data were consistent throughout, and the inter-laboratory validation helped to standardise the conversion of raw data to repeat numbers with at least two countries updating their internal procedures. However, differences in assigned MLVA profiles remain between well-established protocols and should be taken into account when exchanging data. This article is copyright of The Authors, 2017.

  16. Interpretations of antibody responses to Salmonella enterica serotype enteritidis gm flagellin in poultry flocks are enhanced by a kinetics-based enzyme-linked immunosorbent assay.

    PubMed

    McDonough, P L; Jacobson, R H; Timoney, J F; Mutalib, A; Kradel, D C; Chang, Y F; Shin, S J; Lein, D H; Trock, S; Wheeler, K

    1998-07-01

    Many regulatory and diagnostic programs for the detection of Salmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a human S. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemic S. enterica Enteritidis serotype but which also had multiple other Salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. enterica Enteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).

  17. A hotel-based outbreak of Salmonella enterica subsp. Enterica serovar Enteritidis (Salmonella Enteritidis) in the United Kingdom, 2006.

    PubMed

    Calvert, N; Murphy, L; Smith, A; Copeland, D

    2007-03-01

    An outbreak of food-borne Salmonella Enteritidis PT4 occurred in Cumbria, in north-west England, in the summer of 2006. Fifteen people, all with positive stool samples, met the case definition; three of these were admitted to hospital, including one patient who died. Preliminary investigations suggested a link to a meal served at a local hotel. A case control study was implemented, together with microbiological and environmental investigations. Fifteen microbiologically confirmed cases and 27 unmatched controls were included in the study, controls being randomly selected from people who had eaten at the hotel on the same day. The epidemiological evidence indicated a very strong association between infection and consumption of tiramisu made with raw shell eggs, although none were available for microbiological investigation. These results are in line with other salmonellosis outbreaks that have been associated with the use of raw shell eggs in food manufacturing and production. This paper highlights the continuing need for a greater awareness by those who work in the food industry of the health risks associated with the consumption of raw shell eggs.

  18. The role of home-made ice cream as a vehicle of Salmonella enteritidis phage type 4 infection from fresh shell eggs.

    PubMed Central

    Morgan, D.; Mawer, S. L.; Harman, P. L.

    1994-01-01

    A family outbreak of Salmonella enteritidis PT4 infection is described in which home-made ice cream was identified as the vehicle of infection. The ice cream contained approximately 10(5) S. enteritidis PT4 organisms per gm and was probably contaminated by an infected shell egg containing between 10(5)-10(8) organisms. The continued relevance of the Chief Medical Officer's warning on the use of raw shell eggs is highlighted. Home-made ice cream using the same recipe as ice cream that had been incriminated as the cause of the family outbreak of S. enteritidis PT4 infection was used to study the growth of the organism that might have occurred in the 3-4 h it took to prepare the product. When the inoculum was in the stationary phase, as it would be from shell or other cross contamination, there was a lag phase of 3 h before growth occurred at room temperature. Even when actively multiplying organisms were introduced, as may be found in an infected egg, there was less than 3 log(10) increase in the salmonella count in 4 h at room temperature. It was, therefore, given the high S. enteritidis count, unlikely that the ice cream was cross-contaminated. By contrast, raspberry sorbet at pH 3.73 proved to be lethal to a large inoculum of S. enteritidis and may be a relatively safe raw egg containing product. PMID:8062876

  19. Identification of a novel gene in ROD9 island of Salmonella Enteritidis involved in the alteration of virulence-associated genes expression.

    PubMed

    Das, Susmita; Ray, Shilpa; Ryan, Daniel; Sahu, Bikash; Suar, Mrutyunjay

    2018-01-01

    Salmonella enterica subsp. I serovar Enteritidis (S. Enteritidis), one of the causative agents for non-typhoidal gastrointestinal diseases in humans is an intracellular bacterium and mechanism for its invasion into host cells is critical to cause infection. The virulence of the pathogen is explained by the expression of genes located on its pathogenicity islands, mostly encoded under SPI-1 and SPI-2. However, S. Typhimurium SL1344, despite sharing ∼98% of its genome with S. Enteritidis P125109, lacks few regions of differences (ROD) that are hypothesized to impart virulence potential to S. Enteritidis. In this study, we created different mutants in the ROD9 island of S. Enteritidis, also referred as SPI-19 and identified a novel locus, SEN1005, encoding a hypothetical protein that is involved in its pathogenesis. ΔSEN1005 displayed significantly reduced entry into cultured epithelial cells as well as uptake by macrophages and failed to cause acute colitis in C57BL/6 mice at day 3 post-infection (p.i.). Additionally, the global transcriptome analysis revealed a highly repressed SPI-1 and other down-regulated genes responsible for flagellar assembly, chemotaxis and motility in the mutant which correlated with decreased invasion and abated inflammation as compared to the wild-type. Therefore, our findings revealed that ΔSEN1005 was attenuated in vitro as well as in vivo and we propose this hypothetical protein to play a role in altering the expression of genes involved in Salmonella virulence.

  20. Genetic analysis and CRISPR typing of Salmonella enterica serovar Enteritidis from different sources revealed potential transmission from poultry and pig to human.

    PubMed

    Li, Qiuchun; Wang, Xin; Yin, Kequan; Hu, Yachen; Xu, Haiyan; Xie, Xiaolei; Xu, Lijuan; Fei, Xiao; Chen, Xiang; Jiao, Xinan

    2018-02-02

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the most prevalent serotypes in Salmonella isolated from poultry and the most commonly reported cause of human salmonellosis. In this study, we aimed to assess the genetic diversity of 329 S. Enteritidis strains isolated from different sources from 2009 to 2016 in China. Clustered regularly interspaced short palindromic repeat (CRISPR) typing was used to characterize these 262 chicken clinical isolates, 38 human isolates, 18 pig isolates, six duck isolates, three goose isolates and two isolates of unknown source. A total of 18 Enteritidis CRISPR types (ECTs) were identified, with ECT2, ECT8 and ECT4 as the top three ECTs. CRISPR typing identified ECT2 as the most prevalent ECT, which accounted for 41% of S. Enteritidis strains from all the sources except duck. ECT9 and ECT13 were identified in both pig and human isolates and revealed potential transmission from pig to human. A cluster analysis distributed 18 ECTs, including the top three ECTs, into four lineages with LI as the predominant lineage. Forty-eight out of 329 isolates were subjected to whole genome sequence typing, which divided them into four clusters, with Cluster I as the predominant cluster. Cluster I included 92% (34/37) of strains located in LI identified from the CRISPR typing, confirming the good correspondence between both typing methods. In addition, the CRISPR typing also revealed the close relationship between ECTs and isolated areas, confirming that CRISPR spacers might be obtained by bacteria from the unique phage or plasmid pools in the environment. However, further analysis is needed to determine the function of CRISPR-Cas systems in Salmonella and the relationship between spacers and the environment. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Comparative analysis of core genome MLST and SNP typing within a European Salmonella serovar Enteritidis outbreak.

    PubMed

    Pearce, Madison E; Alikhan, Nabil-Fareed; Dallman, Timothy J; Zhou, Zhemin; Grant, Kathie; Maiden, Martin C J

    2018-06-02

    Multi-country outbreaks of foodborne bacterial disease present challenges in their detection, tracking, and notification. As food is increasingly distributed across borders, such outbreaks are becoming more common. This increases the need for high-resolution, accessible, and replicable isolate typing schemes. Here we evaluate a core genome multilocus typing (cgMLST) scheme for the high-resolution reproducible typing of Salmonella enterica (S. enterica) isolates, by its application to a large European outbreak of S. enterica serovar Enteritidis. This outbreak had been extensively characterised using single nucleotide polymorphism (SNP)-based approaches. The cgMLST analysis was congruent with the original SNP-based analysis, the epidemiological data, and whole genome MLST (wgMLST) analysis. Combination of the cgMLST and epidemiological data confirmed that the genetic diversity among the isolates predated the outbreak, and was likely present at the infection source. There was consequently no link between country of isolation and genetic diversity, but the cgMLST clusters were congruent with date of isolation. Furthermore, comparison with publicly available Enteritidis isolate data demonstrated that the cgMLST scheme presented is highly scalable, enabling outbreaks to be contextualised within the Salmonella genus. The cgMLST scheme is therefore shown to be a standardised and scalable typing method, which allows Salmonella outbreaks to be analysed and compared across laboratories and jurisdictions. Copyright © 2018. Published by Elsevier B.V.

  2. Sugar sulfates are not hydrolyzed by the acid-inducible sulfatase AslA from Salmonella enterica Enteritidis NalR and Kentucky 3795 at pH 5.5.

    PubMed

    Ganguly, Arpeeta; Joerger, Rolf D

    2017-08-01

    The open reading frames SEN0085 and SeKA_A4361, from Salmonella enterica serovar Enteritidis Nal R and serovar Kentucky 3795, respectively, corresponding to the acid-inducible sulfatase gene aslA from Salmonella enterica serovar Typhimurium, were previously suggested by microarray analysis to be differentially expressed under acid conditions. However, growth and enzyme activity tests in the present study demonstrated that both wild-type strains exhibited sulfatase activity with 4-nitrophenyl sulfate and 5-bromo-4-chloro-3 indolyl sulfate at pH 5.5. The acid sulfatase does not appear to be involved in sugar sulfate, tyrosine sulfate, 4-hydroxy-3-methoxyphenylglycol sulfate, heparin sulfate, or chondroitin sulfate hydrolysis at pH 5.5. Adhesion and invasion assays did not reveal differences between the serotypes and their corresponding aslA deletion mutants. Thus, the role and substrate(s) of AslA, a protein unique to salmonella and encoded in all sequenced Salmonella strains, remain elusive.

  3. Salmonella enterica Serovar Enteritidis Core O Polysaccharide Conjugated to H:g,m Flagellin as a Candidate Vaccine for Protection against Invasive Infection with S. Enteritidis▿†

    PubMed Central

    Simon, Raphael; Tennant, Sharon M.; Wang, Jin Y.; Schmidlein, Patrick J.; Lees, Andrew; Ernst, Robert K.; Pasetti, Marcela F.; Galen, James E.; Levine, Myron M.

    2011-01-01

    Nontyphoidal Salmonella enterica serovars Enteritidis and Typhimurium are a common cause of gastroenteritis but also cause invasive infections and enteric fever in certain hosts (young children in sub-Saharan Africa, the elderly, and immunocompromised individuals). Salmonella O polysaccharides (OPS) and flagellar proteins are virulence factors and protective antigens. The surface polysaccharides of Salmonella are poorly immunogenic and do not confer immunologic memory, limitations overcome by covalently attaching them to carrier proteins. We conjugated core polysaccharide-OPS (COPS) of Salmonella Enteritidis lipopolysaccharide (LPS) to flagellin protein from the homologous strain. COPS and flagellin were purified from a genetically attenuated (ΔguaBA) “reagent strain” (derived from an isolate from a patient with clinical bacteremia) engineered for increased flagellin production (ΔclpPX). Conjugates were constructed by linking flagellin monomers or polymers at random COPS hydroxyls with various polysaccharide/protein ratios by 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) or at the 3-deoxy-d-manno-octulosonic acid (KDO) terminus by thioether chemistry. Mice immunized on days 0, 28, and 56 with COPS-flagellin conjugates mounted higher anti-LPS IgG levels than mice receiving unconjugated COPS and exhibited high antiflagellin IgG; anti-LPS and antiflagellin IgG levels increased following booster doses. Antibodies generated by COPS-flagellin conjugates mediated opsonophagocytosis of S. Enteritidis cells into mouse macrophages. Mice immunized with flagellin alone, COPS-CRM197, or COPS-flagellin conjugates were significantly protected from lethal challenge with wild-type S. Enteritidis (80 to 100% vaccine efficacy). PMID:21807909

  4. Evaluation of ompA and pgtE genes in determining pathogenicity in Salmonella enterica serovar Enteritidis.

    PubMed

    Zhou, Y; Zhou, J; Wang, D; Gao, Q; Mu, X; Gao, S; Liu, X

    2016-12-01

    Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) is a major causative agent of gastroenteritis in humans. This important food-borne pathogen also colonises the intestinal tracts of poultry and can spread systemically, especially in chickens. To identify the S. Enteritidis virulence genes involved in infection and colonisation of chickens, chromosomal deletion mutants of the ompA and pgtE genes, which encode essential components of omptins, were constructed. There were no significant differences between the wild-type and ompA and pgtE mutants in a series of in vitro assays, including an intracellular survival assay, survival in specific-pathogen-free (SPF) chicken serum, and in vitro competition assays. In contrast, in vivo competition assays revealed that ompA and pgtE mutants underwent attenuated growth in liver, cardiac blood, spleen, lung, and kidney compared to a wild-type strain (CVCC3378). When tested in SPF chickens, ompA or pgtE gene inactivation substantially reduced organ colonisation and delayed systemic infection compared with the wild-type strain. Colonisation was restored in S. Enteritidis mutants by reintroduction of the whole ompA or pgtE gene with the native promoters. The results of this study demonstrate that ompA and pgtE play an important role in the pathogenesis of S. Enteritidis and its ability to infect chickens. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Dimethyl adenosine transferase (KsgA) deficiency in Salmonella Enteritidis confers susceptibility to high osmolarity and virulence attenuation in chickens

    USDA-ARS?s Scientific Manuscript database

    : Dimethyladenosine transferase (KsgA) performs diverse roles in bacteria including ribosomal maturation, DNA mismatch repair, and synthesis of KsgA is responsive to antibiotics and cold temperature. We previously showed that ksgA mutation in Salmonella Enteritidis results in impaired invasiveness i...

  6. Salmosan, a β-Galactomannan-Rich Product, Protects Epithelial Barrier Function in Caco-2 Cells Infected by Salmonella enterica Serovar Enteritidis.

    PubMed

    Brufau, M Teresa; Campo-Sabariz, Joan; Bou, Ricard; Carné, Sergi; Brufau, Joaquim; Vilà, Borja; Marqués, Ana M; Guardiola, Francesc; Ferrer, Ruth; Martín-Venegas, Raquel

    2016-08-01

    One promising strategy for reducing human salmonellosis induced by Salmonella Enteritidis is to supplement animal diets with natural feed additives such as mannan oligosaccharides (MOSs). We sought to investigate the potential role of Salmosan (S-βGM), an MOS product extremely rich in β-galactomannan, in preventing epithelial barrier function disruption induced by S. Enteritidis colonization in an in vitro model of intestinal Caco-2 cells in culture. Differentiated Caco-2 cells were incubated for 3 h with S. Enteritidis at a multiplicity of infection of 10 in the absence or presence of 500 μg S-βGM/mL. Paracellular permeability (PP) was assessed by transepithelial electrical resistance (TER), d-mannitol, and fluorescein isothiocyanate-dextran (FD-4) flux. Tight junction proteins and cytoskeletal actin were also localized by confocal microscopy. Reactive oxygen species (ROS) and lipid peroxidation products were evaluated. Scanning and transmission electron microscopy were used to visualize S. Enteritidis adhesion to, and invasion of, the Caco-2 cell cultures. Compared with controls, TER was significantly reduced by 30%, and d-mannitol and FD-4 flux were significantly increased by 374% and 54% in S. Enteritidis-infected cultures, respectively. The presence of S-βGM in infected cultures induced total recoveries of TER and FD-4 flux to values that did not differ from the control and a partial recovery of d-mannitol flux. These effects were confirmed by immunolocalization of actin, zonula occludens protein 1, and occludin. Similar results were obtained for Salmonella Dublin. The protection of S-βGM on PP in infected cultures may be associated with a total recovery of ROS production to values that did not differ from the control. Moreover, S-βGM has the capacity to agglutinate bacteria, leading to a significant reduction of 32% in intracellular S Enteritidis. The results demonstrate that S-βGM contributes to protecting epithelial barrier function in a Caco-2 cell

  7. Isolation, Characterization, and Bioinformatic Analyses of Lytic Salmonella Enteritidis Phages and Tests of Their Antibacterial Activity in Food.

    PubMed

    Han, Han; Wei, Xiaoting; Wei, Yi; Zhang, Xiufeng; Li, Xuemin; Jiang, Jinzhong; Wang, Ran

    2017-02-01

    Salmonella Enteritidis remains a major threat for food safety. To take efforts to develop phage-based biocontrol for S. Enteritidis contamination in food, in this study, the phages against S. Enteritidis were isolated from sewage samples, characterized by host range assays, DNA restriction enzyme pattern analyses, and transmission electron microscope observations, and tested for antibacterial activity in food; some potent phages were further characterized by bioinformatic analyses. Results showed that based on the plaque quality and host range, seven lytic phages targeting S. Enteritidis were selected, considered as seven distinct phages through DNA physical maps, and classified as Myoviridae or Siphoviridae family by morphologic observations; the combined use of such seven strain phages as a "food additive" could succeed in controlling the artificial S. Enteritidis contamination in the different physical forms of food at a range of temperatures; by bioinformatic analyses, both selected phage BPS 11 Q 3 and BPS 15 Q 2 seemed to be newfound obligate lytic phage strains with no indications for any potentially harmful genes in their genomes. In conclusion, our results showed a potential of isolated phages as food additives for controlling S. Enteritidis contamination in some salmonellosis outbreak-associated food vehicles, and there could be minimized potential risk associated with using BPS 11 Q 3 and BPS 15 Q 2 in food.

  8. National outbreak of Salmonella Enteritidis phage type 14b in England, September to December 2009: case-control study.

    PubMed

    Janmohamed, K; Zenner, D; Little, C; Lane, C; Wain, J; Charlett, A; Adak, B; Morgan, D

    2011-04-14

    We conducted an unmatched retrospective case–control study to investigate an upsurge of non-travel-related sporadic cases of infection with Salmonella enterica subsp. enterica serotype Enteritidis phage type 14b with antimicrobial resistance to nalidixic acid and partial resistance to ciprofloxacin (S. Enteritidis PT 14b NxCp(L)) that was reported in England from 1 September to 31 December 2009. We analysed data from 63 cases and 108 controls to determine whether cases had the same sources of infection as those found through investigation of 16 concurrent local foodborne outbreaks in England and Wales. Multivariable logistic regression analysis adjusting for age and sex identified food consumption at restaurants serving Chinese or Thai cuisine (odds ratio (OR): 4.4; 95% CI: 1.3–14.8; p=0.02), egg consumed away from home (OR: 5.1; 95% CI: 1.3–21.2; p=0.02) and eating vegetarian foods away from home (OR: 14.6; 95% CI: 2.1–99; p=0.006) as significant risk factors for infection with S. Enteritidis PT 14b NxCp(L). These findings concurred with those from the investigation of the16 outbreaks, which identified the same Salmonella strain in eggs from a specified source outside the United Kingdom. The findings led to a prohibition of imports from this source, in order to control the outbreak.

  9. Public health significance of major genotypes of Salmonella enterica serovar Enteritidis present in both human and chicken isolates in Korea.

    PubMed

    Kang, Min-Su; Oh, Jae-Young; Kwon, Yong-Kuk; Lee, Deog-Yong; Jeong, Ok-Mi; Choi, Byung-Kook; Youn, So-Youn; Jeon, Byung-Woo; Lee, Hye-Jin; Lee, Hee-Soo

    2017-06-01

    Salmonella enterica serovar Enteritidis is one of the most common serotypes implicated in Salmonella infections in both humans and poultry worldwide. It has been reported that human salmonellosis is mainly associated with the consumption of poultry products contaminated with serovar Enteritidis. The present study was to extensively analyze the public health risk of serovar Enteritidis isolates from chickens in Korea. A total of 127 chicken isolates were collected from clinical cases, on-farm feces, and chicken meat between 1998 and 2012 and 20 human clinical isolates were obtained from patients with diarrhea between 2000 and 2006 in Korea. To characterize the isolates from chickens and humans, we compared the pulsed-field gel electrophoresis (PFGE) patterns and multilocus variable-number tandem-repeat analysis (MLVA) profiles of the isolates. We further characterized representative isolates of different genotypes using a DNA microarray. PFGE revealed 28 patterns and MLVA identified 16 allelic profiles. The DNA microarray showed high genetic variability in plasmid regions and other fimbrial subunits of the isolates although the virulence gene contents of isolates from the same source and/or of the same genotype were unrelated. PFGE and MLVA showed that major genotypes were present in both human and chicken isolates. This result suggests that chickens in Korea pose a significant risk to public health as a source of serovar Enteritidis as has been noted in other countries. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Probiotic dahi containing Lactobacillus casei protects against Salmonella enteritidis infection and modulates immune response in mice.

    PubMed

    Jain, Shalini; Yadav, Hariom; Sinha, P R

    2009-06-01

    In the present study, effect of dahi containing probiotic Lactobacillus casei (probiotic dahi) was evaluated to modulate immune response against Salmonella enteritidis infection in mice. Animals were fed with milk products along with standard diet for 2 and 7 days prior to the S. enteritidis challenge and continued on the respective dairy food-supplemented diets during the postchallenge period. Translocation of S. enteritidis in spleen and liver, beta-galactosidase and beta-glucuronidase enzymatic activities and secretory IgA (sIgA) in intestinal fluid, lymphocyte proliferation, and cytokine (interleukin [IL]-2, IL-4, IL-6, and interferon-gamma [IFN-gamma]) production in cultured splenocytes were assessed on day 2, 5, and 8 of the postchallenge period. Colonization of S. enteritidis in liver and spleen was remarkably low in probiotic dahi-fed mice than mice fed milk and control dahi. The beta-galactosidase and beta-glucuronidase activities in intestinal fluid collected from mice prefed for 7 days with probiotic dahi were significantly lower at day 5 and 8 postchallenge than in mice fed milk and control dahi. Levels of sIgA and lymphocyte proliferation rate were also significantly increased in probiotic dahi-fed mice compared with the other groups. Production of IL-2, IL-6, and IFN-gamma increased, whereas IL-4 decreased in splenic lymphocytes collected from probiotic dahi-fed mice. Data showed that dahi prefed for 7 days before S. enteritidis challenge was more effective than when mice were prefed for 2 days with dahi. Moreover, probiotic dahi was more efficacious in protecting against S. enteritidis infection by enhancing innate and adaptive immunity than fermented milk and normal dahi. Results of the present study suggest that prefeeding of probiotic dahi may strengthen the consumer's immune system and may protect infectious agents like S. enteritidis.

  11. Outbreak of Salmonella enteritidis phage type 1B associated with frozen pre-cooked chicken cubes, Finland 2012.

    PubMed

    Huusko, S; Pihlajasaari, A; Salmenlinna, S; Sõgel, J; Dontšenko, I; DE Pinna, E; Lundström, H; Toikkanen, S; Rimhanen-Finne, R

    2017-10-01

    In August to October 2012, a nationwide outbreak of Salmonella enteritidis phase type (PT) 1B with 53 cases occurred in Finland. Hypothesis generating interviews pointed toward ready-to-eat chicken salad from a Finnish company and at the same time Estonian authorities informed of a S. enteritidis PT 1B outbreak linked to chicken wrap prepared at an Estonian restaurant. We found that chicken salad was associated with the infection (odds ratio (OR) 16·1, 95% confidence interval (CI) 1·7-148·7 for consumption and OR 17·5. 95% CI 4·0-76·0 for purchase). The frozen pre-cooked chicken cubes used in Finnish salad and in Estonian wraps were traced back to a production plant in China. Great Britain made two Rapid Alert Systems for Food and Feed notifications on chicken cubes imported to the UK from the same Chinese production plant. Microbiological investigation confirmed that the patient isolates in Estonia and in Finland were indistinguishable from the strains isolated from chicken cubes in Estonia and in the UK. We recommend that despite certificates for tested Salmonella, food items should be analyzed when Salmonella contamination in outbreak investigations is suspected. In outbreak investigations, electronically implemented case-case study saves time, effort, and money compared with case-control study.

  12. Occurrence of plasmid-mediated quinolone resistance determinants and rmtB gene in Salmonella enterica serovar enteritidis and Typhimurium isolated from food-animal products in Tunisia.

    PubMed

    Al-Gallas, Nazek; Abbassi, Mohamed Salah; Gharbi, Becher; Manai, Molka; Ben Fayala, Mohamed N; Bichihi, Raghda; Al-Gallas, Amna; Ben Aissa, Ridha

    2013-09-01

    Four hundred and thirty Salmonella isolates, recovered from various food-animal products, were tested for nalidixic acid resistance, plasmid-mediated quinolone resistance, and genetic relationship. One hundred fifteen isolates (113 Salmonella serovar Enteritidis and two Salmonella serovar Typhimurium isolates) of 430 (26.7%) Salmonella isolates exhibited nalidixic acid resistance. Polymerase chain reaction screening for qnrA, qnrB, qnrS, qepA (encoding fluoroquinolones resistance) and rmtB (encoding aminoglycosides resistance) showed that 5 (1.16%) isolates were positive for qnr- and qepA-type genes, and the aac(6')-Ib-cr gene was observed in two (1.7%) Enteritidis isolates concomitantly with qnrA or qnrB. The co-occurrence of qepA and rmtB in one Typhimurium isolate is noteworthy. Pulsed-field gel electrophoresis revealed a high genetic homogeneity of nalidixic-resistant isolates and the persistence of clonal clusters over 4 years in different regions in Tunisia and from various food-animal products. To the best of our knowledge, this is the first report of co-occurrence of qepA and rmtB in a Salmonella strain.

  13. Emergence of Salmonella enteritidis phage type 4 in the Caribbean: case-control study in Trinidad and Tobago, West Indies.

    PubMed

    Indar-Harrinauth, L; Daniels, N; Prabhakar, P; Brown, C; Baccus-Taylor, G; Comissiong, E; Hospedales, J

    2001-03-15

    A prospective case-control study involving 46 case patients and 92 age- and neighborhood-matched control subjects was conducted in Trinidad and Tobago (T&T) between March 1998 and May 1999 to determine the etiology, sources, and risk factors for Salmonella enteritidis (SE) infection. SE infection in T&T was found to be associated with the consumption of shell eggs, and in particular raw or undercooked eggs. SE isolates from 30 (88%) of 34 patients and from 9 implicated egg or egg-containing food samples were phage type 4. Homemade eggnog and ice cream, cake batter, and egg-containing beverages were the main raw egg-containing foods, reflecting the cultural practices of the people of T&T. Public health education on the risks of eating raw or undercooked eggs, thorough cooking of all egg dishes, and refrigeration of shell eggs and egg dishes; studies tracing infected eggs to their sources; and testing of flocks of layer chickens for SE are needed to reduce the incidence of this infection.

  14. Development of a novel loop-mediated isothermal amplification (LAMP) assay for the detection of Salmonella ser. Enteritidis from egg products

    USDA-ARS?s Scientific Manuscript database

    Salmonella ser. Enteritidis is a major public health concern worldwide. Loop-mediated isothermal amplification (LAMP) is a novel simple, easy-to-operate detection technology that amplifies DNA with high speed, efficiency, and specificity under isothermal conditions. The objective of this study was t...

  15. Interpretations of Antibody Responses to Salmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

    PubMed Central

    McDonough, Patrick L.; Jacobson, Richard H.; Timoney, John F.; Mutalib, Ahmed; Kradel, David C.; Chang, Yung-fu; Shin, Sang J.; Lein, Donald H.; Trock, Susan; Wheeler, Kaye

    1998-01-01

    Many regulatory and diagnostic programs for the detection of Salmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a human S. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemic S. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. enterica Enteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%). PMID:9665965

  16. Pathogenesis of Salmonellosis: Salmonella Exotoxins

    DTIC Science & Technology

    1982-03-08

    Newport; Sal. 9633 - serotype Newport; and Sal. 9186 - serotype Newport. Salmonella enteritidis serotype typhimurium strain 2000 was obtained from...7054 Table 1I CULTURE MEDIA SURVEY Salmonella enteritidis Salmonella typhimurium serotype Javiana #10016 SRlI Culture Media C H 0 Cell Factor C H 0 Cell...C r AD REPORT NUMBER 2 0 Pathogenesis of Salmonellosis: Salmonella Exotoxins Annual Progress Report (9/1/78-9/1/79) Johnny W. Peterson, Ph.D. March 8

  17. Development of a New Molecular Subtyping Tool for Salmonella enterica Serovar Enteritidis Based on Single Nucleotide Polymorphism Genotyping Using PCR

    PubMed Central

    Kelly, Hilary; Dupras, Andrée Ann; Belanger, Sebastien; Devenish, John

    2014-01-01

    The lack of a sufficiently discriminatory molecular subtyping tool for Salmonella enterica serovar Enteritidis has hindered source attribution efforts and impeded regulatory actions required to disrupt its food-borne transmission. The underlying biological reason for the ineffectiveness of current molecular subtyping tools such as pulsed-field gel electrophoresis (PFGE) and phage typing appears to be related to the high degree of clonality of S. Enteritidis. By interrogating the organism's genome, we previously identified single nucleotide polymorphisms (SNP) distributed throughout the chromosome and have designed a highly discriminatory PCR-based SNP typing test based on 60 polymorphic loci. The application of the SNP-PCR method to DNA samples from S. Enteritidis strains (n = 55) obtained from a variety of sources has led to the differentiation and clustering of the S. Enteritidis isolates into 12 clades made up of 2 to 9 isolates per clade. Significantly, the SNP-PCR assay was able to further differentiate predominant PFGE types (e.g., XAI.0003) and phage types (e.g., phage type 8) into smaller subsets. The SNP-PCR subtyping test proved to be an accurate, precise, and quantitative tool for evaluating the relationships among the S. Enteritidis isolates tested in this study and should prove useful for clustering related S. Enteritidis isolates involved in outbreaks. PMID:25297333

  18. Inhibition of the early stage of Salmonella enterica serovar Enteritidis biofilm development on stainless steel by cell-free supernatant of a Hafnia alvei culture.

    PubMed

    Chorianopoulos, Nikos G; Giaouris, Efstathios D; Kourkoutas, Yiannis; Nychas, George-John E

    2010-03-01

    Compounds present in Hafnia alvei cell-free culture supernatant cumulatively negatively influence the early stage of biofilm development by Salmonella enterica serovar Enteritidis on stainless steel while they also reduce the overall metabolic activity of S. Enteritidis planktonic cells. Although acylhomoserine lactones (AHLs) were detected among these compounds, the use of several synthetic AHLs was not able to affect the initial stage of biofilm formation by this pathogen.

  19. Molecular and epidemiologic analysis of a county-wide outbreak caused by Salmonella enterica subsp. enterica serovar Enteritidis traced to a bakery

    PubMed Central

    Lu, Po-Liang; Hwang, In-Jane; Tung, Ya-Lina; Hwang, Shang-Jyh; Lin, Chun-Lu; Siu, LK

    2004-01-01

    Background An increase in the number of attendees due to acute gastroenteritis and fever was noted at one hospital emergency room in Taiwan over a seven-day period from July to August, 2001. Molecular and epidemiological surveys were performed to trace the possible source of infection. Methods An epidemiological investigation was undertaken to determine the cause of the outbreak. Stool and blood samples were collected according to standard protocols per Center for Disease Control, Taiwan. Typing of the Salmonella isolates from stool, blood, and food samples was performed with serotyping, antibiotypes, and pulsed field gel electrophoresis (PFGE) following XbaI restriction enzyme digestion. Results Comparison of the number of patients with and without acute gastroenteritis (506 and 4467, respectively) during the six weeks before the outbreak week revealed a significant increase in the number of patients during the outbreak week (162 and 942, respectively) (relative risk (RR): 1.44, 95% confidence interval (CI): 1.22–1.70, P value < 0.001). During the week of the outbreak, 34 of 162 patients with gastroenteritis were positive for Salmonella, and 28 of these 34 cases reported eating the same kind of bread. In total, 28 of 34 patients who ate this bread were positive for salmonella compared to only 6 of 128 people who did not eat this bread (RR: 17.6, 95%CI 7.9–39.0, P < 0.001). These breads were produced by the same bakery and were distributed to six different traditional Chinese markets., Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) was isolated from the stool samples of 28 of 32 individuals and from a recalled bread sample. All S. Enteritidis isolates were of the same antibiogram. PFGE typing revealed that all except two of the clinical isolates and the bread isolates were of the same DNA macrorestriction pattern. Conclusions The egg-covered bread contaminated with S. Enteritidis was confirmed as the vehicle of infection. Alertness in

  20. Genetic basis for loss of immuno-reactive O-chain in Salmonella enterica serovar Enteritidis veterinary isolates.

    PubMed

    Szabo, Istvan; Grafe, Marianne; Kemper, Nicole; Junker, Ernst; Malorny, Burkhard

    2017-05-01

    Fifty-two rough Salmonella enterica serovar Enteritidis (S. Enteritidis) isolates from broilers and the environment were characterized for their serological and genotypic properties. Under routine diagnostic serotyping methods such isolates lack the immuno-reactivity of the O-chain of the lipopolysaccharide (LPS), and are referred to as non-typeable. Using a modified slide agglutination method, the isolates could be differentiated into three different serological variants. Twenty-six isolates (50%) were defined as semi-rough, nineteen isolates (37%) as deep-rough, four isolates (8%) as rough and three isolates could not be assigned. Genetically, all semi-rough isolates lacked the wzyB gene encoding the O-antigen polymerase. Two isolates carried a frameshift mutation in wzyB. In 15 of 23 cases deep-rough or rough isolates had a single point mutation, a single - or double-nucleotide insert or deletion in the wbaP gene. The mutational changes lead to expression of truncated (premature) protein, resulting in the loss of the immuno-reactive O-chain. Both rough and smooth S. Enteritidis isolates showed identical or highly similar XbaI-PFGE profiles. Our results indicate that the loss of a functional LPS in S. Enteritidis isolates is caused by a variety of different mutation events within the wzyB (semi-rough) or the wbaP (deep-rough) gene and is not a result of a vertical spread of a specific S. Enteritidis subtype. The defect of the LPS may be a common evolutionary mechanism through which host defence can be escaped. Copyright © 2017. Published by Elsevier B.V.

  1. Physiology, pathogenicity and immunogenicity of live, attenuated Salmonella enterica serovar Enteritidis mutants in chicks.

    PubMed

    Si, Wei; Wang, Xiumei; Liu, Huifang; Yu, Shenye; Li, Zhaoli; Chen, Liping; Zhang, Wanjiang; Liu, Siguo

    2015-01-01

    To construct a novel live, attenuated Salmonella vaccine, the lon, cpxR and cpdB genes were deleted from a wild-type Salmonella enterica serovar Enteritidis-6 (SM-6) strain using the phage λ Red homologous recombination system, resulting in SM-△CpxR, SM-△C/Lon and SM-△C/L/CpdB. The growth curves of strain SM-△C/Lon grew more rapidly than the other strains and had OD 600 values higher than the other strains starting at the 4 h time point. The growth curves of strain SM-△C/L/CpdB were relatively flat. The colonization time of SM-△C/L/CpdB is about 8-10 days. Deleting the lon/cpxR/cpdB (SM-6) genes resulted in an approximate 10(3)-fold attenuation in virulence assessed by the analysis of the LD50 of specific pathogen-free (SPF) chicks. This result indicated that the deletion of the lon, cpxR and cpdB genes induced significant virulence attenuation. The protective effects of SM-△C/L/CpdB vaccination in SPF chicks against 5 × 10(9) colony forming units (CFU) of S. Enteritidis were resulted from the induction of an effective immune response. These findings demonstrate the potential of mutant SM-△C/L/CpdB to be used as an effective vaccine. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Thermal inactivation of Salmonella enteritidis PT 30 in almond kernels as influenced by water activity.

    PubMed

    Villa-Rojas, Rossana; Tang, Juming; Wang, Shaojin; Gao, Mengxiang; Kang, Dong-Hyun; Mah, Jae-Hyung; Gray, Peter; Sosa-Morales, Maria Elena; López-Malo, Aurelio

    2013-01-01

    Salmonellosis outbreaks related to consumption of raw almonds have encouraged the scientific community to study the inactivation kinetics of pathogens in this dry commodity. However, the low moisture content of the product presents a challenge for thermal control, because the time required to achieve the desired thermal inactivation of microorganisms increases sharply with reduced moisture content and water activity. In this study, we explored and modeled the heat inactivation of Salmonella enterica serovar Enteritidis PT 30 in almond cultivar 'Nonpareil' kernel flour at four water activity (a(w)) values (0.601, 0.720, 0.888, and 0.946) using four temperatures for each a(w). The results showed that the inactivation was well fitted by both Weibull distribution (R(2) = 0.93 to 1.00) and first-order kinetics (R(2) = 0.82 to 0.96). At higher a(w) values, the rate of inactivation increased and less time was needed to achieve the required population reduction. These results suggest that, to avoid deterioration of product quality, shorter process times at lower temperatures may be used to achieve desired inactivation levels of Salmonella Enteritidis PT 30 by simply increasing the moisture content of almonds. These goals could be achieved with the use of existing procedures already practiced by the food industry, such as washing or prewetting scalding before heat inactivation.

  3. 21 CFR 118.8 - Testing methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Salmonella Web site is located at http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/ucm114716.htm... you may examine a copy at the Center for Food Safety and Applied Nutrition's Library, 5100 Paint... Edition, is located at http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/BacteriologicalAnalytical...

  4. 21 CFR 118.8 - Testing methodology for Salmonella Enteritidis (SE).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Salmonella Web site is located at http://www.fda.gov/Food/ScienceResearch/LaboratoryMethods/ucm114716.htm... Nutrition, Food and Drug Administration, 5100 Paint Branch Pkwy., College Park, MD 20740, 301-436-2364, or... Branch Pkwy., College Park, MD, 301-436-2163, or at the National Archives and Records Administration...

  5. Case-control study of infections with Salmonella enteritidis phage type 4 in England.

    PubMed Central

    Cowden, J. M.; Lynch, D.; Joseph, C. A.; O'Mahony, M.; Mawer, S. L.; Rowe, B.; Bartlett, C. L.

    1989-01-01

    OBJECTIVE--To determine the source of indigenous sporadic infection with Salmonella enteritidis phage type 4. DESIGN--Case-control study of primary sporadic cases identified by the Public Health Laboratory Service between 1 August and 30 September 1988. SETTING--PHLS Communicable Disease Surveillance Centre, Division of Enteric Pathogens, 11 PHLS laboratories, and 42 local authority environmental health departments in England. SUBJECTS--232 Patients (cases) with confirmed primary sporadic infection, for 160 of whom (88 female) (median age 30 years, age range 4 months to 85 years) data were obtained by questionnaire about consumption of fresh eggs, egg products, precooked chicken, and minced meat in the three days and one week before onset of the symptoms. Up to three controls, matched for neighbourhood, age, and sex (if aged greater than 11 years), were asked the same questions for the same calendar period. MAIN OUTCOME MEASURE--Association of primary sporadic infection with consumption of suspected food items. RESULTS--Illness due to S enteritidis phage type 4 was significantly associated with consumption of raw shell egg products (homemade mayonnaise, ice cream, and milk drinks containing eggs) (matched p = 0.02) and shop bought sandwiches containing mayonnaise (matched p = 0.00004) or eggs (matched p = 0.02). Illness was also significantly associated with eating lightly cooked eggs (unmatched p = 0.02), but not soft boiled eggs, and precooked hot chicken (matched p = 0.006). Reported consumption of eggs was not appreciably different between cases and controls before or after the median date of interview. CONCLUSIONS--Fresh shell eggs, egg products, and precooked hot chicken are vehicles of S enteritidis phage type 4 infection in indigenous sporadic cases. Public health education and reduction in contamination of eggs and infection of poultry with S enteritidis are needed to reduce the incidence of human infection. PMID:2508916

  6. Pathogenesis of Salmonellosis: Salmonella Exotoxins

    DTIC Science & Technology

    1982-03-08

    of Salmonella enteritidis , which included 9630 serotype newport, 9136 serotype newport, 10016 serotype javiana, and 8832, serotype javiana were also...supplied by Dr. T. Huber. Additionally, four clinical isolates of Salmonella enteritidis , which included 986 serotype typhimurium, 2000 serotype...77Z7I AD _ REPORT NUMBER 3 0 Pathogenesis of Salmonellosis: Salmonella Exotoxins Annual Progress Report (9/1/79-8/31/80) M Johnny W. Peterson, Ph.D

  7. Visualisation of morphological interaction of diamond and silver nanoparticles with Salmonella Enteritidis and Listeria monocytogenes.

    PubMed

    Sawosz, Ewa; Chwalibog, André; Mitura, Katarzyna; Mitura, Stanisław; Szeliga, Jacek; Niemiec, Tomasz; Rupiewicz, Marlena; Grodzik, Marta; Sokołowska, Aleksandra

    2011-09-01

    Currently, medicine intensively searches for methods to transport drugs to a target (sick) point within the body. The objective of the present investigation was to evaluate morphological characteristics of the assembles of silver or diamond nanoparticles with Salmonella Enteritidis (G-) or Listeria monocytogenes (G+), to reveal possibilities of constructing nanoparticle-bacteria vehicles. Diamond nanoparticles (nano-D) were produced by the detonation method. Hydrocolloids of silver nanoparticles (nano-Ag) were produced by electric non-explosive patented method. Hydrocolloids of nanoparticles (200 microl) were added to bacteria suspension (200 microl) in the following order: nano-D + Salmonella E.; nano-D + Listeria monocytogenes; nano-Ag + Salmonella E; nano-Ag + Listeria monocytogenes. Samples were inspected by transmission electron microscopy. Visualisation of nanoparticles and bacteria interaction showed harmful effects of both nanoparticles on bacteria morphology. The most spectacular effect of nano-D were strong links between nano-D packages and the flagella of Salmonella E. Nano-Ag were closely attached to Listeria monocytogenes but not to Salmonella E. There was no evidence of entering nano-Ag inside Listeria monocytogenes but smaller particles were placed inside Salmonella E. The ability of nano-D to attach to the flagella and the ability of nano-Ag to penetrate inside bacteria cells can be utilized to design nano-bacteria vehicles, being carriers for active substances attached to nanoparticles.

  8. Inactivation of Escherichia coli, Listeria monocytogenes, and Salmonella Enteritidis by Cymbopogon citratus D.C. Stapf. Essential Oil in Pineapple Juice.

    PubMed

    Leite, Caroline Junqueira Barcellos; de Sousa, Jossana Pereira; Medeiros, José Alberto da Costa; da Conceição, Maria Lúcia; dos Santos Falcão-Silva, Vivyanne; de Souza, Evandro Leite

    2016-02-01

    In the present study, the efficacy of Cymbopogon citratus D.C. Stapf. essential oil (CCEO) to provoke a 5-log CFU/ml (5-log) inactivation in a mixed composite of Escherichia coli, Listeria monocytogenes, and Salmonella enterica serovar Enteritidis in pineapple (Ananas comosus (L.) Merril) juice (4°C) was assessed. Moreover, the effects of CCEO on the physicochemical and sensory quality parameters of pineapple juice were evaluated. The MIC of CCEO was 5 μl/ml against the composite mix examined. For L. monocytogenes and E. coli inoculated in juice containing CCEO (5, 2.5, and 1.25 μl/ml), a ≥5-log reduction was detected after 15 min of exposure. This same result was obtained for Salmonella Enteritidis incubated alone in pineapple juice containing CCEO at 5 and 2.5 μl/ml. Overall, Salmonella Enteritidis was the most tolerant and L. monocytogenes was the most sensitive to CCEO. The physicochemical properties (pH, titratable acidic [citric acid per 100 g], and soluble solids) of pineapple juice containing CCEO (2.5 and 1.25 μl/ml) were maintained. Juice containing CCEO (2.5 and 1.25 μl/ml) exhibited similar scores for odor, appearance, and viscosity compared with juice without CCEO. However, unsatisfactory changes in taste and aftertaste were observed in juices containing CCEO. These results suggest that CCEO could be used as an alternative antimicrobial compound to ensure the safety of pineapple juice, although CCEO at the tested concentrations negatively impacted its taste. Therefore, further studies are needed to determine the balance between microbial safety and taste acceptability of pineapple juice containing CCEO.

  9. Impact of Skip a Day and Every Day Feeding Programs on the Colonization Rates of Salmonella Enteritidis in Broiler Breeder Pullets

    USDA-ARS?s Scientific Manuscript database

    The impact of feeding programs in the colonization rate after challenge with Salmonella enteritidis was investigated. Broiler breeder pullets from were vaccinated at 19 d of incubation with herpesvirus of turkeys (HVT) + chicken herpesvirus (SB1), or a vector HVT (vHVT) + Infectious bursal disease (...

  10. Colonization of internal organs by Salmonella Enteritidis in experimentally infected laying hens housed in enriched colony cages at different stocking densities

    USDA-ARS?s Scientific Manuscript database

    Epidemiologic analyses have linked the frequency of human infections with Salmonella enterica subspecies enterica serovar Enteritidis to the consumption of contaminated eggs and thus to the prevalence of this pathogen in commercial egg-laying flocks. Contamination of the edible contents of eggs by S...

  11. Multiple-locus variable-number tandem repeat analysis of Salmonella Enteritidis isolates from human and non-human sources using a single multiplex PCR

    PubMed Central

    Cho, Seongbeom; Boxrud, David J; Bartkus, Joanne M; Whittam, Thomas S; Saeed, Mahdi

    2007-01-01

    Simplified multiple-locus variable-number tandem repeat analysis (MLVA) was developed using one-shot multiplex PCR for seven variable-number tandem repeats (VNTR) markers with high diversity capacity. MLVA, phage typing, and PFGE methods were applied on 34 diverse Salmonella Enteritidis isolates from human and non-human sources. MLVA detected allelic variations that helped to classify the S. Enteritidis isolates into more evenly distributed subtypes than other methods. MLVA-based S. Enteritidis clonal groups were largely associated with sources of the isolates. Nei's diversity indices for polymorphism ranged from 0.25 to 0.70 for seven VNTR loci markers. Based on Simpson's and Shannon's diversity indices, MLVA had a higher discriminatory power than pulsed field gel electrophoresis (PFGE), phage typing, or multilocus enzyme electrophoresis. Therefore, MLVA may be used along with PFGE to enhance the effectiveness of the molecular epidemiologic investigation of S. Enteritidis infections. PMID:17692097

  12. Magnetic nanoparticle-enhanced PCR for the detection and identification of Staphylococcus aureus and Salmonella enteritidis.

    PubMed

    Houhoula, Dimitra; Papaparaskevas, Joseph; Zatsou, Katerina; Nikolaras, Nikolaos; Malkawi, Hanan I; Mingenot-Leclercq, Marie-Paule; Konteles, Spyros; Koussisis, Stamatis; Tsakris, Athanassios; Charvalos, Ekatherina

    2017-07-01

    This paper evaluated magnetic nanoparticle-enhanced PCR for the detection and identification of Staphylococcus aureus and Salmonella enteritidis. Two different types of magnetic nanoparticles designated MPIO (iron concentration 2.5 mg/ml, size 1 µm) and NP (iron concentration 8.7 mg/ml, size 60 nm), both conjugated with S. aureus or S. enteritidis antibodies were evaluated as an enrichment procedure for PCR-detection of the pathogens in Trypticase Soy Broth, milk, blood and meat broth. Bacterial suspensions (1.5x108 cfu/ml) were prepared and serial diluted 10-1. The MPIO and NP nanoparticles were added, followed by incubation for 1 hour at room temperature, magnetic separation of the pellet, DNA extraction and PCR, targeting the femA and invA sequences. The nanoparticle-free and the NP-supplemented dilutions were positive down to the 1.5x102 cfu/ml concentration for both bacteria. The MPIO-supplemented dilutions were positive down to approx. 2x100 cfu/ml concentration, respectively. Bacteria-free TSB was negative by PCR. MPIO nanoparticles (size 1 µm) enhanced the detection of S. aureus and S. enteritidis by PCR, whilst NP nanoparticles (size 60 nm) did not, thus indicating that the size of the magnetic nanoparticles play a significant role in the enrichment procedure.

  13. Identification of risk factors and causes of persistence of Salmonella Gallinarum in laying hens farms from Colombia

    USDA-ARS?s Scientific Manuscript database

    The presence of Salmonella Gallinarum (SG) was recently identified in brown egg layers in Colombia. During 2013, twenty isolates were analyzed using Intergenic Sequence Ribotyping. Eighteen (90%) were SG and two (10%) were Salmonella Enteritidis (SE). SG was isolated mainly from organs of sick birds...

  14. Evaluation of a polymerase chain reaction-based system for detection of Salmonella enteritidis, Escherichia coli O157:H7, Listeria spp., and Listeria monocytogenes on fresh fruits and vegetables.

    PubMed

    Shearer, A E; Strapp, C M; Joerger, R D

    2001-06-01

    A polymerase chain reaction (PCR)-based detection system, BAX, was evaluated for its sensitivity in detecting Salmonella Enteritidis, Escherichia coli O157:H7, Listeria sp., and Listeria monocytogenes on fresh produce. Fifteen different types of produce (alfalfa sprouts, green peppers, parsley, white cabbage, radishes, onions, carrots, mushrooms, leaf lettuce, tomatoes, strawberries, cantaloupe, mango, apples, and oranges) were inoculated, in separate studies, with Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes down to the predicted level of 1 CFU per 25-g sample. Detection by BAX was compared to recovery of the inoculated bacteria by culture methods according to the Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM). BAX was essentially as sensitive as the culture-based method in detecting Salmonella Enteritidis and L. monocytogenes and more sensitive than the culture-based method for the detection of E. coli O157:H7 on green pepper, carrot, radish, and sprout samples. Detection of the pathogenic bacteria in samples spiked with a predicted number of less than 10 CFU was possible for most produce samples, but both methods failed to detect L. monocytogenes on carrot samples and one of two mushroom and onion samples spiked with less than 100 CFU. Both BAX and the culture method were also unable to consistently recover low numbers of E. coli O157:H7 from alfalfa sprouts. The PCR method allowed detection of Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes at least 2 days earlier than the conventional culture methods.

  15. [Food poisoning outbreak due to the consumption of spaghetti a la carbonara caused by Salmonella enteritidis].

    PubMed

    Godoy, P; Artigues, A; Usera, M A; González, J L; Pablo, N; Agustí, M

    2000-01-01

    This paper reports a clinico-epidemiological and microbiological investigation conducted into an outbreak of gastrointestinal infection due to Salmonella enteritidis, where the most likely food vehicle was spaghetti a la carbonara. An historic cohort study was conducted out among persons exposed to menus at a school canteen. Data were gathered on age, sex, foods consumed and clinical symptoms. School premises and menus were inspected, food samples obtained (spaghetti and meat balls), and stool samples taken from 30 affected subjects and 8 food handlers. Isolated strains were studied using pulsed-field electrophoresis. Attack rates were computed, and the odds ratio adjusted for the remaining foodstuffs (ORa) used to calculate the independent contribution made by the respective foods to risk of infection. Study coverage was 75.7% (140/185). The overall attack rate was 72.1% (101/140), with 12.9% of those affected requiring hospitalisation. The multivariate analysis showed that, while the spaghetti maintained its association (ORa = 8.4; 95% CI 1.4-51.8), the meat balls registered a reduction in risk (ORa = 1.8; 95% CI 0.4-7.5). S. enteritidis was isolated in stool cultures from 28 affected subjects, and in 2 blood and 6 stool cultures from food handlers (5 of whom were classed as cases). Moreover, S. enteritidis was also isolated in the food samples. On pulsed-field electrophoresis, the strains registered the same electrophoresis pattern. This outbreak serves to underscore the gravity of Salmonella spp. food poisoning, the danger of using inadequately cooked eggs, and the importance of interviewing food handlers to ensure proper classification (i.e., as patients or carriers). Existing recommendations as to the use of pasteurised egg products ought to be extended in scope.

  16. Large outbreak of Salmonella enteritidis PT8 in Portsmouth, UK, associated with a restaurant.

    PubMed

    Severi, E; Booth, L; Johnson, S; Cleary, P; Rimington, M; Saunders, D; Cockcroft, P; Ihekweazu, C

    2012-10-01

    Seventy-five individuals with Salmonella infection were identified in the Portsmouth area during August and September 2009, predominantly Salmonella Enteritidis phage type 8. Five patients were admitted to hospital. A case-case comparison study showed that a local restaurant was the most likely source of the infection with a risk of illness among its customers 25-fold higher than that of those who did not attend the restaurant. A case-control study conducted to investigate specific risk factors for infection at the restaurant showed that eating salad was associated with a threefold increase in probability of illness. Changing from using ready washed lettuces to lettuces requiring washing and not adhering strictly to the 48 hours exclusion policy for food handlers with diarrhoea were likely to have contributed to the initiation and propagation of this outbreak. Possibilities for cross-contamination and environmental contamination were identified in the restaurant.

  17. Salmonella enterica serovar Enteritidis and Listeria monocytogenes in mango (Mangifera indica L.) pulp: growth, survival and cross-contamination.

    PubMed

    Penteado, Ana L; de Castro, M Fernanda P M; Rezende, Ana C B

    2014-10-01

    This study examined the ability of Salmonella enterica serovar Enteritidis and Listeria monocytogenes to grow or survive in mango pulp stored at -20°C, 4°C, 10°C and 25°C, as well as to cross-contaminate mangoes by means of a knife contaminated with different levels of these pathogens. At 25°C lag phase durations of 19 h and 7.2 h and generation times of 0.66 and 1.44 were obtained, respectively, for S. Enteritidis and L. monocytogenes. At 10°C only the growth of L. monocytogenes was observed. At 4°C both bacteria survived for 8 days. At -20°C S. Enteritidis was able to survive for 5 months while L. monocytogenes survived for 8 months. Cross-contamination was observed for knives contaminated with 10⁶, 10⁵ and 10⁴ CFU mL⁻¹ of S. Enteritidis and 10⁶ and 10⁵ CFU mL⁻¹ of L. monocytogenes. Both microorganisms can grow well in mango pulp at 25°C, thus lower temperatures for the maintenance of the pulps are crucial to avoid growth of these microorganisms. A refrigeration temperature of 10°C will avoid only the growth of S. Enteritidis. Thus good handling practices should be rigidly enforced to avoid any contamination as even at refrigeration and freezing temperatures survival of these pathogens may occur. © 2014 Society of Chemical Industry.

  18. The molecular adjuvant mC3d enhances the immunogenicity of FimA from type I fimbriae of Salmonella enterica serovar Enteritidis.

    PubMed

    Musa, Hassan-Hussein; Zhang, Wei-Juan; Lv, Jing; Duan, Xiao-Li; Yang, Yang; Zhu, Chun-Hong; Li, Hui-Fang; Chen, Kuan-Wei; Meng, Xia; Zhu, Guo-Qiang

    2014-02-01

    The fimbriae of Salmonella enterica serovar Enteritidis are used for colonization and invasion into host cells, and have drawn considerable interest because fimbriae can serve as potential immunogens against many pathogenic bacteria that colonize on epithelial surfaces. The purpose of the study is to use a molecular adjuvant, C3d, to enhance the immunogenicity of FimA proteins against Salmonella enterica serovar Enteritidis. FimA of type I fimbriae from Salmonella enteritidis and FimA with one copy of mC3d, two copies of mC3d2 and three copies of mC3d3 were cloned into the expression vector pCold-TF. Soluble fusion proteins of FimA with different copy of mC3d were induced by IPTG and expressed into Escherichia coli BL21 (DE3). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the recombinant proteins from pCold-TF-fimA, TF-fimA-mC3d, TF-fimA-mC3d2, TF-fimA-mC3d3 were 70 kDa, 100 kDa, 130 kDa and 160 kDa, respectively. The fusion protein was recognized by rabbit anti-fimbriae polyclonal antibodies, and then visualized by goat anti-rabbit polyclonal antibodies with a chrome appearance by enzyme-subtract interaction. The recombinant proteins were purified by Ni-TED (tris-carboxymethyl ethylene diamine), immobilized metal ion affinity chromatography (IMAC). Balb/c mice were subcutaneously immunized with the purified proteins and the immune response was monitored by an enzyme-linked immunosorbent assay (ELISA) for FimA-specific antibody. The immunized mice were challenged with a 10-fold LD50 dose (i.e., 100 CFU) of Salmonella enterica serovar Enteritidis standard strain (SD-2) 1 week after the second immunization. The immunized mice with the fusion proteins FimA-mC3d2 and FimA-mC3d3 had increased levels of ELISA titer of antibody that were 2 and 4 logs, respectively, more immunogenic than the TF-FimA protein alone. The challenge results showed that immune protection rate in the mice immunized with 10 μg of FimA, FimA-mC3d2, and FimA-mC3d3

  19. Comparison of Genotypes of Salmonella enterica Serovar Enteritidis Phage Type 30 and 9c Strains Isolated during Three Outbreaks Associated with Raw Almonds▿

    PubMed Central

    Parker, Craig T.; Huynh, Steven; Quiñones, Beatriz; Harris, Linda J.; Mandrell, Robert E.

    2010-01-01

    In 2000 to 2001, 2003 to 2004, and 2005 to 2006, three outbreaks of Salmonella enterica serovar Enteritidis were linked with the consumption of raw almonds. The S. Enteritidis strains from these outbreaks had rare phage types (PT), PT30 and PT9c. Clinical and environmental S. Enteritidis strains were subjected to pulsed-field gel electrophoresis (PFGE), multilocus variable-number tandem repeat analysis (MLVA), and DNA microarray-based comparative genomic indexing (CGI) to evaluate their genetic relatedness. All three methods differentiated these S. Enteritidis strains in a manner that correlated with PT. The CGI analysis confirmed that the majority of the differences between the S. Enteritidis PT9c and PT30 strains corresponded to bacteriophage-related genes present in the sequenced genomes of S. Enteritidis PT4 and S. enterica serovar Typhimurium LT2. However, PFGE, MLVA, and CGI failed to discriminate between S. Enteritidis PT30 strains related to outbreaks from unrelated clinical strains or between strains separated by up to 5 years. However, metabolic fingerprinting demonstrated that S. Enteritidis PT4, PT8, PT13a, and clinical PT30 strains metabolized l-aspartic acid, l-glutamic acid, l-proline, l-alanine, and d-alanine amino acids more efficiently than S. Enteritidis PT30 strains isolated from orchards. These data indicate that S. Enteritidis PT9c and 30 strains are highly related genetically and that PT30 orchard strains differ from clinical PT30 strains metabolically, possibly due to fitness adaptations. PMID:20363782

  20. Altered virulence potential of Salmonella Enteritidis cultured in different foods: A cumulative effect of differential gene expression and immunomodulation.

    PubMed

    Jaiswal, Sangeeta; Sahoo, Prakash Kumar; Ryan, Daniel; Das, Jugal Kishore; Chakraborty, Eesha; Mohakud, Nirmal Kumar; Suar, Mrutyunjay

    2016-08-02

    Salmonella enterica serovars Enteritidis (S. Enteritidis) is one of the most common causes of food borne illness. Bacterial growth environment plays an important role in regulating gene expression thereby affecting the virulence profile of the bacteria. Different foods present diverse growth conditions which may affect the pathogenic potential of the bacteria. In the present study, the effect of food environments on the pathogenic potential of S. Enteritidis has been evaluated. S. Enteritidis was grown in different foods e.g. egg white, peanut butter and milk, and virulent phenotypes were compared to those grown in Luria Bertani broth. In-vivo experiments in C57BL/6 mice revealed S. Enteritidis grown in egg white did not induce significant (p<0.001) production of proinflammatory cytokines in mice and were unable to cause colitis despite efficient colonization in cecum, mesenteric lymph node, spleen and liver. Further studies revealed that bacteria grown in LB activated MAP Kinase and NFκB pathways efficiently, while those grown in egg white poorly activated the above pathways which can account for the decreased production of proinflammatory cytokines. qRT PCR analysis revealed SPI-1 effectors were downregulated in bacteria grown in egg white. Interestingly, bacteria grown in egg white showed reversal of phenotype upon change in growth media to LB. Additionally, bacteria grown in milk and peanut butter showed different degrees of virulence in mice as compared to those grown in LB media. Thus, the present study demonstrates that, S. Enteritidis grown in egg white colonizes systemic sites without causing colitis in a mouse model, while bacteria grown in milk and peanut butter show different pathogenicity profiles suggesting that food environments significantly affect the pathogenicity of S. Enteritidis. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Cell invasion of poultry-associated Salmonella enterica serovar Enteritidis isolates is associated with pathogenicity, motility and proteins secreted by the type III secretion system

    PubMed Central

    Zhou, Xiaohui; Addwebi, Tarek; Davis, Margaret A.; Orfe, Lisa; Call, Douglas R.; Guard, Jean; Besser, Thomas E.

    2011-01-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major cause of food-borne gastroenteritis in humans worldwide. Poultry and poultry products are considered the major vehicles of transmission to humans. Using cell invasiveness as a surrogate marker for pathogenicity, we tested the invasiveness of 53 poultry-associated isolates of S. Enteritidis in a well-differentiated intestinal epithelial cell model (Caco-2). The method allowed classification of the isolates into low (n = 7), medium (n = 18) and high (n = 30) invasiveness categories. Cell invasiveness of the isolates did not correlate with the presence of the virulence-associated gene spvB or the ability of the isolates to form biofilms. Testing of representative isolates with high and low invasiveness in a mouse model revealed that the former were more invasive in vivo and caused more and earlier mortalities, whereas the latter were significantly less invasive in vivo, causing few or no mortalities. Further characterization of representative isolates with low and high invasiveness showed that most of the isolates with low invasiveness had impaired motility and impaired secretion of either flagella-associated proteins (FlgK, FljB and FlgL) or type III secretion system (TTSS)-secreted proteins (SipA and SipD) encoded on Salmonella pathogenicity island-1. In addition, isolates with low invasiveness had impaired ability to invade and/or survive within chicken macrophages. These data suggest that not all isolates of S. Enteritidis recovered from poultry may be equally pathogenic, and that the pathogenicity of S. Enteritidis isolates is associated, in part, with both motility and secretion of TTSS effector proteins. PMID:21292746

  2. In vivo and in vitro evaluation of tissue colonization and survival capacity of Salmonella Oranienburg in laying hens

    USDA-ARS?s Scientific Manuscript database

    Salmonella Oranienburg (SO) was linked to a human salmonellosis outbreak in the Midwest in 2015 and 2016 from consumption of eggs. However, unlike Salmonella Enteritidis (SE), little is known regarding the potential of SO to colonize in laying hens and contaminate eggs. We used in vivo and in vitr...

  3. Immune protection of chickens conferred by a vaccine consisting of attenuated strains of Salmonella Enteritidis, Typhimurium and Infantis.

    PubMed

    Varmuzova, Karolina; Faldynova, Marcela; Elsheimer-Matulova, Marta; Sebkova, Alena; Polansky, Ondrej; Havlickova, Hana; Sisak, Frantisek; Rychlik, Ivan

    2016-10-15

    The colonization of poultry with different Salmonella enterica serovars poses an issue throughout the world. In this study we therefore tested the efficacy of a vaccine consisting of attenuated strains of Salmonella enterica serovars Enteritidis, Typhimurium and Infantis against challenge with the same serovars and with S. Agona, Dublin and Hadar. We tested oral and aerosol administration of the vaccine, with or without co-administration of cecal microbiota from adult hens. The protective effect was determined by bacterial counts of the challenge strains up to week 18 of life and by characterizing the immune response using real-time PCR specific for 16 different genes. We have shown that a vaccine consisting of attenuated S. Enteritidis, S. Typhimurium and S. Infantis protected chickens against challenge with the wild type strains of the same serovars and partially protected chickens also against challenge with isolates belonging to serovars Dublin or Hadar. Aerosol vaccination was more effective at inducing systemic immunity whilst oral vaccination stimulated a local immune response in the gut. Co-administration of cecal microbiota increased the protectiveness in the intestinal tract but slightly decreased the systemic immune response. Adjusting the vaccine composition and changing the administration route therefore affects vaccine efficacy.

  4. Salmonella enterica Serovar Enteritidis Ghosts Carrying the Escherichia coli Heat-Labile Enterotoxin B Subunit Are Capable of Inducing Enhanced Protective Immune Responses

    PubMed Central

    Jawale, Chetan V.

    2014-01-01

    The Escherichia coli heat-labile enterotoxin B subunit (LTB) is a potent vaccine adjuvant. Salmonella enterica serovar Enteritidis ghosts carrying LTB (S. Enteritidis-LTB ghosts) were genetically constructed using a novel plasmid, pJHL187-LTB, designed for the coexpression of the LTB and E lysis proteins. S. Enteritidis-LTB ghosts were characterized using scanning electron microscopy to visualize their transmembrane tunnel structures. The expression of LTB in S. Enteritidis-LTB ghost preparations was confirmed by immunoblot and enzyme-linked immunosorbent assays. The parenteral adjuvant activity of LTB was demonstrated by immunizing chickens with either S. Enteritidis-LTB ghosts or S. Enteritidis ghosts. Chickens were intramuscularly primed at 5 weeks of age and subsequently boosted at 8 weeks of age. In total, 60 chickens were equally divided into three groups (n = 20 for each): group A, nonvaccinated control; group B, immunized with S. Enteritidis-LTB ghosts; and group C, immunized with S. Enteritidis ghosts. Compared with the nonimmunized chickens (group A), the immunized chickens (groups B and C) exhibited increased titers of plasma IgG and intestinal secretory IgA antibodies. The CD3+ CD4+ subpopulation of T cells was also significantly increased in both immunized groups. Among the immunized chickens, those in group B exhibited significantly increased titers of specific plasma IgG and intestinal secretory IgA (sIgA) antibodies compared with those in group C, indicating the immunomodulatory effects of the LTB adjuvant. Furthermore, both immunized groups exhibited decreased bacterial loads in their feces and internal organs. These results indicate that parenteral immunization with S. Enteritidis-LTB ghosts can stimulate superior induction of systemic and mucosal immune responses compared to immunization with S. Enteritidis ghosts alone, thus conferring efficient protection against salmonellosis. PMID:24671556

  5. Antibiotic Resistance in Salmonella Enteritidis Isolates Recovered from Chicken, Chicken Breast, and Humans Through National Antimicrobial Resistance Monitoring System Between 1996 and 2014.

    PubMed

    Paudyal, Narayan; Pan, Hang; Li, Xiaoliang; Fang, Weihuan; Yue, Min

    2018-06-21

    Salmonella enterica subspecies enterica serotype Enteritidis (S. Enteritidis) is one of the leading causes for human salmonellosis all over the world. We analyzed the surveillance data of 18 years on antimicrobial resistance profiling of S. Enteritidis collected and isolated by the National Antimicrobial Resistance Monitoring System (NARMS) from humans, chicken, and chicken breasts. Statistical tool based on the unique individual antibiotic minimum inhibitory concentration (MIC) profiling was used to compare antimicrobial resistance in the isolates. A machine-learning algorithm, Random Forest matrix, segregated a collection of 6819 S. Enteritidis into multiple populations. The MIC value of 13 common antibiotics to individual isolate when taken as the best classifier, resulted in two distinct groups represented herein as Population-I and Population-II. Population-I, which spread within a small tight cluster, comprised all the chicken and chicken breasts' isolates as well as about 13.4% of the human isolates, whereas the Population-II consisted of the human isolates only, with a larger spread over wider area away from the Population-I (p < 0.001). Few overlapping, yet diverse clusters between humans and chicken as well as higher level of resistance of chicken breast isolate toward third-generation cephalosporins and tetracyclines compared to those from human isolates, highlight differences in their population structure. These findings indicate a complex driver for enriching antibiotic-resistant Salmonella in the food-chain other than those of chicken origin. This warrants for other strategies in addition to the judicious/restricted use of antibiotics to mitigate the threat of antimicrobial resistance.

  6. Colonization of internal organs by Salmonella Enteritidis in experimentally infected laying hens of four commercial genetic lines in conventional cage and enriched colony housing

    USDA-ARS?s Scientific Manuscript database

    Human infections with Salmonella Enteritidis are often attributed to the consumption of contaminated eggs, so the prevalence of this pathogen in commercial egg-laying flocks is a significant public health concern. Internal contamination of the edible contents of eggs results from bacterial colonizat...

  7. Frequency and Duration of Fecal Shedding of Salmonella Enteritidis by Experimentally Infected Laying Hens Housed in Enriched Colony Cages at Different Stocking Densities

    PubMed Central

    Gast, Richard K.; Guraya, Rupa; Jones, Deana R.; Anderson, Kenneth E.; Karcher, Darrin M.

    2017-01-01

    Human infections with Salmonella Enteritidis are often attributed to the consumption of contaminated eggs, so the prevalence of this pathogen in egg-laying poultry is an important public health risk factor. Numerous and complex environmental influences on Salmonella persistence and transmission are exerted by management practices and housing facilities used in commercial egg production. In recent years, the animal welfare implications of poultry housing systems have guided the development of alternatives to traditional cage-based housing, but their food safety consequences are not yet fully understood. The present study assessed the effects of different bird stocking densities on the frequency and duration of fecal shedding of S. Enteritidis in groups of experimentally infected laying hens housed in colony cages enriched with perching and nesting areas. In two trials, groups of laying hens were distributed at two stocking densities (648 and 973 cm2/bird) into enriched colony cages and (along with a group housed in conventional cages at 648 cm2/bird) orally inoculated with doses of 1.0 × 108 cfu of S. Enteritidis. At 10 weekly postinoculation intervals, samples of voided feces were collected from beneath each cage and cultured to detect S. Enteritidis. Fecal shedding of S. Enteritidis was detected for up to 10 weeks postinoculation by hens in all three housing treatment groups. The overall frequency of positive fecal cultures was significantly (P < 0.05) greater from conventional cages than from enriched colony cages (at the lower stocking density) for the total of all sampling dates (45.0 vs. 33.3%) and also for samples collected at 4–9 weeks postinfection. Likewise, the frequency of S. Enteritidis isolation from feces from conventional cages was significantly greater than from enriched colony cages (at the higher hen stocking density) for the sum of all samples (45.0 vs. 36.7%) and at 6 weeks postinoculation. Moreover, the frequency of S

  8. [Serotype and phage type distribution of human Salmonella strains isolated in Spain, 1997-2001].

    PubMed

    Echeita, María Aurora; Aladueña, Ana María; Díez, Rosa; Arroyo, Margarita; Cerdán, Francisca; Gutiérrez, Rafaela; de la Fuente, Manuela; González-Sanz, Rubén; Herrera-León, Silvia; Usera, Miguel Angel

    2005-03-01

    Salmonellosis is one of the most frequent causes of gastroenteritis in Spain. Serotyping is the gold standard epidemiological marker for subdividing Salmonella spp. strains. A small number of serotypes are very frequently isolated, reducing the discriminatory power of serotyping. Thus, to increase our knowledge of Salmonella spp. epidemiology, additional epidemiological markers, such as phage typing, should be used for this purpose. Salmonella spp. strains of human origin sent to the Laboratorio Nacional de Referencia de Salmonella y Shigella (LNRSSE, Spanish Reference Laboratory for Salmonella and Shigella) between 1997 and 2001 were serotyped using conventional agglutination methods, and Enteritidis, Typhimurium, Hadar, Virchow and Typhi serotypes were additionally phage typed according to internationally-developed schemes. A total of 30,856 Salmonella spp. strains, isolated in the majority of Spanish Autonomous Communities, were analyzed. Enteritidis (51%) and Typhimurium (24%) were the most frequently isolated serotypes. The following were the most frequent serotype/phage type combinations: Enteritidis/PT1 (18%), Enteritidis/PT4 (15%), Enteritidis/PT6a (5%), Typhimurium/DT104 (5%) and Enteritidis/PT6 (3%). The serotype Enteritidis/PT1 showed the greatest increase over the period studied, from 11.61% in 1997 to 24.74% in 2001. A hierarchical typing approach for Salmonella spp., using serotyping coupled with phage typing allowed a higher level of discrimination among Salmonella serotypes. Application of this approach in epidemiological studies could be highly useful for early characterization of related strains.

  9. 76 FR 81513 - Guidance for Industry: Prevention of Salmonella

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-28

    ...] Guidance for Industry: Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and... ``Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation.'' The document provides guidance to egg producers on how to comply with certain provisions contained in FDA's...

  10. Clinical and veterinary isolates of Salmonella enterica serovar Enteritidis defective in lipopolysaccharide O-chain polymerization

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Guard-Petter, J.; Parker, C.T.; Asokan, K.

    1999-05-01

    Twelve human and chicken isolates of Salmonella enterica serovar Enteritidis belonging to phage types 4, 8, 13a, and 23 were characterized for variability in lipopolysaccharide (LPS) composition. Isolates were differentiated into two groups, i.e., those that lacked immunoreactive O-chain, termed rough isolates, and those that had immunoreactive O-chain, termed smooth isolates. Isolates within these groups could be further differentiated by LPS compositional differences as detected by gel electrophoresis and gas liquid chromatography of samples extracted with water, which yielded significantly more LPS in comparison to phenol-chloroform extraction. The rough isolates were of two types, the O-antigen synthesis mutants and themore » O-antigen polymerization (wzy) mutants. Smooth isolates were also of two types, one producing low-molecular-weight (LMW) LPS and the other producing high-molecular-weight (HMW) LPS. To determine the genetic basis for the O-chain variability of the smooth isolates, the authors analyzed the effects of a null mutation in the O-chain length determinant gene, wzz (cld) of serovar Typhimurium. This mutation results in a loss of HMW LPS; however, the LMW LPS of this mutant was longer and more glucosylated than that from clinical isolates of serovar Enteritidis. Cluster analysis of these data and of those from two previously characterized isogenic strains of serovar Enteritidis that had different virulence attributes indicated that glucosylation of HMW LPS (via oafR function) is variable and results in two types of HMW structures, one that is highly glucosylated and one that is minimally glucosylated. These results strongly indicate that naturally occurring variability in wzy, wzz, and oafR function can be used to subtype isolates of serovar Enteritidis during epidemiological investigations.« less

  11. Geographical information software and shopper card data, aided in the discovery of a Salmonella Enteritidis outbreak associated with Turkish pine nuts.

    PubMed

    Bedard, B; Kennedy, B S; Weimer, A C

    2014-12-01

    In 2011, from August to November, the Monroe County Department of Public Health (MCDPH) investigated 47 salmonellosis cases. Geographical information software (GIS) was used to map the address locations of these cases. The resulting GIS analysis and culture information indicated that there were two distinct clusters of Salmonella that were geographically different. Pulsed-field gel electrophoresis (PFGE) testing was run at the New York State Department of Health Wadsworth Laboratory and identified S. Enteritidis (23 cases) and S. Typhimurium (10 cases). The epidemiological investigation identified Turkish pine nuts as the link between ill S. Enteritidis cases. Pine nut samples sent for laboratory testing were a PFGE match to human isolates with S. Enteritidis. A national recall of Turkish pine nuts ensued. A multistate outbreak was identified as a result of the initial investigation of MCDPH, in which 43 people were infected with the outbreak strain from five states. GIS software and shopper card data provided important tools in the epidemiological investigation.

  12. Changes in thermo-tolerance and survival under simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 in chicken breast meat after exposure to sequential stresses.

    PubMed

    Melo, Adma Nadja Ferreira de; Souza, Geany Targino de; Schaffner, Donald; Oliveira, Tereza C Moreira de; Maciel, Janeeyre Ferreira; Souza, Evandro Leite de; Magnani, Marciane

    2017-06-19

    This study assessed changes in thermo-tolerance and capability to survive to simulated gastrointestinal conditions of Salmonella Enteritidis PT4 and Salmonella Typhimurium PT4 inoculated in chicken breast meat following exposure to stresses (cold, acid and osmotic) commonly imposed during food processing. The effects of the stress imposed by exposure to oregano (Origanum vulgare L.) essential oil (OVEO) on thermo-tolerance were also assessed. After exposure to cold stress (5°C for 5h) in chicken breast meat the test strains were sequentially exposed to the different stressing substances (lactic acid, NaCl or OVEO) at sub-lethal amounts, which were defined considering previously determined minimum inhibitory concentrations, and finally to thermal treatment (55°C for 30min). Resistant cells from distinct sequential treatments were exposed to simulated gastrointestinal conditions. The exposure to cold stress did not result in increased tolerance to acid stress (lactic acid: 5 and 2.5μL/g) for both strains. Cells of S. Typhimurium PT4 and S. Enteritidis PT4 previously exposed to acid stress showed higher (p<0.05) tolerance to osmotic stress (NaCl: 75 or 37.5mg/g) compared to non-acid-exposed cells. Exposure to osmotic stress without previous exposure to acid stress caused a salt-concentration dependent decrease in counts for both strains. Exposure to OVEO (1.25 and 0.62μL/g) decreased the acid and osmotic tolerance of both S. Enteritidis PT4 and S. Typhimurium PT4. Sequential exposure to acid and osmotic stress conditions after cold exposure increased (p<0.05) the thermo-tolerance in both strains. The cells that survived the sequential stress exposure (resistant) showed higher tolerance (p<0.05) to acidic conditions during continuous exposure (182min) to simulated gastrointestinal conditions. Resistant cells of S. Enteritidis PT4 and S. Typhimurium PT4 showed higher survival rates (p<0.05) than control cells at the end of the in vitro digestion. These results show

  13. Quantification of horizontal transmission of Salmonella enterica serovar Enteritidis bacteria in pair-housed groups of laying hens.

    PubMed

    Thomas, M E; Klinkenberg, D; Ejeta, G; Van Knapen, F; Bergwerff, A A; Stegeman, J A; Bouma, A

    2009-10-01

    An important source of human salmonellosis is the consumption of table eggs contaminated with Salmonella enterica serovar Enteritidis. Optimization of the various surveillance programs currently implemented to reduce human exposure requires knowledge of the dynamics of S. Enteritidis infection within flocks. The aim of this study was to provide parameter estimates for a transmission model of S. Enteritidis in laying-type chicken flocks. An experiment was carried out with 60 pairs of laying hens. Per pair, one hen was inoculated with S. Enteritidis and the other was contact exposed. After inoculation, cloacal swab samples from all hens were collected over 18 days and tested for the presence of S. Enteritidis. On the basis of this test, it was determined if and when each contact-exposed hen became colonized. A transmission model including a latency period of 1 day and a slowly declining infectivity level was fitted. The mean initial transmission rate was estimated to be 0.47 (95% confidence interval [CI], 0.30 to 0.72) per day. The reproduction number R(0), the average number of hens infected by one colonized hen in a susceptible population, was estimated to be 2.8 (95% CI, 1.9 to 4.2). The generation time, the average time between colonization of a "primary" hen and colonization of contact-exposed hens, was estimated to be 7.0 days (95% CI, 5.0 to 11.6 days). Simulations using these parameters showed that a flock of 20,000 hens would reach a maximum colonization level of 92% within 80 days after colonization of the first hen. These results can be used, for example, to evaluate the effectiveness of control and surveillance programs and to optimize these programs in a cost-benefit analysis.

  14. A Mexican restaurant-associated outbreak of Salmonella Enteritidis type 34 infections traced to a contaminated egg farm.

    PubMed Central

    McNeil, M. M.; Sweat, L. B.; Carter, S. L.; Watson, C. B.; Holloway, J. T.; Manning, R.; Altekruse, S. F.; Blake, P. A.

    1999-01-01

    In May 1996, the Georgia Division of Public Health was notified about a cluster of persons with Salmonella Enteritidis (SE) infections in Waycross, Georgia. A matched pair case-control study to determine risk factors for illness found a statistically significant association of SE infection with a history of having eaten at Restaurant A during the 5 days before onset of illness (relative risk = 13 [95% confidence interval (CI) = 3-62, P < 0.01]). In a second case-control study, to determine specific food exposures, consumption of a deep-fried Mexican dish (chile relleno) (4 of 21 cases vs. 0 of 26 controls, odds ratio undefined, 95% CI > 1.46, P = 0.034) was found to be significantly associated with SE infection. An environmental investigation found evidence of suboptimal food storage and cooking temperatures at Restaurant A; cross contamination of foods may have contributed to the low attributable risk identified for chile rellenos. Five of 37 Restaurant A food and environment specimens yielded SE strains. All five positive specimens were from chiles rellenos. Of the seven outbreak-associated strains (six patient isolates and one food isolate from Restaurant A) for which phage typing was conducted, all were phage type 34. A FDA traceback investigation through Restaurant A's single-egg supplier identified the potential source as three interrelated farms in South Carolina. Environmental culture from one of these farms yielded SE phage type 34. As a result of this outbreak, FDA helped institute a statewide egg quality-assurance programme in South Carolina to minimize SE contamination of eggs. PMID:10355784

  15. A Mexican restaurant-associated outbreak of Salmonella Enteritidis type 34 infections traced to a contaminated egg farm.

    PubMed

    McNeil, M M; Sweat, L B; Carter, S L; Watson, C B; Holloway, J T; Manning, R; Altekruse, S F; Blake, P A

    1999-04-01

    In May 1996, the Georgia Division of Public Health was notified about a cluster of persons with Salmonella Enteritidis (SE) infections in Waycross, Georgia. A matched pair case-control study to determine risk factors for illness found a statistically significant association of SE infection with a history of having eaten at Restaurant A during the 5 days before onset of illness (relative risk = 13 [95% confidence interval (CI) = 3-62, P < 0.01]). In a second case-control study, to determine specific food exposures, consumption of a deep-fried Mexican dish (chile relleno) (4 of 21 cases vs. 0 of 26 controls, odds ratio undefined, 95% CI > 1.46, P = 0.034) was found to be significantly associated with SE infection. An environmental investigation found evidence of suboptimal food storage and cooking temperatures at Restaurant A; cross contamination of foods may have contributed to the low attributable risk identified for chile rellenos. Five of 37 Restaurant A food and environment specimens yielded SE strains. All five positive specimens were from chiles rellenos. Of the seven outbreak-associated strains (six patient isolates and one food isolate from Restaurant A) for which phage typing was conducted, all were phage type 34. A FDA traceback investigation through Restaurant A's single-egg supplier identified the potential source as three interrelated farms in South Carolina. Environmental culture from one of these farms yielded SE phage type 34. As a result of this outbreak, FDA helped institute a statewide egg quality-assurance programme in South Carolina to minimize SE contamination of eggs.

  16. Comparative analysis of virulence and resistance profiles of Salmonella Enteritidis isolates from poultry meat and foodborne outbreaks in northern Jordan

    PubMed Central

    Jaradat, Ziad W; Abedel Hafiz, Leena; Ababneh, Mustafa M; Ababneh, Qotaibah O; Al Mousa, Waseem; Al-Nabulsi, Anas; Osaili, Tareq M; Holley, Richard

    2014-01-01

    This study was conducted to isolate Salmonella Enteritidis from poultry samples and compare their virulence and antibiotic resistance profiles to S. Enteritidis isolated from outbreaks in northern Jordan. Two hundred presumptive isolates were obtained from 302 raw poultry samples and were subjected to further analysis and confirmation. A phylogenic tree based on 16S rRNA sequencing was constructed and selected isolates representing each cluster were further studied for their virulence in normal adult Swiss white mice. The most virulent strains were isolated from poultry samples and had an LD50 of 1.55 × 105 CFU, while some of the outbreak isolates were avirulent in mice. Antibiotic resistance profiling revealed that the isolates were resistant to seven of eight antibiotics screened with each isolate resistant to multiple antibiotics (from two to six). Of the poultry isolates, 100%, 88.9%, 77.8%, 66.7%, and 50% showed resistance to nalidixic acid, ciprofloxacin, ampicillin, cephalothin, and cefoperazone, respectively. Two outbreak isolates were sensitive to all tested antibiotics, while 71.4% were resistant to cefoperazone and only 28.6% showed resistance to nalidixic acid. Salmonella outbreak isolates were genetically related to poultry isolates as inferred from the 16S rRNA sequencing, yet were phenotypically different. Although outbreak strains were similar to poultry isolates, when tested in the mouse model, some of the outbreak isolates were highly virulent while others were avirulent. This might be due to a variation in susceptibility of the mouse to different S. Enteritidis isolates. PMID:24780883

  17. The epidemiology of travel-related Salmonella Enteritidis in Ontario, Canada, 2010–2011

    PubMed Central

    2012-01-01

    Background Increases in the number of salmonellosis cases due to Salmonella Enteritidis (SE) in 2010 and 2011 prompted a public health investigation in Ontario, Canada. In this report, we describe the current epidemiology of travel-related (TR) SE, compare demographics, symptoms and phage types (PTs) of TR and domestically-acquired (DA) cases, and estimate the odds of acquiring SE by region of the world visited. Methods All incident cases of culture confirmed SE in Ontario obtained from isolates and specimens submitted to public health laboratories were included in this study. Demographic and illness characteristics of TR and DA cases were compared. A national travel survey was used to provide estimates for the number of travellers to various destinations to approximate rates of SE in travellers. Multivariate logistic regression was used to estimate the odds of acquiring SE when travelling to various world regions. Results Overall, 51.9% of SE cases were TR during the study period. This ranged from 35.7% TR cases in the summer travel period to 65.1% TR cases in the winter travel period. Compared to DA cases, TR cases were older and were less likely to seek hospital care. For Ontario travellers, the adjusted odds of acquiring SE was the highest for the Caribbean (OR 37.29, 95% CI 17.87-77.82) when compared to Europe. Certain PTs were more commonly associated with travel (e.g., 1, 4, 5b, 7a, Atypical) than with domestic infection. Of the TR cases, 88.9% were associated with travel to the Caribbean and Mexico region, of whom 90.1% reported staying on a resort. Within this region, there were distinct associations between PTs and countries. Conclusions There is a large burden of TR illness from SE in Ontario. Accurate classification of cases by travel history is important to better understand the source of infections. The findings emphasize the need to make travellers, especially to the Caribbean, and health professionals who provide advice to travellers, aware of this

  18. Acyl homoserine lactone changes the abundance of proteins and the levels of organic acids associated with stationary phase in Salmonella Enteritidis.

    PubMed

    de Almeida, Felipe Alves; Pimentel-Filho, Natan de Jesus; Carrijo, Lanna Clícia; Bento, Cláudia Braga Pereira; Baracat-Pereira, Maria Cristina; Pinto, Uelinton Manoel; de Oliveira, Leandro Licursi; Vanetti, Maria Cristina Dantas

    2017-01-01

    Quorum sensing (QS) is cell-cell communication mechanism mediated by signaling molecules known as autoinducers (AIs) that lead to differential gene expression. Salmonella is unable to synthesize the AI-1 acyl homoserine lactone (AHL), but is able to recognize AHLs produced by other microorganisms through SdiA protein. Our study aimed to evaluate the influence of AI-1 on the abundance of proteins and the levels of organic acids of Salmonella Enteritidis. The presence of N-dodecyl-homoserine lactone (C12-HSL) did not interfere on the growth or the total amount of extracted proteins of Salmonella. However, the abundance of the proteins PheT, HtpG, PtsI, Adi, TalB, PmgI (or GpmI), Eno, and PykF enhanced while the abundance of the proteins RplB, RplE, RpsB, Tsf, OmpA, OmpC, OmpD, and GapA decreased when Salmonella Enteritidis was anaerobically cultivated in the presence of C12-HSL. Additionally, the bacterium produced less succinic, lactic, and acetic acids in the presence of C12-HSL. However, the concentration of extracellular formic acid reached 20.46 mM after 24 h and was not detected when the growth was in the absence of AI-1. Considering the cultivation period for protein extraction, their abundance, process and function, as well as the levels of organic acids, we observed in cells cultivated in presence of C12-HSL a correlation with what is described in the literature as entry into the stationary phase of growth, mainly related to nitrogen and amino acid starvation and acid stress. Further studies are needed in order to determine the specific role of the differentially abundant proteins and extracellular organic acids secreted by Salmonella in the presence of quorum sensing signaling molecules. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Effects of 5-hydroxytryptophan and m-hydroxybenzylhydrazine associated to Lactobacillus spp. on the humoral response of broilers challenged with Salmonella Enteritidis.

    PubMed

    Donato, T C; Baptista, A A S; Garcia, K C O D; Smaniotto, B D; Okamoto, A S; Sequeira, J L; Andreatti Filho, R L

    2015-09-01

    This study investigates the effects of different doses of serotonin, its precursor 5-hydroxytry-ptophan (5HTP), and m-hydroxybenzylhydrazine inhibitor (NSD1015), administered via intraperitoneal for 5 consecutive days, on behavior and average body weight of broilers. We also measured the humoral immune response and quantification of Salmonella Enteritidis in broilers chickens that received the drugs evaluated and a Lactobacillus pool. The study was divided into 3 experiments: Experiment 1--administration of pharmaceuticals with choice of dosage; Experiment 2--administration of pharmaceuticals and a Lactobacillus pool in birds that were not challenged with S. Enteritidis, and Experiment 3--administration of pharmaceuticals and a Lactobacillus pool in birds challenged with S. Enteritidis. The ELISA was used to scan dosages of intestinal IgA and serum IgY. We used colony-forming units to quantify S. Enteritidis. The concentrations of IgA and IgY did not show significant differences (P>0.05) in Experiment 2. In Experiment 3, NSD1015 associated with Lactobacillus determined higher IgA concentrations, promoting greater stimulus to the immune system than 5HTP. Regarding quantification of S. Enteritidis in the cecal content of birds, 5HTP associated to Lactobacillus determined the smallest number of bacteria, showing possible interaction of 5-hydroxytryptophan and Lactobacillus spp. with the immune system of broiler chickens. © 2015 Poultry Science Association Inc.

  20. The effect of dietary fructooligosaccharide supplementation on growth performance, intestinal morphology, and immune responses in broiler chickens challenged with Salmonella Enteritidis lipopolysaccharides.

    PubMed

    Shang, Yue; Regassa, Alemu; Kim, Ji Hyuk; Kim, Woo Kyun

    2015-12-01

    This study was conducted to examine the effects of fructooligosaccharide (FOS) supplementation on growth performance, lymphoid organ weight, intestinal morphology, and immunological status in broilers (n=180) challenged with Salmonella Enteritidis lipopolysaccharides (LPS). Birds were randomly assigned into a 3×2 factorial arrangement that included 1) 3 dietary treatments from d one to 21: positive control (PC), wheat-corn-soybean meal based diet contained antibiotics (virginiamycin and monensin); negative control (NC), as PC without antibiotics; and NC+FOS, as NC supplemented with 0.5% FOS, and 2) 2 intraperitoneal injections: 2 mg/kg Salmonella Enteritidis LPS or sterile phosphate buffered saline (PBS) on d 21. Growth performance and relative lymphoid organ weight were not significantly different among the treatments. Villus height, crypt depth, and total mucosa thickness were significantly increased (P<0.05) in the ileum of broiler chickens fed NC+FOS when compared to PC and NC. Birds in NC+FOS treatment had reduced heterophil but increased monocyte count when compared to NC (P<0.05). Significant diet×challenge interaction was observed on natural IgY levels (P<0.0001), and a significant dietary effect was observed on specific IgY levels in chickens fed NC+FOS (P=0.003). Supplementation of FOS also increased the expression of interleukin (IL)-1ß, -10, and interferon (IFN)-γ mRNA in the ileum of the birds. In summary, Salmonella Enteritidis LPS challenge established significant differences in the immune responses in broiler chickens. FOS supplementation increased ileal mucosa thickness and elevated the expressions of certain cytokine genes. It also led to the alteration of leukocyte compositions and serum IgY levels in response to LPS challenge, suggesting FOS supplementation may be effective to induce protective outcomes in gut health and immunity of broiler chickens. © 2015 Poultry Science Association Inc.

  1. Salmonella enterica Serovar Enteritidis Antimicrobial Peptide Resistance Genes Aid in Defense against Chicken Innate Immunity, Fecal Shedding, and Egg Deposition

    PubMed Central

    McKelvey, Jessica A.; Yang, Ming; Jiang, Yanhua

    2014-01-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major etiologic agent of nontyphoid salmonellosis in the United States. S. Enteritidis persistently and silently colonizes the intestinal and reproductive tract of laying hens, resulting in contaminated poultry products. The consumption of contaminated poultry products has been identified as a significant risk factor for human salmonellosis. To understand the mechanisms S. Enteritidis utilizes to colonize and persist in laying hens, we used selective capture of transcribed sequences to identify genes overexpressed in the HD11 chicken macrophage cell line and in primary chicken oviduct epithelial cells. From the 15 genes found to be overexpressed in both cell types, we characterized the antimicrobial peptide resistance (AMPR) genes, virK and ybjX, in vitro and in vivo. In vitro, AMPR genes were required for natural morphology, motility, secretion, defense against detergents such as EDTA and bile salts, and resistance to antimicrobial peptides polymyxin B and avian β-defensins. From this, we inferred the AMPR genes play a role in outer membrane stability and/or modulation. In the intestinal tract, AMPR genes were involved in early intestinal colonization and fecal shedding. In the reproductive tract, virK was required in early colonization whereas a deletion of ybjX caused prolonged ovary colonization and egg deposition. Data from the present study indicate that AMPR genes are differentially utilized in various host environments, which may ultimately assist S. Enteritidis in persistent and silent colonization of chickens. PMID:25267840

  2. 75 FR 48973 - Draft Guidance for Industry: Prevention of Salmonella

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-12

    ...] Draft Guidance for Industry: Prevention of Salmonella Enteritidis in Shell Eggs During Production... entitled ``Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation'' (the draft guidance). The draft guidance, when finalized, will provide guidance to egg producers...

  3. Application of predictive models to assess the influence of thyme essential oil on Salmonella Enteritidis behaviour during shelf life of ready-to-eat turkey products.

    PubMed

    Possas, Arícia; Posada-Izquierdo, Guiomar D; Pérez-Rodríguez, Fernando; Valero, Antonio; García-Gimeno, Rosa M; Duarte, Marta C T

    2017-01-02

    Consumers' demand for ready-to-eat (RTE) turkey meat is attributed to its convenience and healthy properties. However, as cooked meat product it is subjected to post-process contamination, thus allowing presence and growth of microbial pathogens, such as Salmonella spp.. The aim of this study was to include a natural antimicrobial, thyme essential oil (TEO), on RTE turkey products in order to evaluate its effectiveness throughout the shelf life. To do so, the effect of four different formulations of cooked RTE turkey products on Salmonella Enteritidis behaviour was investigated. Products' slices were surface inoculated with S. Enteritidis (ca. 4 to 5logcfu/g), subsequently stored at 10 and 25°C and microbiologically analysed during 18 and 12days, respectively. Predictive microbiology models fitted to count data were used to evaluate microbial behaviour. Results showed that S. Enteritidis behaviour on RTE turkey products slices during storage was strongly dependent on temperature. The pathogen was able to grow on slices at all tested conditions during storage at 25°C and no statistical differences were detected (p>0.05) between growth parameters. At 10°C, different behaviour patterns were observed. The application of TEO led to higher Salmonella inactivation rates on a product exempt of chemical preservatives. The addition of this novel antimicrobial on meat products or its incorporation on meat active packaging systems as a part of hurdle technology could increase RTE turkey products safety while satisfying the demand of more natural foods. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. The Effect of Lactobacillus acidophilus PTCC 1643 on Cultured Intestinal Epithelial Cells Infected with Salmonella enterica serovar Enteritidis.

    PubMed

    Moshiri, Mona; Dallal, Mohammad Mehdi Soltan; Rezaei, Farhad; Douraghi, Masoumeh; Sharifi, Laleh; Noroozbabaei, Zahra; Gholami, Mehrdad; Mirshafiey, Abbas

    2017-02-01

    Gastrointestinal disorders caused by Salmonella enterica serovar Enteritidis ( Se sE) are a significant health problem around the globe. Probiotic bacteria have been shown to have positive effects on the immune responses. Lactobacillus acidophilus was examined for its capability to influence the innate immune response of HT29 intestinal epithelial cells towards Se sE. The purpose of this work was to assess the effect of L. acidophilus PTCC 1643 on cultured intestinal epithelial cells infected with Se sE. HT29 cells were cultured in Roswell Park Memorial Institute medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were treated with L. acidophilus PTCC 1643 after or before challenge with Se sE. At 2 and 4 hours post-infection, we measured changes in the expression levels of TLR2 and TLR4 via real-time polymerase chain reaction. Treatment with L. acidophilus inhibited Se sE-induced increases in TLR2 and TLR4 expression in the infected HT29 cells. Moreover, the expression of TLR2 and TLR4 in cells that were pretreated with L. acidophilus and then infected with Se sE was significantly higher than that in cells infected with Se sE without pretreatment. Taken together, the results indicated that L. acidophilus had an anti-inflammatory effect and modulated the innate immune response to Se sE by influencing TLR2 and TLR4 expression. Our findings suggested that L. acidophilus PTCC 1643 was able to suppress inflammation caused by Se sE infection in HT29 cells and reduce TLR2 and TLR4 expression. Additional in vivo and in vitro studies are required to further elucidate the mechanisms underlying this anti-inflammatory effect.

  5. Development of a Loop Mediated Isothermal Amplification (LAMP) - Surface Enhanced Raman spectroscopy (SERS) Assay for the Detection of Salmonella Enterica Serotype Enteritidis.

    PubMed

    Draz, Mohamed Shehata; Lu, Xiaonan

    2016-01-01

    As a major foodborne pathogen, Salmonella enterica serotype Enteritidis is increasingly rising as a global health concern. Here, we developed an integrated assay that combines loop mediated isothermal amplification (LAMP) and surface enhanced Raman spectroscopy (SERS) for DNA detection of S. Enteritidis using specifically designed Raman active Au-nanoprobes. The target DNA was amplified by LAMP and then labeled with Au-nanoprobes comprised of gold nanoparticle-modified with specific cy5/DNA probes to allow the detection by SERS. The sensitivity of the developed LAMP-SERS detection assay (66 CFU/mL) was ~100-fold higher than the conventional polymerase chain reaction (PCR) method. Significantly, this technique allowed highly specific detection of the target DNA of S. Enteritidis and could differentiate it from the DNA of closely related bacterial species or non-specific contamination, making it more accurate and reliable than the standard LAMP technique. The applicability of detection of S. Enteritidis in milk samples using LAMP-SERS assay was validated as well. In sum, the developed LAMP-SERS assay is highly specific and sensitive, and has the potential to be applied for rapid detection of different foodborne pathogens and other microbial contaminants.

  6. A label-free ultrasensitive fluorescence detection of viable Salmonella enteritidis using enzyme-induced cascade two-stage toehold strand-displacement-driven assembly of G-quadruplex DNA.

    PubMed

    Zhang, Peng; Liu, Hui; Ma, Suzhen; Men, Shuai; Li, Qingzhou; Yang, Xin; Wang, Hongning; Zhang, Anyun

    2016-06-15

    The harm of Salmonella enteritidis (S. enteritidis ) to public health mainly by contaminating fresh food and water emphasizes the urgent need for rapid detection techniques to help control the spread of the pathogen. In this assay, an newly designed capture probe complex that contained specific S. enteritidis-aptamer and hybridized signal target sequence was used for viable S. enteritidis recognition directly. In the presence of the target S. enteritidis, single-stranded target sequences were liberated and initiated the replication-cleavage reaction, producing numerous G-quadruplex structures with a linker on the 3'-end. And then, the sensing system took innovative advantage of quadratic linker-induced strand-displacement for the first time to release target sequence in succession, leading to the cyclic reuse of the target sequences and cascade signal amplification, thereby achieving the successive production of G-quadruplex structures. The fluorescent dye, N-Methyl mesoporphyrin IX, binded to these G-quadruplex structures and generated significantly enhanced fluorescent signals to achieve highly sensitive detection of S. enteritidis down to 60 CFU/mL with a linear range from 10(2) to 10(7)CFU/mL. By coupling the cascade two-stage target sequences-recyclable toehold strand-displacement with aptamer-based target recognition successfully, it is the first report on a novel non-label, modification-free and DNA extraction-free ultrasensitive fluorescence biosensor for detecting viable S. enteritidis directly, which can discriminate from dead S. enteritidis. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Quantitative studies of the distribution pattern for Salmonella Enteritidis in the internal organs of chicken after oral challenge by a real-time PCR.

    PubMed

    He, G Z; Tian, W Y; Qian, N; Cheng, A C; Deng, S X

    2010-12-01

    This research was undertaken to identify and understand the regular distribution pattern for Salmonella Enteritidis (S. enteritidis) in the internal organs of chicken after oral challenge over a 3 wk period. We used a real-time, fluorescence-based quantitative polymerase chain reaction (FQ-PCR) to detect genomic DNA of S. enteritidis in the blood and the internal organs, including heart, liver, spleen, kidney, pancreas, and gallbladder, from chicken after oral challenge at different time points. The results showed that the spleen was positive at 12 h post inoculation (PI), and the blood was at 14 h PI. The organism was detected in the liver and heart at 16 h PI, pancrea was positive at 20 h PI, and the final organ to show a positive results were the kidney and gallbladder at 22 h PI. The copy number of S. enteritidis DNA in each tissue reached a peak at 24 h-36 h PI, with the liver and spleen containing high concentrations of S. enteritidis, whereas the blood, heart, kidney, pancreas, and gallbladder had low concentrations. S. enteritidis populations began to decrease and were not detectable at 3 d PI, but were still present up to 12 d PI in the gallbladder, 2 wk for the liver, and 3 wk for the spleen without causing apparent symptoms. The results showed that the liver and spleen may be the primary sites for S. enteritidis setting itself up as a commensa over a long time after oral challenge. Interestingly, it may be the first time reported that the gallbladder is a site of carriage for S. enteritidis over a 12 d period. This study will help to understand the mechanisms of action of S. enteritidis infection in vivo.

  8. Effect of high pressure processing on the survival of Salmonella Enteritidis and shelf-life of chicken fillets.

    PubMed

    Argyri, Anthoula A; Papadopoulou, Olga S; Nisiotou, Aspasia; Tassou, Chrysoula C; Chorianopoulos, Nikos

    2018-04-01

    High pressure processing (HPP) is a preservation technology alternative to heat treatment that is mild for food, but effectively inactivates the spoilage microbiota and foodborne pathogens of several foods. The purpose of the current study was to evaluate the effect of HPP on Salmonella ser. Enteritidis, indigenous microbiota and shelf-life of chicken fillets. Chicken fillets were inoculated with S. Enteritidis at three different initial inocula (3, 5, 7 log CFU/g), packed under vacuum, treated or not with HPP (500 MPa/10 min) and stored at 4 and 12 °C. Total viable counts, S. Enteritidis, pseudomonads, Brochothrix thermosphacta, lactic acid bacteria, Enterobacteriaceae and yeasts/molds populations were determined in parallel with sensory analysis of non-inoculated samples. The HPP resulted in the reduction of the pathogen population below the detection limit of the enumeration method (0.48 log CFU/g), irrespective of the inoculum. During the shelf life of the HPP samples, the pathogens population remained below or near the detection limit of the enumeration method at both temperatures, except from the high inoculum case that an increase was observed at 12 °C. At the low inoculum level, the pathogen could not be detected with the enrichment method after the first storage days (2nd day for 4 °C and 0 day for 12 °C). The survival of Salmonella strains was assessed by pulsed field gel electrophoresis and it was shown that the survival of the different strains depended on the inoculum and storage temperature. Regarding the indigenous microbiota, Br. thermosphacta was reported for the first time to be the main spoilage microorganism that survived and dominated after the HPP. From the results it was evident that, HPP may enhance the safety and increase the shelf life (6 at 4 °C and 2 days at 12 °C) of chicken meat. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Cytokine Signaling in Splenic Leukocytes from Vaccinated and Non-Vaccinated Chickens after Intravenous Infection with Salmonella Enteritidis

    PubMed Central

    Matulova, Marta; Stepanova, Hana; Sisak, Frantisek; Havlickova, Hana; Faldynova, Marcela; Kyrova, Kamila; Volf, Jiri; Rychlik, Ivan

    2012-01-01

    In order to design a new Salmonella enterica vaccine, one needs to understand how naive and immune chickens interact differently when exposed to S. enterica. In this study we therefore determined the immune response of vaccinated and non-vaccinated chickens after intravenous infection with Salmonella enterica serovar Enteritidis (S. Enteritidis). Using flow cytometry we showed that 4 days post infection (DPI), counts of CD4 and B-lymphocytes did not change, CD8 and γδ T-lymphocytes decreased and macrophages and heterophils increased in the spleen. When vaccinated and non-vaccinated chickens were compared, only macrophages and heterophils were found in significantly higher counts in the spleens of the non-vaccinated chickens. The non-vaccinated chickens also expressed higher anti-LPS antibodies than the vaccinated chickens. The expression of interleukin (IL)1β, IL6, IL8, IL18, LITAF, IFNγ and iNOS did not exhibit any clear pattern in the cells sorted from the spleens of vaccinated or non-vaccinated chickens. Only IL17 and IL22 showed a differential expression in the CD4 T-lymphocytes of the vaccinated and non-vaccinated chickens at 4 DPI, both being expressed at a higher level in the non-vaccinated chickens. Due to a similar IFNγ expression in the CD4 T-lymphocytes in both the vaccinated and non-vaccinated chickens, and a variable IL17 expression oscillating around IFNγ expression levels, the IL17∶IFNγ ratio in CD4 T-lymphocytes was found to be central for the outcome of the immune response. When IL17 was expressed at higher levels than IFNγ in the non-vaccinated chickens, the Th17 immune response with a higher macrophage and heterophil infiltration in the spleen dominated. However, when the expression of IL17 was lower than that of IFNγ as in the vaccinated chickens, the Th1 response with a higher resistance to S. Enteritidis infection dominated. PMID:22384225

  10. KH-type splicing regulatory protein is regulated by nuclear factor-κB signaling to mediate innate immunity in Caco-2 cells infected by Salmonella enteritidis.

    PubMed

    Nie, Yuanyang; Cao, Mei; Wu, Daoyan; Li, Ningzhe; Peng, Jingshan; Yi, Sijun; Yang, Xiaofan; Zhang, Mao; Hu, Guoku; Zhao, Jian

    2018-05-04

    Salmonella enteritidis infection occurs in enterogenous diseases, such as gastroenteritis and parenteral focal infection, which often involve inflammation of intestinal epithelial cells. The nuclear factor kappa B (NF-κB) pathway participates in the innate immune response to many gram-negative pathogenic bacteria and initiates inflammation in epithelial cells. KH-type splicing regulatory protein (KSRP) is a multi-domain RNA-binding protein that recruits the exosome-containing mRNA degradation complex to mRNAs coding for inflammatory response factors. However, it remains unclear whether KSRP is regulated by NF-κB signaling pathway in response to S. enteritidis infection and affects the development of inflammation. Accordingly, in this study, we investigated the role of KSRP in mediating the response to S. enteritidis in Caco-2 cells. The data revealed that S. enteritidis infection decreased KSRP expression, which was suppressed by blocking the NF-κB pathway. Additionally, S. enteritidis infection significantly increased the expression of inducible nitric oxide synthase and cyclooxygenase-2. Overexpression of KSRP reduced the expression levels of inflammatory factors in Caco-2 cells. KSRP was regulated by the NF-κB signaling pathway and participated in mediating the innate immune response to S. enteritidis infection in Caco-2 cells, and KSRP acted as a negative regulator of inflammatory gene expression.

  11. Mannan- and xylooligosaccharides modulate caecal microbiota and expression of inflammatory-related cytokines and reduce caecal Salmonella Enteritidis colonisation in young chickens.

    PubMed

    Pourabedin, Mohsen; Chen, Qiaoling; Yang, MingMing; Zhao, Xin

    2017-01-01

    Salmonella Enteritidis is a pathogen, which can infect humans and chickens. This study was designed to address the impact of two potential prebiotics, mannanoligosaccharides (MOS) and xylooligosaccharides (XOS), on the caecal microbiota and expression of cytokines in chickens infected with S. Enteritidis. Newly hatched chicks were assigned to one of five groups: (1) uninfected control, (2) infected control, (3) infected + XOS, (4) infected + MOS and (5) infected + virginiamycin. The number of S. Enteritidis recovered from the caecum was significantly lower, by 1.6 log, in the MOS, and to a less extent (1.0 log) in the XOS-fed birds compared to the infected control. Coprococcus, Ruminococcus and Enterococcus genera were increased in response to MOS, whereas XOS enriched Clostridium, Lactobacillus and Roseburia MOS, but not XOS, lessened the increase of lipopolysaccharide-induced tumour necrosis factor alpha factor and interferon-γ in caecal tonsils after challenge. The canonical correspondence analysis for cytokine genes showed a correlation with the composition of the microbial community at the genus level. Thus, MOS and XOS differently changed the relative abundance of specific microbial genera and the immune response during infection, and these changes were correlated with their abilities to reduce S. Enteritidis colonisation. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Whole genome sequencing reveals an outbreak of Salmonella Enteritidis associated with reptile feeder mice in the United Kingdom, 2012-2015.

    PubMed

    Kanagarajah, Sanch; Waldram, Alison; Dolan, Gayle; Jenkins, Claire; Ashton, Philip M; Carrion Martin, Antonio Isidro; Davies, Robert; Frost, Andrew; Dallman, Timothy J; De Pinna, Elizabeth M; Hawker, Jeremy I; Grant, Kathie A; Elson, Richard

    2018-05-01

    Analysis of whole genome sequencing data uncovered a previously undetected outbreak of Salmonella Enteritidis that had been on-going for four years. Cases were resident in all countries of the United Kingdom and 40% of the cases were aged less than 11 years old. Initial investigations revealed that 30% of cases reported exposure to pet snakes. A case-control study was designed to test the hypothesis that exposure to reptiles or their feed were risk factors. A robust case-definition, based on the single nucleotide polymorphism (SNP) profile, increased the power of the analytical study. Following univariable and multivariable analysis, exposure to snakes was the only variable independently associated with infection (Odds ratio 810 95% CI (85-7715) p < 0.001). Isolates of S. Enteritidis belonging to the outbreak profile were recovered from reptile feeder mice sampled at the retail and wholesale level. Control measures included improved public health messaging at point of sale, press releases and engagement with public health and veterinary counterparts across Europe. Mice destined to be fed to reptiles are not regarded as pet food and are not routinely tested for pathogenic bacteria. Routine microbiological testing to ensure feeder mice are free from Salmonella is recommended. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  13. A novel prophage identified in strains from Salmonella enterica serovar Enteritidis is a phylogenetic signature of the lineage ST-1974

    PubMed Central

    D'Alessandro, Bruno; Pérez Escanda, Victoria; Balestrazzi, Lucía; Iriarte, Andrés; Pickard, Derek; Yim, Lucía; Chabalgoity, José Alejandro; Betancor, Laura

    2018-01-01

    Salmonella enterica serovar Enteritidis is a major agent of foodborne diseases worldwide. In Uruguay, this serovar was almost negligible until the mid 1990s but since then it has become the most prevalent. Previously, we characterized a collection of strains isolated from 1988 to 2005 and found that the two oldest strains were the most genetically divergent. In order to further characterize these strains, we sequenced and annotated eight genomes including those of the two oldest isolates. We report on the identification and characterization of a novel 44 kbp Salmonella prophage found exclusively in these two genomes. Sequence analysis reveals that the prophage is a mosaic, with homologous regions in different Salmonella prophages. It contains 60 coding sequences, including two genes, gogB and sseK3, involved in virulence and modulation of host immune response. Analysis of serovar Enteritidis genomes available in public databases confirmed that this prophage is absent in most of them, with the exception of a group of 154 genomes. All 154 strains carrying this prophage belong to the same sequence type (ST-1974), suggesting that its acquisition occurred in a common ancestor. We tested this by phylogenetic analysis of 203 genomes representative of the intraserovar diversity. The ST-1974 forms a distinctive monophyletic lineage, and the newly described prophage is a phylogenetic signature of this lineage that could be used as a molecular marker. The phylogenetic analysis also shows that the major ST (ST-11) is polyphyletic and might have given rise to almost all other STs, including ST-1974. PMID:29509137

  14. Factors affecting thermal resistance of Salmonella enterica serovar enteritidis ODA 99-30581-13 in shell egg contents and use of heat-ozone combinations for egg pasteurization.

    PubMed

    Perry, Jennifer J; Yousef, Ahmed E

    2013-02-01

    Infection of laying hens with Salmonella enterica serovar Enteritidis leads to deposition of the pathogen into the albumen or yolk of forming eggs. Heat treatment can inactivate internalized Salmonella Enteritidis in shell eggs, but factors such as the nature and location of contamination may influence the efficacy of thermal treatments. In the current research, natural contamination was mimicked by introducing small inocula of Salmonella Enteritidis into different locations of shell eggs and incubating inoculated eggs. These pathogen-containing eggs were heated at 57°C for 40 min, and temperature within eggs was monitored at the locations of inocula. Comparison of inactivation at equivalent internal temperatures revealed similar levels of lethality regardless of inoculum location. Refrigeration between incubation and heat treatment did not increase thermal resistance of cells in albumen but decreased cell inactivation in yolk. Sequential application of heat and gaseous ozone allows for the development of a process capable of decontaminating shell eggs with minimal thermal treatment and impact on egg quality. Inoculated eggs were subjected to (i) an immersion heating process similar to that used in commercial pasteurization or (ii) immersion heating, at reduced duration, followed by vacuum (50.8 kPa) and treatment with ozone gas (maximum 160 g/m(3)) under pressure (∼187.5 kPa). All treatments tested produced greater than 5-log inactivation, which is required for "pasteurization" processes. Differences were observed in the visual quality of eggs depending on treatment parameters. Application of ozone subsequent to heating allows for a significant reduction in heating time without decreasing process lethality.

  15. Immunogenicity and efficacy following sequential parenterally-administered doses of Salmonella Enteritidis COPS:FliC glycoconjugates in infant and adult mice.

    PubMed

    Baliban, Scott M; Curtis, Brittany; Toema, Deanna; Tennant, Sharon M; Levine, Myron M; Pasetti, Marcela F; Simon, Raphael

    2018-05-23

    In sub-Saharan Africa, invasive nontyphoidal Salmonella (iNTS) infections with serovars S. Enteritidis, S. Typhimurium and I 4,[5],12:i:- are widespread in children < 5 years old. Development of an efficacious vaccine would provide an important public health tool to prevent iNTS disease in this population. Glycoconjugates of S. Enteritidis core and O-polysaccharide (COPS) coupled to the homologous serovar phase 1 flagellin protein (FliC) were previously shown to be immunogenic and protected adult mice against death following challenge with a virulent Malian S. Enteritidis blood isolate. This study extends these observations to immunization of mice in early life and also assesses protection with partial and full regimens. Anti-COPS and anti-FliC serum IgG titers were assessed in infant and adult mice after immunization with 1, 2 or 3 doses of S. Enteritidis COPS:FliC alone or co-formulated with aluminum hydroxide or monophosphoryl lipid A (MPL) adjuvants. S. Enteritidis COPS:FliC was immunogenic in both age groups, although the immune responses were quantitatively lower in infants. Kinetics of antibody production were similar for the native and adjuvanted formulations after three doses; conjugates formulated with MPL elicited significantly increased anti-COPS IgG titers in adult but not infant mice. Nevertheless, robust protection against S. Enteritidis challenge was seen for all three formulations when three doses were given either during infancy or as adults. We further found that significant protection could be achieved with two COPS:FliC doses, despite elicitation of modest serum anti-COPS IgG antibody titers. These findings guide potential immunization strategies that may be translated to develop a human pediatric iNTS vaccine for sub-Saharan Africa.

  16. Spatial-temporal epidemiology of human Salmonella Enteritidis infections with major phage types (PTs 1, 4, 5b, 8, 13, and 13a) in Ontario, Canada, 2008-2009.

    PubMed

    Varga, Csaba; Pearl, David L; McEwen, Scott A; Sargeant, Jan M; Pollari, Frank; Guerin, Michele T

    2015-12-17

    In Ontario and Canada, the incidence of human Salmonella enterica serotype Enteritidis (S. Enteritidis) infections have increased steadily during the last decade. Our study evaluated the spatial and temporal epidemiology of the major phage types (PTs) of S. Enteritidis infections to aid public health practitioners design effective prevention and control programs. Data on S. Enteritidis infections between January 1, 2008 and December 31, 2009 were obtained from Ontario's disease surveillance system. Salmonella Enteritidis infections with major phage types were classified by their annual health region-level incidence rates (IRs), monthly IRs, clinical symptoms, and exposure settings. A scan statistic was employed to detect retrospective phage type-specific spatial, temporal, and space-time clusters of S. Enteritidis infections. Space-time cluster cases' exposure settings were evaluated to identify common exposures. 1,336 cases were available for analysis. The six most frequently reported S. Enteritidis PTs were 8 (n = 398), 13a (n = 218), 13 (n = 198), 1 (n = 132), 5b (n = 83), and 4 (n = 76). Reported rates of S. Enteritidis infections with major phage types varied by health region and month. International travel and unknown exposure settings were the most frequently reported settings for PT 5b, 4, and 1 cases, whereas unknown exposure setting, private home, food premise, and international travel were the most frequently reported settings for PT 8, 13, and 13a cases. Diarrhea, abdominal pain, and fever were the most commonly reported clinical symptoms. A number of phage type-specific spatial, temporal, and space-time clusters were identified. Space-time clusters of PTs 1, 4, and 5b occurred mainly during the winter and spring months in the North West, North East, Eastern, Central East, and Central West regions. Space-time clusters of PTs 13 and 13a occurred at different times of the year in the Toronto region. Space-time clusters of PT 8

  17. Characterization of Foodborne Outbreaks of Salmonella enterica Serovar Enteritidis with Whole-Genome Sequencing Single Nucleotide Polymorphism-Based Analysis for Surveillance and Outbreak Detection.

    PubMed

    Taylor, Angela J; Lappi, Victoria; Wolfgang, William J; Lapierre, Pascal; Palumbo, Michael J; Medus, Carlota; Boxrud, David

    2015-10-01

    Salmonella enterica serovar Enteritidis is a significant cause of gastrointestinal illness in the United States; however, current molecular subtyping methods lack resolution for this highly clonal serovar. Advances in next-generation sequencing technologies have made it possible to examine whole-genome sequencing (WGS) as a potential molecular subtyping tool for outbreak detection and source trace back. Here, we conducted a retrospective analysis of S. Enteritidis isolates from seven epidemiologically confirmed foodborne outbreaks and sporadic isolates (not epidemiologically linked) to determine the utility of WGS to identify outbreaks. A collection of 55 epidemiologically characterized clinical and environmental S. Enteritidis isolates were sequenced. Single nucleotide polymorphism (SNP)-based cluster analysis of the S. Enteritidis genomes revealed well supported clades, with less than four-SNP pairwise diversity, that were concordant with epidemiologically defined outbreaks. Sporadic isolates were an average of 42.5 SNPs distant from the outbreak clusters. Isolates collected from the same patient over several weeks differed by only two SNPs. Our findings show that WGS provided greater resolution between outbreak, sporadic, and suspect isolates than the current gold standard subtyping method, pulsed-field gel electrophoresis (PFGE). Furthermore, results could be obtained in a time frame suitable for surveillance activities, supporting the use of WGS as an outbreak detection and characterization method for S. Enteritidis. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. A Weibull model to describe antimicrobial kinetics of oregano and lemongrass essential oils against Salmonella Enteritidis in ground beef during refrigerated storage.

    PubMed

    de Oliveira, Thales Leandro Coutinho; Soares, Rodrigo de Araújo; Piccoli, Roberta Hilsdorf

    2013-03-01

    The antimicrobial effect of oregano (Origanum vulgare L.) and lemongrass (Cymbopogon citratus (DC.) Stapf.) essential oils (EOs) against Salmonella enterica serotype Enteritidis in in vitro experiments, and inoculated in ground bovine meat during refrigerated storage (4±2 °C) for 6 days was evaluated. The Weibull model was tested to fit survival/inactivation bacterial curves (estimating of p and δ parameters). The minimum inhibitory concentration (MIC) value for both EOs on S. Enteritidis was 3.90 μl/ml. The EO concentrations applied in the ground beef were 3.90, 7.80 and 15.60 μl/g, based on MIC levels and possible activity reduction by food constituents. Both evaluated EOs in all tested levels, showed antimicrobial effects, with microbial populations reducing (p≤0.05) along time storage. Evaluating fit-quality parameters (RSS and RSE) Weibull models are able to describe the inactivation curves of EOs against S. Enteritidis. The application of EOs in processed meats can be used to control pathogens during refrigerated shelf-life. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. Genomic Comparison of Non-Typhoidal Salmonella enterica Serovars Typhimurium, Enteritidis, Heidelberg, Hadar and Kentucky Isolates from Broiler Chickens

    PubMed Central

    Dhanani, Akhilesh S.; Block, Glenn; Dewar, Ken; Forgetta, Vincenzo; Topp, Edward; Beiko, Robert G.; Diarra, Moussa S.

    2015-01-01

    Background Non-typhoidal Salmonella enterica serovars, associated with different foods including poultry products, are important causes of bacterial gastroenteritis worldwide. The colonization of the chicken gut by S. enterica could result in the contamination of the environment and food chain. The aim of this study was to compare the genomes of 25 S. enterica serovars isolated from broiler chicken farms to assess their intra- and inter-genetic variability, with a focus on virulence and antibiotic resistance characteristics. Methodology/Principal Finding The genomes of 25 S. enterica isolates covering five serovars (ten Typhimurium including three monophasic 4,[5],12:i:, four Enteritidis, three Hadar, four Heidelberg and four Kentucky) were sequenced. Most serovars were clustered in strongly supported phylogenetic clades, except for isolates of serovar Enteritidis that were scattered throughout the tree. Plasmids of varying sizes were detected in several isolates independently of serovars. Genes associated with the IncF plasmid and the IncI1 plasmid were identified in twelve and four isolates, respectively, while genes associated with the IncQ plasmid were found in one isolate. The presence of numerous genes associated with Salmonella pathogenicity islands (SPIs) was also confirmed. Components of the type III and IV secretion systems (T3SS and T4SS) varied in different isolates, which could explain in part, differences of their pathogenicity in humans and/or persistence in broilers. Conserved clusters of genes in the T3SS were detected that could be used in designing effective strategies (diagnostic, vaccination or treatments) to combat Salmonella. Antibiotic resistance genes (CMY, aadA, ampC, florR, sul1, sulI, tetAB, and srtA) and class I integrons were detected in resistant isolates while all isolates carried multidrug efflux pump systems regardless of their antibiotic susceptibility profile. Conclusions/Significance This study showed that the predominant

  20. Genomic Comparison of Non-Typhoidal Salmonella enterica Serovars Typhimurium, Enteritidis, Heidelberg, Hadar and Kentucky Isolates from Broiler Chickens.

    PubMed

    Dhanani, Akhilesh S; Block, Glenn; Dewar, Ken; Forgetta, Vincenzo; Topp, Edward; Beiko, Robert G; Diarra, Moussa S

    2015-01-01

    Non-typhoidal Salmonella enterica serovars, associated with different foods including poultry products, are important causes of bacterial gastroenteritis worldwide. The colonization of the chicken gut by S. enterica could result in the contamination of the environment and food chain. The aim of this study was to compare the genomes of 25 S. enterica serovars isolated from broiler chicken farms to assess their intra- and inter-genetic variability, with a focus on virulence and antibiotic resistance characteristics. The genomes of 25 S. enterica isolates covering five serovars (ten Typhimurium including three monophasic 4,[5],12:i:, four Enteritidis, three Hadar, four Heidelberg and four Kentucky) were sequenced. Most serovars were clustered in strongly supported phylogenetic clades, except for isolates of serovar Enteritidis that were scattered throughout the tree. Plasmids of varying sizes were detected in several isolates independently of serovars. Genes associated with the IncF plasmid and the IncI1 plasmid were identified in twelve and four isolates, respectively, while genes associated with the IncQ plasmid were found in one isolate. The presence of numerous genes associated with Salmonella pathogenicity islands (SPIs) was also confirmed. Components of the type III and IV secretion systems (T3SS and T4SS) varied in different isolates, which could explain in part, differences of their pathogenicity in humans and/or persistence in broilers. Conserved clusters of genes in the T3SS were detected that could be used in designing effective strategies (diagnostic, vaccination or treatments) to combat Salmonella. Antibiotic resistance genes (CMY, aadA, ampC, florR, sul1, sulI, tetAB, and srtA) and class I integrons were detected in resistant isolates while all isolates carried multidrug efflux pump systems regardless of their antibiotic susceptibility profile. This study showed that the predominant Salmonella serovars in broiler chickens harbor genes encoding adhesins

  1. The Antibacterial Activity of Coriolus versicolor Methanol Extract and Its Effect on Ultrastructural Changes of Staphylococcus aureus and Salmonella Enteritidis.

    PubMed

    Matijašević, Danka; Pantić, Milena; Rašković, Božidar; Pavlović, Vladimir; Duvnjak, Dunja; Sknepnek, Aleksandra; Nikšić, Miomir

    2016-01-01

    The antibacterial activity of methanol extract obtained from fruiting body of industrially grown basidiomycete Coriolus versicolor was examined. The Minimum Inhibitory Concentration (MIC) values against various bacteria ranged from 0.625 to 20 mg mL(-1). C. versicolor expressed bactericidal activity against both Gram-positive and Gram-negative bacteria. The growth curves of Staphylococcus aureus and Salmonella enterica serovar Enteritidis, measured at 630 nm, and confirmed with macrodilution method showed that the obtained extract could inhibit the growth of tested bacteria. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and the loss of 260-nm-absorbing material were used to examine the ultrastructural changes in bacteria induced by the extract. When S. aureus was exposed to the MIC of C. versicolor, elongated and malformed cells were observed by SEM, while S. Enteritidis treated cells appeared shorter and aggregated with ruptured cell walls. TEM revealed the formation of non-membrane-enclosed bodies and depleted inner content of S. aureus. Larger and irregular periplasmic space and deformed and scattered components of the cell envelope were observed in treated S. Enteritidis. The loss of 260-nm-absorbing material indicated that the disruptive action of the extract on cytoplasmic membrane was more pronounced in S. aureus than in S. Enteritidis treated cells. The UV and FTIR spectrophotometric analyses revealed diverse composition of C. versicolor extract and high content of total phenolics. Altogether, mushroom extracts could be used to develop nutraceuticals or drugs effective against pathogenic microorganisms.

  2. The Antibacterial Activity of Coriolus versicolor Methanol Extract and Its Effect on Ultrastructural Changes of Staphylococcus aureus and Salmonella Enteritidis

    PubMed Central

    Matijašević, Danka; Pantić, Milena; Rašković, Božidar; Pavlović, Vladimir; Duvnjak, Dunja; Sknepnek, Aleksandra; Nikšić, Miomir

    2016-01-01

    The antibacterial activity of methanol extract obtained from fruiting body of industrially grown basidiomycete Coriolus versicolor was examined. The Minimum Inhibitory Concentration (MIC) values against various bacteria ranged from 0.625 to 20 mg mL-1. C. versicolor expressed bactericidal activity against both Gram-positive and Gram-negative bacteria. The growth curves of Staphylococcus aureus and Salmonella enterica serovar Enteritidis, measured at 630 nm, and confirmed with macrodilution method showed that the obtained extract could inhibit the growth of tested bacteria. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and the loss of 260-nm-absorbing material were used to examine the ultrastructural changes in bacteria induced by the extract. When S. aureus was exposed to the MIC of C. versicolor, elongated and malformed cells were observed by SEM, while S. Enteritidis treated cells appeared shorter and aggregated with ruptured cell walls. TEM revealed the formation of non-membrane-enclosed bodies and depleted inner content of S. aureus. Larger and irregular periplasmic space and deformed and scattered components of the cell envelope were observed in treated S. Enteritidis. The loss of 260-nm-absorbing material indicated that the disruptive action of the extract on cytoplasmic membrane was more pronounced in S. aureus than in S. Enteritidis treated cells. The UV and FTIR spectrophotometric analyses revealed diverse composition of C. versicolor extract and high content of total phenolics. Altogether, mushroom extracts could be used to develop nutraceuticals or drugs effective against pathogenic microorganisms. PMID:27540376

  3. Salmonella and impact on egg production.

    USDA-ARS?s Scientific Manuscript database

    There is a strong association between the incidence of human illness and the prevalence of Salmonella Enteritidis in commercial egg-producing poultry. Although most egg-associated disease around the world has been attributed to S. Enteritidis, other serovars are sometimes implicated. The deposition ...

  4. Outbreak of Salmonella enteritidis PT14b gastroenteritis at a restaurant in England: the use of molecular typing to achieve a successful prosecution.

    PubMed

    Chatt, C; Nicholds-Trainor, D; Scrivener, A; Suleman, S; Harvey, M; Dallman, T; Hawker, J; Sibal, B

    2017-10-01

    To describe an outbreak of Salmonella enteritidis phage type (PT) 14b in people who had eaten at a restaurant, and the investigation and subsequent prosecution of the food business operator (FBO). The local health protection team and environmental health department formed an outbreak control team to investigate the outbreak. Epidemiological, microbiological, and environmental investigations were undertaken. Epidemiological investigations involved case finding and interviews. Microbiological investigation: stool samples from the suspected cases and environmental samples from the implicated food business were investigated. Salmonella isolates obtained were subjected to multiple locus variable-number tandem repeat analysis (MLVA) profiling and whole genome sequencing. In addition, adenosine triphosphate (ATP) hygiene swab tests were used to verify the quality of cleaning procedures and data loggers were used to determine the water temperature of the mechanical dishwasher. Fifteen cases of illness where the causative agent was shown to be S. enteritidis PT14b were identified, all of whom had eaten at the same restaurant. S. enteritidis PT14b was also identified from three of the 11 food and environmental samples taken at the restaurant and found to have the same MLVA profile as the cases. A case for prosecution was built and the FBO was successfully prosecuted in July 2015. This investigation highlighted that the use of molecular typing as part of thorough epidemiological, microbiological, and environmental investigations can present a robust case for prosecution against restaurants which pose a risk to public health. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  5. 78 FR 32400 - Agency Information Collection Activities; Submission for Office of Management and Budget Review...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-30

    ... Request; Prevention of Salmonella Enteritidis in Shell Eggs During Production--Recordkeeping and... information to OMB for review and clearance. Prevention of Salmonella Enteritidis in Shell Eggs During...-0660)--Extension Shell eggs contaminated with Salmonella Enteritidis (SE) are responsible for more than...

  6. High pressure treatments on the inactivation of Salmonella Enteritidis and the physicochemical, rheological and color characteristics of sliced vacuum-packaged dry-cured ham.

    PubMed

    de Alba, María; Montiel, Raquel; Bravo, Daniel; Gaya, Pilar; Medina, Margarita

    2012-06-01

    The effect of high pressure (HP) on Salmonella Enteritidis in sliced dry-cured ham stored under temperature abuse (8°C) during 60d was investigated. After treatment, reductions of S. Enteritidis were 1.06, 2.54 and 4.32 log units in ham treated at 400, 500 and 600MPa for 5min at 12°C, compared to non-pressurized samples. After 60d, counts of S. Enteritidis in ham treated at 400 and 500MPa were 2.56 and 2.66 log units lower than in non-treated ham, whereas the pathogen was only detected after enrichment in ham treated at 600MPa. Lipid oxidation increased with storage and pressurization, whereas total free amino acid contents were similar in HP and control samples after 60d. Dry-cured ham treated at the highest pressures exhibited lower shear resistance, whereas the maximum force to compress the sample was slightly changed. Color (L*, a* and b*) varied with pressurization and storage. Changes induced by HP in dry-cured ham were attenuated during storage. Copyright © 2012 Elsevier Ltd. All rights reserved.

  7. Investigation of Salmonella Enteritidis Outbreak Associated with Truffle Oil - District of Columbia, 2015.

    PubMed

    Kuramoto-Crawford, S Janet; McGee, Sasha; Li, Keith; Hennenfent, Andrew K; Dassie, Kossia; Carney, Jhetari T; Gibson, Arian; Cooper, Ivory; Blaylock, Morris; Blackwell, Reginald; Fields, Angela; Davies-Cole, John

    2017-03-17

    On September 8, 2015, the District of Columbia Department of Health (DCDOH) received a call from a person who reported experiencing gastrointestinal illness after eating at a District of Columbia (DC) restaurant with multiple locations throughout the United States (restaurant A). Later the same day, a local emergency department notified DCDOH to report four persons with gastrointestinal illness, all of whom had eaten at restaurant A during August 30-September 5. Two patients had laboratory-confirmed Salmonella group D by stool culture. On the evening of September 9, a local newspaper article highlighted a possible outbreak associated with restaurant A. Investigation of the outbreak by DCDOH identified 159 patrons who were residents of 11 states and DC with gastrointestinal illness after eating at restaurant A during July 1-September 10. A case-control study was conducted, which suggested truffle oil-containing food items as a possible source of Salmonella enterica serotype Enteritidis infection. Although several violations were noted during the restaurant inspections, the environmental, laboratory, and traceback investigations did not confirm the contamination source. Because of concern about the outbreak, the restaurant's license was suspended during September 10-15. The collaboration and cooperation of the public, media, health care providers, and local, state, and federal public health officials facilitated recognition of this outbreak involving a pathogen commonly implicated in foodborne illness.

  8. The antimicrobial effect of oregano essential oil, nisin and their combination against Salmonella Enteritidis in minced sheep meat during refrigerated storage.

    PubMed

    Govaris, A; Solomakos, N; Pexara, A; Chatzopoulou, P S

    2010-02-28

    The antimicrobial effect of oregano essential oil (EO) at 0.6 or 0.9%, nisin at 500 or 1000 IU/g, and their combination against Salmonella Enteritidis was studied in minced sheep meat during storage at 4 degrees or 10 degrees C for 12 days. Sensory evaluation showed that the addition of oregano EO at 0.6 or 0.9% in minced sheep meat was organoleptically acceptable, and attribute scores were higher for the EO at 0.6 than 0.9%. According to compositional analysis of the oregano EO, the phenols carvacrol (80.15%) and thymol (4.82%) were the predominant components. Treatment of minced sheep meat with nisin at 500 or 1000 IU/g, proved insufficient to act against S. Enteritidis. The combination of the oregano EO at 0.6% with nisin at 500 IU/g showed stronger antimicrobial activity against S. Enteritidis than the oregano EO at 0.6% but lower than the combination with nisin at 1000 IU/g, which in turn was lower than that of the oregano EO at 0.9%. In its turn, oregano EO at 0.9% showed lower antimicrobial activity than its combinations with nisin at 500 or 1000 IU/g, which showed a bactericidal effect against the pathogen. The inhibition percentages of all treatments against S. Enteritidis at 10 degrees C were higher than those at 4 degrees C. (c) 2009 Elsevier B.V. All rights reserved.

  9. Removal of Salmonella Enteritidis from commercial unpasteurized liquid egg white using pilot scale cross flow tangential microfiltration.

    PubMed

    Mukhopadhyay, Sudarsan; Tomasula, Peggy M; Luchansky, John B; Porto-Fett, Anna; Call, Jeffrey E

    2010-09-01

    Effectiveness of a cross flow microfiltration (MF) process for removal of a cocktail of Salmonella enterica serovar Enteritidis species from commercial unpasteurized liquid egg white (LEW) from a local egg breaking plant, while maintaining its functional properties was evaluated. To facilitate MF, LEW was wedge screened, homogenized and then diluted (1:2 w/w) with distilled water containing 0.5% sodium chloride. Diluted unpasteurized LEW was inoculated with five strains of S. Enteritidis (ATCC 4931, ATCC BAA-708, ATCC 49215, ATCC 49218, and ATCC BAA-1045) to a level of approximately 10(7)CFU/mL of LEW and microfiltered using a ceramic membrane. Process parameters influencing egg white functional properties and pathogen removal efficiency were evaluated. Average permeates flux increased by almost 126% when pH of LEW was adjusted from pH 8 to pH 7 at 25 degrees C. Microbial removal efficiency was at least, on average, 6.8Log(10)CFU/mL (limit of detection < or =0.5Log(10)CFU/mL). Functional property analysis indicated that the MF process did not alter the foaming power of LEW. Published by Elsevier B.V.

  10. Subtyping Salmonella enterica serovar enteritidis isolates from different sources by using sequence typing based on virulence genes and clustered regularly interspaced short palindromic repeats (CRISPRs).

    PubMed

    Liu, Fenyun; Kariyawasam, Subhashinie; Jayarao, Bhushan M; Barrangou, Rodolphe; Gerner-Smidt, Peter; Ribot, Efrain M; Knabel, Stephen J; Dudley, Edward G

    2011-07-01

    Salmonella enterica subsp. enterica serovar Enteritidis is a major cause of food-borne salmonellosis in the United States. Two major food vehicles for S. Enteritidis are contaminated eggs and chicken meat. Improved subtyping methods are needed to accurately track specific strains of S. Enteritidis related to human salmonellosis throughout the chicken and egg food system. A sequence typing scheme based on virulence genes (fimH and sseL) and clustered regularly interspaced short palindromic repeats (CRISPRs)-CRISPR-including multi-virulence-locus sequence typing (designated CRISPR-MVLST)-was used to characterize 35 human clinical isolates, 46 chicken isolates, 24 egg isolates, and 63 hen house environment isolates of S. Enteritidis. A total of 27 sequence types (STs) were identified among the 167 isolates. CRISPR-MVLST identified three persistent and predominate STs circulating among U.S. human clinical isolates and chicken, egg, and hen house environmental isolates in Pennsylvania, and an ST that was found only in eggs and humans. It also identified a potential environment-specific sequence type. Moreover, cluster analysis based on fimH and sseL identified a number of clusters, of which several were found in more than one outbreak, as well as 11 singletons. Further research is needed to determine if CRISPR-MVLST might help identify the ecological origins of S. Enteritidis strains that contaminate chickens and eggs.

  11. Electronic network for monitoring travellers' diarrhoea and detection of an outbreak caused by Salmonella enteritidis among overseas travellers.

    PubMed

    Osaka, K; Inouye, S; Okabe, N; Taniguchi, K; Izumiya, H; Watanabe, H; Matsumoto, Y; Yokota, T; Hashimoto, S; Sagara, H

    1999-12-01

    The Traveller's Diarrhoea Network, by which the Infectious Disease Surveillance Center is electronically connected with two major airport quarantine stations and three infectious disease hospitals, was launched in February 1988 in Japan. The data on travellers' diarrhoea detected is reported weekly by e-mail. Two clusters of infection among travellers returning from Italy were reported by two airport quarantine stations at the end of September 1998. A total of 12 salmonella isolates from 2 clusters were examined. All were identified as Salmonella enteritidis, phage type 4 and showed identical banding patterns on pulsed-field gel electrophoresis. A case-control study showed that the scrambled eggs served at the hotel restaurant in Rome were the likely source of this outbreak. This outbreak could not have been detected promptly and investigated easily without the e-mail network. International exchange of data on travellers' diarrhoea is important for preventing and controlling food-borne illnesses infected abroad.

  12. Phage and MLVA typing of Salmonella enteritidis isolated from layers and humans in Belgium from 2000-2010, a period in which vaccination of laying hens was introduced.

    PubMed

    Dewaele, I; Heyndrickx, M; Rasschaert, G; Bertrand, S; Wildemauwe, C; Wattiau, P; Imberechts, H; Herman, L; Ducatelle, R; Van Weyenberg, S; De Reu, K

    2014-09-01

    The aim of the study was to characterize isolates of Salmonella enterica serovar Enteritidis (S. Enteritidis) obtained from humans and layer farms in Belgium collected during 2000-2010. Three periods were compared, namely (i) before implementation of vaccination (2000-2004), (ii) during voluntary vaccination (2005-2006) and (iii) during implementation of the national control program (NCP) for Salmonella including mandatory vaccination against S. Enteritidis (2007-2010). The characteristics compared across time periods were distributions of phage type and multiple-locus variable number tandem-repeat assay (MLVA). While PT4 and PT21 were predominantly isolated in Belgium in layers and humans before 2007, a significant reduction of those PTs was observed in both populations in the period 2007-2010. The relative proportion of PT4b, PT21c and PT6c was found to have increased considerably in the layer population since 2007. In the human population, PT8, PT1 and the group of 'other' PTs were more frequently isolated compared to the previous periods. When comparing the proportion of the predominant MLVA types Q2 and U2, no significant difference was found between the layer and human population in the three periods and between periods within each category (layer and human). A significant difference in isolate distribution among MLVA clusters I and II was found between human and layer isolates recovered during Period 3 and in the human population between Period 1 and 3. Results suggest that the association between S. Enteritidis in layers and the occurrence of the pathogen in humans changed since implementation of the NCP in 2007. © 2013 Blackwell Verlag GmbH.

  13. Nitric Oxide as a Biomarker of Intracellular Salmonella Viability and Identification of the Bacteriostatic Activity of Protein Kinase A Inhibitor H-89

    PubMed Central

    He, Haiqi; Genovese, Kenneth J.; Swaggerty, Christina L.; Nisbet, David J.; Kogut, Michael H.

    2013-01-01

    Salmonella enterica serovar Enteritidis is one of the most prevalent Salmonella serovars in poultry and is often associated with human salmonellosis. S. Enteritidis is known to suppress nitric oxide (NO) production in infected chicken macrophage HD11 cells, while dead S. Enteritidis stimulates a high level of NO production, suggesting a bacterial inhibitory effect on NO production. Based on these observations, the present study was conducted to evaluate whether NO production in S. Enteritidis-infected HD11 cells can be used as a biomarker to identify molecules that kill intracellular Salmonella. Since Salmonella are known to manipulate the host cell kinase network to facilitate intracellular survival, we screened a group of pharmaceutical inhibitors of various kinases to test our hypothesis. A protein kinase A inhibitor, H-89, was found to reverse the suppression of NO production in S. Enteritidis-infected HD11 cells. Production of NO in S. Enteritidis-infected HD11 cells increased significantly following treatment with H-89 at or above 20 µM. Inversely, the number of viable intracellular Salmonella decreased significantly in cells treated with H-89 at or above 30 µM. Furthermore, the growth rate of S. Enteritidis in culture was significantly inhibited by H-89 at concentrations from 20 to 100 µM. Our results demonstrate that NO-based screening using S. Enteritidis-infected HD11 cells is a viable tool to identify chemicals with anti-intracellular Salmonella activity. Using this method, we have shown H-89 has bacteriostatic activity against Salmonella, independent of host cell protein kinase A or Akt1 activity. PMID:23554945

  14. Nitric oxide as a biomarker of intracellular Salmonella viability and identification of the bacteriostatic activity of protein kinase A inhibitor H-89.

    PubMed

    He, Haiqi; Genovese, Kenneth J; Swaggerty, Christina L; Nisbet, David J; Kogut, Michael H

    2013-01-01

    Salmonella enterica serovar Enteritidis is one of the most prevalent Salmonella serovars in poultry and is often associated with human salmonellosis. S. Enteritidis is known to suppress nitric oxide (NO) production in infected chicken macrophage HD11 cells, while dead S. Enteritidis stimulates a high level of NO production, suggesting a bacterial inhibitory effect on NO production. Based on these observations, the present study was conducted to evaluate whether NO production in S. Enteritidis-infected HD11 cells can be used as a biomarker to identify molecules that kill intracellular Salmonella. Since Salmonella are known to manipulate the host cell kinase network to facilitate intracellular survival, we screened a group of pharmaceutical inhibitors of various kinases to test our hypothesis. A protein kinase A inhibitor, H-89, was found to reverse the suppression of NO production in S. Enteritidis-infected HD11 cells. Production of NO in S. Enteritidis-infected HD11 cells increased significantly following treatment with H-89 at or above 20 µM. Inversely, the number of viable intracellular Salmonella decreased significantly in cells treated with H-89 at or above 30 µM. Furthermore, the growth rate of S. Enteritidis in culture was significantly inhibited by H-89 at concentrations from 20 to 100 µM. Our results demonstrate that NO-based screening using S. Enteritidis-infected HD11 cells is a viable tool to identify chemicals with anti-intracellular Salmonella activity. Using this method, we have shown H-89 has bacteriostatic activity against Salmonella, independent of host cell protein kinase A or Akt1 activity.

  15. Antimicrobial Resistance in Nontyphoidal Salmonella Isolated from Human and Poultry-Related Samples in Brazil: 20-Year Meta-Analysis.

    PubMed

    Voss-Rech, Daiane; Potter, Luciana; Vaz, Clarissa Silveira Luiz; Pereira, Daniela Isabel Brayer; Sangioni, Luís Antonio; Vargas, Águeda Castagna; de Avila Botton, Sônia

    2017-02-01

    Nontyphoidal Salmonella are one of the leading causes of foodborne diseases in the world. As poultry products are recognized as main sources of human salmonellosis, nontyphoidal Salmonella control has become a global issue for the poultry industry. The increasing antimicrobial resistance in poultry-related nontyphoidal Salmonella serovars is a global matter of concern. By monitoring the evolution of antimicrobial resistance, alternative treatments can be identified and possible restrictions in the treatment of systemic human salmonellosis foreseen. A meta-analysis was conducted to assess the profile and temporal evolution of the antimicrobial resistance of nontyphoidal Salmonella of poultry and human origin in Brazil, isolated in the period from 1995 to 2014. Four databases were researched; twenty-nine articles met the eligibility criteria and were included in the meta-analysis. In the nontyphoidal isolates of poultry origin, the highest levels of antimicrobial resistance were verified for sulfonamides (44.3%), nalidixic acid (42.5%), and tetracycline (35.5%). In the human-origin isolates, the resistance occurred mainly for sulfonamides (46.4%), tetracycline (36.9%), and ampicillin (23.6%). Twenty-two articles described results of antimicrobial resistance specifically for Salmonella Enteritidis, also enabling the individual meta-analysis of this serovar. For most antimicrobials, the resistance levels of Salmonella Enteritidis were lower than those found when considering all the nontyphoidal serovars. In the poultry-origin isolates, a quadratic temporal distribution was observed, with reduced resistance to streptomycin in Salmonella Enteritidis and in all nontyphoidal serovars, and a linear increase of resistance to nalidixic acid in Salmonella Enteritidis. In the human-origin isolates, a linear increase was identified in the resistance to nalidixic acid in Salmonella Enteritidis and in all the nontyphoidal isolates, and to gentamicin in Salmonella Enteritidis

  16. Antimicrobial drug resistance and genetic properties of Salmonella enterica serotype Enteritidis circulating in chicken farms in Tunisia.

    PubMed

    Ben Salem, Rakia; Abbassi, Mohamed S; García, Vanesa; García-Fierro, Raquel; Fernández, Javier; Kilani, Hajer; Jaouani, Imen; Khayeche, Monia; Messadi, Lilia; Rodicio, María R

    This study focused on 77 isolates of Salmonella enterica serotype Enteritidis collected during 2009 to 2013 from healthy and sick chickens and environmental farm samples in Tunisia. Resistance to 14 antimicrobials and the encoding genes were analyzed. 66, 26, 6.5, 3.9 and 1.3% were pan-susceptible or showed resistance to nalidixic acid (Asp87 to Tyr and Asp87 to Asn substitutions in GyrA), ampicillin (bla TEM-1-like and bla SHV ), sulfonamides (sul1and sul3) and streptomycin (strB), respectively. A single isolate with intermediate susceptibility to ciprofloxacin was positive for qnrB, whereas qnrA, qnrS or aac(6')-Ib-cr were not detected. The virulotype of the isolates was established by testing ten virulence genes. The orgA, ssaQ, mgtC, siiD, sopB genes, located on Salmonella pathogenicity islands, and spvC of the serotype-specific virulence plasmid, were common to all isolates. In contrast, the prophage-associated sopE-1, sodC1 and gipA genes and the fimbrial bcfC gene were variably represented. All isolates except one contained the virulence plasmid, which appeared either alone or together with one or more additional plasmids. One isolate carried a single plasmid of ca. 90Kb which may be derived from the virulence plasmid (60Kb). Overall, seven resistotypes, six virulotypes and six plasmid profiles were identified. XbaI-PFGE revealed four related pulsotypes (X1-X4), with 80% of the isolates sharing the X1 pattern. The latter isolates exhibited different resistance, virulence and plasmid profiles, suggesting that mobile genetic elements, particularly prophages and plasmids, are of central importance for the evolution and adaptation of S. Enteritidis circulating in chicken farms in Tunisia. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  17. The effect of milk fat globules on adherence and internalization of Salmonella Enteritidis to HT-29 cells.

    PubMed

    Guri, A; Griffiths, M; Khursigara, C M; Corredig, M

    2012-12-01

    Milk fat globules were extracted from bovine and goat milk and incubated with HT-29 human adenocarcinoma cells to assess the attachment and internalization of Salmonella Enteritidis. Because the expression of bacterial adhesins is highly affected by the presence of antibiotic, the attachment was studied with and without antibiotic in the cell growth medium. Although no inhibitory effect of the fat globules was observed in the presence of the antibiotic, milk fat globules significantly inhibited the binding and internalization of Salmonella in medium free of antibiotic. The fat globules from both bovine and goat milk markedly reduced bacterial binding and invasion compared with controls, and the cells treated with goat milk-derived fat globules demonstrated greater protective properties than those derived from bovine milk. The effect of heat treatment on bovine fat globules was also investigated, and it was shown that the fat globules from heated milk had a higher degree of inhibition than those from unheated milk. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  18. Persistence and clearance of different Salmonella serovars in buildings housing laying hens.

    PubMed

    Carrique-Mas, J J; Breslin, M; Snow, L; McLaren, I; Sayers, A R; Davies, R H

    2009-06-01

    We investigated factors associated with persistence of different Salmonella serovars in buildings housing laying hens in Great Britain using survival analysis. A total of 264 incidents of Salmonella detection occurring between July 1998 and August 2007 in 152 houses were recorded. For incidents involving Salmonella Enteritidis (SE), both the rodent score of the house and the type of house were positively associated with persistence. For non-SE serovars, only the type of house was associated with persistence. Persistence of SE in the houses was longest (>15 months) in step-cage and cage-scraper houses when high levels of rodents were present, and lowest in non-cage and cage-belt houses. We estimated that 42% (95% CI 23.3-63.1) of SE incidents may be cleared during the lay period, and this was related to elimination of rodents from the houses. From January 2009, EU legislation will ban the sale of fresh eggs from SE-positive and S. Typhimurium-positive flocks over their remaining lifespan. If infection is eliminated from such flocks, they would cease to represent a public health risk.

  19. Adhesion and growth inhibitory effect of chicken egg yolk antibody (IgY) on Salmonella enterica serovars Enteritidis and Typhimurium in vitro.

    PubMed

    Chalghoumi, Raja; Théwis, André; Beckers, Yves; Marcq, Christopher; Portetelle, Daniel; Schneider, Yves-Jacques

    2009-06-01

    The protective effects of powder preparation of egg yolk immunoglobulin Y (IgY), specific to Salmonella Enteritidis and Salmonella Typhimurium outer membrane proteins (OMP), against these two Salmonella sp. serovars were investigated in vitro in two different assays: adhesion-prevention and growth-inhibition. The adhesion-prevention assay was conducted using polarized monolayers of the human intestinal epithelial Caco-2 cell line. First, the conditions of Salmonella adherence to Caco-2 cells were optimized, and interferences of bacteria with the transepithelial electrical resistance (TER) of fully differentiated Caco-2 cell monolayers and the lactate dehydrogenase release upon exposure of the cells to Salmonella were evaluated. Both Salmonella sp. serovars were able to adhere to Caco-2 cells and decreased TER. Results from the adhesion-prevention assay demonstrated that specific IgY reduced the decrease in TER of the infected Caco-2 cell monolayers and blocked the Salmonella sp. adhesion in a concentration-dependent manner (p < 0.05). Nonspecific IgY also exhibited an inhibitory effect on these two parameters, but to a lesser extent than that of the specific IgY (p < 0.05). The protective effect of nonspecific IgY could be attributed to the low-density lipoprotein component of the water-soluble fraction of egg yolks that may not have been eliminated during ultrafiltration. The growth-inhibition assay revealed that specific IgY had an inhibitory effect on the bacterial growth, markedly during the late exponential phase, whereas nonspecific IgY failed to do so. Taken together, these results suggest that the in vitro growth inhibitory effect of specific IgY on Salmonella spp. resulted from the specific binding activity of these IgY to Salmonella sp. OMP. Passive immunization with Salmonella sp. OMP-specific IgY could thus be useful to prevent Salmonella colonization in broiler chickens and the subsequent carcass contamination during processing.

  20. Global Gene-expression Analysis of the Response of Salmonella Enteritidis to Egg White Exposure Reveals Multiple Egg White-imposed Stress Responses

    PubMed Central

    Baron, Florence; Bonnassie, Sylvie; Alabdeh, Mariah; Cochet, Marie-Françoise; Nau, Françoise; Guérin-Dubiard, Catherine; Gautier, Michel; Andrews, Simon C.; Jan, Sophie

    2017-01-01

    Chicken egg white protects the embryo from bacterial invaders by presenting an assortment of antagonistic activities that combine together to both kill and inhibit growth. The key features of the egg white anti-bacterial system are iron restriction, high pH, antibacterial peptides and proteins, and viscosity. Salmonella enterica serovar Enteritidis is the major pathogen responsible for egg-borne infection in humans, which is partly explained by its exceptional capacity for survival under the harsh conditions encountered within egg white. However, at temperatures up to 42°C, egg white exerts a much stronger bactericidal effect on S. Enteritidis than at lower temperatures, although the mechanism of egg white-induced killing is only partly understood. Here, for the first time, the impact of exposure of S. Enteritidis to egg white under bactericidal conditions (45°C) is explored by global-expression analysis. A large-scale (18.7% of genome) shift in transcription is revealed suggesting major changes in specific aspects of S. Enteritidis physiology: induction of egg white related stress-responses (envelope damage, exposure to heat and alkalinity, and translation shutdown); shift in energy metabolism from respiration to fermentation; and enhanced micronutrient provision (due to iron and biotin restriction). Little evidence of DNA damage or redox stress was obtained. Instead, data are consistent with envelope damage resulting in cell death by lysis. A surprise was the high degree of induction of hexonate/hexuronate utilization genes, despite no evidence indicating the presence of these substrates in egg white. PMID:28553268

  1. Genetic Control of the Innate Resistance of Mice to Salmonella typhimurium: Expression of the Ity Gene in Peritoneal Macrophages Isolated In Vitro

    DTIC Science & Technology

    1984-07-20

    Carter and Collins, 1974a; Collins and Carter, 1978; O’Brien, 1982a). S. typhimurium and certain strains of Salmonella enteritidis are facultative...tested by S. enteritidis challenge of mice chronically infected with an antigenically dissimilar Salmonella species, S. montevideo. These mice were...given 5 X lO"’’ S. enteritidis . Crosses of resistant and susceptible strains resulted in Fl progeny of a Salmonella resistant phenotype. Backcross

  2. Outbreak of Salmonella Enteritidis phage type 13a: case-control investigation in Hertsmere, United Kingdom.

    PubMed

    Morgan, O; Milne, L; Kumar, S; Murray, D; Man, W; Georgiou, M; Verlander, N Q; de Pinna, E; McEvoy, M

    2007-07-01

    Cases of illness were reported to Hertsmere Borough Council among attendees of a children's charity event in June 2006. Initial laboratory investigation identified Salmonella Enteritidis PT13a as a possible cause of the outbreak. We carried out an unmatched case-control investigation. The population at risk included all individuals who attended the event. Self-completion questionnaires were sent to 53 presumptive cases and 212 randomly selected potential controls. Information was available for 49 cases and 128 controls (overall response rate=75%). We calculated odds ratios from single and multivariable analysis and tested for all two-way interactions. Risk factors for diarrhoea were eating egg mayonnaise bagels (OR=34.1, 95%CI 10.5 - 111.3) and drinking apple juice (OR=16.1, 95% CI 3.5 - 74.2). There was weak statistical evidence to suggest that the risk of diarrhoea after eating egg mayonnaise bagels was greater in the afternoon. No food samples were available to confirm which food item might have caused this outbreak. Eggs from Spain were used by the caterer. The ecology of salmonella, experience from previous outbreaks and epidemiological findings from this case-control investigation suggest that the most likely cause of the outbreak was contaminated eggs.

  3. Prevalence of gyrA Mutations in Nalidixic Acid-Resistant Strains of Salmonella Enteritidis Isolated from Humans, Food, Chickens, and the Farm Environment in Brazil.

    PubMed

    Campioni, Fábio; Souza, Roberto Antonio; Martins, Vinicius Vicente; Stehling, Eliana Guedes; Bergamini, Alzira Maria Morato; Falcão, Juliana Pfrimer

    2017-06-01

    Salmonella Enteritidis strains that are resistant to nalidixic acid and exhibit reduced susceptibility to fluoroquinolones have been increasing worldwide. In Brazil, few studies have been conducted to elucidate the quinolone resistance mechanisms of S. Enteritidis strains. This study analyzed the profile of gyrA, gyrB, parC, and parE mutations and plasmid-mediated quinolone resistance (PMQR) mechanisms in S. Enteritidis Nal R strains isolated in Brazil. Moreover, the minimum inhibitory concentrations (MICs) of ciprofloxacin were evaluated in 84 Nal R strains and compared with 20 Nal S strains. The mutation profiles of the gyrA gene were accessed by high-resolution melting analysis and gyrB, parC, and parE by quinolone resistance-determining region sequencing. The MICs of ciprofloxacin were accessed with Etest ® . The strains were divided into five gyrA melting profiles. The Nal R strains exhibited the following amino acid substitutions: Ser97→Pro, Ser83→Phe, Asp87→Asn, or Asp87→Tyr. The average MICs of ciprofloxacin was 0.006 μg/ml in the Nal S and 0.09 μg/ml in the Nal R strains. No points of mutation were observed in the genes gyrB, parC, and parE. The qnrB gene was found in two strains. In conclusion, the reduced susceptibility to ciprofloxacin observed in Nal R strains may cause treatment failures once this drug is commonly used to treat Salmonella infections. Moreover, this reduced susceptibility in these Brazilian strains was provided by target alteration of gene gyrA and not by mobile elements, such as resistance plasmids.

  4. The "decline and fall" of nontyphoidal salmonella in the United kingdom.

    PubMed

    O'Brien, Sarah J

    2013-03-01

    Remarkable changes in the epidemiology of human nontyphoidal salmonellosis have occurred in the United Kingdom over the last century. Between 1981 and 1991, the incidence of nontyphoidal salmonellosis in the United Kingdom rose by >170%, driven primarily by an epidemic of Salmonella enterica subspecies enterica serovar Enteritidis phage type (PT) 4, which peaked in 1993. Measures introduced to control this epidemic included legislation, food safety advice, and an industry-led vaccination program in broiler-breeder and laying poultry flocks. The incidence of Salmonella Enteritidis has been falling since 1997, and levels of Salmonella Enteritidis PT4 have fallen to preepidemic levels and have stayed low. The temporal relationship between vaccination programs and the reduction in human disease is compelling and suggests that these programs have made a major contribution to improving public health.

  5. Outbreak of Salmonella Enteritidis linked to the consumption of frozen beefburgers received from a food bank and originating from Poland: northern France, December 2014 to April 2015

    PubMed Central

    Jones, Gabrielle; Pihier, Nathalie; Vanbockstael, Caroline; Le Hello, Simon; Cadel Six, Sabrina; Fournet, Nelly; Jourdan-da Silva, Nathalie

    2016-01-01

    A prolonged outbreak of Salmonella enterica serotype Enteritidis occurred in northern France between December 2014 and April 2015. Epidemiological investigations following the initial notification on 30 December 2014 of five cases of salmonellosis (two confirmed S. Enteritidis) in young children residing in the Somme department revealed that all cases frequented the same food bank A. Further epidemiological, microbiological and food trace-back investigations indicated frozen beefburgers as the source of the outbreak and the suspected lot originating from Poland was recalled on 22 January 2015. On 2 March 2015 a second notification of S. Enteritidis cases in the Somme reinitiated investigations that confirmed a link with food bank A and with consumption of frozen beefburgers from the same Polish producer. In the face of a possible persistent source of contamination, all frozen beefburgers distributed by food bank A and from the same origin were blocked on 3 March 2015. Microbiological analyses confirmed contamination by S. Enteritidis of frozen beefburgers from a second lot remaining in cases’ homes. A second recall was initiated on 6 March 2015 and all frozen beefburgers from the Polish producer remain blocked after analyses identified additional contaminated lots over several months of production. PMID:27748250

  6. Comparison of a real-time PCR method with a culture method for the detection of Salmonella enterica serotype enteritidis in naturally contaminated environmental samples from integrated poultry houses.

    PubMed

    Lungu, Bwalya; Waltman, W Douglas; Berghaus, Roy D; Hofacre, Charles L

    2012-04-01

    Conventional culture methods have traditionally been considered the "gold standard" for the isolation and identification of foodborne bacterial pathogens. However, culture methods are labor-intensive and time-consuming. A Salmonella enterica serotype Enteritidis-specific real-time PCR assay that recently received interim approval by the National Poultry Improvement Plan for the detection of Salmonella Enteritidis was evaluated against a culture method that had also received interim National Poultry Improvement Plan approval for the analysis of environmental samples from integrated poultry houses. The method was validated with 422 field samples collected by either the boot sock or drag swab method. The samples were cultured by selective enrichment in tetrathionate broth followed by transfer onto a modified semisolid Rappaport-Vassiliadis medium and then plating onto brilliant green with novobiocin and xylose lysine brilliant Tergitol 4 plates. One-milliliter aliquots of the selective enrichment broths from each sample were collected for DNA extraction by the commercial PrepSEQ nucleic acid extraction assay and analysis by the Salmonella Enteritidis-specific real-time PCR assay. The real-time PCR assay detected no significant differences between the boot sock and drag swab samples. In contrast, the culture method detected a significantly higher number of positive samples from boot socks. The diagnostic sensitivity of the real-time PCR assay for the field samples was significantly higher than that of the culture method. The kappa value obtained was 0.46, indicating moderate agreement between the real-time PCR assay and the culture method. In addition, the real-time PCR method had a turnaround time of 2 days compared with 4 to 8 days for the culture method. The higher sensitivity as well as the reduction in time and labor makes this real-time PCR assay an excellent alternative to conventional culture methods for diagnostic purposes, surveillance, and research studies

  7. Effects of drinking water treatment on susceptibility of laying hens to Salmonella enteritidis during forced molt.

    PubMed

    Kubena, L F; Byrd, J A; Moore, R W; Ricke, S C; Nisbet, D J

    2005-02-01

    Feed deprivation is used in the layer industry to induce molting and stimulate multiple egg-laying cycles in laying hens. Unfortunately, the stress involved increases susceptibility to Salmonella enteritidis (SE), the risk of SE-positive eggs, and incidence of SE in internal organs. Leghorn hens over 50 wk of age were divided into 4 treatment groups of 12 hens each in experiment 1 and 3 treatment groups of 12 hens in experiments 2 and 3; hens were placed in individual laying hen cages. Treatment groups were 1) nonmolted (NM) and received feed and distilled water for 9 d, 2) force molted by feed removal for 9 d and received distilled water, 3) force molted by feed removal for 9 d and received 0.5% lactic acid (LA) in distilled water. An additional group (4) in experiment 1 only was force molted by feed removal for 9 d and received 0.5% acetic acid in distilled water. Seven days before feed removal hens were exposed to an 8L:16D photoperiod, which was continued throughout the experiment. Individual hens among all treatments were challenged orally with 10(4) SE on d 4 of feed removal. When compared with the NM treatments, weight losses were significantly higher in the M treatments, regardless of water treatments. When compared with NM treatments, crop pH was significantly higher in the M treatment receiving distilled water. Crop pH was reduced to that of the NM controls by 0.5% acetic acid in the drinking water. No consistent significant changes were observed for volatile fatty acids. The number of hens positive for SE in crop and ceca after culture and the number of SE per crop and per gram of cecal contents were higher in the M treatments, when compared with the NM treatments, but there was no effect of addition of either of the acids to the drinking water. Additional research using different acid treatment regimens may provide a tool for reducing the incidence of SE in eggs and internal organs during and following molting of laying hens.

  8. Pathogenesis of Salmonellosis: Salmonella Exotoxins

    DTIC Science & Technology

    1982-03-08

    membrane-as3ociated enterotowin produced by S. enteritidis and by S. typhimurium ; however they could find no similarities between their Salmonella ...AD. . 0 REPORT NUJMBER 1 Pathogenesis of Salmoneiliosis: Salmonella Exotoxins Annual Progress Report (12/1/77-9/1/78) Johnny W. Peterson. Ph.D. March...TYPE OF REPORT & PERIOD COVEREOD",- Uathogenesis of ,Salmonellosils: Salmonella Annual Progress Report Exotoxins 12/T/77 9/1/78 C. PERFORMCNG ORG

  9. Re-evaluation of a 2014 multi-country European outbreak of Salmonella Enteritidis phage type 14b using recent epidemiological and molecular data

    PubMed Central

    Hörmansdorfer, Stefan; Messelhäußer, Ute; Rampp, Albert; Schönberger, Katharina; Dallman, Tim; Allerberger, Franz; Kornschober, Christian; Sing, Andreas; Wallner, Peter; Zapf, Andreas

    2017-01-01

    A European multi-country outbreak of Salmonella Enteritidis phage type (PT) 14b occurred from March to November 2014 associated with the consumption of eggs. The outbreak involved more than 400 human cases from France, Luxembourg, Austria and the United Kingdom. In 2016–2017, it has been re-evaluated combining recent epidemiological results with latest molecular data. The outbreak was traced back to one large Bavarian egg producer with four distinct premises, three located in Bavaria, one in the Czech Republic. The outbreak isolates of S. Enteritidis PT 14b were grouped into three closely related clades by whole genome sequencing. Two of these clades could be referred to two Bavarian premises of the egg producer on the basis of epidemiological and molecular data, while epidemiological data presumably linked the third clade to another premises of the egg producer. Interestingly and in contrast to the situation in other European countries where several outbreaks were documented, all notified 91 laboratory-confirmed cases of S. Enteritidis PT 14b from Bavaria were sporadic, singular cases not belonging to any epidemiological outbreaks. In conclusion, as demonstrated here, the resolution of food-related outbreaks with such a high discriminatory power is rare in outbreak investigation. PMID:29258650

  10. Re-evaluation of a 2014 multi-country European outbreak of Salmonella Enteritidis phage type 14b using recent epidemiological and molecular data.

    PubMed

    Hörmansdorfer, Stefan; Messelhäußer, Ute; Rampp, Albert; Schönberger, Katharina; Dallman, Tim; Allerberger, Franz; Kornschober, Christian; Sing, Andreas; Wallner, Peter; Zapf, Andreas

    2017-12-01

    A European multi-country outbreak of Salmonella Enteritidis phage type (PT) 14b occurred from March to November 2014 associated with the consumption of eggs. The outbreak involved more than 400 human cases from France, Luxembourg, Austria and the United Kingdom. In 2016-2017, it has been re-evaluated combining recent epidemiological results with latest molecular data. The outbreak was traced back to one large Bavarian egg producer with four distinct premises, three located in Bavaria, one in the Czech Republic. The outbreak isolates of S. Enteritidis PT 14b were grouped into three closely related clades by whole genome sequencing. Two of these clades could be referred to two Bavarian premises of the egg producer on the basis of epidemiological and molecular data, while epidemiological data presumably linked the third clade to another premises of the egg producer. Interestingly and in contrast to the situation in other European countries where several outbreaks were documented, all notified 91 laboratory-confirmed cases of S. Enteritidis PT 14b from Bavaria were sporadic, singular cases not belonging to any epidemiological outbreaks. In conclusion, as demonstrated here, the resolution of food-related outbreaks with such a high discriminatory power is rare in outbreak investigation.

  11. The Salmonella Pathogenicity Island 13 contributes to pathogenesis in streptomycin pre-treated mice but not in day-old chickens

    USDA-ARS?s Scientific Manuscript database

    Salmonella Enteritidis (S. Enteritidis) is a human and animal pathogen that causes gastroenteritis characterized by inflammatory diarrhea and occasionally an invasive systemic infection. Salmonella pathogenicity islands (SPIs) are horizontally acquired genomic segments known to contribute to Salmone...

  12. Detection and enumeration of Salmonella enteritidis in homemade ice cream associated with an outbreak: comparison of conventional and real-time PCR methods.

    PubMed

    Seo, K H; Valentin-Bon, I E; Brackett, R E

    2006-03-01

    Salmonellosis caused by Salmonella Enteritidis (SE) is a significant cause of foodborne illnesses in the United States. Consumption of undercooked eggs and egg-containing products has been the primary risk factor for the disease. The importance of the bacterial enumeration technique has been enormously stressed because of the quantitative risk analysis of SE in shell eggs. Traditional enumeration methods mainly depend on slow and tedious most-probable-number (MPN) methods. Therefore, specific, sensitive, and rapid methods for SE quantitation are needed to collect sufficient data for risk assessment and food safety policy development. We previously developed a real-time quantitative PCR assay for the direct detection and enumeration of SE and, in this study, applied it to naturally contaminated ice cream samples with and without enrichment. The detection limit of the real-time PCR assay was determined with artificially inoculated ice cream. When applied to the direct detection and quantification of SE in ice cream, the real-time PCR assay was as sensitive as the conventional plate count method in frequency of detection. However, populations of SE derived from real-time quantitative PCR were approximately 1 log higher than provided by MPN and CFU values obtained by conventional culture methods. The detection and enumeration of SE in naturally contaminated ice cream can be completed in 3 h by this real-time PCR method, whereas the cultural enrichment method requires 5 to 7 days. A commercial immunoassay for the specific detection of SE was also included in the study. The real-time PCR assay proved to be a valuable tool that may be useful to the food industry in monitoring its processes to improve product quality and safety.

  13. The “Decline and Fall” of Nontyphoidal Salmonella in the United Kingdom

    PubMed Central

    O'Brien, Sarah J.

    2013-01-01

    Remarkable changes in the epidemiology of human nontyphoidal salmonellosis have occurred in the United Kingdom over the last century. Between 1981 and 1991, the incidence of nontyphoidal salmonellosis in the United Kingdom rose by >170%, driven primarily by an epidemic of Salmonella enterica subspecies enterica serovar Enteritidis phage type (PT) 4, which peaked in 1993. Measures introduced to control this epidemic included legislation, food safety advice, and an industry-led vaccination program in broiler-breeder and laying poultry flocks. The incidence of Salmonella Enteritidis has been falling since 1997, and levels of Salmonella Enteritidis PT4 have fallen to preepidemic levels and have stayed low. The temporal relationship between vaccination programs and the reduction in human disease is compelling and suggests that these programs have made a major contribution to improving public health. PMID:23166188

  14. Promoting crystallisation of the Salmonella enteritidis fimbriae 14 pilin SefD using deuterium oxide

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liu, Bing; Garnett, James A.; Lee, Wei-chao

    Highlights: Black-Right-Pointing-Pointer The benefits of D{sub 2}O in screening for crystallisation was explored. Black-Right-Pointing-Pointer The crystal structures of the SefD pilin in both H{sub 2}O and D{sub 2}O reveal differences. Black-Right-Pointing-Pointer Crystallisation improvements are explained by altered interactions in D{sub 2}O crystals. Black-Right-Pointing-Pointer D{sub 2}O is useful additive in sparse-matrix screening for crystallisation. -- Abstract: The use of heavy water (D{sub 2}O) as a solvent is commonplace in many spectroscopic techniques for the study of biological macromolecules. A significant deuterium isotope effect exists where hydrogen-bonding is important, such as in protein stability, dynamics and assembly. Here we illustrate the usemore » of D{sub 2}O in additive screening for the production of reproducible diffraction-quality crystals for the Salmonella enteritidis fimbriae 14 (SEF14) putative tip adhesin, SefD.« less

  15. Applications of immunomagnetic capture and time-resolved fluorescence detection for Salmonella enteriditis in liquid eggs

    NASA Astrophysics Data System (ADS)

    Tu, Shu-I.; Gehring, Andrew; Paoli, George

    2008-04-01

    An immuno sandwich method was evaluated for the detection of Salmonella in liquid eggs. Liquid eggs spiked with different out-break strains of Salmonella were mixed with proper enrichment media and incubated at 37 C for 4 to 20 h. After enrichment, immunomagnetic beads (IMB) coated with anti Salmonella antibodies were used to capture the bacteria. Samarium (Sm) labeled anti Salmonella antibodies were then used to form sandwiched complexes with IMB captured bacteria. Sandwiched Salmonella were then treated with Sm-chelator to allow the measurement of the released Sm by time-resolved fluorescence (TRF). The processes ranging from IMB capture to Sm chelation were performed using an automated KingFisher apparatus. With this approach, the presence of ~ 1 CFU of outbreak strains of Salmonella Enteritidis per egg (~50 g of liquid eggs) could be detected after enrichment for 20 h at 37 C. For higher levels of Salmonella Enteritidis contamination, e.g., 10 CFU per 50 g of liquid eggs, the enrichment time could be reduced to 5 h at 37 C. The results demonstrated that a combination of IMB capture and TRF measurement could be a rapid and sensitive method for Salmonella Enteritidis detection in liquid eggs.

  16. A mutation in the NLRC5 promoter limits NF-κB signaling after Salmonella Enteritidis infection in the spleen of young chickens.

    PubMed

    Chang, Guobin; Liu, Xiangping; Ma, Teng; Xu, Lu; Wang, Hongzhi; Li, Zhiteng; Guo, Xiaomin; Xu, Qi; Chen, Guohong

    2015-09-01

    To date, the functions of the NLRC5 in chickens remain undefined. In the current study, chicken NLRC5 was cloned and an A1017G mutation was detected in its promoter region. The relative expression levels of the NLRC5 and key NF-κB pathway genes, IKKα, IKKβ, NF-κB, IL-6, IL-1β and IFN-γ, in the spleens of wild and mutant type birds, AA and GG, were determined using FQ-PCR at 7 day post-infection (DPI) with Salmonella Enteritidis. Additionally, the bacterial burden in the caecum and various immune response parameters were measured to evaluate immune responses. All of the examined immune response parameters were significantly different between the AA chickens and the GG chickens. Specifically, the mRNA expression levels of IKKα, NF-κB, IL-6, IL-1β and IFN-γ were higher in AA chickens than those in GG chickens, while the mRNA expression levels of NLRC5 were lower in AA chickens than those in GG chickens (P<0.05). Moreover, the mRNA expression levels of TLR4 and MyD88 were not affected in either group. Collectively, considering former NLRC5 functional study in vitro, the wild genotype birds presented with better resistance to Salmonella Enteritidis through the actions of the NLRC5 and subsequent inhibition of the NF-κB pathway in chickens. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Highly expressed amino acid biosynthesis genes revealed by global gene expression analysis of Salmonella enterica serovar Enteritidis during growth in whole egg are not essential for this growth.

    PubMed

    Jakočiūnė, Džiuginta; Herrero-Fresno, Ana; Jelsbak, Lotte; Olsen, John Elmerdahl

    2016-05-02

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is the most common cause of egg borne salmonellosis in many parts of the world. This study analyzed gene expression of this bacterium during growth in whole egg, and whether highly expressed genes were essential for the growth. High quality RNA was extracted from S. Enteritidis using a modified RNA-extraction protocol. Global gene expression during growth in whole egg was compared to growth in LB-medium using DNA array method. Twenty-six genes were significantly upregulated during growth in egg; these belonged to amino acid biosynthesis, di/oligopeptide transport system, biotin synthesis, ferrous iron transport system, and type III secretion system. Significant downregulation of 15 genes related to formate hydrogenlyase (FHL) and trehalose metabolism was observed. The results suggested that S. Enteritidis is starved for amino-acids, biotin and iron when growing in egg. However, site specific mutation of amino acid biosynthesis genes asnA (17.3 fold upregulated), asnB (18.6 fold upregulated), asnA/asnB and, serA (12.0 fold upregulated) and gdhA (3.7 fold upregulated), did not result in growth attenuation, suggesting that biosynthesis using the enzymes encoded from these genes may represent the first choice for S. Enteritidis when growing in egg, but when absent, the bacterium could use alternative ways to obtain the amino acids. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Survey of Salmonella contamination of non-United Kingdom-produced raw shell eggs on retail sale in the northwest of England and London, 2005 to 2006.

    PubMed

    Little, C L; Walsh, S; Hucklesby, L; Surman-Lee, S; Pathak, K; Gatty, Y; Greenwood, M; De Pinna, E; Threlfall, E J; Maund, A; Chan, C H

    2007-10-01

    This survey was prompted by a change in the epidemiology of Salmonella Enteritidis infections in England and Wales and elsewhere in Europe and, to our knowledge, is the first survey to provide information on Salmonella contamination of non-United Kingdom eggs on retail sale. Based on 10,464 non-United Kingdom eggs (1744 pooled samples of six eggs) purchased between March 2005 and July 2006, the total weighted prevalence estimate for all Salmonella detected in non-United Kingdom eggs was 3.3%. Of the eggs sampled, most were produced in Spain (66.3%), France (20.0%), or The Netherlands (7.4%). Salmonella was detected from 4.4 and 0.3% of eggs produced in Spain and France, respectively, with weighted prevalence estimates. Eight different Salmonella serotypes were recovered from non-United Kingdom eggs, of which Salmonella Enteritidis predominated, with an estimated prevalence of 2.6%. Salmonella Enteritidis was obtained only from Spanish eggs. Nine different phage types of Salmonella Enteritidis were identified, with phage type 1 found to be the predominant phage type. Most of the Salmonella Enteritidis isolates obtained from Spanish eggs in the survey were resistant to nalidixic acid with concomitant decreased susceptibility to ciprofloxacin (0.125 to 1.0 mg/liter) or ampicillin (8.0 mg/liter). Salmonella Enteritidis phage type 1 until now had not been detected in eggs examined as part of previous United Kingdom egg surveys but has been detected in eggs of Spanish origin examined during recent national outbreaks of Salmonella Enteritidis non-phage type 4 infections in England and Wales. Eggs are a commonly consumed food that may occasionally be contaminated with Salmonella. The rates of contamination may be linked to the origin of the eggs. Consumers and caterers need to be aware of this continuing hazard, adopt appropriate control measures, and follow advice provided by national food agencies in order to reduce the risk of infection.

  19. Prevalence, Virulence Genes and Antimicrobial Resistance Profiles of Salmonella Serovars from Retail Beef in Selangor, Malaysia

    PubMed Central

    Thung, Tze Y.; Radu, Son; Mahyudin, Nor A.; Rukayadi, Yaya; Zakaria, Zunita; Mazlan, Nurzafirah; Tan, Boon H.; Lee, Epeng; Yeoh, Soo L.; Chin, Yih Z.; Tan, Chia W.; Kuan, Chee H.; Basri, Dayang F.; Wan Mohamed Radzi, Che W. J.

    2018-01-01

    The aim of the present study was to investigate the prevalence of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in retail beef from different retail markets of Selangor area, as well as, to assess their pathogenic potential and antimicrobial resistance. A total of 240 retail beef meat samples (chuck = 60; rib = 60; round = 60; sirloin = 60) were randomly collected. The multiplex polymerase chain reaction (mPCR) in combination with the most probable number (MPN) method was employed to detect Salmonella spp., S. Enteritidis and S. Typhimurium in the meat samples. The prevalence of Salmonella spp., S. Enteritidis and S. Typhimurium in 240 beef meat samples were 7.50, 1.25, and 0.83%, respectively. The microbial loads of total Salmonella was found in the range of <3 to 15 MPN/g. Eight different serovars of Salmonella were identified among the 23 isolates, and S. Agona was the predominant serovar (26.09%). Interestingly, all the Salmonella isolates were resistant to penicillin, erythromycin and vancomycin, but the sensitivity was observed for tetracycline, gentamicin and amoxicillin/clavulanic acid. All 23 isolates were resistant to at least three antibiotics. Two S. Typhimurium isolates (8.70%) exhibited the highest multiple antibiotic resistance (MAR) index value of 0.56 which shown resistance to nine antibiotics. PCR analysis of virulence genes showed that all Salmonella isolates (100%) were positive for the invA gene. Meanwhile, pefA was only identified in S. Enteritidis and S. Typhimurium. The findings in this study indicate that retail beef products tested were widely contaminated with multi-drug resistant (MDR) Salmonella and various virulence genes are present among the isolated Salmonella serovars. PMID:29379488

  20. Prevalence, Virulence Genes and Antimicrobial Resistance Profiles of Salmonella Serovars from Retail Beef in Selangor, Malaysia.

    PubMed

    Thung, Tze Y; Radu, Son; Mahyudin, Nor A; Rukayadi, Yaya; Zakaria, Zunita; Mazlan, Nurzafirah; Tan, Boon H; Lee, Epeng; Yeoh, Soo L; Chin, Yih Z; Tan, Chia W; Kuan, Chee H; Basri, Dayang F; Wan Mohamed Radzi, Che W J

    2017-01-01

    The aim of the present study was to investigate the prevalence of Salmonella spp., Salmonella Enteritidis and Salmonella Typhimurium in retail beef from different retail markets of Selangor area, as well as, to assess their pathogenic potential and antimicrobial resistance. A total of 240 retail beef meat samples (chuck = 60; rib = 60; round = 60; sirloin = 60) were randomly collected. The multiplex polymerase chain reaction (mPCR) in combination with the most probable number (MPN) method was employed to detect Salmonella spp., S . Enteritidis and S . Typhimurium in the meat samples. The prevalence of Salmonella spp., S . Enteritidis and S . Typhimurium in 240 beef meat samples were 7.50, 1.25, and 0.83%, respectively. The microbial loads of total Salmonella was found in the range of <3 to 15 MPN/g. Eight different serovars of Salmonella were identified among the 23 isolates, and S . Agona was the predominant serovar (26.09%). Interestingly, all the Salmonella isolates were resistant to penicillin, erythromycin and vancomycin, but the sensitivity was observed for tetracycline, gentamicin and amoxicillin/clavulanic acid. All 23 isolates were resistant to at least three antibiotics. Two S . Typhimurium isolates (8.70%) exhibited the highest multiple antibiotic resistance (MAR) index value of 0.56 which shown resistance to nine antibiotics. PCR analysis of virulence genes showed that all Salmonella isolates (100%) were positive for the invA gene. Meanwhile, pefA was only identified in S . Enteritidis and S . Typhimurium. The findings in this study indicate that retail beef products tested were widely contaminated with multi-drug resistant (MDR) Salmonella and various virulence genes are present among the isolated Salmonella serovars.

  1. Outbreak of Salmonella Enteritidis linked to the consumption of frozen beefburgers received from a food bank and originating from Poland: northern France, December 2014 to April 2015.

    PubMed

    Jones, Gabrielle; Pihier, Nathalie; Vanbockstael, Caroline; Le Hello, Simon; Cadel Six, Sabrina; Fournet, Nelly; Jourdan-da Silva, Nathalie

    2016-10-06

    A prolonged outbreak of Salmonella enterica serotype Enteritidis occurred in northern France between December 2014 and April 2015. Epidemiological investigations following the initial notification on 30 December 2014 of five cases of salmonellosis (two confirmed S. Enteritidis) in young children residing in the Somme department revealed that all cases frequented the same food bank A. Further epidemiological, microbiological and food trace-back investigations indicated frozen beefburgers as the source of the outbreak and the suspected lot originating from Poland was recalled on 22 January 2015. On 2 March 2015 a second notification of S. Enteritidis cases in the Somme reinitiated investigations that confirmed a link with food bank A and with consumption of frozen beefburgers from the same Polish producer. In the face of a possible persistent source of contamination, all frozen beefburgers distributed by food bank A and from the same origin were blocked on 3 March 2015. Microbiological analyses confirmed contamination by S. Enteritidis of frozen beefburgers from a second lot remaining in cases' homes. A second recall was initiated on 6 March 2015 and all frozen beefburgers from the Polish producer remain blocked after analyses identified additional contaminated lots over several months of production. This article is copyright of The Authors, 2016.

  2. Detection and classification of salmonella serotypes using spectral signatures collected by fourier transform infrared (FT-IR) spectroscopy

    USDA-ARS?s Scientific Manuscript database

    Spectral signatures of Salmonella serotypes namely Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Infantis, Salmonella Heidelberg and Salmonella Kentucky were collected using Fourier transform infrared spectroscopy (FT-IR). About 5-10 µL of Salmonella suspensions with concentrations of 1...

  3. The effects of polymorphisms in IL-2, IFN-γ, TGF-β2, IgL, TLR-4, MD-2, and iNOS genes on resistance to Salmonella enteritidis in indigenous chickens.

    PubMed

    Tohidi, Reza; Idris, Ismail Bin; Panandam, Jothi Malar; Bejo, Mohd Hair

    2012-12-01

    Salmonella Enteritidis is a major cause of food poisoning worldwide, and poultry products are the main source of S. Enteritidis contamination for humans. Among the numerous strategies for disease control, improving genetic resistance to S. Enteritidis has been the most effective approach. We investigated the association between S. Enteritidis burden in the caecum, spleen, and liver of young indigenous chickens and seven candidate genes, selected on the basis of their critical roles in immunological functions. The genes included those encoding interleukin 2 (IL-2), interferon-γ (IFN-γ), transforming growth factor β2 (TGF-β2), immunoglobulin light chain (IgL), toll-like receptor 4 (TLR-4), myeloid differentiation protein 2 (MD-2), and inducible nitric oxide synthase (iNOS). Two Malaysian indigenous chicken breeds were used as sustainable genetic sources of alleles that are resistant to salmonellosis. The polymerase chain reaction restriction fragment-length polymorphism technique was used to genotype the candidate genes. Three different genotypes were observed in all of the candidate genes, except for MD-2. All of the candidate genes showed the Hardy-Weinberg equilibrium for the two populations. The IL-2-MnlI polymorphism was associated with S. Enteritidis burden in the caecum and spleen. The TGF-β2-RsaI, TLR-4-Sau 96I, and iNOS-AluI polymorphisms were associated with the caecum S. Enteritidis load. The other candidate genes were not associated with S. Enteritidis load in any organ. The results indicate that the IL-2, TGF-β2, TLR-4, and iNOS genes are potential candidates for use in selection programmes for increasing genetic resistance against S. Enteritidis in Malaysian indigenous chickens.

  4. Determination of an effective sampling regime to detect salmonella enteritidis in the environment of poultry units.

    PubMed

    Davies, R H; Wray, C

    1996-05-01

    A study of the dissemination of Salmonella enteritidis in the poultry breeder industry in the UK showed that the choice of sites for sampling the environment of occupied houses and empty houses which had been disinfected after depopulation had a significant influence on the outcome. Increased isolation rates could be achieved by sampling nest box floors and dust in open slave feed hoppers in occupied poultry houses. Nest box floors were the most sensitive sites for detection of residual environmental contamination in poultry houses where enrofloxacin treatment had been used. Floor sweepings, nest box floors, slave feed hoppers, hydrated wall fabric junctions and high beams and pipes were the most sensitive sample sites in cleansed and disinfected poultry houses. The use of universal disinfectant neutralisers gave good results in laboratory trials but appeared to reduce the isolation rate from field samples.

  5. Identification and characterization of salmonella serotypes using DNA spectral characteristics by fourier transform infrared (FT-IR) spectroscopy

    USDA-ARS?s Scientific Manuscript database

    Analysis of DNA samples of Salmonella serotypes (Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Infantis, Salmonella Heidelberg and Salmonella Kentucky) were performed using Fourier transform infrared spectroscopy (FT-IR) spectrometer by placing directly in contact with a diamond attenua...

  6. Energy partitioning and thyroid hormone levels during Salmonella enteritidis infections in pullets with high or low residual feed intake.

    PubMed

    Van Eerden, E; Van Den Brand, H; Heetkamp, M J W; Decuypere, E; Kemp, B

    2006-10-01

    This experiment was conducted to investigate whether feed efficiency, as measured by residual feed intake as a phenotypic trait, affects energy partitioning in pullets that have received Salmonella inoculation as an immune challenge. In each of 8 trials, energy partitioning was measured during 5 wk in 15-wk-old efficient (R-) and nonefficient (R+) pullets, which were housed per efficiency group in 2 identical climate respiration chambers. After 1 wk of adaptation, the pullets in 4 trials were orally inoculated with 10(8) cfu of Salmonella enteritidis; pullets in the remaining trials were not inoculated and served as controls. Heat production was calculated from continuous recordings of O(2) consumption and CO(2) production. Energy and N partitioning were recorded on a weekly basis. Blood samples for analyses on thyroid hormones were taken at 16, 17, and 19 wk of age. There were no interactions between efficiency type and Salmonella treatment or Salmonella treatment effects in energy partitioning, except for a short-term increase in heat production in inoculated pullets. Nonefficient pullets had higher gross energy and ME intake, higher estimated ME for maintenance, lower ME:gross energy ratio, and higher total heat production and nonactivity-related heat production compared with R- pullets. Triiodothyronine levels in R+ pullets were higher at 16 and 17 wk but were lower at 19 wk of age compared with R- pullets. Thyroxine levels were higher in R- at 16 wk and showed interactions between efficiency type and Salmonella treatment at 17 and 19 wk of age. Body weights and spleen weights did not differ between efficiency groups. Nonefficient pullets had higher heart, liver, and ovary weights and more large yellow follicles than R- pullets. There were no Salmonella effects on body and organ weights. We conclude that R+ pullets have a faster running energy metabolism and that they put more resources into organ development than R- pullets. Inoculation with Salmonella has a

  7. Inactivation of Staphylococcus aureus and Salmonella enteritidis in tryptic soy broth and caviar samples by high pressure processing.

    PubMed

    Fioretto, F; Cruz, C; Largeteau, A; Sarli, T A; Demazeau, G; El Moueffak, A

    2005-08-01

    We studied the action of high pressure processing on the inactivation of two foodborne pathogens, Staphylococcus aureus ATCC 6538 and Salmonella enteritidis ATCC 13076, suspended in a culture medium and inoculated into caviar samples. The baroresistance of the two pathogens in a tryptic soy broth suspension at a concentration of 10(8)-10(9) colony-forming units/ml was tested for continuous and cycled pressurization in the 150- to 550-MPa range and for 15-min treatments at room temperature. The increase of cycle number permitted the reduction of the pressure level able to totally inactivate both microorganisms in the tryptic soy broth suspension, whereas the effect of different procedure times on complete inactivation of the microorganisms inoculated into caviar was similar.

  8. Descriptive survey and Salmonella surveillance of pastured poultry layer farms in California.

    PubMed

    Dailey, Naomi; Niemeier, Deb; Elkhoraibi, Carine; Sentíes-Cué, C Gabriel; Pitesky, Maurice

    2017-04-01

    While pasture-raised poultry comprises a small portion of the commercial poultry industry in North America, these alternative rearing systems have become increasingly popular. As such, it is critical to improve our understanding of husbandry practices and prevalence of zoonotic and epizoonotic diseases in these systems. This research reviews the results of a survey sent to 82 commercial pastured poultry farms in California. While the survey response was low (13.4%), it was enhanced by detailed in-person interviews and farm visits. In addition, we conducted drag swabs for Salmonella Enteritidis. On average, farms utilized 12.3% of their total farmland for pastured poultry operations, which often coexisted with other livestock (45%), touch crops (27%), and non-touch crops (45%). While the mean (44.6 sq. ft./hen) and median (22.2 sq. ft./hen) pasture stocking densities were within auditing guidelines, the mean (1.2 sq. ft./hen) and median (0.5 sq. ft./hen) coop stocking densities were below the pending USDA (2016) guidelines recommended in 7 CFR Part 205. Drag swab results showed the presence of Salmonella Enteritidis (SE) in the environment of one of the 11 farms (9.1%). In addition, Salmonella Pullorum (SP) whole blood agglutination tests were used to understand the prevalence of Salmonella spp. in laying hens within the studied farms. Results showed the presence of antibodies in flocks at six of the seven non-SE vaccinated farms, with a mean on-farm prevalence of 25.6% in laying hens. Logistic regression was used to determine risk factors for Group D Salmonella exposure in non-vaccinated flocks, using the SP blood agglutination data as the dependent variable and the survey questions as the independent variables. Statistically significant (P < 0.05) risk factors included exposed wire floors and flock size. These results improve our understanding of Salmonella prevalence and husbandry practices on commercial pastured poultry farms in California. © 2016 Poultry

  9. Subtyping of Canadian isolates of Salmonella Enteritidis using Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) alone and in combination with Pulsed-Field Gel Electrophoresis (PFGE) and phage typing.

    PubMed

    Ziebell, Kim; Chui, Linda; King, Robin; Johnson, Suzanne; Boerlin, Patrick; Johnson, Roger P

    2017-08-01

    Salmonella enterica subspecies enterica serovar Enteritidis (SE) is one of the most common causes of human salmonellosis and in Canada currently accounts for over 40% of human cases. Reliable subtyping of isolates is required for outbreak detection and source attribution. However, Pulsed-Field Gel Electrophoresis (PFGE), the current standard subtyping method for Salmonella spp., is compromised by the high genetic homogeneity of SE. Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) was introduced to supplement PFGE, although there is a lack of data on the ability of MLVA to subtype Canadian isolates of SE. Three subtyping methods, PFGE, MLVA and phage typing were compared for their discriminatory power when applied to three panels of Canadian SE isolates: Panel 1: 70 isolates representing the diversity of phage types (PTs) and PFGE subtypes within these PTs; Panel 2: 214 apparently unrelated SE isolates of the most common PTs; and Panel 3: 27 isolates from 10 groups of epidemiologically related strains. For Panel 2 isolates, four MLVA subtypes were shared among 74% of unrelated isolates and in Panel 3 isolates, one MLVA subtype accounted for 62% of the isolates. For all panels, combining results from PFGE, MLVA and PT gave the best discrimination, except in Panel 1, where the combination of PT and PFGE was equally as high, due to the selection criteria for this panel. However, none of these methods is sufficiently discriminatory alone for reliable outbreak detection or source attribution, and must be applied together to achieve sufficient discrimination for practical purposes. Even then, some large clusters were not differentiated adequately. More discriminatory methods are required for reliable subtyping of this genetically highly homogeneous serovar. This need will likely be met by whole genome sequence analysis given the recent promising reports and as more laboratories implement this tool for outbreak response and surveillance. Copyright © 2017

  10. Differences in expression of genes in the MyD88 and TRIF signalling pathways and methylation of TLR4 and TRIF in Tibetan chickens and DaHeng S03 chickens infected with Salmonella enterica serovar enteritidis.

    PubMed

    Li, Xiaocheng; Zhang, Peng; Jiang, Xiaosong; Du, Huarui; Yang, Chaowu; Zhang, Zengrong; Men, Shuai; Zhang, Zhikun; Jiang, Wei; Wang, Hongning

    2017-07-01

    Salmonella enterica serovar (S. enteritidis) is a pathogenic bacterium that can cause symptoms of food poisoning, leading to death of poultry, resulting in serious economic losses. The MyD88 and TRIF signalling pathways play important roles in activating innate and adaptive immunity in chickens infected with S. enteritidis. The objective of the present study was to characterize in vivo mRNA expressions, protein levels and methylation levels of genes in the above two pathways in both Tibetan chickens and DaHeng S03 chickens infected with S. enteritidis. MyD88-dependent and TRIF-dependent signalling pathway were activated by infection, and the MyD88 signalling pathway induced cytokines LITAF and IL-8 played important roles in fighting against the S. enteritidis infection in vivo. The TLR4 methylation might alter expression of genes involved in the MyD88 signalling pathway, and thus different breeds of chickens might show differences in susceptibility to the S. enteritidis. The increased expression of INF β was activated by S. enteritidis, but its expressions were different in levels of mRNA and protein in Tibetan chickens and DaHeng chickens, suggesting its functions on the resistance to S. enteritidis infection in chickens. This study contributes to the understanding of two pathways activated in response to S. enteritidis infection, and gives indications on the mechanisms underlying resistance of Tibetan chickens and DaHeng chickens to S. enteritidis. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Comparative study of all Salmonella enterica serovar Enteritidis strains isolated from food and food animals in Greece from 2008 to 2010 with clinical isolates.

    PubMed

    Papadopoulos, T; Petridou, E; Zdragas, A; Mandilara, G; Nair, S; Peters, T; Chattaway, M; de Pinna, E; Passiotou, M; Vatopoulos, A

    2016-05-01

    The aim of the present work was to study the epidemiology of Salmonella enterica serovar Enteritidis (S. Enteritidis) in Greece, comparing all the food and food animal isolates during a 3-year period with clinical isolates. Submission of the generated data to the PulseNet Europe database was carried out in order to study the population structure of this particular serovar and indicate possible connections with European strains. One hundred and sixty-eight (168) S. Enteritidis strains of human, animal, and food origin, isolated during the period 2008-2010 in Greece, were studied. Strains were characterized by phenotypic (antibiotic resistance) and molecular [pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST)] methods. PFGE revealed 39 XbaI, 48 BlnI, and 80 XbaI-BlnI distinct pulsotypes, suggesting several clones circulating through the food chain and multiple sources of transmission. Submission to the PulseNet Europe database indicated that PFGE profile SENTXB.0001, the most common PFGE profile in Europe, was also predominant in Greece (33.3 %). MLST showed that all the strains studied shared the same sequence type (ST11), representing the most common ST in Europe. High rates of resistance to nalidixic acid were observed among human and poultry isolates (~25 %), indicating the potential fluoroquinolone treatment failure. Our data suggest that strains originating from multiple reservoirs circulated in Greece through the food chain during the study period. Predominant profiles in Greece were common to PulseNet Europe profiles, indicating similarities between the S. Enteritidis populations in Greece and Europe.

  12. Simultaneous Detection of Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes at a Very Low Level Using Simultaneous Enrichment Broth and Multichannel SPR Biosensor.

    PubMed

    Zhang, Xiaoguang; Tsuji, Sachiko; Kitaoka, Hayato; Kobayashi, Hiroshi; Tamai, Mitsuru; Honjoh, Ken-Ichi; Miyamoto, Takahisa

    2017-10-01

    Detection of foodborne pathogens at very low levels is still a challenge. A custom-built multichannel surface plasmon resonance (SPR) biosensor and simultaneous enrichment broth (SEB) were used to develop a simultaneous detection method for 3 important foodborne pathogens, Escherichia coli O157:H7 (O157:H7), Salmonella enteritidis, and Listeria monocytogenes, at a very low level. These 3 foodborne pathogens at a very low level (14, 6, and 28 CFU/25 g (mL) for O157:H7, S. enteritidis, and L. monocytogenes, respectively) were inoculated in SEB and incubated at 37 ˚C for 24 h. Sample prepared from the simultaneous enrichment culture was analyzed using the multichannel SPR biosensor and sensor chip immobilized with polyclonal antibodies specific to each of the target pathogens. O157:H7, S. enteritidis, and L. monocytogenes in chicken were detected simultaneously at an inoculum dose of 14, 6, and 28 CFU/25 g, respectively. Our method using a custom-built multichannel SPR biosensor and enrichment in SEB is expected as a rapid and simultaneous detection method for low levels of O157:H7, S. enteritidis, and L. monocytogenes in food. Our method is expected as a rapid and simultaneous detection method for pathogens at very low levels. It has great potential for safety control of food and microbiological detection applications. © 2017 Institute of Food Technologists®.

  13. The extradomain a of fibronectin enhances the efficacy of lipopolysaccharide defective Salmonella bacterins as vaccines in mice

    PubMed Central

    2012-01-01

    The Extradomain A from fibronectin (EDA) has an immunomodulatory role as fusion protein with viral and tumor antigens, but its effect when administered with bacteria has not been assessed. Here, we investigated the adjuvant effect of EDA in mice immunizations against Salmonella enterica subspecies enterica serovar Enteritidis (Salmonella Enteritidis). Since lipopolysaccharide (LPS) is a major virulence factor and the LPS O-polysaccharide (O-PS) is the immunodominant antigen in serological diagnostic tests, Salmonella mutants lacking O-PS (rough mutants) represent an interesting approach for developing new vaccines and diagnostic tests to differentiate infected and vaccinated animals (DIVA tests). Here, antigenic preparations (hot-saline extracts and formalin-inactivated bacterins) from two Salmonella Enteritidis rough mutants, carrying either intact (SEΔwaaL) or deep-defective (SEΔgal) LPS-Core, were used in combination with EDA. Biotinylated bacterins, in particular SEΔwaaL bacterin, decorated with EDAvidin (EDA and streptavidin fusion protein) improved the protection conferred by hot-saline or bacterins alone and prevented significantly the virulent infection at least to the levels of live attenuated rough mutants. These findings demonstrate the adjuvant effect of EDAvidin when administered with biotinylated bacterins from Salmonella Enteritidis lacking O-PS and the usefulness of BEDA-SEΔwaaL as non-live vaccine in the mouse model. PMID:22515195

  14. Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples.

    PubMed

    Kasturi, Kuppuswamy N; Drgon, Tomas

    2017-07-15

    The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA , group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non- Salmonella organisms. The invA - and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella -differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the V itek i mmuno d iagnostic a ssay s ystem (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. The receiver operating characteristic (ROC) analysis documented excellent agreement between the results from the culture and PCR methods (area under the curve, 0.90; 95% confidence interval of 0.76 to 1.0) confirming the validity of the PCR method. IMPORTANCE This validated PCR method detects 55% more positives for Salmonella in half the time required for the reference method, VIDAS. The validated PCR method will help to strengthen public health efforts through rapid screening of Salmonella spp. in environmental samples.

  15. Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples

    PubMed Central

    Drgon, Tomas

    2017-01-01

    ABSTRACT The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA, group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non-Salmonella organisms. The invA- and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella-differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S. Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the Vitek immunodiagnostic assay system (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. The receiver operating characteristic (ROC) analysis documented excellent agreement between the results from the culture and PCR methods (area under the curve, 0.90; 95% confidence interval of 0.76 to 1.0) confirming the validity of the PCR method. IMPORTANCE This validated PCR method detects 55% more positives for Salmonella in half the time required for the reference method, VIDAS. The validated PCR method will help to strengthen public health efforts through rapid screening of Salmonella spp. in environmental samples. PMID:28500041

  16. Isolation and molecular characterization of Salmonella enterica serovar Enteritidis from poultry house and clinical samples during 2010.

    PubMed

    Mezal, Ezat H; Sabol, Ashley; Khan, Mariam A; Ali, Nawab; Stefanova, Rossina; Khan, Ashraf A

    2014-04-01

    A total of 60 Salmonella enterica serovar (ser.) Enteritidis isolates, 28 from poultry houses and 32 from clinical samples, were isolated during 2010. These isolates were subjected to testing and analyzed for antibiotic resistance, virulence genes, plasmids and plasmid replicon types. To assess genetic diversity, pulsed-field gel electrophoresis (PFGE) fingerprinting, using the XbaI restriction enzyme, Multiple-Locus Variable-Number Tandem Repeat Analysis (MLVA) and plasmid profiles were performed. All isolates from poultry, and 10 out of 32 clinical isolates were sensitive to ampicillin, chloramphenicol, gentamicin, kanamycin, nalidixic acid, sulfisoxazole, streptomycin, and tetracycline. Twenty-one of thirty-two clinical isolates were resistant to ampicillin and tetracycline, and one isolate was resistant to nalidixic acid. PFGE typing of sixty ser. Enteritidis isolates by XbaI resulted in 10-12 bands and grouped into six clusters each with similarity from 95% to 81%. The MLVA analysis of sixty isolates gave 18 allele profiles with the majority of isolates displayed in three groups, and two clinical isolates found to be new in the PulseNet national MLVA database. All isolates were positive for 12 or more of the 17 virulence genes mostly found in S. enterica (spvB, spiA, pagC, msgA, invA, sipB, prgH, spaN, orgA, tolC, iroN, sitC, IpfC, sifA, sopB, and pefA) and negative for one gene (cdtB). All isolates carried a typical 58 kb plasmid, type Inc/FIIA. Three poultry isolates and one clinical isolate carried small plasmids with 3.8, 6, 7.6 and 11.5 kb. Ten of the clinical isolates carried plasmids, with sizes 36 and 38 kb, types IncL/M and IncN, and one isolate carried an 81 kb plasmid, type IncI. Southern hybridization of a plasmid with an Inc/FIIA gene probe hybridized one large 58 kb plasmid in all isolates. Several large and small plasmids from poultry isolates were not typed by our PCR-based method. These results confirmed that PFGE fingerprinting has

  17. Antimicrobial susceptibility of Salmonella enterica isolates from healthy breeder and broiler flocks in Portugal.

    PubMed

    Clemente, Lurdes; Correia, Ivone; Themudo, Patrícia; Neto, Isabel; Caniça, Manuela; Bernardo, Fernando

    2014-05-01

    Three hundred and thirty-three isolates representing 40 different serotypes of Salmonella enterica, recovered from environmental and faecal samples of breeder and broiler flocks from 2009 to 2011, were studied. Antimicrobial susceptibility was determined by measuring the minimal inhibitory concentration of 11 antimicrobials using the agar dilution method. Salmonella Havana, S. Enteritidis and S. Mbandaka were the most common serotypes isolated from broiler flocks, while S. Enteritidis was the common isolate from breeder flocks. The frequency of non-wild-type Salmonella isolates (those with decreased susceptibility to the different antimicrobials) varied according to serotype. S. Mbandaka in broilers and S. Enteritidis in both breeders and broilers showed higher frequencies of reduced susceptibility to quinolones, but clinical resistance towards ciprofloxacin was not observed. Reduced susceptibility to sulfamethoxazole, tetracycline, ampicillin and streptomycin were common in Salmonella Typhimurium isolates. Two isolates of S. Havana from broilers were resistant to cefotaxime and phenotypically categorised as extended-spectrum β-lactamase producers. The results presented in this study provide useful data on the antimicrobial susceptibility of different Salmonella serotypes and highlight the high diversity of multi-drug resistance patterns present. Copyright © 2014 Elsevier Ltd. All rights reserved.

  18. Occurrence and characterization of Salmonella from chicken nuggets, strips, and pelleted broiler feed.

    PubMed

    Bucher, O; Holley, R A; Ahmed, R; Tabor, H; Nadon, C; Ng, L K; D'Aoust, J Y

    2007-10-01

    Raw, frozen chicken nuggets and strips have been identified as a significant risk factor in contracting foodborne salmonellosis. Cases of salmonellosis as a result of consuming partly cooked chicken nuggets may be due in part to Salmonella strains originating in broiler feed. This study was undertaken to determine the occurrence and characterize the strains of Salmonella contaminating chicken nuggets, strips, and pelleted feeds, in an attempt to demonstrate whether the same Salmonella strains present in broiler feed could be isolated from raw, frozen chicken nuggets and strips available for human consumption. Salmonellae were recovered using the Health Canada MFHPB-20 method for the isolation and identification of Salmonella from foods. Strains were characterized by serotyping, phage typing, antimicrobial resistance typing (R-typing), and by pulsed-field gel electrophoresis (PFGE). Salmonellae were isolated from 25-g samples in 27% (n=92) of nugget and strip samples, 95% (n=20) of chicken nugget meat samples, and from 9% (n=111) of pelleted feed samples. Salmonella Heidelberg, Salmonella Enteritidis, and Salmonella Orion were the most commonly isolated serovars from chicken nuggets and strips, nugget and strip meat, and pelleted broiler feeds, respectively. Salmonella Enteritidis phage type (PT) 13a with PFGE pattern SENXAI.0006 and R-type sensitive as well as Salmonella Enteritidis PT13a with PFGE pattern SENXAI.0068 and R-type sensitive were isolated from pelleted feed, and chicken nugget and strip meat in two separate instances. Data showed that Salmonella strains isolated from broiler feed were indistinguishable from strains isolated from packaged raw, frozen chicken nuggets and strips. However, results did not rule out the possibility that breeding stock or contamination during processing may have contributed to chicken meat contamination by Salmonella.

  19. Inactivation of Salmonella on Eggshells by Chlorine Dioxide Gas

    PubMed Central

    Yum, Bora; Yoon, Sung-Sik; Song, Kyoung-Ju; Kim, Jong-Rak

    2016-01-01

    Microbiological contamination of eggs should be prevented in the poultry industry, as poultry is one of the major reservoirs of human Salmonella. ClO2 gas has been reported to be an effective disinfectant in various industry fields, particularly the food industry. The aims of this study were to evaluate the antimicrobial effect of chlorine dioxide gas on two strains of Salmonella inoculated onto eggshells under various experimental conditions including concentrations, contact time, humidity, and percentage organic matter. As a result, it was shown that chlorine dioxide gas under wet conditions was more effective in inactivating Salmonella Enteritidis and Salmonella Gallinarum compared to that under dry conditions independently of the presence of organic matter (yeast extract). Under wet conditions, a greater than 4 log reduction in bacterial populations was achieved after 30 min of exposure to ClO2 each at 20 ppm, 40 ppm, and 80 ppm against S. Enteritidis; 40 ppm and 80 ppm against S. Gallinarum. These results suggest that chlorine dioxide gas is an effective agent for controlling Salmonella, the most prevalent contaminant in the egg industry. PMID:27499670

  20. Quantitative Tracking of Salmonella Enteritidis Transmission Routes Using Barcode-Tagged Isogenic Strains in Chickens: Proof-of-Concept Study

    PubMed Central

    Yang, Yichao; Ricke, Steven C.; Tellez, Guillermo; Kwon, Young Min

    2017-01-01

    Salmonella is an important foodborne bacterial pathogen, however, a fundamental understanding on Salmonella transmission routes within a poultry flock remains unclear. In this study, a series of barcode-tagged strains were constructed by inserting six random nucleotides into a functionally neutral region on the chromosome of S. Enteritidis as a tool for quantitative tracking of Salmonella transmission in chickens. Six distinct barcode-tagged strains were used for infection or contamination at either low dose (103 CFUs; three strains) or high dose (105 CFUs; three strains) in three independent experiments (Experiment 1 oral gavage; Experiment 2 contaminated feed; Experiment 3 contaminated water). For all chick experiments, cecal and foot-wash samples were collected from a subset of the chickens at days 7 or/and 14, from which genomic DNA was extracted and used to amplify the barcode regions. After the resulting PCR amplicons were pooled and analyzed by MiSeq sequencing, a total of approximately 1.5 million reads containing the barcode sequences were analyzed to determine the relative frequency of every barcode-tagged strain in each sample. In Experiment 1, the high dose of oral infection was correlated with greater dominance of the strains in the ceca of the respective seeder chickens and also in the contact chickens yet at lesser degrees. When chicks were exposed to contaminated feed (Experiment 2) or water (Experiment 3), there were no clear patterns of the barcode-tagged strains in relation to the dosage, except that the strains introduced at low dose required a longer time to colonize the ceca with contaminated feed. Most foot-wash samples contained only one to three strains for the majority of the samples, suggesting potential existence of an unknown mechanism(s) for strain exclusion. These results demonstrated the proof of concept of using barcode tagged to investigate transmission dynamics of Salmonella in chickens in a quantitative manner. PMID:28261587

  1. Bactericidal effects of negative air ions on airborne and surface Salmonella enteritidis from an artificially generated aerosol.

    PubMed

    Seo, K H; Mitchell, B W; Holt, P S; Gast, R K

    2001-01-01

    The bactericidal effect of high levels of negative ions was studied using a custom-built electrostatic space charge device. To investigate whether the ion-enriched air exerted a bactericidal effect, an aerosol containing Salmonella Enteritidis (SE) was pumped into a sealed plastic chamber. Plates of XLT4 agar were attached to the walls, top, and bottom of the chamber and exposed to the aerosol for 3 h with and without the ionizer treatment. The plates were then removed from the chamber, incubated at 37 degrees C for 24 h, and colonies were counted. An average of greater than 10(3) CFU/plate were observed on plates exposed to the aerosol without the ionizer treatment (control) compared with an average of less than 53 CFU/plate on the ionizer-treated plates. In another series of experiments, the SE aerosol was pumped for 3 h into an empty chamber containing only the ionizer and allowed to collect on the internal surfaces. The inside surfaces of the chamber were then rinsed with 100 ml phosphate-buffered saline that was then plated onto XLT4 plates. While the rinse from the control chamber contained colony counts greater than 400 CFU/ml of wash, no colonies were found in the rinse from the ionizer-treatment chamber. These results indicate that high levels of negative air ions can have a significant impact on the airborne microbial load, and that most of this effect is through direct killing of the organisms. This technology, which also causes significant reduction in airborne dust, has already been successfully applied for poultry hatching cabinets and caged layer rooms. Other potential applications include any enclosed space such as food processing areas, medical institutions, the workplace, and the home, where reduction of airborne and surface pathogens is desired.

  2. Effect of gamma irradiation on commercial eggs experimentally inoculated with Salmonella enteritidis

    NASA Astrophysics Data System (ADS)

    Tellez, I. G.; Trejo, R. M.; Sanchez, R. E.; Ceniceros, R. M.; Luna, Q. P.; Zazua, P.; Hargis, B. M.

    1995-02-01

    Using intact, fresh shell eggs, inoculated with 10 8 colony-forming units (cfu) of S. enteritidis, the effect of three doses of gamma irradiation on bacteriologic population and physical characteristics (Haugh units and yolk color) of the eggs was determinated. Penetration test area was picked at random just off the air cell of each egg. Aluminum cylinders were attached to the egg surface with a rim of molten paraffin, and 10 8S. enteritidis was then applied to inoculate the egg. Eggs were then irradiated within 2 hours using a Cobalt-60 gamma source at either 1, 2, or 3 kGy. A second set of inoculated, non-irradiated was used as controls. Following irradiation, eggs were maintained at 4°C for 42 hours prior culture. Irradiation with 1 kGy resulted in a significant (P < .05), 3.9 log reduction in detectable S. enteritidis in the shell and a higly significant (P < .025) 95% reduction in detectable S. enteritidis in the internal shell membranes. Irradiation of eggs with either 2 or 3 kGy reduced bacterial contamination to non-detectable levels in both the shell and internal membranes. However, irradiation at either 1, 2 or 3 kGy resulted in a significant (P <- .05) decrease (approximately 50%) in Haugh units. Additionally, irradiation of intact shell eggs at 2 or 3 Kgy significantly (P ≤ .05) reduced yolk color regardless of the level of irradiation exposure implemented. This data indicates that gamma irradiation of intact raw eggs is effective in reducing (1 kGy) or eliminating (2 or more kGy) S. enteritidis contamination. However, each of the levels of irradiation used in the present experiments caused marked reduction of selected measures of egg quality.

  3. Detection of Salmonella sp in chicken cuts using immunomagnetic separation

    PubMed Central

    de Cássia dos Santos da Conceição, Rita; Moreira, Ângela Nunes; Ramos, Roberta Juliano; Goularte, Fabiana Lemos; Carvalhal, José Beiro; Aleixo, José Antonio Guimarães

    2008-01-01

    The immunomagnetic separation (IMS) is a technique that has been used to increase sensitivity and specificity and to decrease the time required for detection of Salmonella in foods through different methodologies. In this work we report on the development of a method for detection of Salmonella in chicken cuts using in house antibody-sensitized microspheres associated to conventional plating in selective agar (IMS-plating). First, protein A-coated microspheres were sensitized with polyclonal antibodies against lipopolysacharide and flagella from salmonellae and used to standardize a procedure for capturing Salmonella Enteritidis from pure cultures and detection in selective agar. Subsequently, samples of chicken meat experimentally contaminated with S. Enteritidis were analyzed immediately after contamination and after 24h of refrigeration using three enrichment protocols. The detection limit of the IMS-plating procedure after standardization with pure culture was about 2x10 CFU/mL. The protocol using non-selective enrichment for 6-8h, selective enrichment for 16-18h and a post-enrichment for 4h gave the best results of S. Enteritidis detection by IMS-plating in experimentally contaminated meat. IMS-plating using this protocol was compared to the standard culture method for salmonellae detection in naturally contaminated chicken cuts and yielded 100% sensitivity and 94% specificity. The method developed using in house prepared magnetic microespheres for IMS and plating in selective agar was able to diminish by at least one day the time required for detection of Salmonella in chicken products by the conventional culture method. PMID:24031199

  4. Induction of Viable but Nonculturable Salmonella in Exponentially Grown Cells by Exposure to a Low-Humidity Environment and Their Resuscitation by Catalase.

    PubMed

    Morishige, Yuta; Koike, Atsushi; Tamura-Ueyama, Ai; Amano, Fumio

    2017-02-01

    Salmonella is a major cause of foodborne disease that sometimes occurs in massive outbreaks around the world. This pathogen is tolerant of low-humidity conditions. We previously described a method for induction of viable but nonculturable (VBNC) Salmonella enterica serovar Enteritidis by treatment with hydrogen peroxide (H 2 O 2 ) and subsequent resuscitation with 0.3 mM sodium pyruvate. Here, we report a new method for the induction of the VBNC state in Salmonella Enteritidis cells, one involving dehydration. Exposure of Salmonella Enteritidis cells to dehydration stress under poor nutritional conditions (0.9% [wt/vol] NaCl) and 10 to 20% relative humidity at room temperature decreased the presence of culturable population to 0.0067%, but respiratory and glucose uptake active populations were maintained at 0.46 and 1.12%, respectively, meaning that approximately 1% may have entered the VBNC state. Furthermore, these VBNC cells could be resuscitated to acquire culturability by incubation with catalase in M9 minimal medium without glucose in a manner dependent on the dose of catalase but not sodium pyruvate. These results suggest that a low-humidity environment could cause Salmonella Enteritidis cells to enter the VBNC state and the cells could then be resuscitated for growth by treatment with catalase, suggesting a potential risk of Salmonella Enteritidis to survive in low water activity foods in the VBNC state and to start regrowth for foodborne illness.

  5. A multi-country Salmonella Enteritidis phage type 14b outbreak associated with eggs from a German producer: 'near real-time' application of whole genome sequencing and food chain investigations, United Kingdom, May to September 2014.

    PubMed

    Inns, T; Lane, C; Peters, T; Dallman, T; Chatt, C; McFarland, N; Crook, P; Bishop, T; Edge, J; Hawker, J; Elson, R; Neal, K; Adak, G K; Cleary, P

    2015-04-23

    We report an outbreak of Salmonella Enteritidis phage type 14b (PT14b) in the United Kingdom (UK) between May and September 2014 where Public Health England launched an investigation to identify the source of infection and implement control measures. During the same period, outbreaks caused by a Salmonella Enteritidis strain with a specific multilocus variable-number tandem repeat analysis (MLVA) profile occurred in other European Union Member States. Isolates from a number of persons affected by the UK outbreak, who had initially been tested by MLVA also shared this particular profile. Cases were defined as any person infected with S. Enteritidis PT14b, resident in England or Wales and without history of travel outside of this geographical area during the incubation period, reported from 1 June 2014 onwards, with a MLVA profile of 2–11–9-7–4-3–2-8–9 or a single locus variant thereof. In total, 287 cases met the definition. Food traceback investigations in the UK and other affected European countries linked the outbreaks to chicken eggs from a German company. We undertook whole genome sequencing of isolates from UK and European cases, implicated UK premises, and German eggs: isolates were highly similar. Combined with food traceback information, this confirmed that the UK outbreak was also linked to a German producer.

  6. Differentiation of Salmonella enteritidis phage type 8 strains: evaluation of three additional phage typing systems, plasmid profiles, antibiotic susceptibility patterns, and biotyping.

    PubMed Central

    Stubbs, A D; Hickman-Brenner, F W; Cameron, D N; Farmer, J J

    1994-01-01

    Three additional phage typing systems for Salmonella enteritidis, plasmid analysis, biochemical tests, and antimicrobial susceptibility tests, were used in an attempt to subdivide 30 phage type 8 (phage typing system used by the WHO International Center for Enteric Phage Typing, London, England) isolates. These isolates represented 18 different egg-related outbreaks (21 strains) and 9 reference strains or strains that were not egg-associated. Only 7 of the 30 strains (28%) were subdivided by one or more of the methods used; this included 3 of the 21 strains from egg-related outbreaks. Twenty-seven strains contained a 55-kb plasmid that is associated with S. enteritidis. Of 65 additional phages tested, 2 from the phage typing system obtained from the Pasteur Institute, Paris, France, were useful in differentiating the three strains that lacked the 55-kb plasmid. Although the results obtained for the 21 strains from egg-related outbreaks showed that the strains had minor phenotypic differences, the overall results suggested that the strains may represent a single clone. Studies are planned to test additional phages and other typing methods to see whether strains of phage type 8 can be further differentiated. PMID:8126179

  7. Molecular typing, antibiotic resistance, virulence gene and biofilm formation of different Salmonella enterica serotypes.

    PubMed

    Turki, Yousra; Mehr, Ines; Ouzari, Hadda; Khessairi, Amel; Hassen, Abdennaceur

    2014-01-01

    Salmonella enterica isolates representing commonly isolated serotypes in Tunisia were analyzed using genotyping and phenotyping methods. ERIC and ITS-PCR applied to 48 Salmonella spp. isolates revealed the presence of 12 and 10 different profiles, respectively. The distribution of profiles among serotypes demonstrated the presence of strains showing an identical fingerprinting pattern. All Salmonella strains used in this study were positive for the sdiA gene. Three Salmonella isolates belonging to serotypes Anatum, Enteritidis and Amsterdam were negative for the invA gene. The spvC gene was detected in thirteen isolates belonging to serotypes Anatum, Typhimurium, Enteritidis, Gallinarum and Montevideo. Antibiotic resistance was frequent among the recovered Salmonella isolates belonging to serotypes Anatum, Typhimurium, Enteritidis, Zanzibar and Derby. The majority of these isolates exhibited resistance to at least two antibiotic families. Four multidrug-resistant isolates were recovered from food animals and poultry products. These isolates exhibited not only resistance to tetracycline, sulphonamides, and ampicillin, but also have shown resistance to fluoroquinolones. Common resistance to nalidixic acid, ciprofloxacin and ofloxacin in two S. Anatum and S. Zanzibar strains isolated from raw meat and poultry was also obtained. Furthermore, wastewater and human isolates exhibited frequent resistance to nalidixic acid and tetracycline. Of all isolates, 33.5% were able to form biofilm.

  8. phoP, SPI1, SPI2 and aroA mutants of Salmonella Enteritidis induce a different immune response in chickens.

    PubMed

    Elsheimer-Matulova, Marta; Varmuzova, Karolina; Kyrova, Kamila; Havlickova, Hana; Sisak, Frantisek; Rahman, Masudur; Rychlik, Ivan

    2015-09-17

    Poultry is the most frequent reservoir of non-typhoid Salmonella enterica for humans. Understanding the interactions between chickens and S. enterica is therefore important for vaccine design and subsequent decrease in the incidence of human salmonellosis. In this study we therefore characterized the interactions between chickens and phoP, aroA, SPI1 and SPI2 mutants of S. Enteritidis. First we tested the response of HD11 chicken macrophage-like cell line to S. Enteritidis infection monitoring the transcription of 36 genes related to immune response. All the mutants and the wild type strain induced inflammatory signaling in the HD11 cell line though the response to SPI1 mutant infection was different from the rest of the mutants. When newly hatched chickens were inoculated, the phoP as well as the SPI1 mutant did not induce an expression of any of the tested genes in the cecum. Despite this, such chickens were protected against challenge with wild-type S. Enteritidis. On the other hand, inoculation of chickens with the aroA or SPI2 mutant induced expression of 27 and 18 genes, respectively, including genes encoding immunoglobulins. Challenge of chickens inoculated with these two mutants resulted in repeated induction of 11 and 13 tested genes, respectively, including the genes encoding immunoglobulins. In conclusion, SPI1 and phoP mutants induced protective immunity without inducing an inflammatory response and antibody production. Inoculation of chickens with the SPI2 and aroA mutants also led to protective immunity but was associated with inflammation and antibody production. The differences in interaction between the mutants and chicken host can be used for a more detailed understanding of the chicken immune system.

  9. Microbiological, clinical and molecular findings of non-typhoidal Salmonella bloodstream infections associated with malaria, Oriental Province, Democratic Republic of the Congo.

    PubMed

    Falay, Dadi; Kuijpers, Laura Maria Francisca; Phoba, Marie-France; De Boeck, Hilde; Lunguya, Octavie; Vakaniaki, Emmanuel; Bertrand, Sophie; Mattheus, Wesley; Ceyssens, Pieter-Jan; Vanhoof, Raymond; Devlieger, Hugo; Van Geet, Chris; Verheyen, Erik; Ngbonda, Dauly; Jacobs, Jan

    2016-06-10

    In sub-Saharan Africa, non-typhoidal Salmonella (NTS) can cause bloodstream infections, referred to as invasive non-typhoidal Salmonella disease (iNTS disease); it can occur in outbreaks and is often preceded by malaria. Data from Central Africa is limited. Clinical, microbiological and molecular findings of NTS recovered in a blood culture surveillance project (2009-2014) were analyzed. In March-July 2012 there was an epidemic increase in malaria infections in the Oriental Province of the Democratic Republic of the Congo (DRC). In one referral hospital, overall hospital admissions in June 2012 were 2.6 times higher as compared to the same period in the years before and after (336 versus an average of 128 respectively); numbers of malaria cases and blood transfusions were nearly three- and five-fold higher respectively (317 versus 112 and 250 versus 55). Case fatality rates (in-hospital deaths versus all admissions) peaked at 14.6 %. Salmonella Typhimurium and Salmonella Enteritidis together accounted for 88.9 % of pathogens isolated from blood cultures collected during an outreach visit to the affected districts in June 2012. Children infected with Salmonella Enteritidis (33 patient files available) tended to be co-infected with Plasmodium falciparum more often than children infected with Salmonella Typhimurium (40 patients files available) (81.8 % versus 62.5 %). Through the microbiological surveillance project (May 2009-May 2014) 113 unique NTS isolates were collected (28.5 % (113/396) of pathogens); most (95.3 %) were recovered from children < 15 years. Salmonella Typhimurium (n = 54) and Salmonella Enteritidis (n = 56) accounted for 47.8 % and of 49.6 % NTS isolates respectively. Multilocus variable-number tandem-repeat analysis (MLVA) revealed more heterogeneity for Salmonella Typhimurium than for Salmonella Enteritidis. Most (82/96, 85.4 %) NTS isolates that were available for antibiotic susceptibility testing were multidrug resistant

  10. Salmonella and raw shell eggs: results of a cross-sectional study of contamination rates and egg safety practices in the United Kingdom catering sector in 2003.

    PubMed

    Elson, R; Little, C L; Mitchell, R T

    2005-02-01

    This study was prompted by epidemiological investigations of the unusual number of Salmonella Enteritidis outbreaks associated with the use of eggs in catering premises in England and Wales during 2002. The aims of the study, performed between April and May 2003, were to establish the rate of Salmonella contamination in raw shell eggs from catering premises, investigate any correlation between the origin and type of eggs and the presence of particular serotypes or phage types (PTs) of Salmonella, and examine the use of raw shell eggs in catering premises in the United Kingdom. A total of 34,116 eggs (5,686 pooled samples of six eggs) were collected from 2,104 catering premises, most of which were eggs produced in the United Kingdom (88%). Salmonella was isolated from 17 pools (0.3%) of eggs. Of these, 15 were Salmonella Enteritidis, which were further characterized to PTs as follows: PT6 (0.1%), PT4 (0.07%), PT12 (0.04%), PT1 (0.04%), and PT14b (0.02%). Salmonella Livingstone and Salmonella Typhimurium definitive type 7 resistant to ampicillin, streptomycin, sulfonamides, and tetracycline were also isolated. The Salmonella contamination rate of eggs produced in the United Kingdom appears to have decreased significantly since 1995 and 1996. This trend is reflected in the decrease of Salmonella Enteritidis and, in particular, Salmonella Enteritidis PT4. The impact of the United Kingdom Food Standards Agency's advice on the use of eggs, issued in January 2003, is discussed.

  11. Immunization with Salmonella Enteritidis secreting mucosal adjuvant labile toxin confers protection against wild type challenge via augmentation of CD3+CD4+ T-cell proliferation and enhancement of IFN-γ, IL-6 and IL-10 expressions in chicken.

    PubMed

    Lalsiamthara, Jonathan; Lee, John Hwa

    2017-02-01

    The protective efficacy and immunological profiles of chickens immunized with an attenuated Salmonella Enteritidis (SE) constitutively secreting double mutant heat labile enterotoxin (dmLT) were investigated. The dmLT is a detoxified variant of Escherichia coli heat labile toxin and is a potent mucosal adjuvant capable of inducing both humoral and cell-mediated immunity. In this study, four-week-old chickens were inoculated with SE-dmLT strain JOL1641, parental SE strain JOL1087 or phosphate buffered saline control. Peripheral blood mononuclear cells of SE-dmLT inoculated birds showed significant proliferation upon stimulation with SE antigens as compared to the control and JOL1087 groups (P⩽0.05). One week post-challenge, the ratio of CD3 + CD4 + to CD3 + CD8 + T-cells showed a significant increase in the immunized groups. Significant increases in IFN-γ levels were observed in JOL1641 birds immunized via oral and intramuscular routes. While immunizations with the JOL1087 strain via the intramuscular route also induced significant increases in IFN-γ, immunization via the oral route did not trigger significant changes. Pro-inflammatory cytokine IL-6 was also elevated significantly in immunized birds; a significant elevation of IL-10 was observed only in oral immunization with JOL1641 (P⩽0.05). JOL1641 immunized birds showed significant reduction of challenge bacterial-organ recovery as compared to JOL1087 and non-immunized birds. Collectively, our results revealed that immunization with the adjuvant-secreting S. Enteritidis confers protection against wild type SE challenge via induction of strong cell proliferative response, augmentation of CD3 + CD4 + : CD3 + CD8 + T-cells ratio and enhancement of IFN-γ, IL-6 and IL-10 cytokine secretion. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Mathematical modeling of growth Salmonella and spoilage microorganisms in raw oysters

    USDA-ARS?s Scientific Manuscript database

    The main objective of this study was to develop primary and secondary models to describe the growth of Salmonella as well as background microorganisms in fresh shucked oysters. The cocktail of two Salmonella serotypes, S. Typhimurium (CICC22956) and S. Enteritidis (CICC21482), was inoculated to raw...

  13. Differential antibacterial response of chicken granulosa cells to invasion by Salmonella serovars.

    PubMed

    Babu, Uma S; Harrison, Lisa M; Patel, Isha R; Ramirez, Gerardo A; Williams, Kristina M; Pereira, Marion; Balan, Kannan V

    2016-06-01

    In the United States, Salmonella enterica ser. Enteritidis (SE) is among the leading bacterial cause of foodborne illness via consumption of raw or undercooked eggs. The top Salmonella serovars implicated in U.S. foodborne outbreaks associated with chicken consumption include SE, Typhimurium (ST), Heidelberg (SH), Montevideo, Mbandka, Braenderup, and Newport. While enforcement actions target the eradication of SE from layer hens, there is a growing concern that other serovars could occupy this niche and be a cause of egg-transmitted human salmonellosis. Therefore, we tested the invasion and survival of SE, SH, ST, and Salmonella enterica ser. Hadar (S. Hadar) at 4 and 20 h post infection (hpi) in chicken ovarian granulosa cells (cGC); a cellular layer which surrounds the previtelline layer and central yolk in egg-forming follicles. We also evaluated cGC transcriptional changes, using an antibacterial response PCR array, to assess host response to intracellular SalmonellaWe observed that invasion of cGC by SE, SH, and ST was significantly higher than invasion by S. Hadar, with ST showing the highest level of invasion. The Bacterial Survival Index, defined as the ratio of intracellular bacteria at 20 and 4 h, were 18.94, 7.35, and 15.27 for SE, SH, and ST, respectively, with no significant difference in survival between SE or ST compared to SH. Evaluation of cGC anti-Salmonella gene responses indicated that at 4 hpi there was a significant decrease in Toll-like receptor (TLR)-4 mRNA in cGC infected with SE, whereas TLR5 and myeloid differentiation primary response gene 88 were significantly down regulated across all serovars. At 4 hpi, invasion by Salmonella serovars resulted in significant upregulation of several antimicrobial genes, and proinflammatory cytokines and chemokines (PICs). At 20 hpi, all the serovars induced PICs with SH being the strongest inducer. Additionally, SE, SH and ST differentially induced signal transduction pathways. Although only a single

  14. Evaluating risk factors for endemic human Salmonella Enteritidis infections with different phage types in Ontario, Canada using multinomial logistic regression and a case-case study approach

    PubMed Central

    2012-01-01

    Background Identifying risk factors for Salmonella Enteritidis (SE) infections in Ontario will assist public health authorities to design effective control and prevention programs to reduce the burden of SE infections. Our research objective was to identify risk factors for acquiring SE infections with various phage types (PT) in Ontario, Canada. We hypothesized that certain PTs (e.g., PT8 and PT13a) have specific risk factors for infection. Methods Our study included endemic SE cases with various PTs whose isolates were submitted to the Public Health Laboratory-Toronto from January 20th to August 12th, 2011. Cases were interviewed using a standardized questionnaire that included questions pertaining to demographics, travel history, clinical symptoms, contact with animals, and food exposures. A multinomial logistic regression method using the Generalized Linear Latent and Mixed Model procedure and a case-case study design were used to identify risk factors for acquiring SE infections with various PTs in Ontario, Canada. In the multinomial logistic regression model, the outcome variable had three categories representing human infections caused by SE PT8, PT13a, and all other SE PTs (i.e., non-PT8/non-PT13a) as a referent category to which the other two categories were compared. Results In the multivariable model, SE PT8 was positively associated with contact with dogs (OR=2.17, 95% CI 1.01-4.68) and negatively associated with pepper consumption (OR=0.35, 95% CI 0.13-0.94), after adjusting for age categories and gender, and using exposure periods and health regions as random effects to account for clustering. Conclusions Our study findings offer interesting hypotheses about the role of phage type-specific risk factors. Multinomial logistic regression analysis and the case-case study approach are novel methodologies to evaluate associations among SE infections with different PTs and various risk factors. PMID:23057531

  15. Estimation of the rate of egg contamination from Salmonella-infected chickens.

    PubMed

    Arnold, M E; Martelli, F; McLaren, I; Davies, R H

    2014-02-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the most prevalent causes for human gastroenteritis and is by far the predominant Salmonella serovar among human cases, followed by Salmonella Typhimurium. Contaminated eggs produced by infected laying hens are thought to be the main source of human infection with S. Enteritidis throughout the world. Although previous studies have looked at the proportion of infected eggs from infected flocks, there is still uncertainty over the rate at which infected birds produce contaminated eggs. The aim of this study was to estimate the rate at which infected birds produce contaminated egg shells and egg contents. Data were collected from two studies, consisting of 15 and 20 flocks, respectively. Faecal and environmental sampling and testing of ovaries/caeca from laying hens were carried out in parallel with (i) for the first study, testing 300 individual eggs, contents and shells together and (ii) for the second study, testing 4000 eggs in pools of six, with shells and contents tested separately. Bayesian methods were used to estimate the within-flock prevalence of infection from the faecal and hen post-mortem data, and this was related to the proportion of positive eggs. Results indicated a linear relationship between the rate of contamination of egg contents and the prevalence of infected chickens, but a nonlinear (quadratic) relationship between infection prevalence and the rate of egg shell contamination, with egg shell contamination occurring at a much higher rate than that of egg contents. There was also a significant difference in the rate of egg contamination between serovars, with S. Enteritidis causing a higher rate of contamination of egg contents and a lower rate of contamination of egg shells compared to non-S. Enteritidis serovars. These results will be useful for risk assessments of human exposure to Salmonella-contaminated eggs. © 2013 Crown copyright. This article is published with the

  16. An outbreak of Salmonella Enteritidis phage type 34a infection associated with a Chinese restaurant in Suffolk, United Kingdom

    PubMed Central

    Badrinath, Padmanabhan; Sundkvist, Torbjorn; Mahgoub, Hamid; Kent, Richard

    2004-01-01

    Background On 30th July 2002, the Suffolk Communicable Disease Control Team received notifications of gastrointestinal illness due to Salmonella Enteritidis in subjects who had eaten food from a Chinese restaurant on 27th July. An Outbreak Control Team was formed resulting in extensive epidemiological, microbiological and environmental investigations. Methods Attempts were made to contact everybody who ate food from the restaurant on 27th July and a standard case definition was adopted. Using a pre-designed proforma information was gathered from both sick and well subjects. Food specific attack rates were calculated and two-tailed Fisher's exact test was used to test the difference between type of food consumed and the health status. Using a retrospective cohort design univariate Relative Risks and 95% Confidence Intervals were calculated for specific food items. Results Data was gathered on 52 people of whom 38 developed gastrointestinal symptoms; 16 male and 22 female. The mean age was 27 years. The mean incubation period was 30 hours with a range of 6 to 90 hours. Food attack rates were significantly higher for egg, special and chicken fried rice. Relative risk and the Confidence interval for these food items were 1.97 (1.11–3.48), 1.56 (1.23–1.97) and 1.48 (1.20–1.83) respectively. Interviews with the chef revealed that many eggs were used in the preparation of egg-fried rice, which was left at room temperature for seven hours and was used in the preparation of the other two rice dishes. Of the 31 submitted stool specimens 28 tested positive for S Enteritidis phage type 34a and one for S Enteritidis phage type 4. Conclusion In the absence of left over food available for microbiological examination, epidemiological investigation strongly suggested the eggs used in the preparation of the egg-fried rice as the vehicle for this outbreak. This investigation highlights the importance of safe practices in cooking and handling of eggs in restaurants. PMID

  17. [Study on simultaneous contamination of Salmonella and Campylobacter in retail chicken carcasses in Beijing].

    PubMed

    Hu, Yujie; Wang, Yeru; Li, Fengqin

    2015-01-01

    To elucidate the simultaneous contamination of Salmonella and Campylobacter in retail chicken carcasses in Beijing and to carry out the serological typing of all Salmonella isolates as well as the identification of Campylobacter at the species level. A total of 33 chicken carcasses were collected from Beijing supermarkets and farm's trade markets from May to July. All samples were enumerated for Salmonella and Campylobacter. All Salmonella isolates obtained were further serotyped and Campylobacter were identified at the species level. Totally, 19 samples (19/33, 57.6%) and 5 samples (5/33, 15.2%) were positive for Salmonella with the mean level of 119.4 MPN/100g and Campylobacter with the mean level of 58.6 CFU/g, respectively. In terms of Salmonella, 166 isolates with 14 serotypes were obtained. Salmonella Enteritidis was the most common serovar detected followed by S. Indiana. Serovar diversity was very high in all Salmonella isolates and various Salmonella serovars were detected in the same chicken carcass. A total of 11 serovar distribution spectrums were found and S. Enteritidis in combination with S. Indiana was the predominant. The retail chicken carcasses in Beijing collected from May to July were heavily contaminated by Salmonella with high serovar diversity.

  18. Salmonella spp. on chicken carcasses in processing plants in Poland.

    PubMed

    Mikołajczyk, Anita; Radkowski, Mieczysław

    2002-09-01

    Chickens at selected points in the slaughter process and after slaughter on the dressing line in poultry plants were sampled and analyzed for Salmonella. These chickens came from the northeast part of Poland. The examinations were carried out in quarters I, II, III, and IV of 1999. All the birds were determined to be healthy by a veterinary inspection. Swab samples were taken from the cloaca after stunning and from the skin surface and body cavity of the whole bird after evisceration, after rinsing at the final rinse station but before chilling in the spin-chiller, and after cooling in the continuous cooling plant at the end of the production day. In 1999, 400 whole chickens were examined. The percentage of these 400 chickens from which Salmonella spp. were isolated was relatively high (23.75%; Salmonella-positive results were observed in 95 cases). Salmonella spp. were found after stunning in 6% of the chickens (6 of 100 samples), after evisceration in 24% (24 of 100), before cooling in 52% (52 of 100), and after cooling in 13% (13 of 100). These results show that Salmonella spp. were found more often at some processing points than at others. The lowest Salmonella spp. contamination rate (6%) for slaughter birds was found after stunning, and the highest contamination rate was found before chilling (52%). The serological types of Salmonella spp. isolated from whole chickens were Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Saintpaul, Salmonella Agona, and Salmonella Infantis. The results of these investigations indicate that Salmonella Enteritidis is the dominant serological type in infections of slaughter chickens, as it is in many countries.

  19. Salmonella meningoencephalomyelitis in a northern fur seal (Callorhinus ursinsus)

    USGS Publications Warehouse

    Stroud, R.K.; Roelke, M.E.

    1980-01-01

    Salmonella enteritidis was isolated from the brain of a neonatal northern fur seal (Callorhinus ursinus) with gross and microscopic lesions of meningoencephalomyelitis. Microscopic lesions in the liver and lung suggested septicemia.

  20. Assessment of 2 Salmonella enterica serovar Typhimurium-based vaccines against necrotic enteritis in reducing colonization of chickens by Salmonella serovars of different serogroups.

    PubMed

    Jiang, Yanfen; Kulkarni, Raveendra R; Parreira, Valeria R; Poppe, Cornelius; Roland, Kenneth L; Prescott, John F

    2010-10-01

    This study assessed the protective efficacy of oral vaccination with 2 experimental attenuated Salmonella Typhimurium-vectored vaccines for necrotic enteritis in protecting chickens against intestinal colonization by common serovars of Salmonella belonging to the 4 major serogroups affecting chickens. Birds were vaccinated orally with 1 × 10⁸ colony-forming units (CFU) of 1 of the vaccine strains χ9241 and χ9352, which express a plasmid-encoded partial recombinant hypothetical protein gene (tHP) of Clostridium perfringens, at days 1 and 7 of age, and then were challenged at 14 d of age with 10⁶ CFU of Salmonella serovars Anatum, Enteritidis, Heidelberg, Kentucky, or Typhimurium (representative serovars of serogroups B, C, D, and E). Birds were necropsied at 4 wk of age, and samples were collected to determine reduction in tissue and intestinal colonization. The chickens vaccinated with χ9241-tHP showed reduced colonization by Salmonella Enteritidis (serogroup D) and by Salmonella Heidelberg and Salmonella Typhimurium (serogroup B) compared with the control birds. No reduction in colonization was observed in the chickens vaccinated with χ9352-tHP. There was an association between the efficacy of these vaccine strains in protecting against necrotic enteritis, assessed on an earlier occasion, and their efficacy in protecting against Salmonella colonization. Thus, the choice of an attenuated Salmonella Typhimurium vaccine vector for delivery of heterologous antigens to chickens should be based partly on the vaccine's value in protecting against colonization by serovars within serogroups B and D. Such vectors would have the additional benefit of reducing colonization of important Salmonella serovars.

  1. Assessment of 2 Salmonella enterica serovar Typhimurium-based vaccines against necrotic enteritis in reducing colonization of chickens by Salmonella serovars of different serogroups

    PubMed Central

    Jiang, Yanfen; Kulkarni, Raveendra R.; Parreira, Valeria R.; Poppe, Cornelius; Roland, Kenneth L.; Prescott, John F.

    2010-01-01

    This study assessed the protective efficacy of oral vaccination with 2 experimental attenuated Salmonella Typhimurium-vectored vaccines for necrotic enteritis in protecting chickens against intestinal colonization by common serovars of Salmonella belonging to the 4 major serogroups affecting chickens. Birds were vaccinated orally with 1 × 108 colony-forming units (CFU) of 1 of the vaccine strains χ9241 and χ9352, which express a plasmid-encoded partial recombinant hypothetical protein gene (tHP) of Clostridium perfringens, at days 1 and 7 of age, and then were challenged at 14 d of age with 106 CFU of Salmonella serovars Anatum, Enteritidis, Heidelberg, Kentucky, or Typhimurium (representative serovars of serogroups B, C, D, and E). Birds were necropsied at 4 wk of age, and samples were collected to determine reduction in tissue and intestinal colonization. The chickens vaccinated with χ9241-tHP showed reduced colonization by Salmonella Enteritidis (serogroup D) and by Salmonella Heidelberg and Salmonella Typhimurium (serogroup B) compared with the control birds. No reduction in colonization was observed in the chickens vaccinated with χ9352-tHP. There was an association between the efficacy of these vaccine strains in protecting against necrotic enteritis, assessed on an earlier occasion, and their efficacy in protecting against Salmonella colonization. Thus, the choice of an attenuated Salmonella Typhimurium vaccine vector for delivery of heterologous antigens to chickens should be based partly on the vaccine’s value in protecting against colonization by serovars within serogroups B and D. Such vectors would have the additional benefit of reducing colonization of important Salmonella serovars. PMID:21197226

  2. Detection of cell surface hydrophobicity, biofilm and fimbirae genes in salmonella isolated from tunisian clinical and poultry meat.

    PubMed

    Ben Abdallah, Fethi; Lagha, Rihab; Said, Khaled; Kallel, Héla; Gharbi, Jawhar

    2014-04-01

    The aim of this study was to evaluate the ability of 15 serotypes of Salmonella to form biofilm on polystyrene, polyvinyl chloride (PVC) and glass surfaces. . Initially slime production was assessed on CRA agar and hydrophobicity of 20 Salmonella strains isolated from poultry and human and two Salmonella enterica serovar Typhimurium references strains was achieved by microbial adhesion to n-hexadecane. In addition, biofilm formation on polystyrene, PVC and glass surfaces was also investigated by using MTT and XTT colorimetric assay. Further, distribution of Salmonella enterotoxin (stn), Salmonella Enteritidis fimbrial (sef) and plasmid encoded fimbrial (pef) genes among tested strains was achieved by PCR. Salmonella strains developed red and white colonies on CRA and they are considered as hydrophilic with affinity values to n-hexadecane ranged between 0.29% and 29.55%. Quantitative biofilm assays showed that bacteria are able to form biofilm on polystyrene with different degrees and 54.54% of strains produce a strong biofilm on glass. In addition, all the strains form only a moderate (54.54%) and weak (40.91%) biofilm on PVC. PCR detection showed that only S. Enteritidis harbour Sef gene, whereas Pef and stn genes were detected in S. Kentucky, S. Amsterdam, S. Hadar, S. Enteritidis and S. Typhimurium. Salmonella serotypes are able to form biofilm on hydrophobic and hydrophilic industrial surfaces. Biofilm formation of Salmonella on these surfaces has an increased potential to compromise food safety and potentiate public health risk.

  3. Detection of Cell Surface Hydrophobicity, Biofilm and Fimbirae Genes in Salmonella Isolated from Tunisian Clinical and Poultry Meat

    PubMed Central

    BEN ABDALLAH, Fethi; LAGHA, Rihab; SAID, Khaled; KALLEL, Héla; GHARBI, Jawhar

    2014-01-01

    Abstract Background The aim of this study was to evaluate the ability of 15 serotypes of Salmonella to form biofilm on polystyrene, polyvinyl chloride (PVC) and glass surfaces. . Methods Initially slime production was assessed on CRA agar and hydrophobicity of 20 Salmonella strains isolated from poultry and human and two Salmonella enterica serovar Typhimurium references strains was achieved by microbial adhesion to n-hexadecane. In addition, biofilm formation on polystyrene, PVC and glass surfaces was also investigated by using MTT and XTT colorimetric assay. Further, distribution of Salmonella enterotoxin (stn), Salmonella Enteritidis fimbrial (sef) and plasmid encoded fimbrial (pef) genes among tested strains was achieved by PCR. Results Salmonella strains developed red and white colonies on CRA and they are considered as hydrophilic with affinity values to n-hexadecane ranged between 0.29% and 29.55%. Quantitative biofilm assays showed that bacteria are able to form biofilm on polystyrene with different degrees and 54.54% of strains produce a strong biofilm on glass. In addition, all the strains form only a moderate (54.54%) and weak (40.91%) biofilm on PVC. PCR detection showed that only S. Enteritidis harbour Sef gene, whereas Pef and stn genes were detected in S. Kentucky, S. Amsterdam, S. Hadar, S. Enteritidis and S. Typhimurium. Conclusion Salmonella serotypes are able to form biofilm on hydrophobic and hydrophilic industrial surfaces. Biofilm formation of Salmonella on these surfaces has an increased potential to compromise food safety and potentiate public health risk. PMID:26005652

  4. Effectiveness of superheated steam for inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Enteritidis phage type 30, and Listeria monocytogenes on almonds and pistachios.

    PubMed

    Ban, Ga-Hee; Kang, Dong-Hyun

    2016-03-02

    This study was undertaken to evaluate the effectiveness of superheated steam (SHS) on the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, Salmonella Enteritidis phage type (PT) 30 and Listeria monocytogenes on almonds and in-shell pistachios and to determine the effect of superheated steam heating on quality by measuring color and texture changes. Almonds and in-shell pistachios inoculated with four foodborne pathogens were treated with saturated steam (SS) at 100 °C and SHS at 125, 150, 175, and 200 °C for various times. Exposure of almonds and pistachios to SHS for 15 or 30s at 200 °C achieved >5l og reductions among all tested pathogens without causing significant changes in color values or texture parameters (P>0.05). For both almonds and pistachios, acid and peroxide values (PV) following SS and SHS treatment for up to 15s and 30s, respectively, were within the acceptable range (PV<1.0 meq/kg). These results show that thermal application of 200 °C SHS treatment for 15s and 30s did not affect the quality of almonds and pistachios, respectively. Therefore, SHS treatment is a very promising alternative technology for the tree nuts industry by improving inactivation of foodborne pathogens on almonds and pistachios while simultaneously reducing processing time. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. The Early Innate Response of Chickens to Salmonella enterica Is Dependent on the Presence of O-Antigen but Not on Serovar Classification

    PubMed Central

    Varmuzova, Karolina; Matulova, Marta Elsheimer; Sebkova, Alena; Sekelova, Zuzana; Havlickova, Hana; Sisak, Frantisek; Babak, Vladimir; Rychlik, Ivan

    2014-01-01

    Salmonella vaccines used in poultry in the EU are based on attenuated strains of either Salmonella serovar Enteritidis or Typhimurium which results in a decrease in S. Enteritidis and S. Typhimurium but may allow other Salmonella serovars to fill an empty ecological niche. In this study we were therefore interested in the early interactions of chicken immune system with S. Infantis compared to S. Enteritidis and S. Typhimurium, and a role of O-antigen in these interactions. To reach this aim, we orally infected newly hatched chickens with 7 wild type strains of Salmonella serovars Enteritidis, Typhimurium and Infantis as well as with their rfaL mutants and characterized the early Salmonella-chicken interactions. Inflammation was characterized in the cecum 4 days post-infection by measuring expression of 43 different genes. All wild type strains stimulated a greater inflammatory response than any of the rfaL mutants. However, there were large differences in chicken responses to different wild type strains not reflecting their serovar classification. The initial interaction between newly-hatched chickens and Salmonella was found to be dependent on the presence of O-antigen but not on its structure, i.e. not on serovar classification. In addition, we observed that the expression of calbindin or aquaporin 8 in the cecum did not change if inflammatory gene expression remained within a 10 fold fluctuation, indicating the buffering capacity of the cecum, preserving normal gut functions even in the presence of minor inflammatory stimuli. PMID:24763249

  6. Salmonella induces prominent gene expression in the rat colon

    PubMed Central

    Rodenburg, Wendy; Keijer, Jaap; Kramer, Evelien; Roosing, Susanne; Vink, Carolien; Katan, Martijn B; van der Meer, Roelof; Bovee-Oudenhoven, Ingeborg MJ

    2007-01-01

    Background Salmonella enteritidis is suggested to translocate in the small intestine. In vivo it induces gene expression changes in the ileal mucosa and Peyer's patches. Stimulation of Salmonella translocation by dietary prebiotics fermented in colon suggests involvement of the colon as well. However, effects of Salmonella on colonic gene expression in vivo are largely unknown. We aimed to characterize time dependent Salmonella-induced changes of colonic mucosal gene expression in rats using whole genome microarrays. For this, rats were orally infected with Salmonella enteritidis to mimic a foodborne infection and colonic gene expression was determined at days 1, 3 and 6 post-infection (n = 8 rats per time-point). As fructo-oligosaccharides (FOS) affect colonic physiology, we analyzed colonic mucosal gene expression of FOS-fed versus cellulose-fed rats infected with Salmonella in a separate experiment. Colonic mucosal samples were isolated at day 2 post-infection. Results Salmonella affected transport (e.g. Chloride channel calcium activated 6, H+/K+ transporting Atp-ase), antimicrobial defense (e.g. Lipopolysaccharide binding protein, Defensin 5 and phospholipase A2), inflammation (e.g. calprotectin), oxidative stress related genes (e.g. Dual oxidase 2 and Glutathione peroxidase 2) and Proteolysis (e.g. Ubiquitin D and Proteosome subunit beta type 9). Furthermore, Salmonella translocation increased serum IFNγ and many interferon-related genes in colonic mucosa. The gene most strongly induced by Salmonella infection was Pancreatitis Associated Protein (Pap), showing >100-fold induction at day 6 after oral infection. Results were confirmed by Q-PCR in individual rats. Stimulation of Salmonella translocation by dietary FOS was accompanied by enhancement of the Salmonella-induced mucosal processes, not by induction of other processes. Conclusion We conclude that the colon is a target tissue for Salmonella, considering the abundant changes in mucosal gene expression

  7. Salmonella induces prominent gene expression in the rat colon.

    PubMed

    Rodenburg, Wendy; Keijer, Jaap; Kramer, Evelien; Roosing, Susanne; Vink, Carolien; Katan, Martijn B; van der Meer, Roelof; Bovee-Oudenhoven, Ingeborg M J

    2007-09-12

    Salmonella enteritidis is suggested to translocate in the small intestine. In vivo it induces gene expression changes in the ileal mucosa and Peyer's patches. Stimulation of Salmonella translocation by dietary prebiotics fermented in colon suggests involvement of the colon as well. However, effects of Salmonella on colonic gene expression in vivo are largely unknown. We aimed to characterize time dependent Salmonella-induced changes of colonic mucosal gene expression in rats using whole genome microarrays. For this, rats were orally infected with Salmonella enteritidis to mimic a foodborne infection and colonic gene expression was determined at days 1, 3 and 6 post-infection (n = 8 rats per time-point). As fructo-oligosaccharides (FOS) affect colonic physiology, we analyzed colonic mucosal gene expression of FOS-fed versus cellulose-fed rats infected with Salmonella in a separate experiment. Colonic mucosal samples were isolated at day 2 post-infection. Salmonella affected transport (e.g. Chloride channel calcium activated 6, H+/K+ transporting Atp-ase), antimicrobial defense (e.g. Lipopolysaccharide binding protein, Defensin 5 and phospholipase A2), inflammation (e.g. calprotectin), oxidative stress related genes (e.g. Dual oxidase 2 and Glutathione peroxidase 2) and Proteolysis (e.g. Ubiquitin D and Proteosome subunit beta type 9). Furthermore, Salmonella translocation increased serum IFN gamma and many interferon-related genes in colonic mucosa. The gene most strongly induced by Salmonella infection was Pancreatitis Associated Protein (Pap), showing >100-fold induction at day 6 after oral infection. Results were confirmed by Q-PCR in individual rats. Stimulation of Salmonella translocation by dietary FOS was accompanied by enhancement of the Salmonella-induced mucosal processes, not by induction of other processes. We conclude that the colon is a target tissue for Salmonella, considering the abundant changes in mucosal gene expression.

  8. Isolation and characterization of Salmonella enterica in day-old ducklings in Egypt

    PubMed Central

    Osman, Kamelia M; Marouf, Sherif H; Zolnikov, Tara R; AlAtfeehy, Nayerah

    2014-01-01

    Importing day-old ducklings (DOD) unknowingly infected with non-typhoid Salmonella (NTS) may be associated with disease risk. Domestic and international trade may enhance this risk. Salmonella enterica serovars, their virulence genes combinations and antibiotic resistance, garner attention for their potentiality to contribute to the adverse health effects on populations throughout the world. The aim of this study was to estimate the risk of imported versus domestic DOD as potential carriers of NTS. The results confirm the prevalence of salmonellosis in imported ducklings was 18.5% (25/135), whereas only 12% (9/75) of cases were determined in the domestic ducklings. Fourteen serovars (Salmonella enteritidis, Salmonella kisii, Salmonella typhimurium, Salmonella gaillac, Salmonella uno, Salmonella eingedi, Salmonella shubra, Salmonella bardo, Salmonella inganda, Salmonella kentucky, Salmonella stanley, Salmonella virchow, Salmonella haifa, and Salmonella anatum) were isolated from the imported ducklings, whereas only S. enteritidis, S. typhimurium, S. virchow, and S. shubra were isolated from the domestic ducklings. The isolated Salmonella serovars were 100% susceptible to only colistin sulphate and 100% resistant to lincomycin. The 14 Salmonella serovars were screened for 11 virulence genes (invA, avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, and bcfC) by PCR. The invA, sopB, and bcfC genes were detected in 100% of the Salmonella serovars; alternatively, the gipA gene was absent in all of the isolated Salmonella serovars. The 11 virulent genes were not detected in either of S. stanley or S. haifa serovars. The results confirm an association between antibiotic resistance and virulence of Salmonella in the DOD. This study confirms the need for a country adherence to strict public health and food safety regimes. PMID:24548159

  9. IL-12p40-dependent agonistic effects on the development of protective innate and adaptive immunity against Salmonella enteritidis.

    PubMed

    Lehmann, J; Bellmann, S; Werner, C; Schröder, R; Schütze, N; Alber, G

    2001-11-01

    To study a potential IL-12p40-dependent but IL-12p75-independent agonistic activity regulating the immune response against Salmonella Enteritidis, the course of infection in IL-12p35-deficient mice (IL-12p35(-/-), capable of producing IL-12p40) was compared with that of IL-12p40(-/-) mice. Mice lacking IL-12p40 revealed a higher mortality rate and higher bacterial organ burden than mice capable of producing IL-12p40. This phenotype was found in both genetically susceptible (BALB/c, Ity(s)) and resistant mice (129Sv/Ev, Ity(r)) indicating Ity-independent mechanisms. The more effective control of bacteria in the IL-12p35(-/-) mice was associated with elevated serum IFN-gamma and TNF-alpha levels. In contrast, IL-12p40(-/-) mice showed reduced IFN-gamma production, which was associated with significantly elevated serum IgE levels. Early during infection (days 3 and 4 postinfection), as well as late (day 20 postinfection), the number of infected phagocytes was strongly increased in the absence of IL-12p40 indicating impaired bactericidal activity when IL-12p40 was missing. Liver histopathology revealed a decreased number of mononuclear granulomas in IL-12p40(-/-) mice. Depletion of CD4(+) or CD8(+) T lymphocytes in vivo suggested that both T cell subpopulations contribute to the IL-12p40-dependent protective functions. Analysis of IL-12p40 vs IL-23p19 mRNA expression revealed an up-regulation of only IL-12p40 mRNA during Salmonella infection. Together these data indicate that IL-12p40 can induce protective mechanisms during both the innate and the adaptive type 1 immune response in Salmonella infection. This novel activity of IL-12p40 complements the well described dominant and essential role of IL-12p75 in protective immunity to Salmonella infection.

  10. Evaluation of yeast dietary supplementation in broilers challenged or not with Salmonella on growth performance, cecal microbiota composition and Salmonella in ceca, cloacae and carcass skin.

    PubMed

    Mountzouris, K C; Dalaka, E; Palamidi, I; Paraskeuas, V; Demey, V; Theodoropoulos, G; Fegeros, K

    2015-10-01

    The dietary supplementation of Saccharomyces cerevisiae var. boulardii was evaluated in broilers challenged or not challenged with Salmonella Enteritidis (SE) using a 2 × 2 factorial arrangement. Depending on yeast inclusion at 0 (C) or 1 × 10⁹ cfu/kg diet (Y) and SE challenge (0 or log 6.3 cfu/bird) on d 15, the experiment had four treatments C, Y, C-SE, and Y-SE, respectively. Each treatment had seven replicate floor pens with 15 broilers. Growth performance responses were determined weekly and overall for the 5 week experimental period. Salmonella levels and prevalence in ceca, cloacae, and carcass skin were determined by culture procedures, while cecal microbiota was determined by real time PCR. Yeast supplementation had no effect (PY > 0.05) on growth performance. For the overall post SE-challenge period (i.e., wk 3 to wk 5), Salmonella reduced body weight gain (BWG) (PSE < 0.001), feed intake (FI) (PSE = 0.032), and the European production efficiency (EPEF) factor (PSE = 0.005). Broilers Y-SE had higher (P < 0.001) overall BW gain compared to C-SE ones. Overall mortality was 2.14% and did not differ (P > 0.05) between treatments. Reduced Salmonella levels in the cloacae (P = 0.014) and on the breast skin (P = 0.006) and lower prevalence on the neck skin (P = 0.007) were noted for treatment Y-SE compared to C-SE. Yeast supplementation did not have an effect (P > 0.05) on cecal microbiota composition at d 1 and d 21 post SE-challenge. On the contrary, SE-challenge reduced cecal levels of total bacteria (PSE = 0.002), E. coli (PSE = 0.006), Bifidobacterium spp. (PSE = 0.006), Bacteroides spp. (PSE = 0.010), and Clostridial populations belonging to cluster I and cluster XIVa, (PSE = 0.047 and PSE = 0.001, respectively) on d 1 post SE-challenge. At 21 d post SE-challenge, only the levels of cecal Lactobacillus spp. (PSE = 0.001) and Bifidobacterium spp. (PSE = 0.049) were reduced compared to the non SE-challenged groups. In conclusion, yeast supplementation in

  11. Immune Response of Chicken Gut to Natural Colonization by Gut Microflora and to Salmonella enterica Serovar Enteritidis Infection ▿

    PubMed Central

    Crhanova, Magdalena; Hradecka, Helena; Faldynova, Marcela; Matulova, Marta; Havlickova, Hana; Sisak, Frantisek; Rychlik, Ivan

    2011-01-01

    In commercial poultry production, there is a lack of natural flora providers since chickens are hatched in the clean environment of a hatchery. Events occurring soon after hatching are therefore of particular importance, and that is why we were interested in the development of the gut microbial community, the immune response to natural microbial colonization, and the response to Salmonella enterica serovar Enteritidis infection as a function of chicken age. The complexity of chicken gut microbiota gradually increased from day 1 to day 19 of life and consisted of Proteobacteria and Firmicutes. For the first 3 days of life, chicken cecum was protected by increased expression of chicken β-defensins (i.e., gallinacins 1, 2, 4, and 6), expression of which dropped from day 4 of life. On the other hand, a transient increase in interleukin-8 (IL-8) and IL-17 expression could be observed in chicken cecum on day 4 of life, indicating physiological inflammation and maturation of the gut immune system. In agreement, the response of chickens infected with S. Enteritidis on days 1, 4, and 16 of life shifted from Th1 (characterized mainly by induction of gamma interferon [IFN-γ] and inducible nitric oxide synthase [iNOS]), observed in younger chickens, to Th17, observed in 16-day-old chickens (characterized mainly by IL-17 induction). Active modification of chicken gut microbiota in the future may accelerate or potentiate the maturation of the gut immune system and increase its resistance to infection with different pathogens. PMID:21555397

  12. Effect of freezing, irradiation, and frozen storage on survival of Salmonella in concentrated orange juice.

    PubMed

    Niemira, Brendan A; Sommers, Christopher H; Boyd, Glenn

    2003-10-01

    Six strains of Salmonella (Anatum F4317, Dublin 15480, Enteritidis 13076, Enteritidis WY15159, Stanley H0588, and Typhimurium 14028) were individually inoculated into orange juice concentrate (OJC) and frozen to -20 degrees C. The frozen samples were treated with 0 (nonirradiated), 0.5, 1.0, or 2.0 kGy of gamma radiation and held frozen for 1 h, and the surviving bacterial population was assessed. The strains showed significant variability in their response to freezing and to freezing in combination with irradiation. The response was dose dependent. Relative to the nonfrozen, nonirradiated control, the reduction following the highest dose (2.0 kGy) ranged from 1.29 log CFU/ml (Salmonella Typhimurium) to 2.17 log CFU/ml (Salmonella Stanley). Samples of OJC inoculated with Salmonella Enteritidis WY15159 and irradiated were stored at -20 degrees C for 1, 2, 7, or 14 days, and the surviving population was determined. Relative to the nonfrozen, nonirradiated control, after 14 days, the population was reduced by 1.2 log CFU/ml in the nonirradiated samples and by 3.3 log CFU/ml following treatment with 2.0 kGy. The combination of frozen storage plus irradiation resulted in greater overall reductions than either process alone.

  13. 75 FR 18751 - Prevention of Salmonella

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-13

    ...-0190] Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation... information: (1) The date by which egg producers must register their farm with FDA, (2) the address and telephone number for requesting copies of Form 3733, and (3) the address to which egg producers must send...

  14. Effect of Salmonella infection on cecal tonsil regulatory T cell properties in chickens

    USDA-ARS?s Scientific Manuscript database

    Two experiments were conducted to study Regulatory T cell (Treg) properties post-Salmonella infection in broiler birds. Four-day-old broiler chicks were orally infected with 5x106 CFU/ml Salmonella enteritidis or sterile PBS (control). Samples were collected at 4, 7, 10, and 14 d post-infection. ...

  15. Salmonella species isolated from animal feed in Iraq.

    PubMed Central

    Al-Hindawi, N; Taha, R R

    1979-01-01

    Of 700 animal feed samples, 32 (4.5%) harbored Salmonella. The highest percentage of contamination was found in sheep feed and local protein. A total of 17 Salmonella serotypes were identified. The most frequent serotypes were Salmonella meleagridis. S. bornum, S. montevideo, and S. drypool. S. bornum was isolated for the first time in Iraq and from both local feed and its ingredients. The common somatic group found was that of Salmonella group C; then came groups E, G, B, and D. Three serotypes (S. enteritidis, S. california, and S. muenchen) seemed to form a link of infection among feed, food, patients, and carriers. PMID:453836

  16. Inactivation by lemon juice of Escherichia coli O157:H7, Salmonella Enteritidis, and Listeria monocytogenes in beef marinating for the ethnic food kelaguen.

    PubMed

    Yang, Jian; Lee, Delores; Afaisen, Shayna; Gadi, Rama

    2013-01-01

    Lemon juice, a major source of acidulant citric acid, is frequently used in the preparation of ethnic foods. Raw or partially cooked meats are marinated with lemon juice in the preparation of a popular Chamorro dish called kelaguen, which is, unfortunately, strongly associated with foodborne illness outbreaks in Guam. We investigated the efficacy of lemon juice in reducing numbers of Escherichia coli O157:H7, Salmonella Enteritidis, and Listeria monocytogenes at stationary phase during marination. Beef inoculated with a three-strain mixture of E. coli O157:H7, S. Enteritidis, or L. monocytogenes at 10(6)CFU/mL was marinated with lemon juice from 0.2 to 10mL/g for 48h at 28°C. The decline of the pathogens during marination exhibited various degrees of deviation from first-order kinetics. Based on calculations with both linear regression and Weibull models, the decimal reduction time (4-D values) over the range of lemon concentrations was 366-5.1h for E. coli O157:H7, 282-2.4h for S. Enteritidis, and 104-2.4h for L. monocytogenes, indicating that E. coli O157:H7 was the most lemon-juice-resistant of the three. The pathogen reduction time (log 4-D values) plotted against undissociated titratable citric acid exhibited a biphasic pattern. The pathogen reduction time (log 4-D or δ values) was linearly correlated with the pH of the marinating beef (R(2)=0.92 to 0.98). The Z(pH) values (pH dependence of death rate) with beef marination were 1.03 for E. coli O157:H7, 0.92 for S. Enteritidis, and 1.29 for L. monocytogenes, indicating that L. monocytogenes was the most pH resistant of the three. L. monocytogenes exhibited less resistance to lemon juice than S. Enteritidis at pH of 3.5-4.4 but more resistance at pH of 2.6-2.8. In addition, at 4°C, all three pathogens exhibited 4-D values 1.7-4.1 times greater than those at 24°C at 5mL lemon juice/g beef. In conclusion, the usual beef marinating practice for kelaguen preparation (<0.5mL lemon juice/g beef for 1-12h) did not

  17. A dual mechanism involved in membrane and nucleic acid disruption of AvBD103b, a new avian defensin from the king penguin, against Salmonella enteritidis CVCC3377.

    PubMed

    Teng, Da; Wang, Xiumin; Xi, Di; Mao, Ruoyu; Zhang, Yong; Guan, Qingfeng; Zhang, Jun; Wang, Jianhua

    2014-10-01

    The food-borne bacterial gastrointestinal infection is a serious public health threat. Defensins are evolutionarily conserved innate immune components with broad-spectrum antibacterial activity that do not easily induce resistance. AvBD103b, an avian defensin with potent activity against Salmonella enteritidis, was isolated from the stomach contents of the king penguin (Aptenodytes patagonicus). To elucidate further the antibacterial mechanism of AvBD103b, its effect on the S. enteritidis CVCC3377 cell membrane and intracellular DNA was researched. The cell surface hydrophobicity and a N-phenyl-1-naphthylamine uptake assay demonstrated that AvBD103b treatment increased the cell surface hydrophobicity and outer membrane permeability. Atomic absorption spectrometry, ultraviolet spectrophotometry, flow cytometry, and transmission electron microscopy (TEM) indicated that AvBD103b treatment can lead to the release of the cellular contents and cell death through damage of the membrane. DNA gel retardation and circular dichroism analysis demonstrated that AvBD103b interacted with DNA and intercalated into the DNA base pairs. A cell cycle assay demonstrated that AvBD103b affected cellular functions, such as DNA synthesis. Our results confirmed that AvBD103b exerts its antibacterial activity by damaging the cell membrane and interfering with intracellular DNA, ultimately causing cell death, and suggested that AvBD103b may be a promising candidate as an alternative to antibiotics against S. enteritidis.

  18. Prevalence of Salmonella enterica and Shiga toxin-producing Escherichia coli in zoo animals from Chile.

    PubMed

    Marchant, Paulina; Hidalgo-Hermoso, Ezequiel; Espinoza, Karen; Retamal, Patricio

    2016-12-30

    Salmonella (S.) enterica and Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens. Here, we report the prevalence of S. enterica and STEC in feces of 316 zoo animals belonging to 61 species from Chile. S. enterica and STEC strains were detected in 7.5% and 4.4% of animals, respectively. All Salmonella isolates corresponded to the serotype Enteritidis. To the best of our knowledge, this is the first report of S. Enteritidis in the culpeo fox ( Lycalopex culpaeus ), black-capped capuchin ( Sapajus apella ) and Peruvian pelican ( Pelecanus thagus ) and the first STEC report in Thomson's gazelle ( Eudorcas thomsonii ).

  19. Prevalence of Salmonella enterica and Shiga toxin-producing Escherichia coli in zoo animals from Chile

    PubMed Central

    Marchant, Paulina; Hidalgo-Hermoso, Ezequiel; Espinoza, Karen

    2016-01-01

    Salmonella (S.) enterica and Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens. Here, we report the prevalence of S. enterica and STEC in feces of 316 zoo animals belonging to 61 species from Chile. S. enterica and STEC strains were detected in 7.5% and 4.4% of animals, respectively. All Salmonella isolates corresponded to the serotype Enteritidis. To the best of our knowledge, this is the first report of S. Enteritidis in the culpeo fox (Lycalopex culpaeus), black-capped capuchin (Sapajus apella) and Peruvian pelican (Pelecanus thagus) and the first STEC report in Thomson's gazelle (Eudorcas thomsonii). PMID:27030195

  20. Persistence of Salmonella on egg conveyor belts is dependent on the belt type but not on the rdar morphotype.

    PubMed

    Stocki, S L; Annett, C B; Sibley, C D; McLaws, M; Checkley, S L; Singh, N; Surette, M G; White, A P

    2007-11-01

    Commercial caged layer flocks in Alberta, Canada, are commonly monitored for Salmonella enterica serovar Enteritidis (SE) and S. enterica serovar Typhimurium (ST) by environmental sampling. In one recent case, a SE strain isolated from the egg conveyor belt was a source of persistent infection for the flock. This study was undertaken to examine Salmonella colonization on egg conveyor belts and to determine whether the rdar morphotype, a conserved physiology associated with aggregation and long-term survival, contributed to persistence. Four woven belts constructed of natural or nonnatural fibers and a 1-piece belt made of vinyl were tested with rdar-positive ST and SE strains and a rdar-negative ST DeltaagfD reference strain. The type of egg belt was the most important factor influencing Salmonella colonization and persistence. The vinyl belt, with the least surface area available for colonization, had the fewest Salmonella remaining after washing and disinfection, whereas the hemp-plastic belt, with the greatest surface area, had the most Salmonella remaining. Real-time gene expression indicated that the rdar morphotype was involved in colonizing the egg belt pieces; however, it was not essential for persistence. In addition, rdar-positive and rdar-negative strains were equally similarly to disinfection on the egg belt pieces. The results indicate that Salmonella can persist on a variety of egg belts by mechanisms other than the rdar morphotype, and that using egg conveyer belts with reduced surface area for bacterial colonization can lessen contamination problems.

  1. Molecular epidemiology of antibiotic resistance of Salmonella enteritidis during a 7-year period in Greece.

    PubMed Central

    Tassios, P T; Markogiannakis, A; Vatopoulos, A C; Katsanikou, E; Velonakis, E N; Kourea-Kremastinou, J; Legakis, N J

    1997-01-01

    A significant increase in the frequency of isolation of Salmonella enteritidis has been observed during recent years in Greece, parallelled by an increasing rate of resistance of this organism to antibiotics. A substantial proportion of ampicillin- and doxycycline-resistant isolates exhibited cross-resistance to drugs of other classes, such as sulfonamides and streptomycin. Isolates of human origin were overall less resistant than those of animal or food-feed origin. Indeed, strains associated with animal infections were characterized by the highest rates of resistance to several antibiotics. These phenotypic data were correlated with genotypic information concerning two distinct populations: isolates from all sources that were resistant only to ampicillin, the drug toward which resistance rates were highest, and a control group of sensitive isolates. Ampicillin resistance was due to a 34-MDa conjugative plasmid. DNA fingerprinting by macrorestriction of genomic DNA revealed two types, A and B, common to both ampicillin-resistant and -sensitive strains, with 80 to 90% of strains being of type A. However, a third type, C, was specific for the sensitive population, representing 17% of those strains. Therefore, although the majority of resistant isolates were genetically related to sensitive ones, there existed a susceptible clone which had not acquired any resistance traits. PMID:9163436

  2. Prevalence, seasonal occurrence and antimicrobial resistance of Salmonella in poultry retail products in Greece.

    PubMed

    Zdragas, A; Mazaraki, K; Vafeas, G; Giantzi, V; Papadopoulos, T; Ekateriniadou, L

    2012-10-01

    To detect the prevalence, the seasonal occurrence and distribution of Salmonella serotypes in poultry products and to determine the resistance profile of Salmonella isolates. A total of 96 skin-on chicken carcasses and 30 liver samples were analysed between May 2007 and May 2009 from twenty-two different commercial farm brands found in retail market countrywide. Salmonella was isolated from 38 (39·5%) of 96 chicken carcasses and from 10 (33·3%) of 30 liver samples. Higher isolation rate (60·4%) was observed in carcasses detected during summer (May to October), and lower isolation rate (18·7%) was observed in carcasses detected during winter (November to April); in liver samples, the positive rates were 53·4 and 13·2%, respectively. Twelve serotypes were detected with the serotypes Hadar, Enteritidis and Blockley being the most prevalent at 29·2, 22·9 and 12·5%, respectively. Nine of 11 Salm. Enteritidis isolates occurred during summer. Of 48 isolates, 38 (79%) were resistant to one or more of the antimicrobial agents used. The highest resistance rates were found to the following antimicrobials: streptomycin (64·5%), tetracycline (56·2%), nalidixic acid (39·5%), ampicillin and rifampicin (33·3%). The relatively high Salmonella spp. contamination rates of raw chicken meat and liver have been detected. Salm. Enteritidis isolates peaked in summer, increasing the risk to human health. Antibiotic resistance of Salmonella still remains a threat as resistance plasmids may be extensively shared between animal and humans. The study enabled us to improve the data on the seasonal occurrence of Salmonella and to determine the antimicrobial pattern profile and trends in Salmonella strains isolated from poultry retail products in Greece. © 2012 The Authors. Letters in Applied Microbiology © 2012 The Society for Applied Microbiology.

  3. Protracted outbreak of S. Enteritidis PT 21c in a large Hamburg nursing home

    PubMed Central

    Frank, Christina; Buchholz, Udo; Maaß, Monika; Schröder, Arthur; Bracht, Karl-Hans; Domke, Paul-Gerhard; Rabsch, Wolfgang; Fell, Gerhard

    2007-01-01

    Background During August 2006, a protracted outbreak of Salmonella (S.) Enteritidis infections in a large Hamburg nursing home was investigated. Methods A site visit of the home was conducted and food suppliers' premises tested for Salmonella. Among nursing home residents a cohort study was carried out focusing on foods consumed in the three days before the first part of the outbreak. Instead of relying on residents' memory, data from the home's patient food ordering system was used as exposure data. S. Enteritidis isolates from patients and suspected food vehicles were phage typed and compared. Results Within a population of 822 nursing home residents, 94 case patients among residents (1 fatality) and 17 among staff members were counted 6 through 29 August. The outbreak peaked 7 through 9 August, two days after a spell of very warm summer weather. S. Enteritidis was consistently recovered from patients' stools throughout the outbreak. Among the food items served during 5 through 7 August, the cohort study pointed to afternoon cake on all three days as potential risk factors for disease. Investigation of the bakery supplying the cake yielded S. Enteritidis from cakes sampled 31 August. Comparison of the isolates by phage typing demonstrated both isolates from patients and the cake to be the exceedingly rare phage type 21c. Conclusion Cake (various types served on various days) contaminated with S. Enteritidis were the likely vehicle of the outbreak in the nursing home. While the cakes were probably contaminated with low pathogen dose throughout the outbreak period, high ambient summer temperatures and failure to keep the cake refrigerated led to high pathogen dose in cake on some days and in some of the housing units. This would explain the initial peak of cases, but also the drawn out nature of the outbreak with cases until the end of August. Suggestions are made to nursing homes, aiding in outbreak prevention. Early outbreak detection is crucial, such that

  4. Evaluation of Molecular Methods for Identification of Salmonella Serovars

    PubMed Central

    Gurnik, Simone; Ahmad, Aaminah; Blimkie, Travis; Murphy, Stephanie A.; Kropinski, Andrew M.; Nash, John H. E.

    2016-01-01

    Classification by serotyping is the essential first step in the characterization of Salmonella isolates and is important for surveillance, source tracking, and outbreak detection. To improve detection and reduce the burden of salmonellosis, several rapid and high-throughput molecular Salmonella serotyping methods have been developed. The aim of this study was to compare three commercial kits, Salm SeroGen (Salm Sero-Genotyping AS-1 kit), Check&Trace (Check-Points), and xMAP (xMAP Salmonella serotyping assay), to the Salmonella genoserotyping array (SGSA) developed by our laboratory. They were assessed using a panel of 321 isolates that represent commonly reported serovars from human and nonhuman sources globally. The four methods correctly identified 73.8% to 94.7% of the isolates tested. The methods correctly identified 85% and 98% of the clinically important Salmonella serovars Enteritidis and Typhimurium, respectively. The methods correctly identified 75% to 100% of the nontyphoidal, broad host range Salmonella serovars, including Heidelberg, Hadar, Infantis, Kentucky, Montevideo, Newport, and Virchow. The sensitivity and specificity of Salmonella serovars Typhimurium and Enteritidis ranged from 85% to 100% and 99% to 100%, respectively. It is anticipated that whole-genome sequencing will replace serotyping in public health laboratories in the future. However, at present, it is approximately three times more expensive than molecular methods. Until consistent standards and methodologies are deployed for whole-genome sequencing, data analysis and interlaboratory comparability remain a challenge. The use of molecular serotyping will provide a valuable high-throughput alternative to traditional serotyping. This comprehensive analysis provides a detailed comparison of commercial kits available for the molecular serotyping of Salmonella. PMID:27194688

  5. Vaccines against invasive Salmonella disease

    PubMed Central

    MacLennan, Calman A; Martin, Laura B; Micoli, Francesca

    2014-01-01

    Though primarily enteric pathogens, Salmonellae are responsible for a considerable yet under-appreciated global burden of invasive disease. In South and South-East Asia, this manifests as enteric fever caused by serovars Typhi and Paratyphi A. In sub-Saharan Africa, a similar disease burden results from invasive nontyphoidal Salmonellae, principally serovars Typhimurium and Enteritidis. The existing Ty21a live-attenuated and Vi capsular polysaccharide vaccines target S. Typhi and are not effective in young children where the burden of invasive Salmonella disease is highest. After years of lack of investment in new Salmonella vaccines, recent times have seen increased interest in the area led by emerging-market manufacturers, global health vaccine institutes and academic partners. New glycoconjugate vaccines against S. Typhi are becoming available with similar vaccines against other invasive serovars in development. With other new vaccines under investigation, including live-attenuated, protein-based and GMMA vaccines, now is an exciting time for the Salmonella vaccine field. PMID:24804797

  6. Salmonella spp. contamination in commercial layer hen farms using different types of samples and detection methods.

    PubMed

    Soria, M C; Soria, M A; Bueno, D J; Godano, E I; Gómez, S C; ViaButron, I A; Padin, V M; Rogé, A D

    2017-08-01

    The performance of detection methods (culture methods and polymerase chain reaction assay) and plating media used in the same type of samples were determined as well as the specificity of PCR primers to detected Salmonella spp. contamination in layer hen farms. Also, the association of farm characteristics with Salmonella presence was evaluated. Environmental samples (feces, feed, drinking water, air, boot-swabs) and eggs were taken from 40 layer hen houses. Salmonella spp. was most detected in boot-swabs taken around the houses (30% and 35% by isolation and PCR, respectively) follow by fecal samples (15.2% and 13.6% by isolation and PCR, respectively). Eggs, drinking water, and air samples were negative for Salmonella detection. Salmonella Schwarzengrund and S. Enteritidis were the most isolated serotypes. For plating media, relative specificity was 1, and the relative sensitivity was greater for EF-18 agar than XLDT agar in feed and fecal samples. However, relative sensitivity was greater in XLDT agar than EF-18 agar for boot-swab samples. Agreement was between fair to good depending on the sample, and it was good between isolation and PCR (feces and boot-swabs), without agreement for feed samples. Salmonella spp. PCR was positive for all strains, while S. Typhimurium PCR was negative. Salmonella Enteritidis PCR used was not specific. Based in the multiple logistic regression analyses, categorization by counties was significant for Salmonella spp. presence (P-value = 0.010). This study shows the importance of considering different types of samples, plating media and detection methods during a Salmonella spp. monitoring study. In addition, it is important to incorporate the sampling of floors around the layer hen houses to learn if biosecurity measures should be strengthened to minimize the entry and spread of Salmonella in the houses. Also, the performance of some PCR methods and S. Enteritidis PCR should be improved, and biosecurity measures in hen farms must be

  7. Disinfectant susceptibility of different Salmonella serotypes isolated from chicken and egg production chains.

    PubMed

    Long, M; Lai, H; Deng, W; Zhou, K; Li, B; Liu, S; Fan, L; Wang, H; Zou, L

    2016-09-01

    The study aimed to serotype the Salmonella isolates recovered from chicken and egg production chains, and to investigate the disinfectant resistance phenotypes and genotypes of these isolates. The Salmonella isolates were serotyped, and the minimal inhibitory concentrations (MICs) of disinfectants were determined. Results showed that the Salmonella isolates recovered from both chains were diverse, and the serotypes in each part of the production chain and between the two production chains were significantly different. In the chicken production chain, 19 different serotypes were recovered, while only five serotypes were found in the egg production chain. The isolates showed a high susceptibility to didecyldimethylammonium bromide (DDAB) but a low susceptibility to benzalkonium chloride (BC), benzalkonium bromide (BAB) and chlorhexidine (CHX). Salmonella Enteritidis and Salmonella Typhimurium were more resistant to BC and BAB. The qacEΔ1 and qacF resistance genes were detected in 26·7 and 7·7% of the isolates respectively. The qacEΔ1 gene was frequently found in Salmonella Derby and Salm. Enteritidis (P < 0·05). Our findings indicated that Salmonella was commonly present in both chains, and could serve as a critical vector in spreading disinfectant resistance associated with different serotypes. This study first demonstrated disinfectant resistance phenotypes and genotypes of serotyped Salmonella. The study highlights the need for monitoring the disinfectant resistance varied in different Salmonella serotypes. © 2016 The Society for Applied Microbiology.

  8. Impact of egg holding temperatures on the recovery of Salmonella from eggshells and stainless steel coupons

    USDA-ARS?s Scientific Manuscript database

    This experiment was conducted to determine the impact of egg holding temperature on the ability to recover Salmonella from eggshells after 24 h. Salmonella enterica Enteritidis (nalidixic acid resistant marker strain) inoculated eggshells and stainless steel coupons (SSC, 14 mm diameter) were held a...

  9. Surveillance of Salmonella enteritidis in layer houses: a retrospective comparison of the Food and Drug Administration's egg safety rule (2010-2011) and the California Egg Quality Assurance Program (2007-2011).

    PubMed

    Pitesky, Maurice; Charlton, Bruce; Bland, Mark; Rolfe, Dan

    2013-03-01

    Between July 2007 and December 2011, 2660 environmental drag swab samples were collected in total from California layer flocks on behalf of the California Egg Quality Assurance Program (CEQAP), the egg safety rule (21 CFR Parts 16 and 118) of the Food and Drug Administration (FDA), or both. The samples were processed by the California Animal Health and Food Safety Lab, and positive or negative results for Salmonella enterica serovar Enteritidis (SE) were recorded. This study retrospectively compares the differences between the FDA and CEQAP programs with respect to their SE environmental sampling surveillance results. To accomplish this comparison, two different CEQAP (new and old) data sets representing different SE environmental surveillance approaches in the life of the flock were compared against each other and against the FDA's SE environmental testing plan. Significant differences were noted between the CEQAP and FDA programs with respect to the prevalence of SE in the farm environment. Analyses of the prevalence of SE at different stages in the flock's life cycle (chick papers, preproduction, midproduction, postmolt, and premarket) found the highest prevalence of SE in premarket (11.9%), followed by postmolt (3.5%) and midproduction (3.4%), and there was a tie between chick papers and preproduction (2.1%). To assess the main effects of the presence of SE in the farm environment, backwards binary logistic regression was used. Of six independent variables examined (age of flock, year, season, owner, CEQAP membership, and analysis of pooled samples vs. individual swabs), only age of flock, owner, and year were determined to be significant factors in the final model. Although CEQAP membership and pooling vs. individuals swabs were not included in the final model, Pearson chi-square tests did show significantly higher odds of SE for non-CEQAP member farms and higher odds of SE in pooled samples vs. individual swabs.

  10. Immunisation of chickens with live Salmonella vaccines - Role of booster vaccination.

    PubMed

    Methner, U

    2018-05-17

    It is accepted that booster vaccinations of chickens with live Salmonella vaccines are essential part of vaccinations schemes to induce an effective adaptive immune response. As manufacturer of registered live Salmonella vaccines recommend different times of booster the question raises whether the duration between the first and second immunisation might influence the protective effect against Salmonella exposure. Chickens were immunised with a live Salmonella Enteritidis vaccine on day 1 of age followed by a booster vaccination at different intervals (day 28, 35 or 42 of age) to study the effects on the colonisation and invasion of the Salmonella vaccine strain, the humoral immune response and the efficacy against infection with Salmonella Enteritidis on day 56 of age. Immunisation of all groups resulted in a very effective adaptive immune response and a high degree of protection against severe Salmonella exposure, however, the time of booster had only an unverifiable influence on either the colonisation of the vaccine strain, the development of the humoral immune response or the colonisation of the Salmonella challenge strain. Therefore, the first oral immunisation of the chicks on day 1 of age seems to be of special importance and prerequisite for the development of the effective immune response. A booster immunisation should be carried out, however, the time of booster may vary between week 3 and week 7 of age of the chickens without adversely impact on the efficacy of the adaptive immune response or the protective effects. Copyright © 2018 Elsevier Ltd. All rights reserved.

  11. Inactivation of Salmonella in Shell Eggs by Hot Water Immersion and Its Effect on Quality.

    PubMed

    Geveke, David J; Gurtler, Joshua B; Jones, Deana R; Bigley, Andrew B W

    2016-03-01

    Thermal inactivation kinetics of heat resistant strains of Salmonella Enteritidis in shell eggs processed by hot water immersion were determined and the effects of the processing on egg quality were evaluated. Shell eggs were inoculated with a composite of heat resistant Salmonella Enteritidis (SE) strains PT8 C405, 2 (FSIS #OB030832), and 6 (FSIS #OB040159). Eggs were immersed in a circulating hot water bath for various times and temperatures. Come-up time of the coldest location within the egg was 21 min. SE was reduced by 4.5 log at both hot water immersion treatments of 56.7 C for 60 min and 55.6 °C for 100 min. Decimal reduction times (D-values) at 54.4, 55.6, and 56.7 °C were 51.8, 14.6, and 9.33 min, respectively. The z-value was 3.07 °C. Following treatments that resulted in a 4.5 log reduction (56.7 °C/60 min and 55.6 °C/100 min), the surviving population of SE remained static during 4 wk of refrigerated storage. After processing under conditions resulting in 4.5 log reductions, the Haugh unit and albumen height significantly increased (P < 0.01) and yolk index significantly decreased (P < 0.05). The shell dynamic stiffness significantly increased (P < 0.05), while static compression shell strength showed no significant difference (P < 0.05). Vitelline membrane strength significantly increased (P < 0.05); although, no significant difference (P < 0.05) was observed in vitelline membrane elasticity. In summary, the hot water immersion process inactivated heat resistant SE in shell eggs by 4.5 log, but also significantly affected several egg quality characteristics. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  12. Probability of identifying different salmonella serotypes in poultry samples

    USDA-ARS?s Scientific Manuscript database

    Recent work has called attention to the unequal competitive abilities of different Salmonella serotypes in standard broth culture and plating media. Such serotypes include Enteritidis and Typhimurium that are specifically targeted in some regulatory and certification programs because they cause a l...

  13. 77 FR 14022 - Guidance for Industry: Testing for Salmonella Species in Human Foods and Direct-Human-Contact...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-08

    ... not apply to egg producers and other persons who are covered by FDA's final rule ``Prevention of Salmonella Enteritidis in Shell Eggs During Production, Storage, and Transportation.'' The guidance addresses testing procedures for Salmonella species (spp.) in human foods (except shell eggs) and direct-human...

  14. Consecutive salmonella outbreaks traced to the same bakery.

    PubMed Central

    Evans, M. R.; Tromans, J. P.; Dexter, E. L.; Ribeiro, C. D.; Gardner, D.

    1996-01-01

    Two consecutive community outbreaks of Salmonella enteritidis phage type 4 (PT4) traced to the same bakery occurred in Cardiff, Wales during August-September 1992. In the first outbreak, illness was associated with eating custard slices (odds ratio 23.8, 95% confidence interval 6.5-94.4, P < 0.0001), and in the second, with eating fresh cream cakes (odds ratio 15.8, 95% confidence interval 1.6-374, P = 0.004). Environmental investigations implicated cross-contamination during preparation of the cold-custard mix as the cause of the first outbreak, and inadequate cleaning and disinfection of nozzles used for piping cream in the second outbreak. S. enteritidis PT4 was isolated from fresh cream sponge cake retained by a case and from two fresh cream cakes and four environmental swabs obtained at the bakery. This incident illustrates the hazard of widespread environmental contamination with salmonella and the need for thorough environmental cleansing for any premises implicated in an outbreak of food poisoning. PMID:8620907

  15. Prophage Integrase Typing Is a Useful Indicator of Genomic Diversity in Salmonella enterica

    PubMed Central

    Colavecchio, Anna; D’Souza, Yasmin; Tompkins, Elizabeth; Jeukens, Julie; Freschi, Luca; Emond-Rheault, Jean-Guillaume; Kukavica-Ibrulj, Irena; Boyle, Brian; Bekal, Sadjia; Tamber, Sandeep; Levesque, Roger C.; Goodridge, Lawrence D.

    2017-01-01

    Salmonella enterica is a bacterial species that is a major cause of illness in humans and food-producing animals. S. enterica exhibits considerable inter-serovar diversity, as evidenced by the large number of host adapted serovars that have been identified. The development of methods to assess genome diversity in S. enterica will help to further define the limits of diversity in this foodborne pathogen. Thus, we evaluated a PCR assay, which targets prophage integrase genes, as a rapid method to investigate S. enterica genome diversity. To evaluate the PCR prophage integrase assay, 49 isolates of S. enterica were selected, including 19 clinical isolates from clonal serovars (Enteritidis and Heidelberg) that commonly cause human illness, and 30 isolates from food-associated Salmonella serovars that rarely cause human illness. The number of integrase genes identified by the PCR assay was compared to the number of integrase genes within intact prophages identified by whole genome sequencing and phage finding program PHASTER. The PCR assay identified a total of 147 prophage integrase genes within the 49 S. enterica genomes (79 integrase genes in the food-associated Salmonella isolates, 50 integrase genes in S. Enteritidis, and 18 integrase genes in S. Heidelberg). In comparison, whole genome sequencing and PHASTER identified a total of 75 prophage integrase genes within 102 intact prophages in the 49 S. enterica genomes (44 integrase genes in the food-associated Salmonella isolates, 21 integrase genes in S. Enteritidis, and 9 integrase genes in S. Heidelberg). Collectively, both the PCR assay and PHASTER identified the presence of a large diversity of prophage integrase genes in the food-associated isolates compared to the clinical isolates, thus indicating a high degree of diversity in the food-associated isolates, and confirming the clonal nature of S. Enteritidis and S. Heidelberg. Moreover, PHASTER revealed a diversity of 29 different types of prophages and 23

  16. Prophage Integrase Typing Is a Useful Indicator of Genomic Diversity in Salmonella enterica.

    PubMed

    Colavecchio, Anna; D'Souza, Yasmin; Tompkins, Elizabeth; Jeukens, Julie; Freschi, Luca; Emond-Rheault, Jean-Guillaume; Kukavica-Ibrulj, Irena; Boyle, Brian; Bekal, Sadjia; Tamber, Sandeep; Levesque, Roger C; Goodridge, Lawrence D

    2017-01-01

    Salmonella enterica is a bacterial species that is a major cause of illness in humans and food-producing animals. S. enterica exhibits considerable inter-serovar diversity, as evidenced by the large number of host adapted serovars that have been identified. The development of methods to assess genome diversity in S. enterica will help to further define the limits of diversity in this foodborne pathogen. Thus, we evaluated a PCR assay, which targets prophage integrase genes, as a rapid method to investigate S. enterica genome diversity. To evaluate the PCR prophage integrase assay, 49 isolates of S. enterica were selected, including 19 clinical isolates from clonal serovars (Enteritidis and Heidelberg) that commonly cause human illness, and 30 isolates from food-associated Salmonella serovars that rarely cause human illness. The number of integrase genes identified by the PCR assay was compared to the number of integrase genes within intact prophages identified by whole genome sequencing and phage finding program PHASTER. The PCR assay identified a total of 147 prophage integrase genes within the 49 S. enterica genomes (79 integrase genes in the food-associated Salmonella isolates, 50 integrase genes in S . Enteritidis, and 18 integrase genes in S . Heidelberg). In comparison, whole genome sequencing and PHASTER identified a total of 75 prophage integrase genes within 102 intact prophages in the 49 S. enterica genomes (44 integrase genes in the food-associated Salmonella isolates, 21 integrase genes in S . Enteritidis, and 9 integrase genes in S . Heidelberg). Collectively, both the PCR assay and PHASTER identified the presence of a large diversity of prophage integrase genes in the food-associated isolates compared to the clinical isolates, thus indicating a high degree of diversity in the food-associated isolates, and confirming the clonal nature of S . Enteritidis and S . Heidelberg. Moreover, PHASTER revealed a diversity of 29 different types of prophages and 23

  17. Application of Molecular Typing Results in Source Attribution Models: The Case of Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) of Salmonella Isolates Obtained from Integrated Surveillance in Denmark.

    PubMed

    de Knegt, Leonardo V; Pires, Sara M; Löfström, Charlotta; Sørensen, Gitte; Pedersen, Karl; Torpdahl, Mia; Nielsen, Eva M; Hald, Tine

    2016-03-01

    Salmonella is an important cause of bacterial foodborne infections in Denmark. To identify the main animal-food sources of human salmonellosis, risk managers have relied on a routine application of a microbial subtyping-based source attribution model since 1995. In 2013, multiple locus variable number tandem repeat analysis (MLVA) substituted phage typing as the subtyping method for surveillance of S. Enteritidis and S. Typhimurium isolated from animals, food, and humans in Denmark. The purpose of this study was to develop a modeling approach applying a combination of serovars, MLVA types, and antibiotic resistance profiles for the Salmonella source attribution, and assess the utility of the results for the food safety decisionmakers. Full and simplified MLVA schemes from surveillance data were tested, and model fit and consistency of results were assessed using statistical measures. We conclude that loci schemes STTR5/STTR10/STTR3 for S. Typhimurium and SE9/SE5/SE2/SE1/SE3 for S. Enteritidis can be used in microbial subtyping-based source attribution models. Based on the results, we discuss that an adjustment of the discriminatory level of the subtyping method applied often will be required to fit the purpose of the study and the available data. The issues discussed are also considered highly relevant when applying, e.g., extended multi-locus sequence typing or next-generation sequencing techniques. © 2015 Society for Risk Analysis.

  18. Bacteriophage P22 to challenge Salmonella in foods.

    PubMed

    Zinno, Paola; Devirgiliis, Chiara; Ercolini, Danilo; Ongeng, Duncan; Mauriello, Gianluigi

    2014-11-17

    In this study we considered the influence of phage addition on the fate of Salmonella enterica serovar Typhimurium in different foods. Phage P22 was applied to the following: liquid eggs, energy drinks, whole and skimmed milk, apple juice, chicken breast and chicken mince all spiked with its host, whose growth was monitored for 24 and 48 h at 4 °C. Appreciable host inactivation, generally in the order of 2 log cycles, was achieved compared to phage-free controls in all food matrices when 10(4) UFC/g host inoculum was used. Furthermore, wild food strains belonging to the serotypes Typhimurium, Enteritidis, Derby Give, Newport, Muenchen and Muenster were assayed towards phage P22. Only isolates of Salmonella Typhimurium as well as Salmonella Derby and Salmonella Enteritidis was inhibited by the presence of P22 phage. Additional challenge experiments were carried out by spiking liquid-eggs, chicken breast and chicken mince with mixes of wild Salmonella Typhimurium (at concentration of about 10(4) UFC/g) strains along with their relative phage P22. The results showed a reduction of 2-3 log cycles after 48 h at 4 °C depending on both mix of strains and the specific food. Overall, the results indicate that phages may be useful in the control of food-borne pathogens. The food matrices considered, the liquid more than the solid, do not seem to affect the phage ability of infection compared to similar tests performed in vitro. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Molecular Epidemiology of Nontyphoidal Salmonella in Poultry and Poultry Products in India: Implications for Human Health.

    PubMed

    Saravanan, Sellappan; Purushothaman, Venketaraman; Murthy, Thippichettypalayam Ramasamy Gopala Krishna; Sukumar, Kuppannan; Srinivasan, Palani; Gowthaman, Vasudevan; Balusamy, Mohan; Atterbury, Robert; Kuchipudi, Suresh V

    2015-09-01

    Human infections with non-typhoidal Salmonella (NTS) serovars are increasingly becoming a threat to human health globally. While all motile Salmonellae have zoonotic potential, Salmonella Enteritidis and Salmonella Typhimurium are most commonly associated with human disease, for which poultry are a major source. Despite the increasing number of human NTS infections, the epidemiology of NTS in poultry in India has not been fully understood. Hence, as a first step, we carried out epidemiological analysis to establish the incidence of NTS in poultry to evaluate the risk to human health. A total of 1215 samples (including poultry meat, tissues, egg and environmental samples) were collected from 154 commercial layer farms from southern India and screened for NTS. Following identification by cultural and biochemical methods, Salmonella isolates were further characterized by multiplex PCR, allele-specific PCR, enterobacterial repetitive intergenic consensus (ERIC) PCR and pulse field gel electrophoresis (PFGE). In the present study, 21/1215 (1.73 %) samples tested positive for NTS. We found 12/392 (3.06 %) of tissue samples, 7/460 (1.52 %) of poultry products, and 2/363 (0.55 %) of environmental samples tested positive for NTS. All the Salmonella isolates were resistant to oxytetracycline, which is routinely used as poultry feed additive. The multiplex PCR results allowed 16/21 isolates to be classified as S. Typhimurium, and five isolates as S. Enteritidis. Of the five S. Enteritidis isolates, four were identified as group D Salmonella by allele-specific PCR. All of the isolates produced different banding patterns in ERIC PCR. Of the thirteen macro restriction profiles (MRPs) obtained by PFGE, MRP 6 was predominant which included 6 (21 %) isolates. In conclusion, the findings of the study revealed higher incidence of contamination of NTS Salmonella in poultry tissue and animal protein sources used for poultry. The results of the study warrants further investigation

  20. Effectiveness of a spontaneous carvacrol nanoemulsion against Salmonella enterica Enteritidis and Escherichia coli O157:H7 on contaminated broccoli and radish seeds.

    PubMed

    Landry, Kyle S; Micheli, Sean; McClements, David Julian; McLandsborough, Lynne

    2015-10-01

    The incidence of foodborne illness associated with the consumption of fresh produce has continued to increase over the past decade. Sprouts, such as mung bean, alfalfa, radish, and broccoli, are minimally processed and have been sources for foodborne illness. Currently, a 20,000 ppm calcium hypochlorite soak is recommended for the treatment of sprouting seeds. In this study, the efficacy of an antimicrobial carvacrol nanoemulsion was tested against Salmonella enterica subspecies enterica serovar Enteritidis (ATCC BAA-1045) or EGFP expressing Escherichia coli O157:H7 (ATCC 42895) contaminated sprouting seeds. Antimicrobial treatments were performed by soaking inoculated seeds in nanoemulsions (4000 or 8000 ppm) for 30 or 60 min. Following treatment, surviving cells were determined by performing plate counts and/or Most Probable Number (MPN) enumeration. Treated seeds were sprouted and tested for the presence of pathogens. Treatment successfully inactivated low levels (2 and 3 log CFU/g) of S. Enteritidis and E. coli on radish seeds when soaked for 60 min at concentrations ≥4000 (0.4%) ppm carvacrol. This treatment method was not affective on contaminated broccoli seeds. Total sprout yield was not influenced by any treatments. These results show that carvacrol nanoemulsions may be an alternative treatment method for contaminated radish seeds. Copyright © 2015 Elsevier Ltd. All rights reserved.