Prevalence, numbers and antimicrobial susceptibilities of Salmonella serovars and Campylobacter spp. in retail poultry in Phnom Penh, Cambodia

Salmonella and Campylobacter are common bacterial pathogens associated with human gastro-enteritis; and raw poultry is considered to be an important source of these bacteria. To evaluate whether the Salmonella serovars and Campylobacter spp. bacteria could be monitored for the purpose of microbial presence, enumeration and antimicrobial resistance in raw poultry, 152 poultry carcasses were randomly selected from 10 markets in retail outlets of Phnom Penh during March 2006 to February 2007. The majority of poultry samples was contaminated by Salmonella serovars (88.2%) and Campylobacter spp. (80.9%). A very high contamination of Salmonella was found at 3-4 log?? CFU/g for 22.4% of samples and of Campylobacter at 7-8 log?? CFU/g for 1.3% of samples. Fifty nine different Salmonella serovars contaminated 134 poultry carcasses; five most prevalent serovars covered 29.1% of serovars isolates (Anatum, Typhimurium, Corvallis, Stanley and Enteritidis). Three Campylobacter species contaminating 123 raw poultry were Campylobacter jejuni (50.0%), Campylobacter coli (29.0%) and Campylobacter lari (21.0%). High antibiotic resistance percentages were found among Salmonella serovars and Campylobacter spp. isolates. This study revealed that raw poultry at the retail outlets in Phnom Penh markets are contaminated with high prevalences of food-borne pathogens, and communicating the importance of minimizing this risk in reducing human infections.     

Microbiological baseline study of beef and pork carcasses from provincially inspected abattoirs in Alberta, Canada

In 2006 and 2007 beef and pork carcass swabs from provincially inspected abattoirs in Alberta, Canada were tested to determine the levels of total aerobic bacteria, coliform bacteria, and generic Escherichia coli, and the prevalence of Salmonella spp., Campylobacter spp., and Shiga toxin-producing E. coli (STEC). Swabs from beef and pork carcasses from 48 and 34 facilities, respectively, were analyzed. All samples tested were positive for aerobic bacteria with 99.8% of beef and 96.0% of pork samples, having total counts of ≤ 100 000 CFU/cm(2). Coliform bacteria were isolated from 22.4% and 42.0% of beef and pork carcass samples, respectively. Generic E. coli were recovered from 14.6% of beef and 33.7% of pork carcass samples. For beef carcasses, positive tests were obtained for 0.1% of 1036 samples tested for Salmonella spp., 1.5% of 1022 samples tested for Campylobacter spp. and 5.4% of 1018 samples tested for STEC. For pork carcasses, positive tests were obtained for 1.6 % of 1076 samples tested for Salmonella spp., 8.8% of 1070 samples tested for Campylobacter spp. and 4.8% of 1067 samples tested for STEC.     

Identification of Salmonella spp. isolates from chicken abattoirs by multiplex-PCR

The present study was carried out to report the occurrence Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in chicken abattoirs. Samples of feces; feathers; scald, evisceration, and chiller water; and rinse water of non-eviscerated, eviscerated, and chilled carcass were collected from six chicken abattoirs. Salmonella isolates were identified by a multiplex-PCR using three sets of primers targeting the invA, pefA, and sefA gene sequences from Salmonella spp., S. Typhimurium and S. Enteritidis, respectively. Salmonella spp. was detected in 10% (29/288) of the samples, whereas serovars Enteritidis and Typhimurium were identified in 62% (7/288), respectively. The results indicate the need to improve hygiene and sanitary standards in poultry slaughter lines, besides the education of food handlers and information to consumers.     

Microbiological evaluation of groups of beef carcasses: heifers and steers.

Numbers of mesophilic bacteria were estimated on carcasses of 25 heifers and 25 steers of beef breeds in a modern, high-line-speed abattoir. One side of each carcass from each sex was sampled at the end of the kill-floor, before the carcass wash, on each of 25 visits. Two adjacent excision samples (5 x 5 x 0.5 cm) were taken from each of ten sites and processed for automatic enumeration of aerobic bacteria on hydrophobic grid membrane filters. The effects of sex and carcass weight on bacterial counts were examined. Groups of carcasses were examined to determine the sample size required for future assessments of kill-floor hygiene. The log10 of the most probable number of growth units (MPNGU)/cm2 did not differ significantly between heifers and steers (average over the ten sites of 2.2) and there was no effect of carcass weight on bacterial counts for nine of the ten sites. There were, however, highly significant (p < 0.001) differences in the counts between sites and the counts from the ten sites clustered into five homogenous groups. The between-carcass component of variation at a site was generally larger than the within-carcass component. We conclude that, to estimate the mean log10 MPNGU/cm2 at a site to within +/- 0.5 units, future group-carcass evaluations require about 200 samples from 10 (two adjacent samples/site) or 20 carcasses (one sample/site).     

Effect of boiling water carcass immersion on aerobic bacteria counts of poultry skin and processed ground poultry meat

This study was conducted to determine the relationship between bacteria destruction on poultry carcass skin and bacteria in raw ground poultry meat from the same carcasses. Immersion time in boiling water of broiler chicken whole carcasses required for maximum reduction of naturally occurring aerobic bacterial count on skin was measured. Treatments for chicken carcasses consisted of immersion in boiling water (approximately 95 degrees C) for 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4 min. Four skin samples taken following treatment and three taken from subsequently ground carcass meat were analyzed for total aerobic plate counts (APC). Analysis of the data indicated a linear increase in bacterial destruction on skin with increased boiling water immersion time from 0 to 4 min. Reduction of skin bacteria to less than 1 log10 occurred at 3 min carcass immersion or longer. The analysis also indicated that treatment with boiling water and removal of skin was effective in reducing bacterial counts in ground meat to similar levels at all treatment times from 0.5 to 4.0 min. Findings from this study indicated that a boiling water immersion intervention and removal of skin could reduce subsequent bacteria contamination of ground meat. This intervention could minimize the risk of pathogen-contaminated primary processed poultry carcasses used in further processing.     

新疆拜城赛里木镇传统发酵酸乳中优势菌株的胃肠道耐受特性

通过体外耐酸、耐胆盐实验,从新疆拜城赛里木镇传统拉丝酸乳中的优势乳酸菌中筛选出耐受性和优良益生特性较强的乳酸菌菌株.结果表明:筛选出来的9株乳酸菌,在pH 2.0的条件下表现出-强的耐受能力,S5-8和S2-1L用人工胃液处理后的活菌数量在108 CFU/mL以上,其中S5-8有明显的增长趋势,其他7株菌株活菌数量在107CFU/mL以上,远远超过了106 CFU/mL以上的临界值,对9株强耐酸性乳酸菌进行耐胆盐实验.     

Improved diagnostic and real-time pcr in rapid screening for Salmonella in the poultry food chain

The goal of this study was to improve the diagnostic applicability of genus- and serovar- (S. Enteritidis and S. Typhimurium) specific PCR systems in screening faecal and caecal samples of poultry, poultry feed and poultrymeat for Salmonella, by keeping the opportunity to obtain Salmonella cultures from positive samples. Peptone broth pre-enrichment cultures of the samples were tested by PCR. In faecal and caecal samples from broiler chicks a strong inhibitory action was frequently observed. This could be reduced markedly by the addition of bovine serum albumin (BSA) acting as amplification facilitator. The results of testing pre-enrichment cultures from artificially contaminated faecal, poultry feed and poultrymeat samples (using S. Enteritidis, S. Typhimurium and S. Hadar as contaminants) suggest that the sensitivity of the above systems is 10(1)-10(2) CFU g(-1) sample. The testing of 95 caecal samples from slaughtered chicks resulted in 49% culture-positive and 76% PCR-positive samples. The suitability of a generic real-time PCR for testing faecal samples of poultry was also studied. Its detection limit for these samples was found to be lower than that of the diagnostic PCR system. Both methods reduced the time required for Salmonella detection to 24-30 h, and the advantage of the real-time PCR was its increased sensitivity. We have established a diagnostic and a real-time PCR system for rapid and reliable genus- and serovar- (S. Enteritidis and S. Typhimurium) specific detection of Salmonella for monitoring purposes in the poultry food chain. Sensitivity is equal to, or higher than, that of the standard bacterial culture method, and the method still provides the Salmonella culture if needed.     

The effect of commercial steam pasteurization on the levels of Enterobacteriaceae and Escherichia coli on naturally contaminated beef carcasses

The aim of the study was to assess the reduction achieved by steam pasteurization of beef carcasses of Escherichia coli, Enterobacteriaceae and total aerobic mesophilic plate counts (APCs). In total, 30 carcass halves were exposed to steam pasteurization (90 degrees C, 10 s exposure time) and the 30 corresponding carcass halves remained as untreated controls. The neck, midline and rump were sampled on each carcass half. Significant reductions in E. coli incidence (P < 0.05) and counts, 0.5 log10 CFU 1000 cm(-2) (P < 0.05), were observed on rump sites only. Significant reductions (>0.8 log10 CFU 1000 cm(-2)) of Enterobacteriaceae were observed at all carcass sites sampled (P < 0.05). Enterobacteriaceae reductions (>2 log10 CFU 1000 cm(-2)) were highly significant at the more contaminated sites (P < 0.001). Reductions in total APCs were inconsistent. Steam pasteurization significantly reduced the level of E. coli and Enterobacteriaceae at more contaminated sites, but did not result in complete decontamination. Therefore, steam pasteurization should be classed as an aid to hygienic beef processing, but not as a critical control point.     

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

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.     

Immune response to liposome-associated recombinant SEF21 following oral immunization in chickens

In order to generate Salmonella enterica serovar Enteritidis fimbriae antigens (rSEF21), the intact region encoding SEF21 was amplified from Salmonella Enteritidis by PCR and subcloned into a prokaryotic expression vector pET-28a(+) to yield pET-28a(+)-SEF21. The rSEF21 protein was highly expressed and purified by nickel affinity chromatography. Liposomeassociated rSEF21 was prepared for oral immunization to seek protective efficacy for intestinal infection with Salmonella Enteritidis. Evidence of IgA and IgG responses were found in the intestinal tracts and in the sera of a group of chickens immunized. Two weeks after the booster immunization, the chickens were challenged orally with 2 x 10(6) colony-forming units of live Salmonella Enteritidis, and fecal samples were examined for bacterial excretion from the intestinal tract. Significantly less fecal excretion of bacteria was observed in immunized chickens for 4 wk after challenge. The numbers of bacteria in the intestinal contents (cecum and rectum) were also significantly lower in immunized chickens than in unimmunized controls. Therefore, oral immunization with liposome-associated rSEF21 elicits both systemic and mucosal antibody responses, leading to a reduction in bacterial colonization in the intestinal tract and excretion of Salmonella Enteritidis in the feces.     

Survival of Salmonella typhimurium and Staphylococcus aureus in Genoa salami of varying salt concentration.

Genoa salami prepared using three different salt concentrations (2.0, 2.75 and 3.3%) were inoculated with 2.0 x 10(3) and 1.1 x 10(3) bacteria/g of Salmonella typhimurium and Staphylococcus aureus respectively. Over a period of 74 days samples were taken and analyzed for water activity and pH, counts of S. aureus and presence of Salmonella. After 11 days of dry-curing Salmonella could no longer be detected by preenrichment followed by selective enrichment procedures. Viable S. aureus were still found after 74 days of dry-curing. The results of this study would suggest that water activity and pH measurements are useful in evaluating the safety of dry-cured products.     

Detection and identification of salmonellas from poultry-related samples by PCR

A polymerase chain reaction (PCR) assay was developed for the generic detection of Salmonella sp. and the identification of S. Enteritidis (SE), S. Gallinarum (SG), S. Pullorum (SP) and S. Typhimurium (ST) in material collected in the field from poultry. The specificity and sensitivity of the assay combined with Rappaport-Vassiliadis selective enrichment broth (PCR-RV) were determined, and field samples were analyzed to verify the validity of the method application. Specificity of the assay was tested using 29 SE, 11 SG, 10 ST and 10 SP strains, along with 75 strains of 28 other Salmonella serovars and 21 strains of other bacterial genera. The assay was 100% specific for Salmonella detection and ST identification. The primer pair for SE, SG and SP also detected S. Berta. PCR detection limits for Salmonella at the genus level were 2 ST, 8 SE, 1.1x10(3) SG and 1.8x10(5) SP cells. At the serovar level, detection limits were 7 ST, 1.2x10(3) SE, 4.4x10(7) SG and 1.8x10(6) SP cells. At the genus level, PCR-RV detected approximately 128% more positive field samples than the standard microbiological techniques and results were ready in 48h instead of 7 days. PCR-RV method is diagnostic of Salmonella at the genus level and ST at the serovar level, although other tests are needed to identify SE, SG and SP to serovar level.     

Salmonella enterica serovar typhimurium colonization of the crop in the domestic turkey: influence of probiotic and prebiotic treatment (Lactobacillus acidophilus and lactose)

Acute colonization of the crop of the domestic turkey by Salmonella enterica serovar typhimurium (ST) was examined. The influences of preharvest probiotic and prebiotic treatment with lactobaccilli and lactose on crop colonization with ST were also investigated. Prior to Salmonella challenge, poults received 2.5% lactose and Lactobacillus acidophilus (1.9 x 10(9) organisms/liter) in the only source of drinking water from 1 day old to termination. At 3-wk-old, turkey poults were challenged with ST (1.7 X 10(8) colony-forming units [CFU]/ml) before their natural nocturnal fast to determine the potential effects of supplementation on crop colonization when the crop was engorged and subsequently undergoing emptying. Crop ingesta and tissue were collected at time points 30 min and 4, 8, and 24 hr postchallenge and ST levels were determined. High levels of ST were detected in the crop. For instance, for the poults not receiving lactose or lactobacilli, 30 min after ST challenge, there were 4.4 x 10(7) CFU in the crop ingesta and 5.3 x 10(5) CFU in the crop wall. Ingesta ST levels dropped dramatically to 1.0 x 10(6) CFU after 4 hr as the crop emptied. Crop wall ST levels were steady during the nocturnal crop evacuation. Immunohistochemical staining demonstrated ST in close association with the crop epithelium. Treatment with lactose and L. acidophilus supplementation did not reduce ST colonization.     

Major microbiota of lactic acid bacteria from Matsoni, a traditional Georgian fermented milk

A total of 26 samples of Matsoni were collected in Georgia. From these samples 80 strains of lactic acid cocci and 173 strains of lactobacilli were isolated. The number of lactic acid bacteria varied between 10⁵ and 10¹⁰ colony forming unit (CFU)/mL. All the isolated lactic acid bacteria were thermophilic bacteria that could grow at 45°C. The predominant lactic acid bacteria were Streptococcus thermophilus and Lactobacillus delbrueckii spp. bulgaricus in 25 samples of Matsoni, while Lactobacillus helveticus was also a predominant species, together with the two previous species in one sample of Matsoni. We showed there was diversity in both S. thermophilus and L. delbrueckii spp. bulgaricus at the strain level by randomly amplified polymorphic DNA analysis.     

Effects of chlorination of chill water on the bacteriologic profile of raw chicken carcasses and giblets.

In March 1989, the USDA Food Safety and Inspection Service sampled raw chicken carcasses and giblets at a federally inspected slaughter establishment in Puerto Rico to determine the effects of adding chlorine to carcass and giblet chill water on bacterial contents of raw poultry products. Over four 8-hour workdays, 200 carcass rinse samples were collected at 3 sites in the establishment; 39 giblet rinse samples were collected at 1 site. Analyses of the carcass rinse samples indicated that carcasses had average aerobe plate counts of log10 3.20 before chilling and 2.51 after chilling; Enterobacteriaceae counts of log10 2.57 before chilling and 1.75 after chilling; and Escherichia coli counts of log10 2.04 before chilling and 1.20 after chilling. Salmonellae were found on 43% of the carcasses before chilling and on 46% after chilling. Analyses of the giblet and neck rinse samples indicated that raw giblets and necks after chilling had average aerobe plate count of log10 3.49, Enterobacteriaceae count of log10 2.57, and E coli count of log10 1.06. Salmonellae were found on 12% of the giblets and necks sampled. Results compared favorably with giblet and neck rinse sample results obtained during a baseline sampling study in November and December 1987. The baseline results indicated aerobe plate count of log10 3.72; Enterobacteriaceae count of log10 2.90; E coli count of log10 1.14; and salmonellae on 69% of the giblets and necks sampled. Placing raw chicken carcasses in chlorinated chill water reduced aerobe, Enterobacteriaceae, and E coli plate counts. Prevalence of carcasses with salmonellae remained nearly the same.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative Study of Microbiological Quality of Raw Poultry Meat at Various Seasons and for Different Slaughtering Processes in Casablanca (Morocco)

The aim of the present study was to evaluate the bacterial quality of chicken (n = 96) and turkey (n = 96) marketed in Casablanca, Morocco. Poultry samples (n = 192) were collected randomly from traditional shops and supermarkets during 2 sampling periods (hot and cold seasons). The samples were analyzed for the presence and counts of various bacteria. Results indicated that aerobic plate counts and fecal coliforms were particularly high in all the samples analyzed. Escherichia coli, coagulase-positive Staphylococcus, Clostridium perfringens, Salmonella, and Listeria monocytogenes were detected, respectively, in 48.4, 10.4, 7.2, 1.6, and 0.5% of the poultry meat samples. The chicken and turkey samples contained, respectively, 25 and 33.3% of bacteria above the maximum limits established by the Moroccan regulatory standards. The highest bacterial counts in poultry meat product samples were recorded with the traditional slaughtering process during the hot season (P < 0.05). These high levels of microbial contamination and occurrence of pathogenic bacteria reflect the poor hygienic quality of poultry meat under these conditions.     

Isolation and characterisation of Salmonella in a turkey production facility

1. A comprehensive ecological survey was conducted from April 1997 to June 1999 on 4 turkey flocks (F1 to F4) to identify key pre-harvest sources/vectors of Salmonella colonisation. 2. Turkey caecal and crop content, litter, drinker, air, feed, feeder and environmental swab samples were collected. Conventional microbiological and serological procedures were used to isolate, identify, and confirm the presence or absence of Salmonella. 3. Salmonella was isolated from 13% of litter, 11% of turkey caeca, 10% of drinker, 5% of environmental swab, 3% of feed and 1% of feeder samples. Salmonella heidelberg (65%), S. senftenberg (19%), S. muenster (10%), S. anatum (3%), and S. worthington (3%) were identified. 4. Identifying environmental sources associated with Salmonella colonisation and characterising serotypes would assist in designing pre-harvest controls for this poultry-borne pathogen. Integrators and poultry producers may be able to design hazard analysis and critical control point (HACCP) protocols to reduce the incidence of Salmonella arriving at the processing plant.     

Lactic acid as a decontaminant in slaughter and processing procedures

An attempt was made to interrelate the data obtained in experiments conducted by our Department along beef, veal and pig slaughter lines, using lactic acid (LA) for the decontamination of carcasses, cold and hot boned primal cuts, slaughter byproducts, and butcher's knives. First and foremost it was observed, that provided Good Manufacturing Practices are strictly followed, the microbial load of carcass surfaces will be substantially reduced. LA-decontamination may result in an additional reduction. Since in the early post-mortem period bacteria are not yet attached to the meat surface, LA-decontamination should preferably be applied to the hot carcass. It was demonstrated that, dependent on mode and duration of application, LA sprays not exceeding 1% v/v (beef), 1.25% v/v (veal) and 1.5% v/v (pork) resulted in acceptable carcass colour scores. Blood spots, which are particularly prone to discolouration by lactic acid application, should be removed at an early post-mortem stage e.g. by strong showering. The difference in surface pH between LA-treated and control carcasses disappeared within 72 hours post-mortem. Veal longissimus chops treated with LA solutions up to 2% v/v were not identified by a consumer taste panel as significantly different from controls. The 'immediate' bactericidal effect of LA-decontamination for beef, veal and pig carcasses, as well as for pig liver and veal brain, amounted to approximately 1.5 log cycles for the aerobic colony counts, strongly dependent on substrate and conditions of decontamination. In addition, a 'delayed' bacteriostatic effect was observed during storage, which is probably the result of a prolonged lag phase of acid-injured micro-organisms surviving lactic acid decontamination.(ABSTRACT TRUNCATED AT 250 WORDS)     

Protection conferred by a live Salmonella Enteritidis vaccine against fowl typhoid in laying hens

Fowl typhoid is under control in poultry farms of developed countries, but it still endemically subsists in commercial laying hen farms of some countries. It has been demonstrated that Salmonella live vaccines can elicit cross-immunity against members of the same Kauffmann-White scheme serogroup. In this work, we explored the protection conferred by TAD Salmonella vac E, a live Salmonella enterica serovar Enteritidis vaccine, against fowl typhoid. Three groups of laying hens were vaccinated with different vaccination schedules starting on the first day of life, and afterwards were infected with 2 x 10(5) CFU of a virulent Salmonella Gallinarum strain, either at wk 28 or wk 52. Mortality, fecal shedding, and organ invasion of Salmonella Gallinarum were assessed. In this work we demonstrated that this Salmonella Enteritidis vaccine is able to cross-immunize against Salmonella Gallinarum. At wk 28, hens vaccinated with three oral doses or with two oral doses combined with one subcutaneous dose were protected by the vaccine. At wk 52, when hens were infected 36 wk after the final immunization, the vaccine was not able to confer protection. Thus, revaccination every 3 mo would be highly recommended. In countries where Salmonella Gallinarum subsists together with Salmonella Enteritidis, control programs should include vaccination of laying hens using safe attenuated Salmonella strains.     

Comparison of real-time PCR with conventional PCR and culture to assess the efficacy of a live attenuated Salmonella enterica serovar Typhimurium vaccine against Salmonella enterica serovar Enteritidis in commercial leghorn chicks vaccinated under field and laboratory conditions

The efficacy of a live attenuated Salmonella Typhimurium Megan Vac 1 vaccine (MV1) was evaluated against Salmonella Enteritidis in chicken pullets with the use of PCR and culture methods. Two hundred Hyline W-32 white leghorn chicks were obtained from a local hatchery and divided into four treatment groups. Two of the groups served as positive and negative controls. The MV1 vaccine was administered to the chicks in the remaining two groups at 1 and 35 days old by either the coarse spray (field) or the oral route (laboratory) method. The chicks were challenged with a high dose of a Salmonella Enteritidis strain at 10 wk old and euthanatized 3 days postinoculation. Samples for PCR analysis were collected prior to enrichment, after pre-enrichment in buffered peptone water (BPW) and after primary enrichment from the ceca, liver, and spleen. None of the samples tested yielded positive results for the Salmonella Typhimurium vaccine strain by either the culture or PCR methods. Results from the standard culture method showed that vaccinating the birds with MV1 reduced the counts of Salmonella Enteritidis recovered from the challenged birds. In addition, fewer pre-enriched samples tested positive for Salmonella Enteritidis among the challenged groups that were vaccinated when compared to the unvaccinated challenged group. Under the conditions of this study, MV1 was unable to prevent colonization of other internal organs such as the liver and spleen. Real-time PCR was significantly more sensitive than conventional PCR (C-PCR) prior to enrichment, but after enrichment the sensitivities of the two methods were similar. Enrichment significantly increased the sensitivity of both PCR methods for the detection of Salmonella Enteritidis in cecal samples, but did not significantly increase the sensitivity for detection of Salmonella Enteritidis in liver and spleen samples that were pre-enriched in BPW. There was no significant difference between the laboratory or field vaccination methods with respect to either the prevalence of Salmonella Enteritidis isolation or the bacterial loads in culture-positive samples. Collectively, the data suggest that MV1 offered some protection against Salmonella Enteritidis in commercial layer chick pullets under the conditions of this study. Given the labor and time required to perform the C-PCR and culture methods, the real-time PCR method may prove to be a more useful method to use in diagnostics.