Observations on Salmonella contamination of commercial laying farms before and after cleaning and disinfection

Little is known about the effectiveness of the cleaning and disinfection methods in use on commercial laying farms in Great Britain. Samples were taken from poultry house structures and equipment of five cage layer flocks, five barn egg production flocks and two free-range flocks. In the free-range houses there was a decrease in Salmonella after cleaning and disinfection, although the soil in the paddocks remained contaminated. In the barn and especially the cage layer houses, significant residual contamination remained on the surfaces of buildings and equipment. Wildlife pests were also found to be carrying Salmonella in the disinfected houses and free-range paddocks.     

Detection of specific antibodies against deflagellated Salmonella Enteritidis and S. Enteritidis FliC-specific 9kDa polypeptide

Specific antibody levels of laying hens and young chickens experimentally infected with Salmonella Enteritidis and vaccinated farm flocks were evaluated by enzyme-linked immunosorbent assays (ELISAs) with two different antigens, deflagellated S. Enteritidis whole cell (DEWC) and S. Enteritidis FliC-specific 9kDa polypeptide (SEP9). Infected laying hens excreted S. Enteritidis throughout the experimental period, and the specific antibody titers in DEWC-ELISA, were significantly higher than the uninfected group. It suggests that this DEWC-specific antibody will serve as an effective indicator of S. Enteritidis infection, especially for non-vaccinated laying flocks. SEP9-specific antibodies were detected in spray-inoculated young chickens but not in oral-inoculated young chickens. Compared with greatly high SEP9-specific antibody levels of vaccinated farm flocks, no response was observed in orally infected hens. These results indicate that S. Enteritidis discontinues expressing SEP9 once it has crossed the intestinal barrier, and that SEP9-ELISA will serve as a valuable monitoring tool for the status of S. Enteritidis vaccination on a flockwide basis, independent of stable S. Enteritidis infections.     

Occurence of salmonellae in laying hens in different housing systems and inferences for control

As eggs represent now as ever the most important source for Salmonella infection in human beings and because of the currently occurring shift in housing conditions for laying hens from conventional cages to alternative systems it was studied whether the Salmonella prevalence in layers is influenced by the housing system. Following systems were considered: organic farming with free range management systems, floor management systems with free range, floor management systems without free range, conventional cages. 453 pooled faecal samples as single or double examination per herd from 329 flocks in different housing systems for table egg production from three Federal Lander were examined bacteriologically. The share of layer flocks which were Salmonella positive at least once independently of the housing system amounted to 32.2%. Analysis of the Salmonella findings in the single housing systems revealed that the share of Salmonella positive flocks was higher in conventional cage systems (46.3%) than in alternative housing systems (32.996% in organic farming with free range management systems, 21.9% in floor management systems with free range, 23.4% in floor management systems without free range).The results of the study clearly show that Salmonella Enteritidis (mostly phage type 4, other phage types rarely) presents with a share of 78% the dominant serovar in laying hens.The total number of all other serovars (apart from Salmonella Enteritidis and subspecies I rough) reached a share of ca. 14%, however, no other single serovar was dominant. As Salmonella Enteritidis is the predominant serovar in laying hens it is strongly recommended to use Salmonella Enteritidis vaccines for immunisation programmes of chickens during the rearing period. Because of the high prevalence of Salmonella organisms in the different housing systems, detailed information on the epidemiology of Salmonella in laying hens are needed to introduce effective control measures. Of particular interest is the question whether the Salmonella findings in laying flocks are the result of multiplication of already existing Salmonella organisms in the animals or whether the bacteria are introduced only during the laying period.     

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.     

Bayesian estimation of flock-level sensitivity of detection of Salmonella spp., Enteritidis and Typhimurium according to the sampling procedure in French laying-hen houses

A study was carried out to estimate the prevalence of flocks infected by Salmonella spp., S. Enteritidis and S. Typhimurium in 521 French laying-hen farms from October 1st 2004 to September 30th 2005 as part of a European Union-wide baseline study to define targets for Salmonella reduction in member states. The sampling scheme prescribed and financed by the European Commission to detect Salmonella in laying-hen flocks was based on 2 dust-samples and 5 faeces-samples per farm. A latent-class Bayesian approach for correlated tests was used to estimate the sensitivity of detection of reduced sampling schemes corresponding to the 16 combinations of 2 dust- and 5 faeces-samples. For each model the full sampling scheme (7 samples) and the reduced protocol were considered as two correlated tests, the biological principle being identical and the reduced protocol being a subset of the full sampling scheme. As the observed apparent prevalence in cage flocks was higher than in other systems (barns, outdoor, or organic) these two sub-populations were considered separately. Bayesian estimation of posterior medians with 95% probability intervals for true prevalence in cage flocks were 0.34 (0.29; 0.39) and 0.13 (0.10; 0.18) for Salmonella spp. and Salmonella Enteritidis+Typhimurium respectively. In alternative flocks posterior medians with 95% probability intervals for true prevalence were 0.09 (0.06; 0.13) and 0.05 (0.03; 0.08) for Salmonella spp. and Salmonella Enteritidis+Typhimurium, respectively. In cage flocks Bayesian estimation of posterior distributions for sensitivity indicated that at least 5 samples, including 2 dust samples were necessary to attain comparable sensitivity levels to the full sampling scheme. In alternative flocks and for Salmonella spp. 6 samples were required to ensure a comparable sensitivity level to the full sampling scheme. Detection sensitivity was improved by increasing the number of dust samples in cage farms and by increasing the total number of samples whatever their type in alternative farms.     

Comparison of the prevalence of Salmonella infection in layer hens from commercial layer farms with high and low rodent densities

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

On-farm risk factors for Salmonella Enteritidis contamination

Forty layer farms from 2 states participated in a study to examine the risk factors and incidence of Salmonella Enteritidis from multiple samples, including environmental drag swabs from the bird areas, feed, water, flies, rodents, live rodent traps, and environmental swabs from areas occupied by other livestock. Twenty-four of these farms had between 3,000 and 31,000 bird flocks (medium-sized flocks) and 16 had less than 3,000 birds (small-sized flocks). All were housed in cage-free production systems. Twenty-two farms included outside pasture areas for the birds. Most of the participants had just come under the FDA Egg Rule and had not yet tested their flocks (flocks under 3,000 birds are exempt) for Salmonella Enteritidis. Many, however, obtained their pullets from commercial Salmonella Enteritidis-clean breeder sources hatched in National Poultry Improvement Plan hatcheries. Vaccination against Salmonella Enteritidis was performed on 21 of the 40 farms (combination of live and killed vaccines). Salmonella Enteritidis was detected on 7 out of the 40 farms, primarily in rodents, their feces, or from swabs taken inside live traps. Of these 7 Salmonella Enteritidis-positive farms, 3 farms that had vaccinated their pullets with live Salmonella Typhimurium vaccine and killed-Salmonella Enteritidis vaccine; no Salmonella Enteritidis was isolated from the environmental drag swabs taken from the bird area or from the eggs on these farms. However, on the farms that had not vaccinated for Salmonella Enteritidis, the organism was isolated from 4 environmental drag swabs and 3 egg pools. The last 4 farms had flocks under 3,000 birds. No Salmonella Enteritidis was isolated from any of the samples of feed, flies, water, or swabs taken from other livestock areas. Based on the initial findings in this study, we suggest the 2 most important risk factors for Salmonella Enteritidis contamination inside the bird area and in the eggs in these small- and medium-sized flocks are the presence of infected rodents and the absence of an Salmonella Enteritidis vaccination program.     

Assessment of cleaning and disinfection in Salmonella-contaminated poultry layer houses using qualitative and semi-quantitative culture techniques

Salmonella infection of laying flocks in the UK is predominantly a problem of the persistent contamination of layer houses and associated wildlife vectors by Salmonella Enteritidis. Methods for its control and elimination include effective cleaning and disinfection of layer houses between flocks, and it is important to be able to measure the success of such decontamination. A method for the environmental detection and semi-quantitative enumeration of salmonellae was used and compared with a standard qualitative method, in 12 Salmonella-contaminated caged layer houses before and after cleaning and disinfection. The quantitative technique proved to have comparable sensitivity to the standard method, and additionally provided insights into the numerical Salmonella challenge that replacement flocks would encounter. Elimination of S. Enteritidis was not achieved in any of the premises examined although substantial reductions in the prevalence and numbers of salmonellae were demonstrated, whilst in others an increase in contamination was observed after cleaning and disinfection. Particular problems with feeders and wildlife vectors were highlighted. The use of a quantitative method assisted the identification of problem areas, such as those with a high initial bacterial load or those experiencing only a modest reduction in bacterial count following decontamination.     

Observations on the distribution and persistence of Salmonella enterica serovar enteritidis phage type 29 on a cage layer farm before and after the use of competitive exclusion treatment

1. A continuously occupied cage layer house, which had been linked with a human outbreak of Salmonella Enteritidis PT29, was investigated to assess the distribution of contamination and the options for control. 2. The presence and persistence of Salmonella before and after application of competitive exclusion (CE) treatment was investigated by culturing samples from faeces, the environment, spent hens and eggs, and use of an ELISA to detect egg yolk antibodies. 3. A high prevalence of Salmonella was found in faecal and environmental samples before CE treatment was used but this reduced to minimal levels after treatment. 4. Egg yolk antibody assay suggested that although treated birds showed reduced excretion of Salmonella there was no difference between these and non-treated birds in terms of seroprevalence. 5. Contamination of the egg packing plant disappeared following CE. 6. Chicks and pullets in separate accommodation on site remained Salmonella free throughout despite no precautions being taken to avoid transmission of infection from the laying flock. 7. The rapid and substantial reduction in Salmonella in faeces, birds and the environment following the introduction of CE treatment suggests that further controlled field studies would be justified.     

Salmonella in poultry flocks and humans--S. enterica subspecies enterica serovar Enteritidis in the past

After importing of breeder lines for laying flocks from Canada into the former GDR in 1966 the egg industry in this country was completely isolated from that in Western Germany or other Western European countries until opening the border in Germany in 1989. Because of this isolation from other countries, an analysis of the clonal diversity of Salmonella (S.) Enteritidis isolates originated from humans, chickens and food in the former GDR during the 1980s would provide a unique opportunity to obtain new insight into factors that may have triggered the S. Enteritidis epidemic. While isolates had previously been typed by the phage typing scheme of Lalko and Laszlo we applied for the first time the extended phage typing scheme by Ward for the retrospective analysis of the S. Enteritidis strains. Furthermore, isolates of phage type (PT) 4/6 (Ward / Lalko and Laszlo) from different livestocks were investigated by ribotyping. Although in total the PT4/6 dominated between 1986 and 1989 in poultry, other phage types have prevailed in the early 1980s and represented a considerable fraction of isolates until 1989. For instance, PT8/7 was isolated from one large layer farm (district Halle) from 1988 until 1989. During that time in another farm (district Cottbus) only PT8/7 was detected too. PT4/6 was isolated from neither of these two laying hen farms. The strains of PT4/6 could be distinguished by ribotyping in 19 different subtypes. The strains from the northern farms were distinct from those isolated in the southern regions. As farms which were harbouring either PT4/6 or PT8/7 had obtained laying hens from the same sources (breeder lines in Deersheim and Spreenhagen) it is highly probable that S. Enteritidis infection was acquired from the environment at each individual farm. This conclusion is also supported by the presence of different PT4/6 ribotypes in different farms. The presence of different phage types or PT4/6 ribotypes at different farms of laying hens suggests that in each case the S. Enteritidis strains present in the environment were able to enter chicken flocks.     

A case study on Salmonella enteritidis (SE) origin at three egg-laying farms and its control with an S. enteritidis bacterin

In the early 1990s, three egg-laying farms (farms S, T, and B) were thought to have the possibility of Salmonella enteritidis (SE) contamination because positive liquid egg samples originated from those farms. The present study was therefore conducted. The first clarification for SE contamination was the study on the origin of SE contamination including its vertical transmission. The results of SE contamination profiling with dust and manure, food materials, dead embryos, and residual yolks on hatch day in regular monitoring over a few years were clearly negative. Therefore, we concluded the SE transmission/infection was attributed to horizontal infection in the egg-laying farms but not vertical transmission from parental stock, hatcheries, growth, or food materials during a 7-yr experimental period. Second, we attempted to clarify if administration of an SE bacterin (Layermune SE) to growth flocks for the egg-laying farms could reduce SE incidence in liquid egg samples from each egg-laying farm. In the first experiment, we compared SE incidence in liquid egg samples from vaccinated and nonvaccinated flocks (similar age flocks). SE incidence from vaccinated and nonvaccinated flocks showed negative and <2 most probable number (MPN)/100 ml for farm B, <2 and >1600 MPN/100 ml for farm S, and negative and >1600 MPN/100 ml for farm T, respectively. In the second experiment, we compared the SE isolation incidence in the liquid egg samples from nonvaccinated and newly replaced vaccinated flocks in the same chicken houses from each of the three egg-laying farms. SE incidence in the liquid egg samples was similar to that in the first experiment. Therefore, the SE bacterin may play an important role in reducing the SE incidence of liquid egg samples.     

Effects of Salmonella enterica subsp. enterica serovar Enteritidis vaccination in layer hens subjected to S. Enteritidis challenge and various feed withdrawal regimens

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

Vaccination against Salmonella enteritidis in Dutch commercial layer flocks with a vaccine based on a live Salmonella gallinarum 9R strain: evaluation of efficacy, safety, and performance of serologic Salmonella tests

This study describes a field trial in which 80 commercial layer flocks, with an increased risk of Salmonella enteritidis (SE) infection and placed on farms with a certified Standardized Biosecurity Programme (SBP) or a request for a SBP certificate, were vaccinated with a vaccine based on a live attenuated Salmonella gallinarum (SG) 9R strain. An evaluation is presented of the efficacy of the vaccine against SE infections, the effect on the performance of serologic Salmonella tests, and the spread of the vaccine strain to the egg content. For the efficacy study, assessment of the flock level occurrence of SE infections in the vaccinated group of 80 flocks was compared with that of a nonvaccinated group of 1854 flocks hatched in the same period. This control group was examined according to the compulsory control programme in The Netherlands. An evaluation was done of the performance of serologic Salmonella tests and the spread of the vaccine strain to the inner egg content of five of the vaccinated flocks. Findings demonstrated the flock level occurrence of SE infections in the vaccinated group (2/80 = 2.5%) to be significantly (P = 0.01) lower than that of the nonvaccinated group (214/1854 = 11.5%). Vaccination resulted in 59.0% positive test results in lipopolysaccharide BD enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against Salmonella serogroups B and D and 0% positive test results in the rapid plate agglutination test for detecting antibodies against S. pullorum (SP)/SG. The mean specificities of two blocking ELISAs (gm- and i-double antibody sandwich ELISAs) based on the flagellar antigen of SE and Salmonella typhimurium (ST) on the same sera were 99.6% and 96.1%, respectively. The vaccine strain could not be isolated from any of the 450 pools of 10 eggs. On the basis of these results, we concluded that vaccination with a vaccine based on an attenuated SG 9R strain contributes to the reduction of SE infections in commercial layer flocks. Furthermore, serologic monitoring of SE, ST, and SP/SG can still be carried out on flocks vaccinated with an attenuated SG 9R strain. Additionally, we found no indication of the spread of the vaccine strain to the egg content.     

Evaluation of a French ELISA for the detection of Salmonella Enteritidis and Salmonella Typhimurium in flocks of laying and breeding hens

In France, the regular and compulsory detection of Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) in flocks of breeding and laying hens is based on bacteriological examination of environmental swabs and faeces samples. The aim of this study was to compare this bacteriological examination with a serological method (ELISA) developed in our laboratory. This ELISA was first evaluated by use of artificially infected hens. During these experimental infection studies, several groups of hens were inoculated with SE, ST, different vaccines and different Salmonella serovars to calculate the experimental parameters of our ELISA. Then, in a field study, 43 flocks were followed monthly using two bacteriological samples (environmental swab and pool of faeces) and 20 serological samples (sera or yolks). Twenty-seven flocks without SE or ST gave a negative serological response throughout their surveillance. Among the 10 various serovars different from SE and ST isolated in this study, S. Heidelberg, S. Agona and S. Hadar gave seropositive results in seven flocks. Consequently, this ELISA was not specific of SE and ST as it detected serovars sharing or not common antigens with SE and ST. Seropositive results were also obtained each month for two flocks where no Salmonella could be isolated. Finally, in seven flocks found infected with SE or ST, the positive ELISA results appeared later than the bacteriological detection. Therefore, for the detection of chicken flocks recently infected with SE or ST, bacteriological examination currently used in France seems to be more appropriate than this ELISA.     

Environmental contamination and detection of Salmonella enterica serovar enteritidis in laying flocks.

Faecal, dust and other environmental samples were collected from the floors, droppings belts, egg-collection systems and other areas of 14 cage-layer flocks, 10 barn egg production flocks and seven free-range flocks, and cultured for Salmonella species. The distribution of the organism varied with its prevalence and with the vaccination status of the birds. No one sample type was found to be suitable for identifying all contaminated houses. Salmonella was also frequently found on egg-packing equipment and in samples from rodents and wild birds.     

Vaccination against infectious anemia of poultry (CAA)--results of field studies]

Infectious anemia of poultry is a disease of high economical significance. Connatal infection of chicks with the chicken anemia agent (CAA) via the embryonated egg causes anemia along with severe immunosuppression, thus rendering the chicken susceptible for secondary infections. In order to prevent infection of young chicks, it is necessary to induce immunity against CAA in parent flocks, with the aim to prevent connatal spread of the infection and provide maternal protection for baby chicks. In this publication, the efficacy and use of a live CAA vaccine is reported. From autumn 1986 until summer 1990, 3 experimental vaccine charges were applied in 85 broiler parent flocks with totally 3.1 million chickens. In addition, totally 293,000 broiler breeder and 171,000 layer breeder chicken were vaccinated in 1989/90. The vaccine was administered between the 13th and 19th week of life by drinking water without adverse effect to the birds. Chicken anemia symptoms were observed only at the begin of laying period in two parent flocks. These flocks had been vaccinated in the 17th and 19th week, respectively. The offsprings of all other vaccinated parent flocks remained free of chicken anemia. Day-old chicks derived from vaccinated parent flocks were protected against CAA challenge infection. It is emphasized, that vaccination should be performed within the 13th to 15th week of life, because according to our observations, this will lead to an immediate seroconversion.     

Infection dynamics of Ascaridia galli in non-caged laying hens

The infection dynamics of Ascaridia galli in laying hens was investigated in six commercial non-caged flocks. Three flocks were managed in accordance with the regulations for organic production and had outdoor access, whereas three flocks were housed indoors in aviaries or traditional floor systems. Faecal egg counts and total worm burdens were determined at specified intervals during the first 50 weeks of the production period. In two conventional flocks the efficacy of flubendazole on lumenal stages was investigated. All flocks became infected following the arrival of the birds (post placement) with residual infective eggs derived from the previous flock. In four flocks (two organic and two conventional) parasite eggs were first detected in faeces 6-7 weeks post placement, whereas parasite eggs were not detected until after 17-18 weeks in two flocks. This delay was observed in two of three flocks that were housed in barns that had been thoroughly cleaned and disinfected by chlorocresol. In three flocks (two conventional and one organic) flubendazole was administered to the birds in the drinking water for approximately one week. Both conventional flocks were dewormed twice approximately 20 weeks apart, whereas the organic flock was dewormed only once about 40 weeks post placement. Parasite eggs reappeared after deworming in all flocks, often within 2-4 weeks, followed by a rapid increase in parasite egg expulsion. Our results suggested impairment of host immunity post treatment, as the egg counts exceeded pre-treatment levels after 7-8 weeks on both conventional farms. Accordingly, the way by which anthelmintics and/or disinfectants are used in non-caged chicken flocks must be refined.     

The spread of Salmonella gallinarum 9R vaccine strain under field conditions.

A live vaccine based on an attenuated Salmonella gallinarum 9R strain is in use in a Salmonella enteritidis control program in commercial layer flocks in The Netherlands. In a field study, the potential spread of the vaccine strain from vaccinated flocks to nonvaccinated flocks has been studied after both the primary and the booster injection at four different rearing farms and at one layer farm. The vaccinated and the nonvaccinated flocks were monitored at regular intervals by bacteriologic and serologic examination. In this field study, no evidence was found for the fecal spread of the vaccine strain.     

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.     

Detection of Salmonella in spent hens and eggs associated with foodborne infections

About 16,000 spent hens from 23 farms in the northern area of Japan were purchased in 1996, 1997, 1998, and 1999 to isolate Salmonella in two poultry processing plants. Salmonella was detected in 12 of 23 farms (52.2%). In particular, the serotypes Enteritidis and Infantis were detected in four and three farms, respectively. The prevalence rates in the hens' ceca, immature eggs, and the yolk of mature eggs in oviducts were 14%, 7.2%, and 6.8%, respectively. A total of 23 serotypes were detected. The major serotypes of the strains were Enteritidis, Corvallis, Typhimurium, and Infantis, but most of the strains were untypable. In the same area during 1992 to 1996, Salmonella was detected in eggs associated with four outbreaks of Salmonella Enteritidis infection and one outbreak of Salmonella Infantis infection. The ratio of contamination was approximately 1%, and the level was estimated to be 93 MPN(most probable number)/100 g in one outbreak. In farms that produced the eggs associated with all of the five outbreaks of Salmonella, the serotype Enteritidis or Infantis was isolated from hens. Farms where Salmonella was not detected were not related to any of the outbreaks.