The role of litter beetles as potential reservoir for Salmonella enterica and thermophilic Campylobacter spp. between broiler flocks

We evaluated the role of beetles infesting broiler chicken rearing facilities as potential reservoirs for Salmonella enterica infections between successive broiler flocks. In addition, their role as potential reservoirs for thermophilic Campylobacter spp. was also investigated. Fourteen broiler houses located at 11 different farms were included in the study. The houses were nonrandomly selected on the basis of their salmonella status; nine were persistently contaminated with salmonella whereas five were salmonella negative. For each broiler house, two consecutive broiler flocks (i.e., 28 broiler flocks in all) as well as beetles collected during both rotations of production and in the empty period (after cleaning and disinfection) between these flocks were monitored for the presence of salmonella. Examinations for the presence of campylobacter in the same sample materials were also performed. Beetles sampled during production were positive for salmonella or campylobacter or both. Furthermore, in one house, the occurrence of Salmonella indiana in two consecutive broiler flocks coincided with the presence of S. indiana-contaminated beetles in the empty period between the flocks. The genotype of the identified S. indiana was in all cases identical when analyzed by pulsed-field gel electrophoresis. However, our results also suggest that salmonella from beetles may not always be transmitted to the chickens and that beetles living in contaminated houses can remain free of infection. All cases of campylobacter-positive beetle samples were detected in connection with a positive chicken flock; in no case was campylobacter isolated from beetles taken from the empty period between rotations. Four beetle species were identified during this study. Alphitobius diaperinus was found in all houses and was relatively abundant in most. Typhaea stercorea and Ahasverus advena were found in eight and nine houses, respectively, and were abundant in most of these. Carcinops pumilio was found in small numbers in eight houses. No other insect species was identified. These investigations have shown that beetles in broiler houses infrequently are positive for salmonella. However, transmission of S. indiana between two consecutive broiler flocks can coincide with the presence of salmonella-contaminated beetles in the empty period, indicating that the beetles were the reservoir of S. indiana between the two flocks. Concerning campylobacter, the results suggest that beetles do not play a significant role as a reservoir of campylobacter from one rotation to the next.     

Detection of antibody to Salmonella enteritidis and S typhimurium in the yolk of hens' eggs

Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect IgG antibodies to Salmonella enteritidis and S typhimurium in the yolk of hens' eggs. Better discrimination and more consistent results were obtained between eggs from experimentally infected and uninfected hens by using saline-dilution of yolk rather than chloroform extraction. Threshold absorbance values were determined in three salmonella-free flocks, and on the basis of these results ELISA optical density values greater than 0.25 were considered to be positive for antibodies to salmonella. Four flocks with a history of salmonella infection were examined; three contained birds which were seropositive for S enteritidis by ELISA and from which S enteritidis was isolated, and a large proportion of eggs from these birds contained antibody to S enteritidis. Eggs from the fourth flock had no detectable antibody, although serum antibody was detected in some birds. No salmonellae were isolated from the yolks of the eggs from any of the four flocks.     

Experimental horizontal transmission of Salmonella enteritidis strains (phage types 4, 8, and 13a) in chicks

The recent isolation of phage type 4 Salmonella enteritidis strains from poultry and humans in the United States has generated considerable concern because this phage type is predominant in both animals and humans in many other nations. Understanding whether the presence of these strains in poultry flocks poses an elevated threat to public health is a critical issue for developing effective disease control programs. The present study evaluated whether S. enteritidis strains of various phage types found in poultry in the United States (phage types 4, 8, and 13a) differed in their potential for horizontal transmission from experimentally infected chicks to uninoculated chicks housed in the same isolator units. Five days after two seeder chicks in each group of 12 were inoculated with oral doses of approximately 10(3) S. enteritidis cells at 8 days of age, ceca and livers were sampled from seeder chicks and from their contact-exposed penmates. On the basis of the detection of S. enteritidis in cecal samples, phage type 4 strains were transmitted horizontally at a significantly lower frequency than were strains of other phage types. Nevertheless, two of three phage type 4 strains evaluated were very highly invasive.     

Salmonella enteritidis and other Salmonella in laying hens and eggs from flocks with Salmonella in their environment.

Seven Canadian layer flocks with Salmonella enteritidis in their environment were investigated to determine the numbers of hens infected with S. enteritidis, the localization of S. enteritidis in organs of infected hens and the numbers of S. enteritidis-infected eggs produced by two affected flocks. By a microagglutination test (MAT) using S. pullorum antigens, these flocks had more seropositive hens (mean 51.9 +/- 16.9%) than two Salmonella-free flocks (mean 13.0 +/- 4.2%). Culture of tissues of 580 hens (433 seropositive) from the seven flocks detected 26 (4.5%) S. enteritidis-infected hens from two flocks. In one flock, 2/150 hens were infected with S. enteritidis phage type (PT) 8, which was confined to the ceca, and no Salmonella spp. were isolated from 2520 eggs (one day's lay). In the second flock, where 24/150 hens were infected with S. enteritidis PT13, extraintestinal infection was found in nine hens and involved the ovaries and/or oviduct in two hens. Salmonella enteritidis PT13 was isolated from one sample of egg contents and from one sample of cracked shells from among 14,040 eggs (one day's lay) from this flock. The overall prevalence of S. enteritidis-contaminated eggs from the two flocks with infected hens was less than 0.06%. Other Salmonella spp. isolated were S. heidelberg from 58 hens (10%), and S. hadar, S. mbandaka and S. typhimurium from one hen (0.2%) each. The MAT with antigens of S. pullorum had a sensitivity of 81% and a specificity of 24% for detecting S. enteritidis-infected hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Control, prevention and eradication of Salmonella enteritidis infection in broiler and broiler breeder flocks

Salmonella enteritidis was identified by serological and bacteriological techniques in two clinically normal breeder flocks in an integrated broiler organisation in Northern Ireland. The organism was transmitted vertically to clinically affected progeny flocks. The infected breeder flocks were slaughtered and the infection throughout the organisation controlled and subsequently eradicated. A working group, consisting of the senior management of the broiler organisation and veterinary staff from the Veterinary Research Laboratories at Stormont, was formed to establish procedures to minimise the risk of the reintroduction of salmonella infection, by preventing vertical transmission from grandparent flocks, or lateral transmission from personnel, other animal species and fomites, or transmission through the feed. All feed was heated to a minimum of 70 degrees C for 12 minutes immediately before it was pelleted and subsequently transported to the flocks through a dedicated system of conveyor belts, bins and lorries. A comprehensive system for monitoring the efficacy of the preventive procedures was established and is now used throughout the poultry industry of Northern Ireland.     

Serological detection of chicken flocks naturally infected with salmonella enteritidis,using an enzyme‐linked immunosorbent assay based on monoclonal antibodies against the flagellar antigen

Summary

The Dutch Salmonella enteritidis monitoring and eradication programme for poultry prescribes a periodic examination of all breeding flocks for the presence of S. enteritidis. For the first years of the programme this was done by bacteriological examination of 50 faecal samples per visit per flock.

In this study we compare the results of bacteriological examination of faecal samples taken at 1580 visits from 545 flocks with those of a S. enteritidis enzyme‐linked immunosorbent assay (ELISA) applied on 24 serum samples per visit per flock. Two flocks were found positive for S. enteritidis by bacteriological examination; both flocks were also detected by ELISA. Ten flocks, bacteriologically negative for S. enteritidis were found positive by ELISA. S. enteritidis was isolated from three of these flocks by repeated and extensive bacteriological examination for verification. Verification was not possible in the fourth EL1SA positive flock. S. enteritidis infections were likely in three other flocks because of the farm histories.

On the basis of the results of this study it was decided to use this ELISA, starting from April 1992, as screening technique in the Dutch S. enteritidis programme instead of bacteriological examination of faecal samples. The ELISA is regarded as a flock test; an extensive, confirmatory bacteriological investigation for S. enteritidis is carried out in ELISA positive flocks to decide whether the flocks are truly infected.     

The role of mice in the epizootiology of Salmonella enteritidis infection on chicken layer farms.

A microbiological survey of 10 mice-infested poultry farms was conducted to determine the role of mice in the epizootiology of S. enteritidis infection. Five of the farms were rated as clean of S. enteritidis and five as contaminated based on culture results of environmental samples for S. enteritidis. Of 2103 environmental samples and 715 mice and rats tested, 5.1% and 16.2%, respectively, were culture-positive for S. enteritidis. On contaminated farms, S. enteritidis was isolated from 24.0% of the mice and 7.5% of the environmental samples, which represented 75.3% of all Salmonella isolations from mice but only 18.0% of Salmonella isolations from environmental samples on these farms. S. enteritidis was not detected in mice on clean farms. Phage types 13a and 14b were the two most frequently isolated phage types from mice and environmental samples. Although only a single phage type was isolated from single free-standing poultry houses, multiple phage types were isolated from multi-house complexes. A bacterial count from the feces of one mouse yielded 2.3 x 10(5) S. enteritidis bacteria per fecal pellet. S. enteritidis persisted at least for 10 months in an infected mouse population.     

Development and application of an ELISA for detecting antibodies to Salmonella enteritidis in chicken flocks.

Enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of IgG antibody to Salmonella enteritidis in poultry flocks. A lipopolysaccharide (LPS) and heat-extracted (HE) antigen for use in the ELISA were evaluated together with the rapid slide test (RST), microagglutination test (MT) and the microantiglobulin (MAG) test. In experimentally infected specific pathogen free chickens, good correlation was seen between all tests although, generally, the MT and MAG detected antibody earlier and titres peaked earlier than the ELISAs. The LPS antigen detected antibody earlier than the HE antigen but the latter gave higher titres in the later stages of infection. Cross reactions were seen between S enteritidis and S typhimurium in the ELISAs although homologous reactions were always much higher. Antisera to S montevideo or S senftenberg gave weak positive reactions in both S enteritidis ELISAs. Serological and bacteriological examinations of representative samples from two commercial chicken flocks were carried out. In flock A the HE-ELISA and MAG test detected antibody in nearly all birds. The LPS-ELISA detected antibody in over 60 per cent of birds, while the MT and RST detected few seropositive birds. The whole blood test using the stained S pullorum antigen on the farm detected antibody in just under 25 per cent of the birds. S enteritidis was isolated from the organs of 25 per cent of the birds. All birds in flock B were seronegative by all tests; no salmonellae were isolated from the organs of these birds.     

Risk factors for Campylobacter spp. contamination in French broiler-chicken flocks at the end of the rearing period.

Our objectives were to identify risk factors for contamination of French broiler flocks by Campylobacter. We used 75 broiler farms in western France. A questionnaire was administered to the farmers and samples of fresh droppings were taken to assess the Campylobacter status of the broiler flocks. 42.7% of the flocks were positive for Campylobacter spp. The risk of contamination of the broiler flocks by Campylobacter was increased in summer/autumn, in houses with static air distribution, when two or more people took care of the flock, in poultry farms with three or more houses and when the drinking water for the chickens was acidified. The presence of litter-beetles in the change room also increased the risk of contamination. The administration of an antibiotic treatment following a disease decreased the risk of a flock being contaminated by Campylobacter.     

Field observations with Salmonella enteritidis bacterins

A study involving 11 commercial layer flocks was conducted to determine the efficacy of Salmonella enteritidis bacterins (autogenous or federally licensed). The criterion for evaluation of vaccine efficacy was the presence or absence of S. enteritidis in the environment, the organs of the bird (including ovary and oviduct), and eggs. Environmental, rodent, and organ specimens from dead birds as well as eggs were cultured throughout the life of the flock. All layers were obtained from pullet sources that were negative for S. enteritidis, as determined by organ and environmental cultures. Despite the use of S. enteritidis vaccination, 63.6% of the houses had S. enteritidis-positive environmental cultures and 100% of the flocks had S. enteritidis organ-culture-positive birds. The range of positive cultures for S. enteritidis in the environment in vaccinated flocks was between 0 and 45.5%. Birds in vaccinated flocks were organ-culture positive for S. enteritidis between 10% and 40% of the time. The unvaccinated portion of flocks in the same house and the unvaccinated flock in a complex had similar results compared with the vaccinated portion of the flocks.     

The prevalence and genetic diversity of Campylobacter spp. in domestic 'backyard' poultry in Canterbury, New Zealand

Campylobacteriosis is the most commonly notified illness in New Zealand. Whilst the importance of commercial poultry in campylobacteriosis is well established, little is known about the possible role of chickens kept at home as a direct animal/faecal contact or consumption exposure pathway. The aim of this study was to determine the prevalence and genetic diversity of Campylobacter spp. in domestic backyard chicken flocks in the Canterbury region of New Zealand. Poultry faecal samples were collected from 35 domestic 'backyard' poultry flocks from urban and rural properties around the Canterbury Region of New Zealand. A total of 291 samples were collected and tested for the presence of thermotolerant Campylobacter spp. and positive isolates were analysed using pulsed-field gel electrophoresis (PFGE) using both SmaI and KpnI enzymes. There was a high prevalence of Campylobacter spp. with 86% of flocks testing positive. Campylobacter jejuni alone, Campylobacter coli alone and both C. jejuni and C. coli were detected in 20 (57%), 2 (6%) and 8 (23%) of the flocks respectively. SmaI/KpnI PFGE analysis identified 50 different genotypes across the 35 flocks. Genotype diversity richness was highest on the lifestyle block and farm properties with 43 different genotypes isolated, whilst urban properties displayed the least richness with 12 genotypes isolated. Rural flocks tended to have more different genotypes in a given flock than urban flocks. Comparison of the genotypes with the PulseNet Aotearoa Campylobacter database showed that 28 of the genotypes had previously been isolated from human cases of campylobacteriosis. Many of these were also indistinguishable from Campylobacter spp. previously isolated from retail chicken. Therefore, contact with backyard poultry or their faecal material is a potential additional infection pathway outside of exposure to the established pathways associated with the consumption of Campylobacter-contaminated commercial meat or foods cross-contaminated from contaminated poultry.     

The occurrence and distribution of Salmonella enteritidis and other serovars on California egg laying premises: a comparison of two sampling methods and two culturing techniques

Between the summer of 1998 and the winter of 2000, Salmonella analysis was performed on 2128 single and 532 pooled manure drag swabs obtained from 133 California commercial egg laying farms. The isolation of Salmonella from all rows and from all flocks using single or pooled swabs was 80% and 92%, respectively. Hence, there was no statistical difference between single vs. pooled swabs in terms of identifying Salmonella on a row or flock basis. A total of 14 serogroups comprising 44 serotypes were isolated from 123 of 133 farms. When the top 10 serotypes were considered, there was no significant difference in the range of serotypes isolated by the two culturing methods. The overall S. enteritidis prevalence for California flocks was 10.5% (14/133). The overall row prevalence for S. enteritidis for all the farms was 1.1% (24/2128), and the overall pool prevalence was 2.4% (13/532). Sixty percent (12/20) of the S. enteritidis isolates from the positive farms were phage type 4, and 40% (8/20) represented five other phage types (1, 6B, 7, 8, and 28).     

Prevalence and risk factors of Campylobacter infection in broiler flocks from southern Spain

An extensive epidemiological study was performed to determine the prevalence and risk factors of Campylobacter infection in broiler farms in Andalusia (southern Spain). A total of 2221 cloacal swabs and 747 environmental swabs from 291 broiler flocks were screened between April 2010 and May 2012. The prevalence of Campylobacter in individual animals was 38.1%, and the flock prevalence was 62.9%. Flocks were predominantly infected by C. jejuni and C. coli but were also infected by untyped Campylobacter spp., and mixed-species infection could be found. Risk factors for Campylobacter infection were assessed from direct interview of the farmers. The number of positive samples by flock was modelled assuming a binomial distribution. Analysis indicated five factors associated with increased intra-flock prevalence: presence of dogs or cats on the farm, older age of the broiler flock, the application of thinning of flocks, the presence of windows with canvas blinds, and the presence of rodents in the poultry house. Two factors were associated with decreased intra-flock prevalence: the treatment of drinking water and having an entrance room for access into the poultry house. This is the first study performed on broilers farms from Spain reporting the risk factors of Campylobacter infection and is the largest study on the prevalence of Campylobacter infection.     

A retrospective study on salmonella infection in Danish broiler flocks

A retrospective longitudinal study was conducted to identify risk factors associated with Salmonella enterica infection in Danish broiler production. The study was based on information in the antemortem database (AM database) where data were available for all broiler flocks slaughtered over the 2-year period from 1992 to 1993 in Denmark. The AM database contains information collected by the ante-mortem veterinarians, from the slaughterhouses, and from the salmonella examinations carried out at the National Veterinary Laboratory. The epidemiological unit was the individual broiler flock. The salmonella status of the flock was determined by examining the caecal tonsils from 16 3-week-old chickens from each flock. This procedure would detect a salmonella-infected flock, with a probability above 95%, if the prevalence is above 20%. Furthermore, the structure and quality of the collected data have been evaluated.

Fourteen variables were selected for analysis by multivariable logistic regression. An increased risk of salmonella infection in the broiler flocks was associated with the biggest hatcheries and feedmill, with an increasing number of houses on the farm, if the preceding flock was infected, and if the flock was reared in the autumn. Additionally, the main variables of the model were analysed by including a random effect at the house level. This resulted only in minor changes of the parameter estimates.     

Salmonella infection sources in poultry flocks in northern Greece]

From 10 egg production poultry farms 1516 samples were collected and examined for the presence of salmonella. The samples were: 201 chicken, 36 sparrows, 35 rats, 35 pools of 20 flies each, 450 eggs, 60 mattresses, 188 feces, 425 feedstuffs and 86 water samples. Salmonellae were isolated only from 163 (10.8%) samples. From the 146 (89.6%) of these S. gallinarum was isolated. From the rest 17 (10.4%) the following mobile salmonella strains were isolated: two strains of S. virchow and Salmonella of subgroup II, four strains of S. typhimurium var. Copenhagen, seven strains of S. Livingstone, one S. enteritidis and one S. infantis The S. gallinarum was isolated from dead or sick chicken (46%), eggs (10.4%), rats Rattus norvegicus (14.3%) and mattresses 1.6%. The mobile salmonellae were isolated from feedstuffs (2%), flies (14.3%), rats (2.8%), feces (1%). From the present study, it seems that rats, chicken and eggs are important for the salmonella dissemination.     

Serologic detection of experimental Salmonella enteritidis infections in laying hens by fluorescence polarization and enzyme immunoassay

Detection of infected poultry flocks is essential for controlling eggborne transmission of Salmonella enteritidis to humans. The present study evaluated the detection of antibodies in the sera of experimentally infected chickens by a fluorescence polarization assay with a tracer prepared from the O-polysaccharide of S. enteritidis and an enzyme-linked immunosorbent assay (ELISA) with an S. enteritidis flagellin antigen. In two trials, groups of specific-pathogen-free laying hens were infected orally with either 10(6) or 10(8) colony-forming units (CFU) of S. enteritidis (phage type 13a) or with 10(8) CFU of Salmonella typhimurium. Serum samples were collected before inoculation and at five subsequent weekly intervals. Both assays successfully detected the majority of hens infected with S. enteritidis at either dose level, but they also identified a substantial number of hens infected with S. typhimurium as seropositive. The fluorescence polarization test detected S. enteritidis infection significantly more often and cross-reacted with sera from hens infected with S. typhimurium significantly less often than the ELISA. The fluorescence polarization assay also offered advantages in terms of speed and methodologic simplicity.     

The relationship between the magnitude of the specific antibody response to experimental salmonella enteritidis infection in laying hens and their production of contaminated eggs.

Detecting infected laying flocks is a vital part of many efforts to control egg-associated transmission of Salmonella enteritidis to humans. The relationship between the development of a specific antibody response in infected hens and the deposition of S. enteritidis in eggs is important for establishing the epidemiologic relevance of serologic testing methods. In two trials, laying hens were infected with large oral doses of phage types 13a and 14b isolates of S. enteritidis. Approximately 38% of all infected hens produced at least one contaminated egg, at an overall incidence of 5.2%, between 3 and 23 days postinoculation. As determined by enzyme-linked immunosorbent assay with an S. enteritidis flagellar antigen, 91.7% of inoculated hens produced specific serum antibodies. Although hens with very high antibody titers were associated with a significantly elevated frequency of egg contamination, a consistently direct relationship was not evident between the magnitude of the antibody responses of individual hens and the frequency at which they laid contaminated eggs. Accordingly, although serologic tests can be valuable screening tools for preliminary detection of S. enteritidis infections in poultry, the magnitude of the antibody responses detected in individual hens may not predict the overall risk of egg contamination associated with particular laying flocks.     

Pathologico-anatomic, bacteriologic and serologic findings in laying hens from small neighborhood flocks with Salmonella enteritidis phage type 4 infection

Two small free range flocks with 32 hens, whose Salmonella enteritidis contaminated eggs caused salmonellosis in man, were investigated for localisation of the agent. Salmonella enteritidis phage type 4 was isolated from 12 of 32 hens (37.5%). Eight hens (25%) harbored the bacterium in the ovary and/or the oviduct. The rapid slide agglutination test with Salmonella pullorum-antigen revealed 21 positive hens (66%). All eight hens with Salmonella enteritidis phage type 4 in the reproductive tract were seropositive. The significance of these findings for the control of Salmonella enteritidis phage type 4-infections in poultry is discussed.     

Sources of Salmonella contamination during broiler production in Eastern Spain

Prevention of Salmonella contamination of poultry products requires detailed knowledge of the main sources associated with its presence in the production system. The aims of this study were to determine the main sources of Salmonella contamination in broiler production during growing, to assess the risk factors for Salmonella contamination at the end of the rearing period and to determine the main serovars involved in broiler production systems in Eastern Spain. A total of 65 different broiler houses from different farms were sampled. Each house was sampled at different times during the rearing period. First, when the previous flock was taken to the slaughterhouse, samples of dust, surfaces and previous flock faeces were collected. After cleaning and disinfection (C&D), samples of dust and surfaces were also taken. On the first day of rearing, samples of water, bedding, farming boots, meconiums, delivery-box liners and feed were collected. During rearing, feed samples were taken directly from the truck and from feeders. On slaughter day, samples of dust, surfaces, water, feed and faeces were also collected. Finally, two days after slaughter, carriers (rodents, flies and beetles) were trapped. All samples collected were analysed according to ISO 6579:2002 (Annex D) and positive samples were serotyped in accordance with Kauffman-White-Le-Minor technique. Our results showed that all different types of samples collected were contaminated with Salmonella (prevalence ranged between 1.5% and 38.6%). The most contaminated samples related with poultry production were: delivery-box liners (32.0%), faeces samples (31.2%), dust samples (25.0%), farming boots (19.7%) and feed from feeders (16.0%). However, the most important risk factors for Salmonella contamination of the flocks at the end of the rearing period were Salmonella status of the house after cleaning and disinfection, Salmonella status of day-old chick flocks and feed from feeders. Twenty-one different serovars were isolated from the samples analysed. The most prevalent were in decreasing order: Salmonella Enteritidis (52.9%), S. Hadar (17.8%), S. Virchow (8.9%) and S. Ohio (5.4%). The study suggested that there are many sources for Salmonella contamination and persistence in broiler production. Hence, the whole production chain needs to be controlled to eradicate the bacteria from primary production.     

Results of a Salmonella enteritidis vaccination field trial in broiler-breeder flocks in The Netherlands

From August 1995 until December 1997, the effect of adding Salmonella enteritidis (SE) vaccination to a certified standardized biosecurity program in a situation of increased infection risk was examined in a field trial in The Netherlands. In this field trial, two groups of broiler-breeder flocks with increased infection risk were vaccinated, one group with VAC-T/TALOVAC logSE(group A) and the second group with SALENVAC (group B). The determination of increased infection risk in groups A and B was based on an SE infection history; flocks were either previously infected and treated (PIT) or had other risk factors than previously infected and treated (OPIT). SE infections in both vaccinated groups were assessed by monitoring according to the Dutch salmonella control program. Under field conditions, designation of a vaccinated and a control group on the farm was not possible. In the same period as the vaccinated groups, 608 nonvaccinated flocks (group C) were hatched and monitored according to the Dutch salmonella control program. The flock level occurrence of SE infection in the vaccinated groups was compared with the flock level occurrence of SE infection in the nonvaccinated group on the basis of comparability of infection risk. In group C, whether or not flocks had infection risk PIT was known and for risk factor OPIT, only whether or not a flock had been placed on a previously contaminated farm (= risk of reinfection) was known. The proportion of SE-infected flocks with risk factor PIT in the vaccinated groups was not significantly different from that in the nonvaccinated group C. Only the proportion of SE-infected flocks with a risk of reinfection in the vaccinated group B (0) was significantly lower (P = 0.02) than in the nonvaccinated group C (18%). The fact that no significant result was found in favor of group A is because of the small number of flocks in this part of the study. On the basis of the conditions of the setup of this trial, it can only be concluded that there is an indication that vaccination contributes in the reduction of SE reinfection in broiler breeder flocks.