Competitive Exclusion Treatment Research Articles

Mechanisms of maintenance of intestinal homeostasis by autochthonic microbiota and probiotics

Intestinal microbes are taxonomically diverse and constitute an ecologically dynamic microbiom interactively performing various physiological and physiopathological processes. It has been proposed that normal intestinal microbiotas play a critical role in the host’s metabolic homeostasis and immune tolerance. The modulation of intestinal microbiota populations by prebiotics, probiotics, and synbiotics may be beneficial for the host’s health. Under certain conditions, the intestinal microbiota and the host’s homeostasis can be restored by introducing bacteria that co-mediate anti-inflammatory responses. Commensal microbes and probiotics exert their beneficial effect by at least three mechanisms. These include - the maintenance of the epithelial barrier function and the attenuation of changes in intestinal permeability through effects on tight junction, decreasing paracellular permeability, providing innate defense against pathogens, and enhancing the physical impediment of the mucous layer, - competitive exclusion by the application of probiotic bacteria stabilizing the indigenous microflora, - immunomodulatory capacity, affecting a variety of signaling pathways with modulation of proper immune, inflammatory and allergic responses. The epithelial gut barrier faces important challenges, since its function is to prevent pathogens and harmful elements of the gut lumen from penetrating into the internal environment. Competitive exclusion treatment can increase resistance to pathogen colonization and control intestinal disturbance. The dominance of symbiotic and probiotic bacteria among the gut microbiota favors a tolerogenic immune response. The release of secretory IgA stabilizes tight junctions between cells of the epithelial layer as well as hampers pathogens and symbionts invading deeper layers. The understanding of these vital processes may help to protect the host against infection, prevent chronic inflammation, and maintain mucosal integrity.

Chicken intestinal microbiota function with a special emphasis on the role of probiotic bacteria.

Bacterial colonization of the chicken gut by environmental microbes begins immediately after hatching. Composition of the intestinal microbiota is dependent on the surrounding environment, diet variation, pathological conditions, antibiotic therapy, and others. The genomes of all these intestinal microbes form a microbiome which by far outnumbers the host's genome. As a consequence, the microbiome provides additional metabolic functions to the host, including nutrient utilization and absorption, fermentation of non-digestible dietary fiber, synthesis of some vitamins, biotransformation of bile acids, and the well-being of their chicken host. Microorganisms can also directly interact with the lining of the gastrointestinal tract, which may alter the physiology and immunological status of the bird. Since newly hatched broiler chickens demonstrate delayed commensal colonization and low bacterial diversity, the most effective and harmless method available to control the development and composition of the intestinal microbiota is a competitive exclusion treatment by applying probiotic bacteria. Additionally, recent research has shown that probiotic bacteria have a variety of beneficial effects, including counteraction of dysbiosis, promotion of gut health and homeostasis, enhancement of immune defenses and antagonization of infectious agents.

Competitive ExclusionEscherichia coliCultures onE. coliO157 Growth in Batch Culture Ruminal or Fecal Microbial Fermentation

The purpose of this study was to determine the efficacy of competitive exclusion (CE) Escherichia coli cultures (ATCC 202018 and 202019) for reducing E. coli O157 in batch culture fermentations with ruminal or fecal microbial inoculum derived from a cannulated feedlot steer fed a corn-grain-based, high-grain diet. A non-CE (NCE) E. coli strain (ATCC 25922) was included as a control. Anaerobic fermentations (50 mL) were set up in serum bottles with a 2:1 mineral buffer to microbial inoculum ratio and with or without gluconic acid, a component of intestinal mucus, which has been shown to be stimulatory to the growth of E. coli O157 in vitro. A five-strain mixture of E. coli O157 made resistant to nalidixic acid (50 microg/mL; Nal(R)) was added to each fermentation, and concentrations of Nal(R) E. coli O157 were determined after 0, 6, 12, and 24 h of fermentation. In ruminal microbial fermentation, the CE treatment had a lower (0.6 log; p < 0.01) concentration of Nal(R) E. coli O157 than the NCE control at 24 h, regardless of gluconic acid inclusion. In fecal microbial fermentation, the CE treatment did not affect Nal(R) E. coli O157 concentrations in fermentations without gluconic acid; but with gluconic acid inclusion, Nal(R) E. coli O157 concentrations were lower in the CE treatment than in NCE controls at 12 h (1.7 log CFU/mL lower) and 24 h (1.9 log CFU/mL lower). In vitro fermentation with ruminal or fecal microbial inoculum could be a simple and inexpensive initial approach to assess efficacy of CE cultures for reducing E. coli O157.

Resistance to Challenge of Breeders and Their Progeny with and Without Competitive Exclusion Treatment to Salmonella Vaccination Programs in Broiler Breeders

3 Abstract: Resistance to Salmonella challenge of breeders under three vaccination programs and of their chicks with and without mucosal Competitive Exclusion (CE) (CHR Hansen) treatment was assessed. Vaccine treatments combined a live Aro-A Salmonella Typhimurium (ST) vaccine and an autogenou s commercially prepared (Lohmann Animal Health) trivalent killed vaccine (serogroups B, C and D). 2 1 Treatments combined: 2 live and 2 killed doses or 3 live and 1 killed dose delivered at 1, 21, 77 and 126 d of age; or 2 killed doses delivered at 77 and 126 of age; and a non-vaccinated control (C). At 3, 6, 11, 17 and 22 wks of age, a portion of breeder pullets was removed and challenged per os with 10 cells of a 3-strain 7 10 significant reductions between (live) vaccinates and non-vaccinates at 3 (0.82 log) and 6 wks (0.85 log) challenges. By 11 wks, there were no differences in Salmonella levels between vaccinates and controls, indicating that 1-d and 3-wk live vaccine protection had diminished with time. All vaccination treatments reduced breeder cecal counts (1.15-1.30 log) by wk 22. Passive immunity from breeder vaccination treatments was not effective in diminishing chick cecal counts as shown by comparable susceptibility of chicks from vaccinated and control breeders, regardless of breeder age. Chick CE treatment consistently diminished cecal (1.41 log) and LHS (0.306 log) counts. These results show that live Aro-A ST vaccination decreases counts during the first 6 wks of age, as do all programs by 22 wks of age and that competitive exclusion is the most effective treatment in reducing hatchling Salmonella counts.

Development of a competitive exclusion product for poultry meeting the regulatory requirements for registration in the European Union

Competitive exclusion treatment is able to increase the pathogen colonization resistance of day-old chicks by applying probiotic bacteria stabilizing the indigenous microflora. In order to develop a safe microbial feed additive, various bacterial strains were isolated out of the gastrointestinal tract of healthy chickens. One hundred twenty-one representatives were selected based on differences in whole-cell protein patterns and screened for antagonistic properties. Five effective strains (Pediococcus acidilactici, Enterococcus faecium, Bifidobacterium animalis ssp. animalis, Lactobacillus reuteri, and Lactobacillus salivarius ssp. salivarius) exhibited in vitro the ability to inhibit a range of common pathogens and were evaluated with regard to the risks associated with genetic transfer of antibiotic resistances from animals to humans via the food chain. The probiotic strains were sensitive to several clinically effective antibiotics, though some of them showed single resistances. None of the vancomycin-resistant (R) strains carried the enterococcal vanA gene. Two tetracycline R strains were shown to harbor a tet(M)-associated resistance. The strains contained no extrachromosomal DNA and were not able to transfer the resistance by means of conjugation. On basis of the collected data the presence of easy transferable resistances was excluded and the chicken strains were considered to be suitable for the use as feed additive.

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.

Effectiveness of an Undefined Mucosal Competitive Exclusion Treatment to Control Salmonella in Turkeys During Brooding

Mucosal competitive exclusion (MCE) cultures generated from the mucosal scrapings of healthy adult turkeys were administered to commercial turkey poults. MCE-treated poults were placed on tom and hen farms with paired untreated control poults in adjacent houses. After 6 wk in the brood house, cecal droppings from control and treated flocks were collected and analyzed for the presence of salmonellae. Salmonellae were detected in 14 of 30 cecal droppings (47%) from control flocks and from only 1 of 30 (3.3%) droppings from the treated poults. This study demonstrated that mucosal competitive exclusion could be used to effectively control salmonellae in a commercial field trial of young turkey flocks.

Comparison of Mucosal Competitive Exclusion and Competitive Exclusion Treatment to Reduce Salmonella and Campylobacter spp. Colonization in Broiler Chickens

Control of Salmonella spp. during the earliest phases of broiler production may provide the best opportunity to reduce human pathogens on processed broiler carcasses. Application of the “Nurmi concept” has been demonstrated to be an effective means in reducing Salmonella colonization among broiler chicks. In 1989, Aho et al. developed a competitive exclusion (CE) culture for control of Salmonella spp., whereas a mucosal competitive exclusion culture (MCE) developed in the United States was originally created to control Campylobacter colonization (Stern et al., 1995). The major differences in the two patents were the higher level of anaerobic culture required, the degree of epithelial scraping and washing of the ceca, media used for subculturing, and the culture incubation temperatures (35 C vs. 42 C). The CE and MCE were compared for efficacy in reducing Salmonella and Campylobacter colonization in broiler chicks. Nine adult birds (three for each of three replicates) were slaughtered, and each of a bird's paired ceca were used to produce corresponding antagonistic microflora, which were administered to day-of-hatch chicks. The chicks (a total of 210) were challenged 24 h later with Salmonella and Campylobacter and were killed 1 wk later, and levels of the pathogens were determined. Ninety CE-treated birds were significantly more colonized by Salmonella typhimurium than those 90 chicks treated with the MCE microflora (3.97 log 10 cfu/g cecal contents vs. 1.25 log 10 cfu/g cecal contents). Also, Campylobacter spp. colonization of these birds was significantly higher for CE-treated birds when compared with MCE-treated birds (6.96 log 10 cfu/g cecal contents vs. 5.03 log 10 cfu/g cecal contents). These results can be useful in developing intervention strategies to reduce chicken colonization by Salmonella and Campylobacter.

Development of a rapid and inexpensive assay for the nonspecific detection of antimicrobial residues in chicken egg yolks and neonatal yolk sacs.

Competitive exclusion of intestinal pathogens by administration of beneficial and defined cultures of normal intestinal microflora is a safe and effective means of reducing the incidence and severity of chick infections with Salmonella and other intestinal pathogens. It is important that competitive exclusion cultures not carry genetic material (e.g., plasmids), which could transfer antibiotic resistance to other microflora, including pathogens. As such, safe and effective competitive exclusion cultures must be sensitive to commonly used antimicrobial agents. By necessity, intentional or accidental exposure of these beneficial microflora to antibiotics will reduce or eliminate the protection provided by competitive exclusion culture establishment. As antibiotic residues can be present from embryonic, hatchling, or maternal administration, a rapid and sensitive assay for the nonspecific detection of residues, which could interfere with competitive exclusion culture establishment, is needed. This study was conducted to develop a rapid and inexpensive bioassay to detect multiple antimicrobial residues in egg yolk and neonatal yolk sacs. Aerobic bacterial strains with known sensitivity to several antibiotics used by the poultry industry were selected and individually compared for sensitivity to enrofloxacin, gentamicin, tetracycline, ceftiofur, and tylosin concentrations in egg yolks. This assay was found to be relatively sensitive for the detection of these antimicrobials, and detection of residues was associated with reduced competitive exclusion culture (PREEMPT) establishment in one experiment. Importantly, this assay can be implemented with minimal training or equipment under commercial hatchery practices and could be used to determine embryo groups, in advance of hatch, that are not suitable candidates for competitive exclusion treatment in the hatchery.

Combined Effect of Antibiotic and Competitive Exclusion Treatment on Salmonella Enteritidis Fecal Shedding in Molted Laying Hens

Salmonella Enteritidis is an important pathogen for the layer industry, primarily because of its ability to infect hens and ultimately contaminate egg contents. Studies have shown that stress situations, such as flock recycling (induced molting), can increase Salmonella Enteritidis problems in the flock. The present study examined the effect of antibiotic treatment and competitive exclusion (CE) on Salmonella Enteritidis shedding in the period following molt and 14-day feed withdrawal. In two separate trials, 48 birds after molt and feed withdrawal were divided into one group that was treated for 10 days with enrofloxacin in water followed by administration of CE culture and a group that was left untreated. Salmonella Enteritidis shedding was significantly reduced in the antibiotic-CE group. The Salmonella Enteritidis shedding rate was 33 and 25% in untreated birds versus 4 and 0% in the enrofloxacin-CE group on the two test days. These results indicate that treatment of Salmonella Enteritidis–positive laying hens after molting with enrofloxacin and CE culture can substantially reduce Salmonella Enteritidis problems due to molting and would be a possible alternative to diverting eggs for pasteurization or slaughtering the infected flock. Possible development of bacterial resistance in conjunction with antibiotic use is also discussed.

Poultry meat and food safety: pre- and post-harvest approaches to reduce foodborne pathogens

Poultry and poultry products have repeatedly been implicated as a source of foodborne infection for humans. The significant increase in the number of reported foodborne disease outbreaks worldwide has altered opinions on food safety. It has become clear that current legislation alone is inadequate, and that industry must accept a greater share of responsibility for the quality and safety of the food it produces. The fact that processing plants are not able to reduce the incidence of pathogenic bacteria effectively in poultry products means that every effort must be made to reduce infection in and/or contamination of the live birds with foodborne pathogens before dispatch to processing plants. The main control strategy should include the following: cleaning the production pyramid from the top, as in the case of Salmonella enteritidis and Salmonella typhimurium, by destroying infected flocks (with compensation), sanitising hatching eggs and limiting the introduction and spread of pathogens at the farm through Good Animal Husbandry Practices (GAHPs). To achieve GAHPs, effective measures of hygiene should be applied to poultry houses and the feed. Reducing salmonella colonisation by using antimicrobial feed additives, competitive exclusion treatment or vaccines is worth consideration. Furthermore, hygienic systems for catching and transporting live poultry must be included. To reduce carcass contamination during processing, thorough schedules for cleaning and disinfection of the equipment and plant are essential. Machine design must permit easy and thorough cleaning. Because the success of any food safety programme on the farm and in the processing plant depends on the hygiene practices of the staff, it is essential to incorporate education programmes for all people involved in the poultry production chain. In the farm-to-fork concept, post-processing food handling is also a very important factor in reducing foodborne infections. Finally, the development and application of acceptable methods for end product decontamination and preservation in the processing plant must be pursued.

Inhibitory Effects of Competitive Exclusion and Fructooligosaccharide, Singly and in Combination, on Salmonella Colonization of Chicks

The inhibitory effects of competitive exclusion (CE) and 0.1% concentration of fructooligosaccharide (FOS), singly and in combination, on Salmonella colonization of chicks were investigated. Moreover, quantitation of the major cecal flora (Bifidobacterium, Bacteroides, Lactobacillus, and Escherichia coli) was performed. One-day-old birds were divided into four groups: (i) control, (ii) CE, (iii) FOS, and (iv) CE plus FOS. Chicks received Salmonella Enteritidis at 7 days (experiment 1) or 21 days (experiment 2). Birds in each group were killed at 1 day, 7 days, and 14 days after inoculation of Salmonella Enteritidis for count of salmonella in cecal contents. In experiment 1, the mean number of Salmonella Enteritidis in the chicks inoculated with CE was significantly decreased compared with the other three groups at 1 day postinoculation. In experiment 2, the mean numbers of Salmonella Enteritidis in the chicks of the FOS group and the FOS plus CE group were significantly decreased compared with the control group at 1 day and 7 days postinoculation. On 7- and 21-day-old chicks, few changes on number of total bacteria, Bifidobacterium, Bacteroides, Lactobacillus, and E. coli were observed in the cecal contents of treated groups compared with the control group. Low-dose feeding of FOS in the diet of chicks with a CE treatment may result in reduced susceptibility to Salmonella colonization but may not lead to a shift in the intestinal gut microflora on 7- and 21-day-old chicks.

Effect of Simultaneous or Delayed Competitive Exclusion Treatment on the Spread of Salmonella in Chicks

Exposure of newly hatched chicks in the hatchery to salmonellae results in the establishment of seeder chicks with fecal shedding and the transmission of salmonellae to other contact chicks in the broiler house during growout. This study investigated the protective effect of treatment with a characterized competitive exclusion culture, CF3 on the transmission of Salmonella from seeder to contact chicks when CF3 treatment occurred simultaneously with seeder challenge or when treatment was delayed until 24 hr after challenge. Delayed treatment failed to prevent the establishment of seeder chicks, but did significantly (P≤.005) reduce the spread of Salmonella from seeder to non-challenged contact chicks. Simultaneous Salmonella challenge and CF3 treatment significantly (P≤.005) decreased the establishment of seeders and was highly effective in preventing the spread of Salmonella from seeder to contact chicks. The results demonstrated that treatment on the day of hatch can significantly help to reduce seeder establishment and the spread of Salmonella from seeders to highly susceptible contact chicks.

Effect of Salmonella in young chicks on competitive exclusion treatment

The potential of Salmonella contamination in hatching cabinets to 1) generate seeder chicks and 2) interfere with the efficacy of competitive exclusion (CE) treatments was investigated in six experiments. Hatchery-generated seeder: chicks were produced by hatching in the same hatcher with inoculated eggs (immersed in a 1.0 x 10(5) cfu/mL suspension of Salmonella typhimurium or inoculated with 10(5) to 10(6) cells on the air cell membrane). Salmonella spread through the hatching cabinet to chicks hatching from uninoculated eggs in adjacent trays. In two experiments, Salmonella was isolated from 31 and 100% of the chick rinses after the birds were held in groups for 3 d in isolation units. When these hatchery-generated seeder chicks were stocked in floor pens at a 1:10 ratio with uncontaminated contact chicks, the pen environment became contaminated to the extent that greater than 50% of the contact chicks became contaminated. In two experiments with only one to three inoculated eggs per 200 egg hatching cabinet, 98% of uninoculated chicks were intestinally colonized with Salmonella after the birds were held 1 wk in isolation cabinets. This hatchery-acquired Salmonella substantially reduced the effectiveness of subsequent CE treatments to prevent Salmonella colonization of the young chicks. These studies demonstrate that control of Salmonella in hatching cabinets is critically important for control of Salmonella in broilers.

The use of competitive exclusion in broilers to reduce the level of Salmonella contamination on the farm and at the processing plant

The effect of a competitive exclusion (CE) product, Broilact, on Salmonella contamination of broiler chickens was studied on the farm and at the processing plant. In the first part of the study, two flocks per week, a CE-treated and an untreated control flock, were placed in similar broiler houses. The CE treatment was administered in the hatchery using a modified spray vaccination cabinet. Salmonella was analyzed from the paper pads of the transport boxes on arrival at the farm and from fecal samples taken 2 wk before slaughter. The results of Salmonella sampling were received for 67 flocks. The other 141 flocks of the company that were reared during the trial period were also sampled for Salmonella and the results were compared to those of treatment and control groups. Broiler performance, including mortality, weight, and feed conversion, was recorded for the trial flocks. In the second part of the study, Salmonella contamination of neck skin samples taken at the processing plant from 18 CE-treated and 28 control flocks was compared. The Broilact-treatment significantly reduced Salmonella contamination both on the farm and at the processing plant. At the level of the farm, the percentage of Salmonella-positive flocks was essentially the same in the control flocks and in other flocks reared during the trial period. An improvement in broiler performance was indicated, although the difference was not significant.

Bacteriological monitoring of Salmonella enteritidis carrier birds after decontamination using enrofloxacin, competitive exclusion and movement of birds

Two hundred and forty, four-week-old laying birds naturally infected with Salmonella enteritidis PT33 (Pasteur Institute phage typing system) were randomly divided twice (before and during the treatments) to obtain four...

Evaluation of combined antibiotic and competitive exclusion treatment in broiler breeder flocks infected with Salmonella enterica serovar Enteritidis

Summary Strategic medication in breeding flocks of domestic fowl (Gallus gallus), using antimicrobial treatment followed by competitive exclusion, was evaluated in 13 trials between February and September 1993. In each trial, the flock had been confirmed as naturally infected with Salmonella enterica serovar Enteritidis and the effect of treatment was determined on salmonella isolation from tissues. Of 11 trials where enrofloxacin was used, a long‐term reduction of salmonella was observed in two and a short‐term reduction was measured in birds from another five trials. Salmonella Enteritidis was isolated from birds after treatment in four other trials with enrofloxacin and in two trials of medication with amoxycillin. After treatment with enrofloxacin significant reductions were found in the prevalence of 5. Enteritidis in tissues from birds, and in levels and prevalence of salmonellas in their environment. No salmonella was identified in statutory meconium samples taken from the hatched chicks derived from the flocks after treatment. The programme of antibiotic treatment and competitive exclusion offers an alternative to slaughter, but the approach must be part of a co‐ordinated programme which will effect a decrease in the prevalence of 5. Enteritidis over time by contemporary use of disease security measures. The risk of development of antimicrobial resistance is also discussed.

Resistance of chicks and poults fed vermicompost to caecal colonization bySalmonella

Vermicompost (VC) was produced by feeding the earthworm Eisenia foetida vegetable matter and fresh faeces from adult specific pathogen-free chickens. This was used as a competitive exclusion treatment for chicks and poults against challenge with Salmonella spp. One-day-old chicks and poults housed in floor pens, were treated with VC sprinkled on feed on days 1 and 2. For studies with Salmonella typhimurium, chicks were challenged on day 5 by either oral inoculation with 10(4) colony-forming units (CFU) or by contact transmission and killed 7 to 9 days later. In four trials, conducted on a total of 412 VC-treated and 275 control chicks, and in one trial with about 40 poults per group, treatment with VC significantly reduced (P<0.01) caecal colonization by 5. typhimurium. For experiments with Salmonella enteritidis, chicks were challenged orally with 10(5) CFU and kept in isolators on wire floors. They were also significantly protected (P<0.01) against caecal colonization by VC-treatment. In addition, in one trial S. enteriditis was isolated by enrichment techniques from the livers of nine of 10 non-treated and one of 39 VC-treated chicks.

Inhibition of Campylobacter jejuni colonization in chicks by defined competitive exclusion bacteria

Campylobacter enteritis in humans has been linked to consumption of chicken. Reducing Campylobacter jejuni colonization in chickens can potentially reduce Campylobacter infections in humans. In this study, the reduction of C. jejuni colonization in chicks by oral administration of defined competitive exclusion (CE) cultures, 2.5% dietary carbohydrates, or CE cultures and carbohydrates was examined. Prevention, elimination, or direct challenge of Campylobacter infection was simulated by administering treatments before, after, or at the same time as that of the Campylobacter inoculation. Additionally, the effect of maintaining high levels of protective bacteria was evaluated by administering CE cultures on days 1 and 4 (booster treatment). All treatments reduced C. jejuni colonization. Protection by aerobically grown CE cultures was not statistically different from that by anaerobically grown CE cultures. A combination of Citrobacter diversus 22, Klebsiella pneumoniae 23, and Escherichia coli 25 (CE 3) was the most effective CE treatment. Maintaining high numbers of CE isolates by administering CE boosters did not increase protection. The greatest reduction of Campylobacter colonization was observed in schemes to prevent or eliminate C. jejuni infection. C. jejuni was not detected in the ceca of birds receiving the prevention treatment, CE 3 with mannose, representing a 62% reduction in the colonization rate. The protection factor (PF), a value combining the colonization rate and the level of infection, for CE 3 with mannose was high (> 13.2). Fructo-oligosaccharides alone strongly prevented Campylobacter colonization. Only 8% of the chicks in this group were colonized, with a PF of > 14.3.(ABSTRACT TRUNCATED AT 250 WORDS)

Experience of a 10-year use of competitive exclusion treatment as part of the Salmonella control programme in Sweden

In Sweden, competitive exclusion (CE) treatment has been used since 1981 as a part of the national control programme for Salmonella. According to the programme all broiler flocks are tested for Salmonella before slaughter thus providing an evaluation of CE treatment. The CE culture used is mainly for the two consecutive flocks raised in a unit after a Salmonella-infected flock has been identified. During the period 1981–1990 CE culture has been given to 179 flocks, involving 3.82 million chickens. Only one of the treated flocks was found to be Salmonella-positive. A special study during a period when Salmonella was spread by contaminated feed demonstrated that CE treatment was effective in controlling Salmonella under field conditions. A likely contributory factor is that the hatcheries in Sweden can be claimed to be Salmonella-free. In addition, the control programme and related action, have led to a very low incidence of Salmonella in broiler chickens. A nationwide study carried out in 1990 demonstrated that less than 1% of broiler chickens was contaminated with Salmonella after slaughter.