Effect of a commerical competitive exclusion product on the colonization of Salmonella infantis in day-old pheasant chicks

One-day-of-age pheasant chicks were treated orally with the competitive exclusion product Broilact in three replicate trials. The following day the treated chicks and untreated control chicks were challenged likewise with approximately 10(3) colony-forming units (cfu) of Salmonella infantis. Five days after challenge the cecal contents of the birds were examined quantitatively and by enrichment for S. infantis. In all three trials Broilact effectively reduced colonization of the challenge organism, the mean infection factor (IF) value (the logarithmic number of colony forming units of salmonella organisms per gram of cecal contents) for the treated groups being 2.9 and that for the salmonella control groups 8.4. Mortality during the 1 week rearing period was 5.0% in the Broilact treated groups and 8.5% in the salmonella control groups.     

The effects of a competitive exclusion product and two probiotics on Salmonella colonization and nutrient digestibility in broiler chickens

Competitive exclusion (CE) cultures, given as a single dose on the day of hatch, together with good hygienic practices has been shown to be a novel approach to control Salmonella in poultry. The ability of the CE product Broilact and 2 probiotics, FloraMax-B11 and Colostrum, to prevent Salmonella colonization in newly hatched chickens was evaluated employing a slightly modified Mead-model chicken assay. In a parallel study the effect of the 3 treatments on the production of volatile fatty acids in the ceca were determined. In the Salmonella study 2 separate experiments were done. In the first experiment all 3 treatment materials were given as a single dose on d 1. In the second experiment, which consisted only of Broilact and FloraMax-B11, the latter was given in the drinking water during the 3 first d after hatch. In both experiments the chicks were challenged with Salmonella enterica serovar Infantis on d 2. The results of the present study show that Broilact was superior to the 2 other treatment materials in protecting the newly hatched chickens against Salmonella colonization. The parallel study showed only minor differences among the different treatments. Based on the results of the Salmonella challenge study, it was concluded that Broilact was the only treatment material that was established in the gut of the newly hatched chickens in such a way that the colonization of Salmonella was prohibited.     

Antimicrobial resistance in fecal Escherichia coli isolated from growing chickens on commercial broiler farms

To investigate the effects of rearing practices of commercial broiler chickens on the incidence of antimicrobial resistance in commensal Escherichia coli isolates, fecal E. coli isolates obtained in 4 farms were screened for anitimicrobial resistance. Ten E. coli isolates were recovered from each of the fecal samples collected from 10 birds in the farms at the ages of 2 days, 14-17 days, and 47-50 days. In 2 out of the 4 farms, no antimicrobials were used during the rearing period. In the other two farms, following collection of the fecal samples at 14 and 15 days of age, oxytetracycline (OTC), sulfadimethoxine (SDMX), and tylosin were given to birds on one farm and SDMX was used in the other. Isolates resistant to ampicillin and OTC that were obtained from an untreated flock at different sampling times were closely related to each other by pulsed-field gel electrophoresis patterns (PFGE) of XbaI-digested chromosomal DNA. PFGE analysis together with in vitro conjugation experiments suggested that diversity of resistance phenotypes within a clone may be resulted from the acquisition and loss of R-plasmids in an untreated and a treated flock. The numbers of resistance phenotypes observed among fecal isolates increased during the growth of the chickens in all the farms. The results in the present study suggest that persistence of commensal E. coli strains resistant to antimicrobials even in the absence of antimicrobial administration. It is also hypothesized that horizontal transmission of resistance determinants resulted in the emergence of different resistance phenotypes in those farms.     

Increased tracheal colonization in chickens without impairing pathogenic properties of avian pathogenic Escherichia coli MT78 with a fimH deletion

Several studies suggest that the expression of F1 fimbriae could be involved in the virulence of Escherichia coli for chickens. F1 fimbriae display multivalent properties such as adhesion to epithelia or interaction with the immune system that imply specific interactions between the adhesin FimH and different cell receptors. We constructed a delta fimH mutant of the avian pathogenic E. coli MT78 and evaluated its in vivo colonization and pathogenicity, as compared to that of the parent strain. The generated mutant PA68 was unable to adhere in vitro to chicken epithelial pharyngeal or tracheal cells; mutant bacteria were mostly afimbriated although a minority of them displayed altered piliation phenotypes. Two inoculation routes were used to compare the ability of MT78 and PA68 to colonize the respiratory tract and to induce colibacillosis in chickens. In the first model, 2-wk-old axenic chickens were inoculated intratracheally with one or both E. coli strains, after primary infection with infectious bronchitis virus. In the second model, 3-wk-old specific-pathogen-free chickens were inoculated via the caudal thoracic air sac. After intratracheal inoculation, the delta fimH mutant was found to be a better colonizer than MT78 in the trachea of inoculated chickens. Furthermore, when both strains were inoculated simultaneously, the delta fimH mutant constituted 98% of the bacterial population in the trachea at day 7 postinoculation. Irrespective to the inoculation route, MT78 and PA68 showed similar abilities to induce macroscopic lesions in chickens, to provoke bacteremia, and to colonize the internal organs. However, 4 days after intra-air sac inoculation, bacterial counts of the mutant were lower in the spleen and liver than those of MT78. Our results show that FimH is not required for colonization of the trachea of axenic chickens by E. coli and that it is not a major determinant of bacterial pathogenicity. On the contrary, the lack of expression of FimH seems to favor the in vivo colonization of the trachea of chickens by E. coli.     

Effect of carbohydrates on Salmonella typhimurium colonization in broiler chickens

The effect of carbohydrates in the drinking water of broiler chickens on Salmonella typhimurium colonization was evaluated. Results indicate that mannose and lactose (2.5%) significantly (P less than 0.05) reduced intestinal colonization of S. typhimurium by at least one-half, as compared with dextrose, maltose, and sucrose. Lactose and mannose also significantly reduced (P less than 0.01) the mean log10 number of S. typhimurium in the cecal contents. Although mannose was the most effective sugar at blocking colonization, lactose may be more practical because it is effective and costs much less than mannose. Provision of carbohydrates in the drinking water had no significant effect on weight gain.     

Carriage of potentially pathogenic Escherichia coli in chickens

DNA-DNA hybridization, cultured cell lines, and transmission electron microscopy were used to study pathogenicity traits of 64 Escherichia coli isolated from apparently healthy chickens from 18 small-scale farms in Thika District, Kenya. A total of 39 (60.9%) isolates hybridized with the eae gene probe for enteropathogenic E. coli (EPEC) whereas another 16 (25%) hybridized with the lt and st gene probes and were categorized as enterotoxigenic E. coli. Electron microscopic examination of the eae probe-positive E. coli cultures with the HT-2919A cell line confirmed that they were able to attach intimately and produced effacement typical of EPEC. In addition, negative stain electron microscopy showed that the EPEC strains produced pili that have previously been associated with increased virulence of E. coli infections in chickens. This study has also demonstrated that apparently healthy chickens may carry enteropathogenic E. coli strains.     

Acute airsacculitis in untreated and cyclophosphamide-pretreated broiler chickens inoculated with Escherichia coli or Escherichia coli cell-free culture filtrate.

Ninety commercial broiler chickens were divided into three equal groups; 30 were injected with brain-heart-infusion broth into the cranial thoracic air sacs (controls), 30 were similarly inoculated with a culture of Escherichia coli, and 30 were similarly inoculated with E. coli cell-free culture filtrate. Birds were examined from 0 to 6 hours post-inoculation. E. coli-inoculated and cell-free culture filtrate-inoculated chickens reacted similarly, with exudation of heterophils into the air sac. Microscopically, heterophils were present in low numbers perivascularly 0.5 hour after inoculation and became more numerous by 3 hours post-inoculation. By 6 hours post-inoculation, there was severe swelling of air sac epithelial cells and thickening of the air sac by proteinaceous fluid and heterophils. Ultrastructurally, air sac epithelial cells were swollen and vacuolated, and interdigitating processes were separated. Histologically and ultrastructurally, all features in control chickens were normal, with only rare heterophils in the air sac interstitium. In E. coli-inoculated and cell-free culture filtrate-inoculated chickens, cell counts (predominantly heterophils) in air sac lavage fluids increased markedly at 3 and 6 hours, with only slight increases in counts from lavages of controls. Heteropenia was observed in E. coli-inoculated chickens, whereas heterophilia was observed in cell-free filtrate chickens and controls. Ninety additional chickens were pretreated with cyclophosphamide, subdivided into three equal groups, and inoculated and examined similarly as above. Cyclophosphamide pretreatment reduced inflammatory changes in air sacs, lowered cell numbers in lavage fluids, and abolished hematologic changes; however, it did not prevent epithelial cell changes. These results indicate that cell-free culture filtrate of E. coli induces changes similar to those induced by cultures of E. coli.     

Bacteriophage treatment of a severe Escherichia coli respiratory infection in broiler chickens

A bacteriophage to a serotype 02, nonmotile Escherichia coli was isolated from municipal waste treatment facilities and poultry processing plants. A study was conducted to determine the efficacy of multiple vs. single intramuscular (i.m.) injections of bacteriophage to treat a severe E. coli respiratory infection. The birds were challenged at 7 days of age by injection of 6 x 10(4) colony-forming units (cfu) of E. coli into the thoracic air sac followed by an i.m. injection into the thigh with either heat-killed or active bacteriophage. There were 16 treatments with three replicate pens of 10 birds. There were four control treatments, which included untreated birds, birds injected with either heat-killed or active bacteriophage, and birds challenged only with E. coli. In the remaining treatments, birds were injected with heat-killed or active bacteriophage either once immediately after E. coli challenge or immediately after challenge and at 8 and 9 days of age, once at 8 days of age or at 8, 9, and 10 days of age, and once at 9 days of age or at 9, 10, and 11 days of age. Mortality was significantly decreased from 57% to 13% in the birds given a single i.m. injection of bacteriophage immediately after E. coli challenge, and there was complete recovery in birds treated immediately after challenge and at 8 and 9 days of age, which was a significant improvement from the single injection treatment. There was a significant reduction in mortality from 57% to 10% in the birds treated with bacteriophage once at 8 days of age and those birds treated at 8, 9, and 10 days of age, with no difference between single or multiple treatments. The mortality in the single or multiple phage treated birds that started at 9 days of age was reduced from 57% to 28% and 27%, respectively, but was not statistically different from the control. These data suggest that bacteriophage can be an effective treatment when administered early in this experimental E. coli respiratory disease and that early multiple treatments are better than a single treatment. The efficacy of bacteriophage treatment diminishes as it is delayed, with no difference between single or multiple treatments. Bacteriophage may provide an effective alternative to antibiotics, but like and biotic therapy, the effectiveness of phage to rescue animals decreases the longer treatment is delayed in the disease process.     

Effect of dietary lactose on Salmonella colonization of market-age broiler chickens

The effect of providing lactose in feed and inoculation with volatile fatty acid-producing anaerobic cultures (AC) of cecal flora on Salmonella typhimurium colonization was evaluated in broilers. One-day-old chicks were divided into four groups and provided 1) no lactose, no AC; 2) AC, no lactose; 3) AC and lactose on days 1-10; or 4) AC and lactose on days 1-40. All groups were challenged per os with 10(6) Salmonella on day 3 and with 10(8) Salmonella on day 33. Salmonella growth in the cecal contents was significantly decreased (P less than 0.01) on day 10 in the chicks provided lactose from day 1-10. However, after the removal of lactose from the diet, the chicks were susceptible to Salmonella colonization. The number of Salmonella in the ceca was significantly reduced (P less than 0.05) in the chicks provided lactose throughout the 40-day growing period. Dietary lactose decreased the pH of the cecal contents and was accompanied by marked increases in the concentrations of undissociated bacteriostatic volatile fatty acids in the cecal contents.     

Prevalence of pathogenic Escherichia coli in the broiler house environment

Matched sampling of Escherichia coli from broiler house litter and bird lesions of either cellulitis or colibacillosis was conducted to investigate the relationship of pathogenic E. coli to those found in the environment. Isolates were collected from six broiler flocks representing six geographically disparate ranches. Isolates were compared by flock for similarity in serotype and genotyped by pulsed-field gel electrophoresis. Serotyping revealed a considerable dissociation between the two groups of isolates. The prevalence of pathogenic E. coli that matched the environmental isolates from the same house was 0 to 3%. Statistical analysis of the serotype data showed a strong dependence of serotype on isolate source, indicating a high probability that a particular serotype would be found among lesions or litter but not in both groups. Genotyping of isolates on two farms supported the results of serotyping and provided differentiation of isolates that could not by typed by serology. These results suggested that the prevalence of pathogenic E. coli in the broiler house was independent of the prevalence of other commensal or environmental E. coli. Understanding the composition of E. coli populations in commercial poultry production may have bearing on the epidemiology and control of E. coli related diseases.     

Epidemiological studies of congo red Escherichia coli in broiler chickens.

This prospective cohort study was designed to confirm the association between Congo red binding Escherichia coli (CREC) and E. coli air sacculitis in commercial broilers. It was also designed to evaluate CREC as an air sacculitis risk factor and to explore the CREC relationship to other air sacculitis risk factors (poultry house temperature, air-ammonia levels, and presence of other diseases). In addition, this study was used to assess a possible role of the broiler-breeder flocks and hatchers in the spread of CREC air sacculitis. Congo red E. coli-associated airsacculitis risk was based on CREC exposure of the chicks in the hatchers. Breeder flocks with greater than 30 CREC colonies/plate from hatcher air sampling tests were placed in the high risk group; flocks with less than five CREC colonies/plate were placed in the low risk group. Increased risks of death due to air sacculitis (RR = 2.26), and increased death rates due to CREC air sacculitis (RR = 9.45) in high-risk flocks, identified CREC as an important air sacculitis risk factor. The attributable risk percent of CREC airsacculitis from hatcher exposure of CREC was 89.4%, pointing to the hatcher as the source of CREC infection. The association of specific broiler-breeder flocks to high levels of CREC in the hatchers, and subsequent air sacculitis, suggests that the broiler-breeders are the ultimate source of CREC.     

Virulence factors associated with Escherichia coli present in a commercially produced competitive exclusion product

In this study, we assessed the pathogenic potential of Escherichia coli associated with a commercial competitive exclusion (CE) product by examining the phenotypic characteristics associated with E. coli virulent for humans and domestic animals. Most E. coli isolates were capable of proliferating in iron-deplete chicken sera. Interestingly, none of the E. coli isolates from the commercial CE product contained the bacterial adhesin Tsh characteristic of avian pathogenic E. coli associated with airsacculitis and colisepticemia. In terms of virulence potential for humans, most E. coli isolates (78%) were sensitive to killing by 12.5% human sera. Because of their sensitivity to human sera, the E. coli in the CE product are not likely to cause a serious systemic infection in humans and, therefore, do not present a risk of causing septicemia in humans. Because these isolates also lack the gene tsh, they are also less likely to cause systemic disease or airsacculitis in poultry than pathogenic strains commonly isolated from diseased birds.     

Movement of broiler chickens reared in commercial conditions

1. Bird movement was studied at various ages in two flocks of 18,200 broiler chickens. 2. One-d-old chicks placed in corners of a 29 x 14 m brooding area dispersed evenly over the whole area in a period of 48 h. 3. At 15 d of age, when released from the brooding area, birds which were among the first to move to the far end of the house did not necessarily remain in that area of the house. 4. Starting at 4 and 5 weeks of age, birds marked individually were variable in their movement throughout the house during a 16- and 9-d period respectively. Even the least mobile individual covered an area of the house containing several thousand birds. 5. Closer scrutiny of movement by individual birds showed that some moved over areas of 20 m2 in an hour. Birds moved further than was necessary simply to reach food and water. No agonistic interactions were observed and there was no evidence that crowding was a physical restraint on bird movement. 6. Results indicated that birds were not generally attached to a particular site in the shed and that social factors did not restrict movement. Some implications of bird movement are discussed with regard to performance, welfare and leg abnormalities.     

Further studies on competitive exclusion for controlling Salmonellae in chickens

A native intestinal microflora of chickens which is protective against salmonella readily transferred to penmates and apparently to birds in adjacent pens. The microflora not only minimized infection resulting from exposure following colonization of the gut with microflora, but significantly abbreviated the period of infection when introduced after a salmonella infection was established in chicks. A microflora with undiminished protective activity, sensitive to only a few commonly used antibacterials, was established in a SPF-Cofal/Marek-negative population. Intestinal microflora from mourning doves was at least partially effective in protecting chicks against a naladixic-acid-resistant strain of Salmonella infantis. In limited tests with 2 of 3 sources of protective microflora, the growth rate of chicks in the absence of salmonellae was significantly improved. A hypothesis involving specificity of attachment between the glycocalyces of the protective microflora and of the intestinal mucosa is offered as the likely mechanism of protection.