Teresa Y. Morishita

Campylobacter colonization in poultry: sources of infection and modes of transmission

Abstract Since its recognition as a human pathogen in the early 1970s, Campylobacter jejuni has now emerged as the leading bacterial cause of food-borne gastroenteritis in developed countries. Poultry, particularly chickens, account for the majority of human infections caused by Campylobacter. Reduction or elimination of this pathogen in the poultry reservoir is an essential step in minimizing the public health problem; however, farm-based intervention measures are still not available because of the lack of understanding of the ecological aspects of C. jejuni on poultry farms. Although Campylobacter is highly prevalent in poultry production systems, how poultry flocks become infected with this organism is still unknown. Many investigations indicate that horizontal transmission from environmental sources is the primary route of flock infections by Campylobacter. However, some recent studies also suggest the possibility of vertical transmission from breeder to progeny flocks. The transmission of the organism is not well understood, but it is likely to be through both vertical and horizontal transmission and may be affected by the immune status of the poultry host and the environmental conditions in the production system. Intervention strategies for Campylobacter infection in poultry should consider the complex nature of its transmission and may require the use of multiple approaches that target different segments of the poultry production system.

Evaluation of an Avian-specific probiotic to reduce the colonization and shedding of Campylobacter jejuni in broilers

Campylobacter jejuni has often been responsible for human gastroenteritis. Poultry have often been implicated as a source for these human infections. Intestinal colonization of C. jejuni in the chicken plays a role in carcass contamination during slaughter. Thus, reducing C. jejuni colonization in chickens can potentially reduce the incidence of C. jejuni infections in humans. The use of probiotics to competitively exclude the colonization of intestinal pathogens has been proposed for poultry. Hence, the purpose of this study was to evaluate the use of an avian-specific probiotic containing Lactobacillus acidophilus and Streptococcus faecium for reducing the shedding and colonization of C. jejuni in the chicken intestinal tract. Day-old chicks were randomly allocated into either a probiotic-treated group or a control group. The treated group was given probiotic from day 1 to day 3, and the control group was not given any probiotic. Six hours after the first oral administration of probiotics (treatment) or double distilled water (control), these chicks were challenged with C. jejuni. The frequency of the C. jejuni shedding was monitored until market age. Intestinal colonization was determined for the two experimental groups at slaughter. Results indicated that chickens given probiotics from day 1 to day 3 had a 70% reduction in the frequency of C. jejuni shedding in colonized chicks (P = 0.0001) and a 27% reduction in jejunal colonization in colonized chicks (P = 0.0001) at slaughter when compared with the control group. Thus, the use of the avian-specific probiotic containing L. acidophilus and S. faecium can reduce the colonization and frequency of fecal shedding of C. jejuni in market-aged broilers.

Prevalence, Antigenic Specificity, and Bactericidal Activity of Poultry Anti-Campylobacter Maternal Antibodies

ABSTRACT Poultry are considered the major reservoir for Campylobacter jejuni, a leading bacterial cause of human food-borne diarrhea. To understand the ecology of C. jejuni and develop strategies to control C. jejuni infection in the animal reservoir, we initiated studies to examine the potential role of anti-Campylobacter maternal antibodies in protecting young broiler chickens from infection by C. jejuni. Using an enzyme-linked immunosorbent assay (ELISA), the prevalence of anti-C. jejuni antibodies in breeder chickens, egg yolks, and broilers from multiple flocks of different farms were examined. High levels of antibodies to the organism were detected in serum samples of breeder chickens and in egg yolk contents. To determine the dynamics of anti-Campylobacter maternal antibody transferred from yolks to hatchlings, serum samples collected from five broiler flocks at weekly intervals from 1 to 28 or 42 days of age were also examined by ELISA. Sera from the 1-day and 7-day-old chicks showed high titers of antibodies to C. jejuni. Thereafter, antibody titers decreased substantially and were not detected during the third and fourth weeks of age. The disappearance of anti-Campylobacter maternal antibodies during 3 to 4 weeks of age coincides with the appearance of C. jejuniinfections observed in many broiler chicken flocks. As shown by immunoblotting, the maternally derived antibodies recognized multiple membrane proteins of C. jejuni ranging from 19 to 107 kDa. Moreover, in vitro serum bactericidal assays showed that anti-Campylobacter maternal antibodies were active in antibody-dependent complement-mediated killing of C. jejuni. Together, these results highlight the widespread presence of functional anti-Campylobacter antibodies in the poultry production system and provide a strong rationale for further investigation of the potential role of anti-C. jejunimaternal antibodies in protecting young chickens from infection byC. jejuni.

SHEDDING AND COLONIZATION OF CAMPYLOBACTER JEJUNI IN BROILERS FROM DAY-OF-HATCH TO SLAUGHTER AGE

Poultry are considered to be the primary reservoirs of Campylobacter jejuni for humans. Campylobacter jejuni can colonize the poultry intestinal tract and its subsequent shedding can result in environmental contamination, resulting in an increased risk of infection for the rest of the flock. At present, there is no information on the daily shedding pattern of C. jejuni in broiler chickens. Thus, the purpose of this study was to determine the daily shedding pattern of C. jejuni in broiler chickens and to correlate intestinal colonization with fecal shedding, which would aid in the development of intervention strategies such as the use of competitive exclusion products at 1 day of age. Twenty-four broiler chicks were orally inoculated with 1.6 x 10(7) colony-forming units of C. jejuni, and the reisolation rate of the organism was determined daily from day 1 to day 43. Fifty percent and 70% of the chicks were shedding C. jejuni within 24 and 48 hr postinoculation, respectively. The group collectively reached a peak excretion on days 13-19 postinoculation. There was a steady decline in fecal shedding after the third week. By market age, on day 43, only 37.5% (9/24) of the birds were shedding C. jejuni in their feces. Throughout the sampling period from days 1 to 43, a cyclic pattern of shedding was observed in individual birds. Individual birds excreted C. jejuni on an average of 25 out of 43 days. The C. jejuni isolate failed to colonize 16.6% (4/24) of the birds. A small percentage of the birds, 12.5% (3/24), were observed to be chronic shedders. Enumeration of C. jejuni in the crop, jejunum, and cecum on day 43 revealed that the cecum was the major colonization site, and 15 out of the 24 birds carried C. jejuni in their intestinal tract.

Fecal shedding and antimicrobial susceptibility of selected bacterial pathogens and a survey of intestinal parasites in free-living waterfowl.

Free-living waterfowl residing in metropolitan parks in central Ohio were surveyed for the fecal shedding and antimicrobial susceptibility patterns of Campylobacter jejuni, Escherichia coli, Salmonella spp., and Pasteurella multocida. In addition, a survey for intestinal parasites was also conducted in these same waterfowl to determine parasite burdens in free-living waterfowl. Prevalences of 67%, 50%, and 0.2% of E. coli, C. jejuni, and Salmonella spp., respectively, were observed for all waterfowl species. Pasteurella multocida was not isolated from the sampled population. Salmonella java was isolated from one mallard duck. Statistically, there was a significantly higher E. coli isolation rate for mallard ducks than for Canada geese, but no difference was observed for C. jejuni isolation rates between waterfowl species. Antimicrobial susceptibility testing was conducted via the disk diffusion method and multidrug resistance was exhibited for penicillin G, lincomycin, vancomycin, erythromycin, and bacitracin. In addition, the prevalence of endoparasites in these sampled waterfowl ranged from 5% to 66%. Protozoan oocysts were most prevalent followed by nematode ova. No trematode or cestode ovum was recovered from this sampled population.

Survey of Pathogens and Blood Parasites in Free-Living Passerines

To determine the disease prevalence of free-living passerines, 1709 passerines were sampled from 38 different field sites in Ohio. Choanal and cloacal swabs were collected from each bird and cultured for the presence of Pasteurella multocida, Salmonella spp., and Escherichia coli by standard microbiologic techniques. In addition, the serum from each bird was analyzed for the presence of antibodies to Mycoplasma gallisepticum, Mycoplasma synoviae, Newcastle disease virus, and avian influenza virus. A blood smear was also made to examine for the presence of blood parasites. Results indicated that the isolation of E. coli varied with bird species, with the European starling having a higher (21.4%) isolation of E. coli. Salmonella spp. were also isolated from these free-living passerines. Pasteurella multocida was not isolated from any of the sampled passerines. These birds did not have antibodies to M. gallisepticum, M. synoviae, Newcastle disease virus, or avian influenza virus. Blood parasites were not detected in any of the birds sampled.

Relative Ammonia Concentrations, Dust Concentrations, and Presence of Salmonella Species and Escherichia coli Inside and Outside Commercial Layer Facilities

Abstract Three air contaminants that may have serious health consequences to humans and poultry are ammonia, dust, and aerosolized bacteria. This study measured ammonia concentrations, dust concentrations, and the presence or absence of aerosolized Salmonella spp. and Escherichia coli inside and outside five commercial layer facilities. The average outside ammonia concentration measurements decreased as the distance from the facility increased from 10 to 40 ft. The measurements at 10 ft from the facilities were consistently higher than the average concentrations inside the facilities. The ammonia measurement trends inside of the facilities were affected by the temperature-dependent ventilation systems. Average dust concentrations inside the five facilities were consistently below 2 mg/m3. Three facilities also experienced average outside dust concentrations at all measured distances below 2 mg/m3. Two facilities had relatively high average dust measurements at 10 ft from ventilation fans (32.12 mg/m3 and 75.18 mg/m3). Escherichia coli and Salmonella were isolated from the air inside all five facilities and outside the facilities up to 40 ft from the ventilation fans. In conclusion, dust concentrations may pose the largest risk to human and animal health at 10 ft away from the poultry facilities; risks associated with ammonia inhalation are greatest inside facilities during the coolest months of the year; and aerosolized bacteria are found inside and outside poultry facilities, but further work is needed to quantify the bacteria to further assess the health risk related to this issue.

Survey of Parasites and Bacterial Pathogens from Free-Living Waterfowl in Zoological Settings

To determine if free-living waterfowl residing in a zoological setting pose health risks for its animal collections, visitors, and employees, 450 fecal samples were collected and cultured for the presence of Campylobacter jejuni, Escherichia coli, Salmonella spp., and Pasteurella multocida. A survey of endoparasites infecting the waterfowl was also conducted. Sixty-seven percent, 42%, and 1.7% of the samples tested positive for E. coli, C. jejuni, and Salmonella spp., respectively. No P. multocida was isolated from the sampled population. Antimicrobial susceptibility testing for the bacterial isolates demonstrated that a majority of the isolates were susceptible to the antibiotics tested. A survey for parasites revealed 16% of the samples had coccidia oocytes; 8% of the sample had spirurid ova; and 17% of the sample had strongylate-type nematode ova. Ascaris spp. ova, Capillaria spp. ova, oxyurid ova, and mites were also noted in some fecal samples.

Prevalence of Salmonella, Campylobacter, and Escherichia coli on ostrich carcasses and the susceptibility of ostrich-origin E. coli isolates to various antibiotics.

There has been limited research on the prevalence of foodborne pathogens such as Escherichia coli O157:H7, Salmonella, and Campylobacter on ostrich carcasses. Likewise, few studies have been done in ostriches to determine the antimicrobial susceptibilities of common bacteria, like E. coli. In this study, ostrich carcasses were sampled from eight slaughterhouses in Ohio and one in Indiana. Although results demonstrated no E. coli O157:H7 from the carcasses sampled, 91% (116/128) of the dressed carcasses sampled had E. coli present. One carcass sample (1/152) was positive for Salmonella. Campylobacter were detected in 10% (19/191) of the carcasses. Antimicrobial susceptibility testing on 93 carcass E. coli isolates showed resistance to erythromycin (99%), neomycin (65%), netilmicin (2%), oxytetracycline (22%), streptomycin (2%), and trimethoprim (3%). All isolates were resistant to bacitracin, lincomycin, penicillin, and vancomycin. For the large intestinal sampling, 149 of the 217 (69%) samples had E. coli present. Fifty of these 149 samples had E. coli levels ranging from 10(2) to 10(5) colony-forming units/g feces. Campylobacter were isolated from 6 of 201 (3%) samples. No Salmonella colony was detected. Antimicrobial susceptibility testing on 131 intestinal E. coli isolates showed resistance to erythromycin (98%), neomycin (66%), netilmicin (34%), oxytetracycline (34%), streptomycin (40%), and trimethoprim (13%). All isolates were resistant to bacitracin, lincomycin, penicillin, and vancomycin.

Experimental oral inoculations in birds to evaluate potential definitive hosts of Neospora caninum

Experimental oral inoculations to evaluate potential definitive hosts of Neospora caninum were conducted by feeding infected rodent tissues to 9 carnivorous birds of 4 species. Birds included 2 red-tailed hawks (Buteo jamaicensis), 2 turkey vultures (Cathartes aura), 2 barn owls (Tyto alba), and 3 American crows (Corvus brachyrhynchus). The rodents (mice or rats) had been inoculated with 100,000 culture-derived tachyzoites of N. caninum 1-6 mo before feeding to the birds. Fecal samples were collected from each bird daily for 1 mo after feeding rodents and examined for oocysts by fecal flotation. In addition, processed aliquots from all avian fecal samples were fed to BALB/c mice. Five weeks after feeding, mice were bled and sera were tested for antibodies against N. caninum. One to two months later, mice were killed and brain tissue was examined microscopically for protozoal cysts. While occasional oocysts were found in avian fecal samples, these were likely not N. caninum because they were not infective to BALB/c mice. It was concluded that the bird species tested are not likely to be definitive hosts of N. caninum.