Epidemiologic features of Campylobacter jejuni isolated from poultry broiler houses and surrounding environments as determined by use of molecular strain typing.

OBJECTIVE: To genetically type Campylobacter jejuni isolates from broiler houses or the external environment to identify the source of Campylobacter organisms in broiler chickens. SAMPLE POPULATION: Environmental samples associated with broiler chickens, in commercial grow-out houses. PROCEDURE: Polymerase chain reaction (PCR) was used to amplify flaB, and the amplicon was digested with Sau3A to create a restriction fragment length polymorphism assay; PCR was also used to detect a transcribed spacer region in the 23S rRNA gene. RESULTS: Isolates possessing a 23S spacer region were more prevalent outside broiler houses than inside. Houses that had previously contained chickens or lacked biosecurity procedures were more likely to contain isolates possessing the 23S spacer. One house contained only isolates possessing the spacer, whereas an adjacent house contained only isolates lacking the spacer. The flaB type detected in broiler houses was different from the type detected in the environment; however, many isolates within the broiler houses contained untypable flaB genotypes. CONCLUSIONS AND CLINICAL RELEVANCE: Most isolates from within houses were genetically distinct from isolates from outside houses that were examined by bacteriologic culture, suggesting an undetected source of C jejuni. Detection of isolates containing the 23S spacer appeared to be an indicator of environmental contamination of the houses. The observation of completely different C jejuni genetic types simultaneously within adjacent houses suggests that some types do not compete successfully during the grow-out period. In addition, the diversity of genotypes identified within broiler houses indicates the complexity of the ecologic features of C jejuni in the chicken environment.

Purified Chick Embryo Cell Culture Rabies Vaccine: interchangeability with Human Diploid Cell Culture Rabies Vaccine and comparison of one versus two-dose post-exposure booster regimen for previously immunized persons.

The equivalence and interchangeability of Purified Chick Embryo Cell Culture Rabies Vaccine (PCECV) to Human Diploid Cell Culture Rabies Vaccine (HDCV) and the immunogenicity of a reduced post-exposure regimen with PCECV was investigated. Statistical analyses revealed no difference (P

Molecular epidemiology of Campylobacter jejuni in broiler flocks using randomly amplified polymorphic DNA-PCR and 23S rRNA-PCR and role of litter in its transmission.

Poultry has long been cited as a reservoir for Campylobacter spp., and litter has been implicated as a vehicle in their transmission. Chicks were raised on litter removed from a broiler house positive for Campylobacter jejuni. Litter was removed from the house on days 0, 3, and 9 after birds were removed for slaughter. Chicks were raised on these three litters under controlled conditions in flocks of 25. None of these birds yielded C. jejuni in their cecal droppings through 7 weeks. Two successive flocks from the same Campylobacter-positive broiler house were monitored for Campylobacter colonization. Campylobacter jejuni prevalence rates were determined for each flock. Randomly amplified polymorphic DNA (RAPD)-PCR and 23S rRNA-PCR typing methods were used to group isolates. A high prevalence (60%) of C. jejuni in flock 1 coincided with the presence of an RAPD profile not appearing in flock 2, which had a lower rate of prevalence (28%). A 23S rRNA-PCR typing method was used to determine if strains with different RAPD profiles and different prevalence rates contained different 23S sequences. RAPD profiles detected with higher prevalence rates contained a spacer in the 23S rRNA region 100% of the time, while RAPD profiles found with lower prevalence rates contained an intervening sequence less than 2% of the time. Data suggest varying colonizing potentials of different RAPD profiles and a source other than previously used litter as a means of transmission of C. jejuni. These molecular typing methods demonstrate their usefulness, when used together, in this epidemiologic investigation.

Current recommendations for the prophylaxis and treatment of rabies.

Once onset of clinical rabies develops in an individual, death is inevitable. Thus, it is imperative that, for persons exposed or potentially exposed to rabies virus, prophylaxis must be instituted as soon as possible following the exposure. Local wound management is an essential part of postexposure rabies prophylaxis. Exposed persons should receive a recommended series of a tissue culture or cell culture origin vaccine. The number of doses and route of vaccination differ in various regions of the world and are discussed in the text. The administration of a rabies immune globulin is generally recommended in conjunction with the first dose of the rabies vaccine. Nerve tissue origin vaccines, although used extensively in some parts of the world, are not recommended if cell or tissue culture vaccines are available. Decision trees are presented in the text to aid in determining if rabies vaccine is necessary following a known or presumed exposure to the virus, along with a table outlining the various rabies vaccines available in the World. Rabies pre-exposure immunisation is recommended for those individuals at risk of exposure to the virus. Pre-exposure prophylaxis consists of 3 doses of an approved rabies vaccine administered either intramuscularly or intradermally on days 0, 7, and 21 or 28 with periodic booster doses or titre determination depending on the level of risk of potential exposure to the virus.

Pathogenesis of experimentally induced rabies in domestic ferrets.

OBJECTIVE: To determine susceptibility, incubation and morbidity periods, clinical signs, serologic response, and excretion of virus in domestic ferrets inoculated with rabies virus. ANIMALS: 55 domestic ferrets. PROCEDURE: 5 groups of 10 ferrets were inoculated with rabies virus, IM, at doses of 10(5.5) to 10(1.5) median mouse intracerebral lethal dose. Ferrets were observed and behavior was recorded. Rectal temperature, body weight, and samples from the oral cavity and samples of saliva and blood were obtained. Virus isolation was attempted, using intracranial mouse inoculation and cell culture. Virus neutralizing antibodies were determined by rapid fluorescent focus inhibition test. Ferrets were euthanatized immediately if clinical signs were severe. Rabies was confirmed by direct immunofluorescent antibody test. RESULTS: Mean incubation period was 33 days (range, 16 to 96 days). Clinical signs included ascending paralysis, ataxia, cachexia, bladder atony, fever, hyperactivity, tremors, and paresthesia. Mean morbidity period was 4 to 5 days (range, 2 to 10 days). Virus antigen was detected in brain tissue from all clinically rabid ferrets. Ferrets given the highest viral dose were euthanatized and had VNA; ferrets receiving the next dilution also were euthanatized, but only 4 had seroconverted. Of 17 ferrets that survived, 5 seroconverted. Survivors remained clinically normal except for 1 that recovered with severe paralytic sequelae. Rabies virus was isolated from the salivary gland of 1 ferret that was euthanatized. CONCLUSIONS AND CLINICAL RELEVANCE: Rabies should be considered as a differential diagnosis in any ferret that has acute onset of paralysis or behavioral changes and a condition that rapidly deteriorates despite intense medical intervention.

A global review of rabies vaccines for human use.

Rabies is one of the oldest known diseases of mankind, yet it has been only slightly more than 100 years since Pasteur developed the first vaccine for post-exposure treatment. Since this first crude nerve tissue vaccine, numerous other rabies vaccines for human use have been developed and used with varying degrees of effectiveness and safety. When used appropriately, new cell culture vaccines provide nearly 100% protection with a high degree of safety: yet over 40,000 people world-wide die from rabies each year. Several pre- and post-exposure controlled vaccine trials and clinical studies have shown that the purified chick embryo cell (PCEC) vaccine, Rabipur, is as safe and effective as the rabies human diploid cell vaccine (HDCV), which is currently considered the gold standard. Additionally, PCEC vaccine does not result in immune-mediated hypersensitivity reactions following booster doses seen in about 6% of those receiving HDCV boosters following an initial series of HDCV.

Epidemiological study of Campylobacter spp. in broilers: source, time of colonization, and prevalence.

From October 1993 to August 1994, broiler chickens in four grow-out houses, two previously used (houses 1 and 2) and two newly constructed (houses 3 and 4), were used in a study to determine the source, time of infection, and prevalence of Campylobacter spp. Cecal droppings and cecal samples were obtained from the broilers. Samples were also obtained from water, feed, litter, soil, fans, and workers' boots. Samples were obtained from domestic animals and wildlife species (rectal swabs), including insects, on or near the premises. Broilers in houses 2, 3, and 4 became infected in the second or third week and were fully colonized by day 42. Campylobacter appeared in house 1 during week 2 in a low percentage of the birds, disappearing by the fourth week. Isolates were also recovered from domestic pigs and water on this farm. In house 3, the organism was isolated from workers' boots and a wild bird prior to isolation from the broilers. Following isolation from cecal droppings, the organism was isolated from water, feed, litter, feathers, flies, cattle, feces, and wild animals. In house 2, Campylobacter was isolated from cattle feces and wild birds prior to week 5, when the broilers first became infected, and thereafter from water, feed, insect, and wildlife, and cecal droppings. It was subsequently isolated from workers' boots, cattle feces, feathers, insects, and other wildlife. All ceca taken from 20 birds each from houses 2 and 3 were positive at time of slaughter (day 49). All ceca from house 1 were negative. No ceca were collected from birds originating in house 4. No specific source could be identified from the samples obtained, although apparently the organism permeates the environment and several potential sources are discussed in this paper.

Flagellin A gene restriction fragment length polymorphism patterns of Campylobacter spp. isolates from broiler production sources.

The purpose of this study was to determine the potential reservoirs for Campylobacter spp. that provide the initial sources involved with broiler chicken colonization during poultry production. We characterized the flagellin A gene (flaA) of the organism by restriction fragment length polymorphism (RFLP) for 59 isolates of the bacterium provided during an epidemiological study. Isolates were obtained from three broiler production houses existing at separate locations. They were cultured and isolated from other (nonbroiler) domestic farm animals, wild birds, rodents, feed, farmers' boots, chicken feathers, and chicken intestinal materials. Eight distinctive flaA types were found in two of the houses. In one house, at least five flaA types (4, 6, 8, 15, and 21) were characterized from the poultry production environment, with three types isolated and identified from the chicken intestinal tract. flaA type 15 was found in flies, on boots, and in chicken intestinal samples. In another house, a distinctive diversity of flaA types existed (4, 7, 43, and 53). At least three flaA types found in samples from chicken intestinal tracts were also found in warm-blooded animals outside of the poultry house (domesticated animals, wild birds, and vermin).

A simulation of microbial competition in the human colonic ecosystem.

Many investigations of the interactions of microbial competitors in the gastrointestinal tract used continuous-flow anaerobic cultures. The simulation reported here was a deterministic 11-compartment model coded by using the C programming language and based on parameters from published in vitro studies and assumptions were data were unavailable. The resource compartments were glucose, lactose and sucrose, starch, sorbose, and serine. Six microbial competitors included indigenous nonpathogenic colonizers of the human gastrointestinal tract (Escherichia coli, Enterobacter aerogenes, Bacteroids ovatus, Fusobacterium varium, and Enterococcus faecalis) and the potential human enteropathogen Salmonella typhimurium. Flows of carbon from the resources to the microbes were modified by resource and space controls. Partitioning of resources to the competitors that could utilize them was calculated at each iteration on the basis of availability of all resources by feeding preference functions. Resources did not accumulate during iterations of the model. The results of the computer simulation of microbial competition model and for various modifications of the model. The results were based on few measured parameters but may be useful in the design of user-friendly software to aid researchers in defining and manipulating the microbial ecology of colonic ecosystems as relates to food-borne disease.

Virulence determinants invA and spvC in salmonellae isolated from poultry products, wastewater, and human sources.

The presence of two virulence foci, invA and spvC, in Salmonella isolates obtained from poultry, wastewater, and human sources was determined. All isolates (n = 245) were positive for the invA gene sequence. Differences in degree of invasiveness were apparent with the Madin Darby canine kidney cell line, as only 79 of 159 randomly selected isolates (49.7%) tested were invasive at > 0.1% of the inoculum. 25% were invasive between 0.1 and 1.0% of the inoculum, and 24.5% were invasive at > 1.0% of the inoculum. There was a significant correlation between degree of invasion and source from which the isolate was recovered but no correlation between geographic origin of poultry isolates and degree of invasion. Only 37 of 245 isolates (15.1%) hybridized with the spvC DNA probe. All isolates that were recovered from a commercial egg production environment and chicken eggs and whose sequences exhibited homology with the spvC gene sequence were determined to be either Salmonella enteritidis PT 23 or PT 13. The sequences of few isolates from ceca and none from wastewater or humans demonstrated homology with the spvC gene.

Safety and immunogenicity of Lyssavac Berna human diploid cell rabies vaccine in healthy adults.

A clinical trial testing the safety and immunogenicity of a newly developed human diploid cell rabies vaccine (Lyssavac-HDC) was conducted on subjects at three colleges of veterinary medicine in the United States. Lyssavac-HDC is a sterile lyophilized vaccine containing no antibiotics or preservatives and is administered intramuscularly as a 0.5 ml dose of vaccine containing at least 2.5 i.u. of rabies inactivated antigen per dose. Subjects were given either a three dose pre-exposure series (days 0, 7, and 28), followed by one booster dose of vaccine (day 360); or a five dose simulated post-exposure series of injections (days 0, 3, 7, 14, and 28). All subjects in the post-exposure and pre-exposure groups possessed adequate levels of rabies neutralizing antibody (> or = 5) when tested on day 14 and day 28, respectively. Subjects in the pre-exposure group demonstrated a vigorous anamnestic response after the administration of one booster dose of vaccine on day 360. The type and severity of local and systemic reactions observed were comparable to other primary cell culture rabies vaccines. Significantly, there were no type III hypersensitivity reactions reported in subjects previously immunized with Lyssavac-HDC after the administration of a booster dose of vaccine on day 360.

Bordetella avium hemagglutination and motility mutants: isolation, characterization, and pathogenicity.

Transposon mutagenesis was used to produce Bordetella avium mutants, which were screened for the lack of potential virulence factors, including a hemagglutinin, flagella, pili, and toxins. A mini-Tn10 transposon containing a kanamycin-resistance gene was introduced into the chromosomal DNA of the virulent 002/S isolate by electroporation. A hemagglutination-negative (HA-) mutant and a motility-negative mutant were obtained. Southern blot analysis showed that only the motility-negative mutant contained the transposon, whereas the HA- mutant was a spontaneous kanamycin-resistant mutant. Both mutants were stable in vitro and in vivo. Following inoculation of 2-week-old poults, the HA- mutant was determined to be less virulent than the 002/S parent, whereas the motility-negative mutant was similar in virulence to the 002/S parent. These results indicate that the hemagglutinin of B. avium is a virulence factor, but motility does not appear to contribute to virulence.

Risk factors for systemic hypersensitivity reactions after booster vaccinations with human diploid cell rabies vaccine: a nationwide prospective study.

To determine the incidence of and risk factors for adverse reactions following the boosters, we conducted a nationwide prospective study of persons receiving pre-exposure booster vaccination with human diploid cell rabies vaccine (HDCV). Persons who had previously received three pre-exposure doses of HDCV and whose rabies neutralizing antibody titres were < or = 1:5 were enrolled in the study if they stated that they intended to receive a booster. Of the 98 persons enrolled in the study, 40 (41%) were in risk groups for whom boosters are not recommended. Three (3%) of 98 developed generalized urticaria or wheezing within 1 day of receiving boosters and three others (3%) developed urticaria 6 to 14 days after the booster. No differences were found between individuals with reactions (either type) and those with no adverse reaction according to age, gender, occupation, history of previous allergies, or time since or route of primary vaccination. Reactions were somewhat more common among persons who received primary vaccinations by the intramuscular route (i.m.) and booster vaccinations by the intradermal route (i.d.) (3/15, 20%) or primary vaccinations i.d. and booster vaccinations i.m. (2/10, 20%), and somewhat less common among persons who received both these vaccinations i.d. (1/52, 2%) or i.m. (0/7). The number of persons who develop allergic reactions may be minimized by administering vaccinations only when vaccination is strictly indicated. The influence of the route of primary and booster vaccinations on the development of reactions deserves further study.

Isolation of Salmonella from ovaries and oviducts from whole carcasses of spent hens.

Spent hens containing hard-shelled eggs were obtained to study the influence of sample collection and sample preparation methodology on the detection of Salmonella in ovaries. Four hundred eighteen birds from 19 flocks were collected, and the carcasses were opened aseptically within 8-12 hours of collection. A sample set containing hard-shelled egg, ovary, and oviduct section were collected from each carcass, and surface contaminants were removed from ovaries with polyoxyethylene ether. Three of 19 flocks (15.8%) and six of 407 ovary samples were positive for Salmonella; two oviduct samples were positive (0.5%). No eggs were Salmonella-positive. Single and multiple serotypes were detected in ovaries. Results indicated that Salmonella recovery rates can be significantly affected by speed in processing samples after collection, by cleanliness of the tissue-collection environment (laboratory vs. slaughter plant), and by removal of surface contaminants.

Frequency of Salmonella enteritidis and other salmonellae in the ceca of spent hens at time of slaughter.

A study was conducted to determine the frequency of Salmonella enteritidis (SE) and other Salmonella serovars in the cecal contents of spent laying hens at a hen-processing plant in the southeastern United States over a 4 1/2-month period, from October 1990 through February 1991. A total of 1920 pooled cecal samples (three ceca per sample) from 38 flocks representing 23 producers were obtained and tested for the presence of SE and other Salmonella serovars. A total of 359 samples (18.7%) from 37 of the 38 flocks (97.4%) showed characteristic reactions for salmonellae on triple sugar iron agar (TSIA) slants. Twenty-nine of the 359 Salmonella-positive samples (8.1%) were Group D-positive, all of which were found to be SE on further serotyping. The SE-positive samples were from seven of the 38 flocks (18.4%); four flocks originated from the USDA/APHIS-designated Northern Region of the United States, and three were from the Southeastern Region. Serotyping of the 330 TSIA-positive Group-D negative Salmonella revealed 37 different serovars. S. heidelberg, the predominant serovar, was identified in 49.1% of these isolates.

Prevalence of toxoplasmosis in swine from Iowa.

Of swine from 104 herds, 2,616 were tested for antibodies against Toxoplasma gondii, using an ELISA. Data were analyzed according to swine type, herd size, facility type, and season. The true prevalence of toxoplasmosis was estimated as 5.4% among finishing swine and 11.4% among sows and gilts. Herds with less than 100 breeding swine were significantly (P less than 0.05) more likely to be infected than were herds with greater than or equal to 100 breeding swine. The rate of seropositivity in breeding swine was approximately the same in infected herds, regardless of herd size. Herds with finishing swine maintained in total confinement were as likely to become infected as were herds maintained in other types of facilities, but infected herds with finishing swine maintained in confinement appeared to have a lower in-herd prevalence than did herds maintained in other types of facilities (P = 0.09). Seasonal effects were not observed, and prevalence remained relatively constant throughout the year.