Summary of the 2001-02 Pennsylvania H7N2 low pathogenicity avian influenza outbreak in meat type chickens.

H7N2 low-pathogenicity (LP) avian influenza (AI) virus was isolated from chickens submitted to the Pennsylvania Animal Diagnostic Laboratory System on December 4 and 5, 2001. The cases were from two broiler breeder flocks in central Pennsylvania that had clinical signs of an acute, rapidly spreading respiratory disease. Seroconversion to AI virus was detected on follow-up sampling. Subsequently, H7N2 LPAI virus was isolated in five different broiler flock cases submitted between December 14, 2001, and January 3, 2002. Clinical signs and lesions in broilers, when present, were compatible with multicausal respiratory disease. With the exception of one broiler flock that was processed, birds from all of the virus positive flocks were euthanatized in-house within 11 days of the original case submission date. Increased surveillance of poultry flocks within 10-mile radius zones centered at the foci of the positive farms continued until March 1, 2002. No additional cases were detected.

Epidemiology, production losses, and control measures associated with an outbreak of avian influenza subtype H7N2 in Pennsylvania (1996-98).

An outbreak of H7N2 low-pathogenicity (LP) avian influenza (AI) occurred in a two-county area in Pennsylvania from December of 1996 through April of 1998. The outbreak resulted in infection of 2,623,116 commercial birds on 25 premises encompassing 47 flocks. Twenty-one (one premise with infection twice) of the twenty-five infected premises housed egg-laying chickens and one premise each had turkeys, layer pullets, quail, and a mixed backyard dealer flock. Despite dose proximity of infected flocks to commercial broiler flocks, no infected broilers were identified. Experimentally, when market age broilers were placed on an influenza-infected premise they seroconverted and developed oviduct lesions. The outbreak was believed to have originated from two separate introductions into commercial layer flocks from premises and by individuals dealing in sales of live fowl in the metropolitan New York and New Jersey live-bird markets. Source flocks for these markets are primarily in the northeast and mid-Atlantic areas, including Pennsylvania. Mixed fowl sold include ducks, geese, guinea hens, quail, chukar partridges, and a variety of chickens grown on perhaps hundreds of small farms. Infections with the H7N2 AI virus were associated with variable morbidity and temporary decreases in egg production ranging from 1.6% to 29.1% in commercial egg-laying chickens. Egg production losses averaged 4.0 weeks duration. Mortality ranged from 1.5 to 18.3 times normal (mean of 4.3 times normal). Duration of mortality ranged from 2 to 13 weeks (average of 3.9 weeks) in flocks not depopulated. Lesions observed were primarily oviducts filled with a mucous and white gelatinous exudates and atypical egg yolk peritonitis. Quarantine of premises and complete depopulation were the early measures employed in control of this outbreak. Epidemiological studies suggested that depopulation furthered the spread of influenza to nearby flocks. Thereafter, later control measures included quarantine, strict biosecurity, and controlled marketing of products.

Lipopolysaccharide O-chain microheterogeneity of Salmonella serotypes Enteritidis and Typhimurium.

Variability in the lipopolysaccharide (LPS) of the two most prevalent Salmonella serotypes causing food-borne salmonellosis was assessed using gas chromatography analysis of neutral sugars from 43 Salmonella enterica serovar Enteritidis (S. Enteritidis) and 20 Salmonella enterica serovar Typhimurium (S. Typhimurium) isolates. Four substantially different types of O-chain chemotypes were detected using cluster analysis of sugar compositions; these were low-molecular-mass (LMM) LPS, glucosylated LMM LPS, high-molecular-mass (HMM) LPS and glucosylated HMM LPS. Nineteen out of 20 S. Typhimurium isolates yielded glucosylated LMM. In contrast, S. Enteritidis produced a more diverse structure, which varied according to the source and history of the isolate: 45.5% of egg isolates yielded glucosylated HMM LPS; 100% of stored strains lacked glucosylation but retained chain length in some cases; and 83.3% of fresh isolates from the naturally infected house mouse Mus musculus produced glucosylated LMM LPS. A chain length determinant (wzz) mutant of S. Enteritidis produced a structure similar to that of S. Typhimurium and was used to define what constituted significant differences in structure using cluster analysis. Fine mapping of the S. Enteritidis chromosome by means of a two-restriction enzyme-ribotyping technique suggested that mouse isolates producing glucosylated LMM LPS were closely related to orally invasive strains obtained from eggs, and that stored strains were accumulating genetic changes that correlated with suppression of LPS O-chain glucosylation. These results suggest that the determination of LPS chemotype is a useful tool for epidemiological monitoring of S. Enteritidis, which displays an unusual degree of diversity in its LPS O-chain.

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.

Management and environmental risk factors for Salmonella enteritidis contamination of eggs.

OBJECTIVE: To analyze data for 60 poultry flocks voluntarily enrolled in the Pennsylvania Salmonella enteritidis Pilot Project and determine management and environmental risk factors associated with production of S enteritidis-contaminated eggs. SAMPLE POPULATION: 60 flocks for which at least 1 environmental sample (manure or egg-handling equipment) was positive for S enteritidis. PROCEDURE: Samples of manure, egg-handling equipment, and mice were submitted for bacterial culture of S enteritidis. When S enteritidis was isolated from environmental samples, 1,000 eggs were collected from the flock every 2 weeks for 8 weeks and submitted for bacterial culture. RESULTS: 18 flocks were found to have produced contaminated eggs. Estimated overall prevalence of contaminated eggs was 2.64/10,000 eggs produced, but flock-specific prevalence ranged from 0 to 62.5/10,000 eggs. Flocks with high levels of manure contamination were 10 times as likely to produce contaminated eggs as were flocks with low levels. However, 5 flocks with low levels of manure contamination produced contaminated eggs. CONCLUSIONS: Evaluation of the level of manure contamination could be used to help identify flocks at risk of producing S enteritidis-contaminated eggs. CLINICAL RELEVANCE: Flocks with high levels of S enteritidis-contaminated manure appeared to pose the greatest public health threat, and on-farm programs to reduce the prevalence of egg contamination should be developed for farms with high levels of manure contamination. Efforts to reduce the overall number of on-farm pathogens should decrease the incidence of foodborne disease in humans.

On-farm monitoring of mouse-invasive Salmonella enterica serovar enteritidis and a model for its association with the production of contaminated eggs.

Mice (Mus musculus) captured in henhouses were assessed for the presence of salmonellae in spleens. Of 621 and 526 spleens cultured during the first and second years of collection, 25.0 and 17.9%, respectively, were positive for Salmonella enterica serovar Enteritidis. Contaminated eggs were cultured from nine houses during the first year of sampling, and for eight of these houses, serovar Enteritidis was recovered from the spleens of mice. Rank sum statistical analysis of positive mouse spleens indicated that three overlapping bacterial populations were present. This pattern of infection was repeated when lipopolysaccharide (LPS) variants were used to infect chicks, and the worst infections were associated with isolates producing high-molecular-weight (HMW) LPS. Mouse isolates were capable of producing unprecedented amounts of HMW LPS as indicated by compositional analysis of six isolates that swarmed across 2% agar, which is a type of bacterial migration dependent upon production of HMW LPS. It is suggested that serovar Enteritidis cultured from the spleens of mice caught on farms will detect strains that are enhanced in their ability to contaminate eggs, in part because they are able to produce HMW LPS.

Salmonella enteritidis in eggs from commercial chicken layer flocks implicated in human outbreaks.

Eggs were cultured from four commercial chicken layer houses implicated in three human outbreaks of Salmonella enteritidis serotype enteritidis infection as part of the activities of the USDA-APHIS, VS, Salmonella enteritidis Task Force. Each house was part of a multiple in-line complex, ranging from three to seven houses. Houses were located on three separate farms, and each house contained between 50,000 and 80,000 chickens. S. enteritidis phage types 8, 13a, and 23 were isolated from samples taken from environmental and organ tissue samples, but only phage type 8 was cultured from eggs. Phage type 8 was isolated from humans in all three S. enteritidis outbreaks. Frequency estimates of contaminated eggs ranged from 0.03% to 0.90%.

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.