Differentiation of vaccine strains and Georgia field isolates of infectious laryngotracheitis virus by their restriction endonuclease fragment patterns

Seven restriction endonucleases (REs) were used to cleave the DNA from seven vaccine strains of infectious laryngotracheitis (ILT) virus and from six Georgia field isolates of ILT virus. After electrophoresis of the resulting RE fragments, the patterns were compared in order to differentiate strains of ILT virus. The six chicken-embryo-origin (CEO) vaccines were identical with each RE, but the tissue-culture-origin (TCO) vaccine strain differed from the CEO vaccines using five of the REs. Four of the six field isolates were identical by each RE, but two field isolates differed from each other and from the four identical field isolates on the basis of patterns produced by some but not all of the REs. The four identical field isolates could not be differentiated from the CEO vaccine strains by any RE, but the other two field isolates were not identical to either strain of vaccine virus. This work demonstrates that differentiable strains of ILT virus exist in the United States and that viruses other than vaccine viruses are involved in field outbreaks of ILT.     

Dietary patterns in middle-aged Irish men and women defined by cluster analysis

OBJECTIVES: To identify and characterise dietary patterns in a middle-aged Irish population sample and study associations between these patterns, sociodemographic and anthropometric variables and major risk factors for cardiovascular disease. DESIGN: A cross-sectional study. SUBJECTS AND METHODS: A group of 1473 men and women were sampled from 17 general practice lists in the South of Ireland. A total of 1018 attended for screening, with a response rate of 69%. Participants completed a detailed health and lifestyle questionnaire and provided a fasting blood sample for glucose, lipids and homocysteine. Dietary intake was assessed using a standard food-frequency questionnaire adapted for use in the Irish population. The food-frequency questionnaire was a modification of that used in the UK arm of the European Prospective Investigation into Cancer study, which was based on that used in the US Nurses' Health Study. Dietary patterns were assessed primarily by K-means cluster analysis, following initial principal components analysis to identify the seeds. RESULTS: Three dietary patterns were identified. These clusters corresponded to a traditional Irish diet, a prudent diet and a diet characterised by high consumption of alcoholic drinks and convenience foods. Cluster 1 (Traditional Diet) had the highest intakes of saturated fat (SFA), monounsaturated fat (MUFA) and percentage of total energy from fat, and the lowest polyunsaturated fat (PUFA) intake and ratio of polyunsaturated to saturated fat (P:S). Cluster 2 (Prudent Diet) was characterised by significantly higher intakes of fibre, PUFA, P:S ratio and antioxidant vitamins (vitamins C and E), and lower intakes of total fat, MUFA, SFA and cholesterol. Cluster 3 (Alcohol & Convenience Foods) had the highest intakes of alcohol, protein, cholesterol, vitamin B(12), vitamin B(6), folate, iron, phosphorus, selenium and zinc, and the lowest intakes of PUFA, vitamin A and antioxidant vitamins (vitamins C and E). There were significant differences between clusters in gender distribution, smoking status, physical activity, body mass index, waist circumference and serum homocysteine concentrations. CONCLUSION: In this general population sample, cluster analysis methods yielded two major dietary patterns: prudent and traditional. The prudent dietary pattern is associated with other health-seeking behaviours. Study of dietary patterns will help elucidate links between diet and disease and contribute to the development of healthy eating guidelines for health promotion.     

Aerosol vaccination against Newcastle disease. I. Studies on particle size.

Studies revealed that the majority of particles emitted by three different aerosol generators were in the size ranges of 0.5-1 and 1-3 mum. In general, the smaller the aperture setting, the larger the total number of particles. The difference between the generators in number of particles emitted were not significant enough to produce appreciable difference in antibody titers obtained from vaccinated chickens. Antibody titers were higher in birds vaccinated with the LaSota strain of Newcastle disease virus than in birds vaccinated with the B-1 strain, although protection against challenge was approximately similar. Aerosol challenge was found as effective as intramuscular challenge.     

Comparative microscopic lesions in reoviral and staphylococcal tenosynovitis

Experimental inoculation of 1-day-old male broiler chickens with avian reo-virus or Staphylococcus aureus caused tenosynovitis of the gastrocnemius and digital flexor tendons. Reovirus inoculation by either the oral or footpad route initiated a diffuse lymphocytic infiltration in the peritendineum, synovial membrane, and epitenon from 1 to 5 weeks postinoculation (PI). Heterophils were not a predominant feature of the inflammatory response, but when present they were localized with fibrin in and around synovial spaces. The prevalence of microscopic tendon lesions was less common with staphylococcal infection than with reovirus infection. With staphylococcus, lesions were localized to the synovial space and membranes and were characterized by heterophils and fibrin but few lymphocytes. Synovial cell hyperplasia and bursal atrophy were common in both groups. From 10 to 20 weeks PI, both groups developed progressive tendon fibrosis. These results indicate that tenosynovitis due to inoculation with reovirus or staphylococcus may be differentiated histologically from 1 through 5 weeks PI. After 10 weeks, this may not be possible, because diffuse fibrosis was the major lesion with both. Perhaps fibrosis predisposes older, heavier broilers to tendon failure and rupture.     

A comparison between the effect of an avian reovirus and infectious bursal disease virus on selected aspects of the immune system of the chicken

Reovirus 81-176 was inoculated subcutaneously into day-old specific-pathogen-free leghorns and evaluated for its effects on the immune system over a 3-week period. Structural criteria included organ weights of the bursa of Fabricius (BF) and spleen (SP), scoring of histological lesions in the BF, SP, and thymus, and hematological analyses of the circulating leukocytes. Alterations in the functional capacity of the immune system were measured using the graft-versus-host reaction, the response of peripheral blood lymphocytes (PBLs) to mitogens, the ability of circulating monocytes to phagocytize latex beads, and the serological responses to Newcastle disease virus, sheep red blood cells, and Brucella abortus antigens. For comparison, infectious bursal disease virus (IBDV) was similarly evaluated by most of the same tests. Structurally, reovirus 81-176 altered BF and SP organ weights, the total numbers of white blood cells in circulation, and the degree of follicular atrophy in the BF. Functionally, reovirus inoculation reduced both the response of PBLs to the phytohemagglutinin-P stimulation and monocyte uptake of latex beads. According to the protocols used here, no significant alteration in B-cell function could be detected in reovirus-infected chicks. With the exception of leukocyte hematology, IBDV-infected chicks had significantly altered responses in all tests used. By way of comparison, the effects of IBDV were more persistent and pronounced than were those seen with reovirus. The graft-versus-host reaction indicated an elevated and/or uninhibited response of T-cells in the blood of IBDV-infected chicks.     

Experimental infection of laying chickens with adenovirus 127 and with a related virus isolated from ducks

Infection of commercially reared white leghorn and white rock hens with adenovirus 127 was associated with decreases in total egg production, external egg quality, egg weight (P less than 0.01), and eggshell thickness (P less than 0.01). The egg-production and egg-quality disturbances were transient, and production returned to normal approximately 4 weeks postinfection. Infection of white leghorns with a hemagglutinating adenovirus isolated from Missouri ducks did not adversely affect egg production, external egg quality, or eggshell thickness, but it was associated with decreased egg weight (P less than 0.01). Prior infection with the duck adenovirus prevented the adverse egg-production effects of adenovirus 127 infection. Mean hemagglutination-inhibition antibody titers of chickens infected with adenovirus 127 or with duck adenovirus ranged from 1:588 to 1:10809, and mean titers of uninfected chickens did not exceed 1:2.     

Evaluation of ciliary movement in tracheal rings to assess immunity against infectious bronchitis virus

Chickens vaccinated at one and 14 days of age with a live vaccine containing the Massachusetts and Connecticut strains of infectious bronchitis virus were protected against challenge with a Massachusetts strain administered at 5 weeks of age. The virus recovery method in which tracheal swabs taken 4 days post-challenge were used to inoculate chicken embryos was compared with the method based on ciliary activity in tracheal explants. The results showed that evaluation of ciliary activity effectively measured immunity to infectious bronchitis virus. Serological results obtained in a microneutralization test using chicken embryo kidney cells correlated with the immune status of the birds as well as with the alpha neutralization results obtained using chicken embryos.     

Molecular characterization of avian infectious bronchitis virus strains isolated in Colombia during 2003

Sixteen infectious bronchitis virus (IBV) isolates were recovered from broilers and layers from five geographic poultry regions in Colombia. The viruses were isolated from tracheas, lungs, and cecal tonsils of birds, previously vaccinated with the Massachusetts strain, that were showing respiratory signs. Further analysis of the IBV isolates was achieved by phylogenetic analysis comparing their deduced amino acid sequences in the hypervariable region 1 of the S1 gene with reference strains. Four unique genotype clusters containing isolates with indigenous genotypes were observed. One isolate was found to be the Connecticut genotype and three isolates were found to be the Massachusetts genotype.     

Characterization of infectious bronchitis virus using monoclonal antibodies

Three monoclonal antibodies (MABs) reactive against two structural proteins--the nucleoprotein (NP) or the surface (S) protein--of avian infectious bronchitis virus (IBV) were produced and characterized. The MABs did not neutralize virus infectivity or inhibit hemagglutination. Their reactivity patterns with the homologous strain and eight heterologous strains of IBV were determined using the indirect immunoperoxidase test, the indirect immunofluorescent test, transfer-immunoblotting of separated proteins, and a dot-immunoblotting assay (DIA). Two MABs, NP- or S-protein-specific, reacted with all nine strains; one (NP-specific) reacted with only two strains. The two MABs reacting with all nine strains of IBV also detected 18 IBV field isolates of unknown serotype in the DIA. The MAB detecting only two strains did not react in the DIA. The diagnostic application of these MABs appears promising.     

Studies of infectious laryngotracheitis vaccines: immunity in layers

Ten-week-old layer chickens obtained from a commercial source were eye-drop vaccinated with chicken-embryo-origin (CEO) or tissue-culture-origin (TCO) vaccines for infectious laryngotracheitis (ILT). Controls were not vaccinated. Approximately one-third of the layers were challenged with virulent ILT virus at 21, 40, or 60 weeks of age. Serum samples taken from the layers before challenge were used in a virus neutralization (VN) test to determine vaccination titers at those three ages. Both vaccines induced low VN titers (geometric mean titer [GMT] less than 6). At 21 weeks of age, the titers produced by the two vaccines were not significantly different, but at 40 and 60 weeks of age the VN GMT of the CEO-vaccinated group was significantly greater than that of the TCO-vaccinated group. The VN GMTs did not drop over time in either group and actually rose between 21 and 60 weeks of age in the CEO group. Both vaccines protected layers against severe challenge with virulent ILT virus, neither being significantly better than the other under these experimental conditions. Unvaccinated sentinel chickens were maintained in contact with the vaccinated layers during three intervals between 1 day and 6 weeks post-vaccination. Diagnostic tests performed on the sentinels to detect lateral spread of vaccine virus from vaccinated to unvaccinated chickens showed scattered positive results.