Outbreak of Newcastle disease in commercial poultry flocks, NSW.
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Bordetella avium is an avian respiratory disease pathogen responsible for substantial economic losses to the turkey industry. The inability to distinguish isolates has hampered outbreak investigations and prevents a complete understanding of transmission mechanisms. Isolates of Bordetella hinzii, often referred to as B. avium-like or as Alcaligenes faecalis type II prior to 1995, have also been acquired from the respiratory tracts of diseased poultry but are not believed to be pathogenic for birds. Therefore, differentiating between B. avium and B. hinzii is of importance for veterinary diagnostic laboratories. It was recently reported that both PvuII ribotyping and HinfI/DdeI restriction endonuclease analysis (REA) show promise for distinguishing isolates of B. avium and B. hinzii. Here we compare the ability of these techniques to discern inter- and intraspecies differences. While both approaches distinguished numerous types within a species, only REA was sufficiently discriminatory for routine use as an epidemiologic tool. Both techniques clearly distinguish between B. avium and B. hinzii, although the results of ribotyping are more easily interpreted. Ribotyping and REA identified numerous, previously unrecognized B. hinzii strains from a collection of bordetella isolates, including one acquired from a rabbit. This is the first report of B. hinzii isolation from a nonhuman mammalian species. At least some of the newly recognized B. hinzii isolates have been previously reported to cause disease in poults, suggesting that the pathogenicity of this agent for poultry should be more rigorously examined.
Conventional diagnostics for livestock and poultry outbreaks commonly rely on culture or targeted PCR panels, which may be too slow or too narrow to guide early control decisions. Portable metagenomics, particularly real-time nanopore sequencing, offers a route to broad pathogen detection, antimicrobial-resistance gene profiling, and outbreak investigation within an integrated workflow. This implementation-focused review evaluates how near-point-of-care metagenomics may support preventive veterinary medicine through earlier detection, surveillance, cohorting, biosecurity decisions, and antimicrobial stewardship. We synthesize sample-to-answer workflows for enteric and respiratory disease in food-producing animals, including sampling, nucleic-acid extraction, host depletion or target enrichment, library preparation, sequencing, bioinformatics, quality control, and interpretation. Applications in calf diarrhea, bovine respiratory disease, poultry outbreaks, mastitis, and resistome monitoring are considered alongside the central limitation that detection alone does not establish causation. Pathogen and resistance-gene signals must therefore be interpreted with clinical signs, lesions, epidemiology, controls, and confirmatory testing. We also propose a minimum reporting checklist, intended as a practical framework rather than a validated consensus standard. Portable metagenomics is not a replacement for conventional diagnostics, but appropriately validated workflows can reduce uncertainty during time-sensitive outbreaks and support more judicious antimicrobial use.
The antibiotic sensitivity patterns of 100 strains of S. pullorum and 24 strains of S. gallinarum against 19 antibiotics and 3 antibiotic mixtures were determined by the broth dilution method. The strains studied were isolated from enzootic outbreaks of pullorum disease and fowl typhoid. It was found that all the strains were sensitive to ampicillin, streptomycin, neomycin, kanamycin, oxytetracycline, chloromycetin, gentamycin, signamycin, polzomycin, and neotarchocin. Besides, S. pullorum was sensitive to aureomycin. In vitro studies showed that polzomycin and neotarchocin appeared to be the most effective drugs in the treatment of poultry diseases caused by S. pullorum or S. gallinarum. The appearance of high percentage of S. pullorum and S. gallinarum strains resistant simultaneously to several antibiotics points to the necessity of the determination of antibiotic sensitivity patterns of salmonellae isolated from birds.
Four flocks of clinically normal turkey breeder hens were shown to have suspect and positive Mycoplasma synoviae (MS) hemagglutination-inhibition (HI), enzyme-linked immunosorbent assay, and, in some cases, serum plate agglutination serology in the absence of MS isolation. In all cases, HI serology for Mycoplasma gallisepticum (MG) and M. meleagridis was negative. Acholeplasma laidlawii was isolated from some hens in each of these MS-seropositive culture-negative flocks. Immunoblotting was used to help determine if this positive MS serology was a result of cross-reactive antibodies to A. laidlawii or to some other Mycoplasma species. When sera from two of the flocks were reacted with MS antigen in immunoblotting, a strong and characteristic MS immunoblot profile was seen. Immunoblotting gave no evidence of a strong antibody response to A. laidlawii, M. iowae, or MG. This suggests the presence (or earlier presence) of MS in these flocks that is difficult to isolate by routine methods. Furthermore, this work shows that immunoblotting can be an important tool in the diagnosis of poultry diseases.
The availability of safe and effective drugs to treat and control poultry diseases and to enhance the efficiency of poultry production depends on the rate at which new drugs are approved by the Food and Drug Administration. This paper briefly describes the components of the regulatory approval process for a new animal drug for use in food-producing animals, with specific reference to some problems encountered in the approval process for poultry drugs.
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Each chicken carcass intended for U.S. consumers is mandated to be inspected by Food Safety and Inspection Service (FSIS) inspectors for its wholesomeness at the processing plants. Fluorescence responses of wholesome and unwholesome chicken carcasses were characterized and further evaluated for potential on-line applications for detection and classification of wholesome and unwholesome chicken carcasses. For this study, unwholesome chicken carcasses included cadaver and those with disease conditions such as airsacculitis and septicemia. Fluorescence characteristics from the epidermal layers in the breast areas of chicken carcasses were dynamic in nature. Emission peaks and ridges (maxima) were observed at 386, 444, 472, 512, and 554 nm and valleys (minima) were observed at 410, 460, 484, and 538 nm. One of the major factors affecting the line shapes of fluorescence responses from chicken carcass skin layers was absorption by hemoglobin. With the use of the normalized ratio spectra (NRS) approach, oxyhemoglobin was shown to be a major constituent in chicken carcasses affecting the fluorescence emission line shapes. Subtle line shape changes in the NRS also provided a qualitative means by which to assess the minute differences in oxy- and deoxyhemoglobin compositions perturbed by poultry diseases such as septicemia and airsacculitis. With the use of simple fluorescence band ratios as a multivariate model, wholesome and unwholesome chicken carcasses were correctly classified with 97.1% and 94.8% accuracies, respectively. On-line implementation of fluorescence techniques for the assessment of chicken carcass wholesomeness appears promising.
A competitive enzyme immunoassay was developed to measure the changes in serum levels of ovotransferrin (OTF) during inflammation and infectious diseases in chickens. The assay is based on the competition of serum OTF with a fixed concentration of biotin-labeled OTF to bind to a rabbit anti-chicken transferrin antibody immobilized on microtiter wells. After several washing steps, the antibody-bound biotinylated OTF is probed with streptavidin-horseradish peroxidase conjugate (HRP) followed by a colorimetric detection of the HRP activity. The relative changes in the optical density of color are plotted against the competing concentrations of OTF with logarithmic regression to generate a standard curve that is used to determine the concentrations of OTF in unknown samples. Serum had no effect on the measurement of OTE By this method, the time course changes of serum OTF levels in 4-wk-old male broiler chickens that were subjected to inflammation by croton oil injection were measured. The results showed croton oil-induced inflammation elevated serum OTF levels at 16 hr postinjection. OTF levels reached a peak by 72 hr, remained high through 120 hr, and returned to a basal level of olive oil-injected controls by 240 hr. There were no changes in serum OTF levels at any of the above time points in olive oil-injected control chickens. For studies with poultry diseases, specific-pathogen-free (SPF) male chickens were challenged with known bacterial and viral pathogens, and serum was collected at the height of the infection, i.e., 7 days after the challenge. Compared with uninjected controls, the SPF chickens challenged with Escherichia coli, fowl poxvirus, respiratory enteric orphan virus, infectious bursal disease virus, infectious bronchitis virus, or infectious laryngotracheitis virus had higher levels of OTF in serum. Inflammation-induced changes in serum OTF levels were also evident in the changes in the density of a 65-kD band protein corresponding to OTF. These results demonstrate that serum OTF may be a nonspecific clinical marker of inflammation associated with traumatic or infectious avian diseases.
The authors describe the source and prevalence of pathogenic Salmonella serovars among poultry farms in Saudi Arabia. A total of 1,052 (4%) Salmonella isolates were recovered from 25,759 samples of poultry (broilers, layers, broiler breeders and layer breeders) and poultry environments (box liner, litter, drag swab, droppings, mice and feed) were examined bacteriologically between 1988 and 1997 at the Poultry Disease Laboratory at the National Agriculture and Water Research Center in Riyadh. Eleven Salmonella serogroups representing 38 different Salmonella serovars were identified by means of antigenic analysis. The majority of the 276 isolates (26.2%) of Salmonella typed, were recovered from liver, heart and intestines of the broilers and layers. The most prominent Salmonella serogroups isolated were as follows: serogroup C1 (392 isolates, 37.26%), B (289 isolates, 27.47%) and D1 (269 isolates, 25.69%). However, untypable and multiple serogroups were also encountered, the most frequent isolates serotyped belonged to groups C1 (97 isolates, 24.7%), D1 (86 isolates, 31.9%), and B (71 isolates, 24.6%). Salmonella Enteritidis (85 isolates, 98.8%), Salmonella Virchow (48 isolates, 57.8%), Salmonella Paratyphi B var. Java (41 isolates, 57.7%) and Salmonella Infantis (30 isolates, 20.6%) were distributed the most widely as all were encountered in poultry and in poultry environments. S. Enteritidis phage type 4 (30 isolates, 35.3%), was the phage type most frequently detected among group D1 phage types, while 39 (45.8%) of the isolates of S. Enteritidis could not be phage typed.
Modern husbandry practices, regional concentration of the industry, high stocking densities, uniform age-distribution of birds and continuous feeding promote the spread of poultry diseases. Moreover, the immature state of the intestinal microflora or disturbance of the developing flora by antibiotics increases susceptibility of chicks to salmonellas. If an estimate of the number of salmonella-positive birds in a flock is needed, then the required number of samples can be assessed by using the binomial distribution function. Whenever a qualitative result is sufficient, the samples can be pooled or the flock litter can be sampled using an 'overshoe method', which is a novel, low-cost and rapid technique. An optimal pooling factor can be assessed at low prevalence levels (less than 10%). Serological methods will only detect the presence of antibodies to invasive strains of Salmonella. The sampling interval depends on the strategy of the Salmonella Control Programme. Breeder flocks should be sampled more frequently than meat flocks and laying flocks. The new salmonella standard, ISO 6579-1990, is applicable in the poultry industry. When bacterial numbers are likely to be low, or the organisms in a stressed condition, a pre-enrichment step should be included. In the case of faecal samples, however, pre-enrichment should be omitted. A whole carcass rinsing and massaging method is preferred for the examination of finished carcasses.
BACKGROUND: Infectious bronchitis is highly contagious and constitutes one of the most common and difficult poultry diseases to control. IBV is endemic in probably all countries that raise chickens. It exists as dozens of serotypes/genotypes. Only a few amino acid differences in the S1 protein of vaccine and challenge strains of IBV may result in poor protection. Tropism of IBV includes the respiratory tract tissues, proventriculus and caecal tonsils of the alimentary tract, the oviduct and the kidney. RESULTS: Infectious bronchitis virus (IBV) strain closely related to Massachusetts (Mass) serotype was isolated from broiler chickens suffering from severe renal and respiratory distresses. The isolate was serologically identified by Dot-ELISA and further characterized by RT-PCR then genotyped using S1 gene sequence analysis. Alignment of the S1 sequence of the isolate with 16 IBV strains revealed high homology to isolates related to Mass serotype. Inoculation with the strain reproduced the disease in experimental 1-day-old chickens and resulted in 20% mortality, severe renal and moderate respiratory distresses. Marked histopathological changes in both kidney and trachea were observed in experimentally infected chickens. A protection study using the H120 live attenuated vaccine showed low protection rate in spite of high S1 sequence homology (97%). Protection based criteria were: virus re-isolation attempts from trachea, tracheal and renal histopathology as well as IBV antigens detection by immunofluorescent antibody technique in kidney sections. CONCLUSION: Periodical evaluation of cross-protective capabilities of IBV vaccine(s) versus recently recovered field isolates should be performed to ensure optimum control of IBV.
Avian pathogenic Escherichia coli strains are associated with a variety of extraintestinal poultry diseases, including airsacculitis, colisepticemia, and cellulitis. A number of E. coli serotypes are associated with these diseases, although the most prevalent serotype is O78. Fimbrial proteins expressed by these strains appear to be important virulence factors, including type 1 fimbriae, P fimbriae, and curli. We have been working to develop an effective vaccine to protect chickens against these diseases. We have previously shown that an attenuated Salmonella typhimurium strain expressing O78 lipopolysaccharide provides protection against challenge with an O78 avian pathogenic E. coli strain. In this work, we have constructed an attenuated S. typhimurium that expresses both the O78 lipopolysaccharide and E. coli-derived type 1 fimbriae. In these studies, chickens were vaccinated at day of hatch and again at 2 wk of age. Birds were challenged at 4 wk of age. We found that the vaccine candidate provided significant protection against airsacculitis as compared to untreated controls or birds vaccinated with an attenuated S. typhimurium that did not express any E. coli antigens. In a separate experiment, challenged vaccinates showed significant weight gain compared to challenged nonvaccinates. We were not able to demonstrate protection against E. coli O1 or O2 serotype challenge, nor against challenge with wild-type S. typhimurium.
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