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Antibiotic susceptibility of campylobacter isolates from sewage and poultry abattoir drain water.

In this study, the in vitro susceptibility of 209 campylobacter strains to the quinolones nalidixic acid, flumequine, ciprofloxacin, enrofloxacin, and to ampicillin, tetracycline and erythromycin was tested by the disk diffusion method. The strains were isolated from poultry abattoir effluent (DWA) and two sewage purification plants (SPA and SPB). Sewage purification plant SPA received mixed sewage, including that from a poultry abattoir, whereas SPB did not receive sewage from any meat-processing industry. The quinolone resistance of the DWA isolates ranged from 28% for enrofloxacin to 50% for nalidixic acid. The strains isolated from the sewage purification plants were more susceptible to the quinolones with a range of 11-18% quinolone resistance for SPB isolates to 17-33% quinolone resistance for SPA isolates. The susceptibility criteria as recommended by National Committee Clinical Laboratory Standards (USA) cannot readily be employed for campylobacter isolates. This investigation shows that the resistance of campylobacter bacteria is highest in the plant receiving sewage from a poultry slaughterhouse. Monitoring of antibiotic resistance of aquatic Campylobacter spp. is important, as surface waters are recognized as possible sources of infection.

Abattoirs↗

The role of management systems in the epidemiology of thermophilic campylobacters among poultry in eastern zone of Tanzania.

A total of 255 samples of droppings collected from a total of 22 different poultry units were examined for the presence of thermophilic campylobacters and the isolates biotyped using Skirrow's protocol. The organisms were isolated from 90 (35.3%) of all samples. Among the 22 units investigated, 13 (59%) were found to have unsatisfactory management systems, while 7 (32%) and 2 (9%) were found to have unsatisfactory and good systems respectively. Significantly large numbers of isolations, 68 of 147 (46.2%), were made from samples collected from poultry units with poor management (P < 0.005), compared with 19 out of 84 (22.6%) samples which were collected from satisfactory units and 3 out of 24 (12.5%) samples collected from units exercising particularly good management. Nineteen of 72 (26.4%) samples collected from broilers, 32 out of 132 (24.2%) samples collected from layers and 39 out of 51 (76.49%) samples collected from indigenous free range poultry were positive for campylobacters. Among the 90 strains isolated from various units, 64 (70.1%) were Campylobacter jejuni, 25 (27.7%) were C. coli, and only 1 (2.2%) was C. laridis.

Animal Husbandry↗

Human salmonellosis associated with young poultry from a contaminated hatchery in Michigan and the resulting public health interventions, 1999 and 2000.

Although approximately 95% of disease caused by nontyphoidal salmonella is transmitted by foodborne vehicles, four documented salmonella outbreaks in the 1990s have been traced to contact with young poultry. No environmental studies of source hatcheries were completed. This case-control study was performed by comparing culture-confirmed Salmonella Infantis in Michigan residents, identified between May and July 1999, with two age- and neighbourhood-matched controls. Eighty environmental and bird tissue samples were collected from an implicated hatchery; all salmonella isolates underwent pulsed-field gel electrophoresis (PFGE) analysis. The study included 19 case-patients sharing the same PFGE subtype and 37 matched controls. Within 5 days before illness onset, 74% of case-patients resided in households raising young poultry compared with 16% of controls (matched OR 19.5; 95% CI 2.9, 378.1). Eight hatchery samples yielded Salmonella Infantis with PFGE subtypes matching the patients' isolates. This investigation identified birds from a single hatchery as the source of human illness and confirmed the link by matching PFGE patterns from humans, birds and the hatchery environment. Subsequent public health interventions reduced, but did not eliminate, transmission of poultry-associated salmonellosis. Five additional PFGE-linked cases were identified in Spring 2000, necessitating quarantine of the hatchery for depopulation, cleaning and disinfection.

Adolescent↗

Potential microbiological contamination of effluents in poultry and swine abattoirs.

Health risks in the effluents of seven swine abattoirs and of seven poultry abattoirs were evaluated with regard to environment degradation and to dissemination of pathogenic microorganisms during the rainy and dry seasons. Supply-water samples from affluents and effluents of the treatment systems at different sites within the abattoir processing system were analysed. Similarly, water samples from the three recipient sites (emission point, 100 m upstream, 100 m downstream) were also analysed. Temperature, free residual chlorine (FRC), total coliform bacteria, Escherichia coli, enterococci, identification and serotyping of salmonellae were assessed. Scalding is the most significant stage in the slaughtering chain (P<0.05) when temperature is taken into account. Temperatures at effluents and at the sampled sites in the water bodies accorded to state and federal legislation standards. Supply waters did not meet the standards for FRC and microbial count standards according to the Ministry of Health and within limits imposed by the Industrial and Sanitary Inspection Regulations for Animal Products. Feather plucking and evisceration in poultry slaughter and the cleansing of carcasses and facilities in poultry and swine slaughtering had the highest contamination impact. The three loci at the water bodies were above the microbiological standards for classes II and III sites, in conformity with Law 8468 of the state of São Paulo, Brazil and Conama. Salmonella was found at several sites during slaughter, at both types of abattoirs, including in the effluent treatment system. This showed that these sites were the dissemination sources of the microorganism.

Abattoirs↗

Environmental fate of roxarsone in poultry litter. I. Degradation of roxarsone during composting.

Roxarsone, 3-nitro-4-hydroxyphenylarsonic acid, is an organoarsenic compound that is used extensively in the feed of broiler poultry to control coccidial intestinal parasites, improve feed efficiency, and promote rapid growth. Nearly all the roxarsone in the feed is excreted unchanged in the manure. Poultry litter composed of the manure and bedding material has a high nutrient content and is used routinely as a fertilizer on cropland and pasture. Investigations were conducted to determine the fate of poultry-litter roxarsone in the environment Experiments indicated that roxarsone was stable in fresh dried litter; the primary arsenic species extracted with water from dried litter was roxarsone. However, when water was added to litter at about 50 wt % and the mixture was allowed to compost at 40 degrees C, the speciation of arsenic shifted from roxarsone to primarily arsenate in about 30 days. Increasing the amount of water increased the rate of degradation. Experiments also suggested that the degradation process most likely was biotic in nature. The rate of degradation was directly proportional to the incubation temperature; heat sterilization eliminated the degradation. Biotic degradation also was supported by results from enterobacteriaceae growth media that were inoculated with litter slurry to enhance the biotic processes and to reduce the concomitant abiotic effects from the complex litter solution. Samples collected from a variety of litter windrows in Arkansas, Oklahoma, and Maryland also showed that roxarsone originally present had been converted to arsenate.

Animal Feed↗

Arsenic speciation and reactivity in poultry litter.

Recent U.S. government action to lower the maximum concentration levels (MCL) of total arsenic (As) (10 ppb) in drinking water has raised serious concerns about the agricultural use of As-containing biosolids such as poultry litter (PL). In this study, solid-state chemical speciation, desorbability, and total levels of As in PL and long-term amended soils were investigated using novel synchrotron-based probing techniques (microfocused (micro) synchrotron X-ray fluorescence (SXRF) and micro-X-ray absorption near-edge structure (XANES) spectroscopies) coupled with chemical digestion and batch experiments. The total As levels in the PL were as high as approximately 50 mg kg(-1), and As(II/III and V) was always concentrated in abundant needle-shaped microscopic particles (approximately 20 microm x 850 microm) associated with Ca, Cu, and Fe and to a lesser extent with S, Cl, and Zn. Post-edge XANES features of litter particles are dissimilar to those of the organo-As(V) compound in poultry feed (i.e., roxarsone), suggesting possible degradation/transformation of roxarsone in the litter and/or in poultry digestive tracts. The extent of As desorption from the litter increased with increasing time and pH from 4.5 to 7, but at most 15% of the total As was released after 5 d at pH 7, indicating the presence of insoluble phases and/or strongly retained soluble compounds. No significant As accumulation (< 15 mg kg(-1)) was found in long-term PL-amended agricultural surface soils. This suggests that As in the PL may have undergone surface and subsurface transport processes. Our research results raise concerns about long-term PL amendment effects on As contamination in surrounding soil-water environments.

Absorptiometry, Photon↗

Village poultry production systems in the central highlands of Ethiopia.

Participatory rural appraisal (PRA), supported by checklists and intensive case studies on individual households, was carried out in three villages at three different altitudes in the central highlands of Ethiopia. The chicken production system in each village is described and the problems are discussed. More than 60% of the families kept chickens, and in most cases the women owned and managed the birds and controlled the cash from the sales. The production systems followed were mainly low-input and small-scale, with 7-10 mature birds per household, reared in the back yards with inadequate housing, feeding and health care. The average egg production per clutch was 15-20, with 3-4 clutches per year. The mean number of eggs set per bird was 12.9 +/- 2.2 (n = 160), depending on the size of the bird and season, and the hatching rate was 80.9% +/- 11.1%, range 44%-100% (n = 160). Poultry meat and eggs were generally accepted and appreciated in all three villages. In addition to the small amount of cash income they provide, scavenging chickens have nutritional, cultural and social functions. The flock composition, price of poultry and poultry products, disease outbreaks and hatching of chicks were strongly affected by season. Disease was cited as the most important problem by most of the members of the community, followed by predation, lack of feed, poor housing, insufficient water and parasites. Disease periodically decimated the flocks, and consequently, about 50% of the eggs produced were incubated in order to replace the birds that had died. The major source of loss in the system was the high mortality of chicks (61%) that occurred between hatching and the end of brooding at 8 weeks of age. The system was characterized by no or few inputs and a low output level. The major input was the cost of foundation stock, but after that virtually no cost was involved. The major source of feed for the birds was from the scavenging feed resource base, which comprised table leftovers, small grain supplements and anything edible from the immediate environment.

Animal Feed↗

Mesophilic biomethanation and treatment of poultry waste-water using pilot scale UASB reactor.

The feasibility of applying the up-flow anaerobic sludge blanket (UASB) treatment for poultry waste (faeces) water was examined. A continuous-flow UASB pilot scale reactor of 3.50 L capacity using mixed culture was operated for 95 days to assess the treatability of poultry waste-water and its methane production. The maximum chemical oxygen demand (COD) removed was found to be 78% when organic loading rate (OLR) was 2.9 kg COD m(-3) day(-1) at hydraulic retention times (HRT) of 13.2 hr. The average biogas recovery was 0.26 m3 CH4 kg COD with an average methane content of 57% at mean temperature of 30 degrees C. Data indicate more rapid methanogenesis with higher loading rates and shorter hydraulic retention times. At feed concentration of 4.8 kg COD m(-3) day(-1), anaerobic digestion was severely retarded at all hydraulic retention time tested. This complication in the reactor operations may be linked to build-up of colloidal solids often associated with poultry waste water and ammonia toxicity. Isolates from granular sludge and effluent were found to be facultative anaerobes most of which were Pseudomonas genera.

Anaerobiosis↗

Identification of unusual Campylobacter-like isolates from poultry products as Helicobacter pullorum.

Twenty-six unclassified Campylobacter-like strains previously isolated from 15 chicken carcasses and caecal contents, together with two more strains isolated from chicken faeces on a different occasion, were identified as Helicobacter pullorum using various phenotypic identification methods. API Campy identification kits and a 16-test identification scheme developed for campylobacters failed to identify these bacteria, or identified them as Campylobacter spp. Eighteen strains (including the two isolated on a different occasion) were chosen for examination using a more comprehensive probabilistic identification scheme. Using this method, 14 of the 18 strains were identified as H. pullorum with ID scores > 95%; two strains were also identified as H. pullorum with lower ID scores. Of the remaining two strains, one was not identified with this scheme and the other was misidentified to the H. acinonyx pylori complex. Whole cell protein profiling by SDS-PAGE confirmed the identity of these isolates as H. pullorum, affirming the value of a polyphasic approach for accurately identifying campylobacteria. The comparatively high prevalence of H. pullorum in poultry determined in this study (60%) suggests that routine isolation and identification methods should be amended to enable a thorough evaluation of its role in human gastroenteritis and avian hepatitis. Some phenotypic characters useful in identifying poultry campylobacteria are presented which could be utilized, along with other technique(s), for improved differentiation of the campylobacteria that are found in poultry.

Animals↗

Laboratory heating studies with Salmonella spp. and Escherichia coli in organic matter, with a view to decontamination of poultry houses.

AIMS: To determine a temperature-humidity-time treatment that eliminates Salmonella and Escherichia coli in substrates representing organic matter in poorly cleaned poultry houses, i.e. worst case scenario laboratory tests. METHODS AND RESULTS: Organic matter (poultry faeces and feed) in a 2.5-cm layer was inoculated with 2 x 10(5)-3 x 10(6) Salmonella g(-1), left undried or dried at ca. 30% relative humidity (RH) during a 10-day period, and temperature increased at 1 degrees C h(-)1 to the final heating temperature of 50, 55, 60, 65 or 70 degrees C and held at 16-30 or 100% RH. All samples were tested for Salmonella according to predetermined sampling time schedules and faecal samples were also tested for naturally occurring E. coli. Overall, humidity was an important factor in the elimination of Salmonella and E. coli. Results for recovery of Salmonella and E. coli were highly associated. CONCLUSIONS: The application of >/=60 degrees C and 100% RH during a 24-h period eliminated Salmonella and E. coli in all samples. Escherichia coli could be used as an indicator bacterium for the elimination of Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: The results from worst case scenario laboratory tests could be applied in steam heating of persistently Salmonella-infected poultry houses. The use of E. coli as an indicator bacterium for the validation of Salmonella results should be considered.

Animal Feed↗

Antibiotic resistance of Escherichia coli isolated from poultry workers, patients and chicken in the eastern province of Saudi Arabia.

The prevalence of antibiotic-resistant Escherichia coli isolates from faecal samples from 117 poultry industry workers, 100 patients and119 healthy chicken were compared. Resistance of E. coli chicken isolates to ampicillin, chloramphenicol, gentamicin, spectinomycin, tetracycline and trimethoprim + sulphamethoxazole (TMP + SMX) (range 57% - 99.1%) were significantly higher than those isolated from patients (range 21.9% - 71.4%) and workers (range 35% - 71.8%). However, for drugs not used in poultry, such as amoxicillin + cluvalanate (AMX + CLV), ceftazidime and nitrofurantoin, resistance rates of chicken isolates (range 0% - 2.6%) were significantly lower than those of patient isolates (range 8.7% - 30%). Resistance to spectinomycin reached 96% in E. coli chicken isolates and 71% in organisms isolated from humans. Use of this drug in Saudi Arabia is mostly limited to veterinary purposes. Multidrug resistance is alarmingly high in all groups but was highest in chicken isolates (77.4%). Serotyping of E. coli isolates showed that 27% of the organisms isolated from patients were overlapping with 10.9% of the chicken isolates, indicating the possibility of chicken being a source of the resistance pool for humans. We therefore call for the banning of antibiotics in the poultry industry as growth promoters and recommend that their use be restricted to treating infections.

Animals↗

Isolation of Salmonella resistant to hypochlorous acid from a poultry abattoir.

Salmonella is a Gram-negative bacterium that has been implicated in several food-borne poisoning outbreaks. Poultry products are considered to be the major vehicles of food-borne poisoning caused by Salmonella. The resistance of Salmonella, isolated from different stages in a local poultry abattoir, to hypochlorous acid (HOCl) was studied. Isolates cultured in quarter strength Tryptone Soya Broth were treated with 72 ppm HOCl, incubated at 30 degrees C with shaking, and absorbance was measured at 660 nm every 20 min. A number of resistant isolates, which carried on growing following HOCl addition, were obtained. All the isolates from the bleeding stage were sensitive to HOCl, whereas those from the scalding stage were resistant to HOCl. Other stages had a population of resistant and sensitive isolates. It is evident that some Salmonella will grow in the presence of concentrations of HOCl deemed to be inhibitory. Hence, an effective concentration of HOCl must be applied in poultry abattoirs to ensure that all the Salmonella are eradicated.

Animals↗

Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry.

AIMS: The present study describes the implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry. METHODS AND RESULTS: Real-time PCR with Salmonella invA-specific primers and a standard bacteriological method was applied to detect Salmonella in tetrathionate enrichment cultures of 492 intestinal homogenates and 27 drag swabs from 47 poultry flocks. The number of positive individual samples by real-time PCR and culture method was 65 (12.5%) and 35 (6.8%), respectively. The number of Salmonella-positive flocks was 13 (27.7%) by both methods. PCR detection required 25 min for up to 32 samples. Melting curve analysis revealed the Tm for Salmonella-specific PCR product as 87 +/- 1 degrees C. CONCLUSIONS: Implementation of real-time PCR to tetrathionate broth enrichment step reduces the Salmonella detection time to 18 h and 25 min. Isolation of Salmonella should be carried out with PCR to determine the serovar. SIGNIFICANCE AND IMPACT OF THE STUDY: Real-time PCR is a powerful tool in rapid and accurate Salmonella monitoring in poultry companies, together with standard bacteriology.

Animals↗

Virulence-associated genes in avian pathogenic Escherichia coli (APEC) isolated from internal organs of poultry having died from colibacillosis.

Escherichia coli infections are responsible for significant losses in the poultry industry in many parts of the world. The pathogenesis and the role of virulence factors are not yet totally elucidated. We, therefore, examined 150 E. coli strains isolated from visceral organs of poultry having died from colibacillosis for the presence of virulence-associated genes by PCR. The E. coli strains were investigated for the presence of a total of 17 virulence-associated genes described for diarrheagenic (stx1/2, eae, hlyEHEC, estl, eltI, astA, cdtb), septicemic (hlyA, papC, cnf1/2, fyuA, irp2) and avian pathogenic E. coli (APEC; iucD, tsh, fimC, and hlyE as well as stx2f). Seven genes were significantly distributed among APEC strains, while most of the other investigated genes could be demonstrated only sporadically or not at all. FimC (Type I fimbriae) was detected with the highest prevalence in 92.7% of the isolates. Most of the strains harboring iucD (88.7%) also gave positive results for tsh (85.3%). Genes fyuA (ferric yersiniabactin uptake) (66.0%) and irp2 (iron-repressible protein) (68.0%), necessary for Yersinia to acquire iron in the mouse infection model, were regularly detected in combination. Moreover, we found papC (pyelonephritis-associated pili) in 30.0% and astA (enteroaggregative heat stable toxin) in 17.3% of the field strains. A significant amount of strains (57.3%) harbored a combination of iucD, tsh, fimC, fyuA and irp2 virulence-associated genes, presumably rendering these strains particularly virulent. These findings provide novel insights into the presence and distribution of virulence-associated genes in avian pathogenic E. coli field strains, which will help to more comprehensively characterize APEC in future epidemiological studies. It is assumed that the existence of two iron acquisition systems points towards their important role in virulence. Furthermore, we suggest that characterization of the respective phenotypes in infection models will provide substantial information to better understand the pathogenesis of colibacillosis in poultry.

Animals↗

Poultry breeding in the post-genomics era.

1. I introduce the problems facing poultry breeding in the future and the role biotechnology may have to play. 2. I then review recent research in the genomics and bioinformatics of poultry and where this may lead in the near future. 3. Finally, I discuss the scientific and commercial opportunities of this research and strategies for implementation into poultry breeding practices.

Animals↗

Poultry science: the next 20 years?

1. The theme of the lecture is that research in poultry science has moved too far in the direction of molecular biology and away from studies with whole animals. This has happened partly because exciting prospects are opening up in the field of gene manipulation but mainly because of the use of inappropriate referees to evaluate research proposals. 2. Agricultural research is defined as work intended to benefit agriculture and directed towards those problems which seem capable of solution. Science research is something else. Too much of the money allocated for agricultural and biotechnology research is being spent on science research. The system of rewarding agricultural scientists needs to be adjusted away from counting papers published. 3. Some examples are given of problems in poultry science which seem likely to be soluble by gene manipulation. These include "essential" amino acid synthesis within the chicken, improvement of shell strength, the prevention of many diseases, but probably not the improvement of quantitative traits or of behavioural adaptation to intensive husbandry. 4. Examples are also given of problems likely to require empirical solutions, such as the benefits of acclimatisation or the long-term response to a lighting programme. Here the need is to develop better theories to guide modelling activities. 5. The author concludes that there is much research that can and should be done in poultry science in the next 20 years but calls for a recognition that some problems cannot be solved by a "fundamental" approach but will need experiments with whole animals coupled with model-building activities.

Animal Husbandry↗

Effects of AvGard treatment on the microbiological flora of poultry carcases.

1. The efficiency of the AvGard (or Assur-Rince in the USA) trisodium phosphate poultry carcase decontamination process was evaluated during both manual and industrial trials against total aerobic mesophilic count (TAMC), thermotolerant coliforms, Pseudomonas, Enterobacteriaceae, Campylobacter, Listeria monocytogenes and Salmonella. 2. The TSP treatment proved to have significant effects on the bacterial decontamination of poultry neck skin, lowering the contamination by a factor of about 10 for TAMC and of 100 for Coliform and Pseudomonas. 3. Numeration of Salmonella with an innovative miniaturised most probable number method has proved that the effect upon these micro-organisms was also close to 2 log10 reduction. 4. The effect of TSP treatment on the ecological balance of psychrotrophic bacterial flora was also investigated to study the origin of the shelf-life flora of treated carcases (Pseudomonas being reduced to the limit of detection) and to ascertain whether L. monocytogenes might gain a competitive advantage. In fact AvGard reduced the number of L. monocytogenes on poultry carcases. 5. As a consequence of the virtual elimination of the Pseudomonas usually present, Brochothrix thermosphacta became the main species responsible for putrefaction. 6. Because the growth rate of Brochothrix thermosphacta was greater than that of L. monocytogenes at refrigeration temperature, it was considered that putrefaction would occur before the emergence of large numbers of L. monocytogenes.

Animals↗

Salmonella contamination of poultry flocks in The Netherlands.

The contamination of poultry in the Netherlands with Salmonella enteritidis was tested. For this, different methods (detection of S. enteritidis in faecal samples of 25 g; detection of S. enteritidis in cloacal swabs; detection of S. enteritidis by serological testing of antibodies in serum) were compared for their efficiency to detect S. enteritidis in flocks of poultry. Testing of faecal samples clearly yielded the best results. This method was used in a transmission study, in which 14 flocks descending from a contaminated primary mother flock were screened for the presence of S. enteritidis. The method was also used for screening 49 flocks of laying hens and 52 flocks of broiler chickens throughout the Netherlands. From the transmission study it became clear that S. enteritidis, phage type 2 (Dutch phage set) was isolated both from the mother flock and from five of the descendent flocks. Screening of poultry flocks for the presence of salmonella revealed that salmonella was present in 47% of the layer flocks and in 94% of the broiler flocks. S. enteritidis was isolated from 15% of the flocks screened.

Animals↗