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Factors regulating bone maturity and strength in poultry.

Adolescent meat-type poultry and cage layers exhibit a high incidence of bone problems that include bone weakness, deformity, breakage, and infection and osteoporosis-related mortalities. These problems include economic and welfare issues. To improve bone quality in poultry, it is essential to understand the physiological basis of bone maturity and strength in poultry. A complex array of factors that include structural, architectural, compositional, physiological, and nutritional factors interactively determine bone quality and strength. Bone is approximately 70% mineral, 20% organic, and 10% water. Collagen is the major organic matrix that confers tensile strength to the bone, whereas hydroxyapatite provides compressional strength. In recent years, the roles of different collagen crosslinks have been shown to be important in the increase of bone mechanical strength. Similarly, age-related glyco-oxidative modifications of collagen have been shown to increase the stiffness of collagen. These posttranslational modifications of matrix can affect bone quality as it would be affected by the changes in the mineralization process. Our studies show that the growth in the tibia continued until 25 wk of age, which correlated with the increase in the content of hydroxylysylpridinoline (HP) and lysylpyridinoline (LP), the collagen crosslinks. The tibia from 5-wk-old chicks were strong but brittle because of low collagen crosslinks and high mineral content. Bone maturity may relate to its crosslink content. Compared to crosslink content, bone density and ash content showed moderate increases during growth. The bones from younger turkeys were more susceptible to corticosteroid-induced stunting of growth, which also resulted in decreased bone strength. This review discusses how different factors can compromise bone strength by reducing growth, altering shape, affecting mineralization, and affecting collagen crosslinking.

Aging↗

Nutrient substrates used by bacterial isolates from the poultry processing environment.

In order to successfully prevent pathogens found in biofilms in poultry processing facilities from contaminating products, knowledge of the sources and properties of the bacteria from these environments is needed. Bacteria were isolated from fresh cut-up meat samples (breast with or without skin, wings, and thighs) that were not stored or were stored at 4 or 13 C (temperatures relevant to poultry processing facilities). Profiles of the nutrient substrates used by individual bacterial species were determined using Biolog microtiter plates with different substrates in 95 wells of each plate. Pure cultures of bacterial isolates were inoculated onto gram-positive (GP) or gram-negative (GN) Biolog plates, and transformation of specific substrates was determined by reduction (indicated by a purple color) of the tetrazolium violet included in each well. Of the 62 substrates common to GP and GN microtiter plates, one-third were used by 50% or more of all bacterial isolates. Similarly, approximately one-third of the substrates were used by gram-negative bacteria but not by gram-positive bacteria. One-fourth of the substrates were also only used by a single isolate in the respective gram-type category. For the remaining 33 substrates on each of the GP and GN plates, a higher percentage of substrates on GN than on GP plates were used by the respective isolates. These substrate utilization profiles of bacteria isolated from the poultry processing environment are a useful reference for selecting nutrients for the growth or control of these bacteria, especially pathogens.

Animals↗

Development of research paper writing skills of poultry science undergraduate students studying food microbiology.

Because food and poultry industries are demanding an improvement in written communication skills among graduates, research paper writing should be an integral part of a senior undergraduate class. However, scientific writing assignments are often treated as secondary to developing the technical skills of the students. Scientific research paper writing has been emphasized in an undergraduate course on advanced food microbiology taught in the Poultry Science Department at Texas A& M University (College Station, TX). Students' opinions suggest that research paper writing as part of a senior course in Poultry Science provides students with scientific communication skills and useful training for their career, but more emphasis on reading and understanding scientific literature may be required.

Agriculture↗

Dose-response relationships between occupational aerosol exposures and cross-shift declines of lung function in poultry workers: recommendations for exposure limits.

Numerous articles have been published regarding the adverse respiratory health consequences of working in intensive livestock and poultry housing. Threshold limit exposure guidelines are not currently applied to this environment, but they are essential to implement and monitor effective environmental controls. Previous dose-response research work with swine workers has resulted in exposure limit recommendations of 2.5 mg/m3 total dust, 0.23 mg/m3 respirable dust, 100 EU/m3 endotoxin, and 7 ppm ammonia. No similar recommendations have been reported previously for poultry workers. Therefore, an industry-wide study was conducted to examine dose-response relationships of bioaerosol exposures and worker respiratory health. A total of 257 poultry workers were studied for respiratory symptoms, pulmonary function, and exposure to dust (total and respirable), endotoxin (respirable and total), and ammonia. Details of the sampling plan and environmental assessment are described elsewhere. Relationships between exposures and response were studied by correlation and multiple regressions. Significant dose-response relationships were observed between exposures and pulmonary function decrements over a work shift. Exposure concentrations associated with significant pulmonary function decrements were as follows: 2.4 mg/m3 total dust, 0.16 mg/m3 respirable dust, 614 EU/m3 endotoxin, and 12 ppm ammonia.

Adult↗

Pseudo-outbreak of tuberculosis in poultry plant workers, Sussex County, Delaware.

Delaware is a leading US poultry-producing state, and foreign-born workers make up a significant percentage of those employed by Delaware's poultry plants. In Sussex County, Delaware, a high percentage of the poultry workers are from two countries with a high incidence of tuberculosis (TB), Mexico and Guatemala, and thus are at risk for TB infection and disease. Furthermore, their risk of TB may be increased because many of these workers live in crowded conditions and lack access to medical care.

Adult↗

Prevalence of 11 pathogenic genes of Campylobacter jejuni by PCR in strains isolated from humans, poultry meat and broiler and bovine faeces.

Although many genes related to the pathogenicity of Campylobacter jejuni have been reported, the relationships between these genes and the sources of strains are not clear. In this study, the presence of 11 pathogenic genes responsible for the expression of adherence, invasion, colonization and cytotoxin production was examined in 111 C. jejuni isolated from human clinical samples, poultry meat, broiler faeces and bovine faeces. For most of the pathogenic genes, no difference in their presence in C. jejuni was found among the sources, but, for racR, wlaN and virB11, there were some variations among sources. The racR gene was present at rates of 98.2 (human clinical samples), 90.5 (poultry meat), 85.7 (broiler faeces) and 76.7 % (bovine faeces). Detection rates for the wlaN gene were 25.0, 23.8, 4.7 and 7.7 % and those for the virB11 gene were 10.7, 9.5, 9.5 and 15.4 % in human clinical samples, poultry meat, broiler faeces and bovine faeces, respectively. One hundred and seven of 111 strains (96.4 %) carried from eight to 10 of the pathogenic genes. These data did not show remarkable differences in the presence of pathogenic genes carried by C. jejuni from various sources.

Animals↗

In vivo characterization of Lactobacillus johnsonii FI9785 for use as a defined competitive exclusion agent against bacterial pathogens in poultry.

AIMS: To test the efficacy of Lactobacillus johnsonii FI9785 in reducing the colonization and shedding of Salmonella enterica serotype Enteritidis, Escherichia coli O78:K80 and Clostridium perfringens in poultry. METHODS AND RESULTS: Specific pathogen-free chicks (1 day old) were dosed with a single oral inoculum of 1x10(9) CFU. Lactobacillus johnsonii FI9785 and 24 h later were challenged in separate experiments with S. Enteritidis (S1400, nalr) and E. coli O78:K80 (EC34195, nalr). There were no significant effects against S. Enteritidis whereas colonization of the small intestine by E. coli O78:K80 was reduced significantly. Both S. Enteritidis and E. coli colonized the caeca and colon to levels equivalent to control birds and there was no reduction in shedding as assessed by a semi-quantitative cloacal swabbing technique. Specific pathogen-free chicks (20 day old) were dosed with a single oral inoculum of 1x10(9) CFU L. johnsonii FI9785 and 24 h later were challenged with C. perfringens. A single oral dose of L. johnsonii FI9785 was sufficient to suppress all aspects of colonization and persistence of C. perfringens. CONCLUSIONS: Lactobacillus johnsonii FI9785 may be given to poultry for use as a competitive exclusion agent to control C. perfringens. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus johnsonii FI9785 may be a valuable tool to control the endemic disease of necrotic enteritis, thereby reducing economic losses associated with reduced use of antimicrobials in the poultry industry.

Animals↗

Importance of pre-enrichment media for isolation of Salmonella spp. from swine and poultry.

The performance of two new (1-day) culture methods, Salmonella Enrichment Broth (SEB) and Revive, and an alternative pre-enrichment broth, designated Universal pre-enrichment broth (UB), was compared to the internationally accepted buffered peptone water (BPW). The study was directed towards detection of Salmonella in 100 faecal samples from porcine and 100 neck-skin samples from poultry. The sensitivity (number of positive cases per method among all the positive cases) of the conventional pre-enrichment in BPW was found to be 0.77 for swine and 0.66 for poultry samples, while a combination of the BPW method with parallel pre-enrichment of the same sample in UB resulted in high sensitivity for swine (0.92) and poultry (0.95) samples. A 2-h pre-enrichment in the non-selective Revive, followed by overnight enrichment in selective broth, resulted in a low sensitivity, particularly for the neck-skin samples (0.16, P = 0.001). The SEB method in the porcine samples resulted in a sensitivity (0.71) comparable to the standard method (P = 0.31). In conclusion, additional pre-enrichment of samples in UB may substantially increase the culture sensitivity. During routine screening of large numbers of samples, it may be advantageous to use SEB rather than standard culturing.

Abattoirs↗

Isolation of Escherichia coli O157:H7 from retail fresh meats and poultry.

A total of 896 samples of retail fresh meats and poultry was assayed for Escherichia coli serogroup O157:H7 by a hydrophobic grid membrane filter-immunoblot procedure developed specifically to isolate the organism from foods. The procedure involves several steps, including selective enrichment, filtration of enrichment culture through hydrophobic grid membrane filters, incubation of each filter on nitrocellulose paper on selective agar, preparation of an immunoblot (by using antiserum to E. coli O157:H7 culture filtrate) of each nitrocellulose paper, selection from the filters of colonies which corresponded to immunopositive sites on blots, screening of isolates by a Biken test for precipitin lines from metabolites and antiserum to E. coli O157:H7 culture filtrate, and confirmation of isolates as Vero cell cytotoxic E. coli O157:H7 by biochemical, serological, and Vero cell cytotoxicity tests. E. coli O157:H7 was isolated from 6 (3.7%) of 164 beef, 4 (1.5%) of 264 pork, 4 (1.5%) of 263 poultry, and 4 (2.0%) of 205 lamb samples. One of 14 pork samples and 5 of 17 beef samples contaminated with the organism were from Calgary, Alberta, Canada, grocery stores, whereas all other contaminated samples were from Madison, Wis., retail outlets. This is the first report of the isolation of E. coli O157:H7 from food other than ground beef, and results indicate that the organism is not a rare contaminant of fresh meats and poultry.

Animals↗

High-resolution genotyping of Campylobacter strains isolated from poultry and humans with amplified fragment length polymorphism fingerprinting.

For epidemiological studies of Campylobacter infections, molecular typing methods that can differentiate campylobacters at the strain level are needed. In this study we used a recently developed genotyping method, amplified fragment length polymorphism (AFLP), which is based on selective amplification of restriction fragments of chromosomal DNA, for genetic typing of Campylobacter jejuni and Campylobacter coli strains derived from humans and poultry. We developed an automated AFLP fingerprinting method in which restriction endonucleases HindIII and HhaI were used in combination with one set of selective PCR primers. This method resulted in evenly distributed band patterns for amplified fragments ranging from 50 to 500 bp long. The discriminatory power of AFLP was assessed with a C. jejuni strain, an isogenic flagellin mutant, and distinct C. jejuni strains having known pulsed-field gel electrophoresis and fla PCR-restriction fragment length polymorphism genotypes. Unrelated C. jejuni strains produced heterogeneous patterns, whereas genetically related strains produced similar AFLP patterns. Twenty-five Campylobacter strains obtained from poultry farms in The Netherlands grouped in three C. jejuni clusters that were separate from a C. coli cluster. The band patterns of 10 C. jejuni strains isolated from humans were heterogeneous, and most of these strains grouped with poultry strains. Our results show that AFLP analysis can distinguish genetically unrelated strains from genetically related strains of Campylobacter species. However, desirable genetically related strains can be differentiated by using other genotyping methods. We concluded that automated AFLP analysis is an attractive tool which can be used as a primary method for subtyping large numbers of Campylobacter strains and is extremely useful for epidemiological investigations.

Animals↗

Assessment of the genetic diversity among arcobacters isolated from poultry products by using two PCR-based typing methods.

In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and randomly amplified polymorphic DNA PCR (RAPD-PCR) were optimized for characterization of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. In addition, a simple and rapid DNA extraction method was tested for use in both typing procedures. Both methods had satisfactory typeability and discriminatory power, but the fingerprints generated with ERIC-PCR were more reproducible and complex than those obtained with RAPD-PCR. The use of nondiluted boiled cell suspensions as DNA templates was found to be very useful in ERIC-PCR. Characterization of large numbers of Arcobacter isolates is therefore preferably performed by the ERIC-PCR procedure. Isolates for which almost identical ERIC fingerprints are generated may subsequently be characterized by RAPD-PCR, although adjustment and standardization of the amount of the DNA template are necessary. In the second part of this study, the genotypic diversity of arcobacters present on broiler carcasses was assessed by using both typing methods. A total of 228 cultures from 24 samples were examined after direct isolation and enrichment. The isolates were identified by using a multiplex PCR as A. butzleri (n = 182) and A. cryaerophilus (n = 46). A total of 131 types (91 A. butzleri types and 40 A. cryaerophilus types) were discerned without discordance between the two typing techniques. The analysis of the poultry isolates showed that poultry products may harbor not only more than one species but also multiple genotypes. All genotypes were confined to one poultry sample, and only three genotypes were found after simultaneous enrichment and direct isolation. These results demonstrate that different outcomes can be obtained in epidemiological studies depending on the isolation procedure used and the number of isolates characterized.

Arcobacter↗

Evaluation of DNA extraction methods for use in combination with SYBR green I real-time PCR to detect Salmonella enterica serotype enteritidis in poultry.

The objective of this study was to develop a rapid, reproducible, and robust method for detecting Salmonella enterica serotype Enteritidis in poultry samples. First, for the extraction and purification of DNA from the preenrichment culture, four methods (boiling, alkaline lysis, Nucleospin, and Dynabeads DNA Direct System I) were compared. The most effective method was then combined with a real-time PCR method based on the double-stranded DNA binding dye SYBR Green I used with the ABI Prism 7700 system. The specificity of the reaction was determined by the melting temperature (T(m)) of the amplicon obtained. The experiments were conducted both on samples of chicken experimentally contaminated with serotype Enteritidis and on commercially available poultry samples, which were also used for comparisons with the standard cultural method (i.e., ISO 6579/2001). The results of comparisons among the four DNA extraction methods showed significant differences except for the results from the boiling and Nucleospin methods (the two methods that produced the lowest threshold cycles). Boiling was selected as the preferred extraction method because it is the simplest and most rapid. This method was then combined with SYBR Green I real-time PCR, using primers SEFA-1 and SEFA-2. The specificity of the reaction was confirmed by the T(m), which was consistently specific for the amplicon obtained; the mean peak T(m) obtained with curves specific for serotype Enteritidis was 82.56 +/- 0.22 degrees C. The standard curve constructed using the mean threshold cycle and various concentrations of serotype Enteritidis (ranging from 10(3) to 10(8) CFU/ml) showed good linearity (R(2) = 0.9767) and a sensitivity limit of less than 10(3) CFU/ml. The results of this study demonstrate that the SYBR Green I real-time PCR constitutes an effective and easy-to-perform method for detecting serotype Enteritidis in poultry samples.

Animals↗

Rapid detection of Campylobacter coli, C. jejuni, and Salmonella enterica on poultry carcasses by using PCR-enzyme-linked immunosorbent assay.

Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 x 10(2) and 4 x 10(1) CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100- to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa = 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for Salmonella. With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry.

Animals↗

Arsenic resistance in Campylobacter spp. isolated from retail poultry products.

Organoarsenicals are commonly used for growth promotion in U.S. poultry production. Susceptibilities to arsenite, arsenate, and the organoarsenical roxarsone were measured in 251 Campylobacter isolates from conventional and antimicrobial-free retail poultry products. Isolates from conventional poultry products had significantly higher roxarsone MICs (z = 8.22; P < 0.0001).

Animals↗

Diversity of strains of Salmonella enterica serotype enteritidis from English poultry farms assessed by multiple genetic fingerprinting.

Reliable and sufficiently discriminative methods are needed for differentiating individual strains of Salmonella enterica serotype Enteritidis beyond the phenotypic level; however, a consensus has not been reached as to which molecular method is best suited for this purpose. In addition, data are lacking on the molecular fingerprinting of serotype Enteritidis from poultry environments in the United Kingdom. This study evaluated the combined use of classical methods (phage typing) with three well-established molecular methods (ribotyping, macrorestriction analysis of genomic DNA, and plasmid profiling) in the assessment of diversity within 104 isolates of serotype Enteritidis from eight unaffiliated poultry farms in England. The most sensitive technique for identifying polymorphism was PstI-SphI ribotyping, distinguishing a total of 22 patterns, 10 of which were found among phage type 4 isolates. Pulsed-field gel electrophoresis of XbaI-digested genomic DNA segregated the isolates into only six types with minor differences between them. In addition, 14 plasmid profiles were found among this population. When all of the typing methods were combined, 54 types of strains were differentiated, and most of the poultry farms presented a variety of strains, which suggests that serotype Enteritidis organisms representing different genomic groups are circulating in England. In conclusion, geographical and animal origins of Salmonella serotype Enteritidis isolates may have a considerable influence on selecting the best typing strategy for individual programs, and a single method cannot be relied on for discriminating between strains.

Animals↗

Pasteurella multocida infection of poultry farm rats.

Thirty-four rat carcases from 11 poultry farms were examined for the presence of Pasteurella multocida; 14 out of 34 (41 per cent) proved positive after mouse inoculation, compared with five out of 34 (14 per cent) using media alone. No salmonellae were recovered from 27 rat carcases using enrichment media. Poultry pasteurellosis was present on two farms with infected rats and the same serotype was present in rats and poultry in those cases.

Animals↗

A major outbreak of botulism in cattle being fed ensiled poultry litter.

Eighty of a group of 150 housed beef cattle showed classical signs of botulism after eating a batch of ensiled poultry litter. Sixty-eight of the animals died and Clostridium botulinum type C toxin was detected in 18 of 22 sera examined. C botulinum organisms were isolated from the ensiled litter and type C toxin was demonstrated in samples of decomposed poultry carcases present in the litter. This outbreak of bovine botulism was the most serious to have been recorded in Europe and was the first associated with feeding ensiled poultry litter.

Animal Feed↗

Phylogrouping and genotyping of mcr-1 postives avian pathogenic Escherichia coli isolates in Algerian poultry farms.

Colibacillosis is a highly prevalent bacterial disease in poultry, resulting in the widespread use of antibiotics for both curative and preventive purposes. Consequently, avian pathogenic Escherichia coli (APEC) continues to act as a reservoir for antibiotic resistance genes, including the mcr-1 gene, which codes for resistance to colistin, a crucial antibiotic in human medicine. The aim of this study was to evaluate the antibiotic resistance pattern of APEC and to investigate the genotyping, phylogrouping, and virulence of mcr-1-positive isolates. A total of 113 APEC were isolated, of which 92% were multidrug resistant (MDR). The mcr-1 gene was detected in 41 isolates originating from turkeys and broilers. Two isolates carried blaTEM, one of which also harboured blaCTX-M encoding beta-lactamases. The Clermont phylogrouping revealed that 76% of the isolates belonged to phylogroup B1. Concerning the detection of the virulence-associated genes, 88% of isolates carried at least 3 genes. The ERIC-PCR classified our isolates into 6 different clusters. Our study highlights the emergence of colistin resistance and MDR, which pose a real threat to poultry production and public health. Control of antibiotic use in the poultry sector is urgent and mandatory.

Animals↗