Kinematic analysis quantifies gait abnormalities associated with lameness in broiler chickens and identifies evolutionary gait differences

This is the first time that gait characteristics of broiler (meat) chickens have been compared with their progenitor, jungle fowl, and the first kinematic study to report a link between broiler gait parameters and defined lameness scores. A commercial motion-capturing system recorded three-dimensional temporospatial information during walking. The hypothesis was that the gait characteristics of non-lame broilers (n = 10) would be intermediate to those of lame broilers (n = 12) and jungle fowl (n = 10, tested at two ages: immature and adult). Data analysed using multi-level models, to define an extensive range of baseline gait parameters, revealed inter-group similarities and differences. Natural selection is likely to have made jungle fowl walking gait highly efficient. Modern broiler chickens possess an unbalanced body conformation due to intense genetic selection for additional breast muscle (pectoral hypertrophy) and whole body mass. Together with rapid growth, this promotes compensatory gait adaptations to minimise energy expenditure and triggers high lameness prevalence within commercial flocks; lameness creating further disruption to the gait cycle and being an important welfare issue. Clear differences were observed between the two lines (short stance phase, little double-support, low leg lift, and little back displacement in adult jungle fowl; much double-support, high leg lift, and substantial vertical back movement in sound broilers) presumably related to mass and body conformation. Similarities included stride length and duration. Additional modifications were also identified in lame broilers (short stride length and duration, substantial lateral back movement, reduced velocity) presumably linked to musculo-skeletal abnormalities. Reduced walking velocity suggests an attempt to minimise skeletal stress and/or discomfort, while a shorter stride length and time, together with longer stance and double-support phases, are associated with instability. We envisage a key future role for this highly quantitative methodology in pain assessment (associated with broiler lameness) including experimental examination of therapeutic agent efficacy.     

Cyclopiazonic acid production by Aspergillus flavus and its effects on broiler chickens

Cyclopiazonic acid (CPA) was purified from cultures of Aspergillus flavus, and ca. 14 g of the toxin was collected for use in feeding studies. Chicken rations were artificially contaminated with purified CPA at concentrations of 10, 50, and 100 ppm (microgram/g) and fed ad libitum to eight groups of chickens for 7 weeks. Chickens receiving feed with 100 ppm of CPA had high mortality, decreased weight gain, and poor feed conversion when compared with birds receiving other doses. Postmortem examination showed that chickens fed the two greatest doses of CPA had proventricular lesions characterized by mucosal erosion and hyperemia (100 ppm) and by thick mucosa and dilated proventricular lumens (50 ppm). Birds given 100 ppm of CPA in feed also had numerous yellow foci in their livers and spleens. Microscopic examination of tissues of birds that received 100 ppm of CPA revealed ulcerative proventriculitis, mucosal necrosis in the gizzard, and hepatic and splenic necrosis and inflammation. Birds given 50 ppm of CPA had hyperplasia of the proventricular mucosal epithelium. Birds given 10 ppm of CPA and control birds had no significant treatment-related lesions.     

Cyclopiazonic acid production by Aspergillus flavus and its effects on broiler chickens.

Cyclopiazonic acid (CPA) was purified from cultures of Aspergillus flavus, and ca. 14 g of the toxin was collected for use in feeding studies. Chicken rations were artificially contaminated with purified CPA at concentrations of 10, 50, and 100 ppm (microgram/g) and fed ad libitum to eight groups of chickens for 7 weeks. Chickens receiving feed with 100 ppm of CPA had high mortality, decreased weight gain, and poor feed conversion when compared with birds receiving other doses. Postmortem examination showed that chickens fed the two greatest doses of CPA had proventricular lesions characterized by mucosal erosion and hyperemia (100 ppm) and by thick mucosa and dilated proventricular lumens (50 ppm). Birds given 100 ppm of CPA in feed also had numerous yellow foci in their livers and spleens. Microscopic examination of tissues of birds that received 100 ppm of CPA revealed ulcerative proventriculitis, mucosal necrosis in the gizzard, and hepatic and splenic necrosis and inflammation. Birds given 50 ppm of CPA had hyperplasia of the proventricular mucosal epithelium. Birds given 10 ppm of CPA and control birds had no significant treatment-related lesions.     

The prevalence of campylobacters and arcobacters in broiler chickens

Chicken carcasses from a supermarket and from a poultry abattoir were examined using methods designed to isolate as many strains of campylobacters and related organisms as possible. Strains of arcobacter, but no campylobacters, were isolated from every carcass after enrichment. Campylobacter jejuni subsp. jejuni was isolated from all carcasses examined by direct plating and other Campylobacter -like strains were isolated from nine out of 15 abattoir carcasses by direct plating but not after enrichment. Only the Camp. jejuni subsp. jejuni strains could be identified to species level using a readily available identification scheme and/or a commercial identification kit. Examination of caecal contents from the 15 abattoir poultry yielded Camp. jejuni subsp. jejuni and Campylobacter -like strains from 15 and eight by direct plating, and from six and nine after enrichment, respectively. Four sites in the intestine of the abattoir birds (60 samples) were examined for arcobacters and only one strain was isolated. This indicates that arcobacters are probably not normal inhabitants of the poultry intestine. Poultry is a rich source of other campylobacteria besides the thermophilic Campylobacter spp.     

Detoxification of mycotoxins by probiotic preparation for broiler chickens

Biological decontamination of mycotoxins using microorganisms is one of the well known strategies for the management of mycotoxins in foods and feeds. Among the different potential decontaminating microorganisms,Saccharomyces cerevisiae and lactic acid bacteria represent unique groups, which are widely used in food fermentation and preservation. The aim of this study was to determine the influence of spontaneous fermentation with the use of probiotic bacteria and yeast (Lactobacillus paracasei/casei ŁOCK 0920,L. brevis ŁOCK 0944,L. plantarum ŁOCK 0945,Saccharomyces cerevisiae ŁOCK 0142), on reduction of sum of aflatoxines (B₁, B₂, G₁, G₂) and ochratoxin A concentration during fermentation and the microflora pattern during fermentaton. The probiotic bacteria and yeast applied creates a starter culture for flour fermentation that has a stable feature of detoxication of aflatoxines and especially ochratoxin A. Presented at the 28^th Mykotoxin-Workshop, Bydgoszcz, Poland, May 29–31, 2006     

Cecal microbiome divergence of broiler chickens by sex and body weight

The divergence of gut bacterial community on broiler chickens has been reported as potentially possible keys to enhancing nutrient absorption, immune systems, and increasing poultry health and performance. Thus, we compared cecal bacterial communities and functional predictions by sex and body weight regarding the association between cecal microbiota and chicken growth performance. In this study, a total of 12 male and 12 female 1-day-old broiler chickens were raised for 35 days in 2 separate cages. Chickens were divided into 3 subgroups depending on body weight (low, medium, and high) by each sex. We compared chicken cecal microbiota compositions and its predictive functions by sex and body weight difference. We found that bacterial 16S rRNA genes were classified as 3 major phyla (Bacteroidetes, Firmicutes, and Proteobacteria), accounting for > 98% of the total bacterial community. The profiling of different bacterial taxa and predictive metagenome functions derived from 16S rRNA genes were performed over chicken sex and bodyweight. Male chickens were related to the enrichment of Bacteroides while female chickens were to the enrichment of Clostridium and Shigella. Male chickens with high body weight were associated with the enrichment of Faecalibacterium and Shuttleworthia. Carbohydrate and lipid metabolisms were suggested as candidate functions for weight gain in the males. This suggests that the variation of cecal bacterial communities and their functions by sex and body weight may be associated with the differences in the growth potentials of broiler chickens.     

Inheritance of fertility in broiler chickens

Background

The fertility of a chicken''s egg is a trait which depends on both the hen that lays the egg and on her mate. It is also known that fertility of an individual changes over the laying period.

Methods

Longitudinal models including both random genetic and permanent environmental effects of both the female and her male mate were used to model the proportion of fertile eggs in a pedigree broiler population over the ages 29-54 weeks.

Results

Both the male and the female contribute to variation in fertility. Estimates of heritability of weekly records were typically 7% for female and 10% for male contributions to fertility. Repeatability estimates ranged from 24 to 33%, respectively. The estimated genetic variance remained almost constant for both sexes over the laying period and the genetic correlations between different ages were close to 1.0. The permanent environment components increased substantially towards the end of the analyzed period, and correlations between permanent environment effects at different ages declined with increasing age difference The heritability of mean fertility over the whole laying period was estimated at 13% for females and 17% for males. A small positive correlation between genetic effects for male and female fertility was found.

Conclusion

Opportunities to improve fertility in broiler stocks by selection on both sexes exist and should have an impact throughout the laying period.     

The aversion of broiler chickens to concurrent vibrational and thermal stressors

The requirement for assessing the effects of multiple concurrent stressors in improving the welfare of broiler chickens during transport has not been widely recognised. A discrete-choice technique was used to investigate the aversion of broiler chickens to concurrent vibrational and thermal transport stressors. In experiment 1, 12 female broiler chickens, aged 42+/-3 days were studied individually using two choice-chambers. Each chamber had four compartments connected via a central zone and offered four treatments; thermal (T: air temperature; 40 degrees C, 21% RH), vibrational (V: frequency; 2Hz, acceleration; 1ms(-2)), concurrent vibrational and thermal (VT) and no applied stressors (N). Coloured compartment wall panels, allocated at random, assisted chickens' identification of compartments. Birds were fasted overnight and were required to make five consecutive choices on each of four consecutive days. A choice was defined as entering a compartment and feeding (5g pellets), whereupon confinement for 60min was initiated. Choices were totalled over all birds and analysed using a log-linear generalised linear model. The vibration was significantly avoided (V and VT versus N and T; P<0.05) but the thermal stressor was not (T and VT versus N and V; P>0.05) and there was no interaction. In experiment 2, the procedure was repeated with 12 more birds and modifications to increase method sensitivity and maximise bird learning. Choices were more disparate than before with vibration avoided to a greater extent (V and VT versus T and N; P<0.01) but there was still no main effect of the thermal treatment or a significant interaction. Substantial differences between individuals were observed in both experiments. The overall response to vibration supported previous findings for short-term exposure, however, non-avoidance of the thermal treatment was unexpected. Possibly, the birds were unable to associate the delayed heat stress with the compartment. Alternatively, the thermal conditions were not perceived as aversive either initially or throughout the 60min confinement. The preference method provides a useful starting point for assessing combinations of stressors which affect broiler welfare, allowing relative ranking of treatments from an animal-centred perspective.     

Contamination of broiler chickens by Staphylococcus aureus during processing; incidence and origin

Staphylococcus aureus was present in only small numbers ( ca 10/g) on the skin of broiler chickens before processing. During processing, contamination of carcases with this organism increased to > 10³/g of skin. Based on the results of phage typing, it was shown that the increase in contamination was due to a strain of Staph. aureus which was indigenous to the processing plant. Plucking and evisceration appeared to be the main stages at which contamination of carcases with Staph. aureus occurred.     

Nutritional value of waxy barleys for broiler chickens

Winter wheat was compared with a barley which has a normal amylopectin/amylose starch ratio, and waxy barleys which have essentially 100% amylopectin starch. In one trial, broiler chicks were fed on one of seven diets which had grain portions of all wheat (100W), 50% wheat–50% Compana (normal) barley (50W–50C), 25% wheat–75% Compana (25W–75C), 100% Compana (0W–100C), 50% wheat–50% Wapana (waxy) barley (50W–50Wa), 25% wheat–75% Wapana (25W–75Wa), or 100% Wapana (0W–100Wa). Body weights (g) at 7 weeks were (100W) 1659, (50W–50C) 1523, (25W–75C) 1401, (0W–100C) 1396, (50W–50Wa) 1503, (25W–75Wa) 1403 and (0W–100Wa) 1293. Dressing percentage was not affected by diet but mortality was greater on 0W–100C. Metabolizable energy (kJ/g dry matter) values determined on non-laying Leghorn hens were 12.93 ± 0.38 and 12.76 ± 0.46 for the Compana and Wapana barley, respectively. In a second trial, addition of a food-grade diastase at 2 g per kg of feed did not affect body weight on wheat-based diets, increased weights over corresponding control diets for Wapana-based diets but gave inconsistent results for Washunopana, a hull-less waxy barley with an active α-amylase-like activity on the starch granules. Group feed values indicated that enzyme supplements improved feed efficiency for barley diets but metabolizable energy values were similar for diets with or without diastase additions.     

Effect of indomethacin on hyperthermia induced by heat stress in broiler chickens

 An investigation was carried out to verify whether the heat stress hyperthermia response of broilers is prostaglandin-dependent. Male broiler chickens of the Hubbard-Petterson strain, aged 35–49 days, were used. Chickens were injected with indomethacin (1 mg/kg intraperitoneally ) 15 min before or 2 h after heat exposure (at 35°C for 4 h), and rectal temperature was measured before injection and up to 4 h thereafter. Birds were separated into two groups with and without access to water during heat stress. The increase in rectal temperature was lower (P<0.05) in birds with access to drinking water during heat exposure. All birds injected with indomethacin exhibited an increase in rectal temperature, irrespective of whether indomethacin was administered before or in the course of the rise in temperature. The results revealed that the increase in rectal temperature during heat exposure is not prostaglandin-dependent, and that the use of cyclooxigenase inhibitors is not recommended to attenuate heat stress hyperthermia in broiler chickens. Received: 15 December 1997 / Revised: 29 June 1998 / Accepted: 31 July 1998     

The effect of site of starch digestion on performance of broiler chickens

A growth trial with 420 broiler chickens (35 per experimental unit) was conducted in order to evaluate whether bird performance (Days 0–38) is affected by site of starch digestion. Two diets were formulated with the same calculated apparent metabolisable energy, digestible lysine, and digestible starch content. The diets contained starch sources with known amounts of rapidly digestible starch (RDS, starch digested until the posterior jejunum) and slowly digestible starch (SDS, starch digested in the ileum). Diets were either high (H) or low (L) in SDS content. RDS and SDS contents were 298 and 52 g/kg for the H diet and 345 and 7 g/kg for the L diet, respectively. Starch in the H diet was supplied by waxy maize, peas and sorghum and starch in the L diet was supplied by tapioca and common maize. Birds receiving the H diet consumed more feed (P<0.10), grew faster (P<0.01) and had a lower feed conversion ratio (P<0.01) than birds receiving the L diet. The results suggest that broiler chickens perform better on diets containing a minimal amount of slowly digestible starch. Future experiments are necessary to investigate the relation between amount of slowly digestible starch and performance of broiler chickens.     

The effect of microbial challenge on the intestinal proteome of broiler chickens

Background

In poultry production intestinal health and function is paramount to achieving efficient feed utilisation and growth. Uncovering the localised molecular mechanisms that occur during the early and important periods of growth that allow birds to grow optimally is important for this species. The exposure of young chicks to used litter from older flocks, containing mixed microbial populations, is a widely utilised model in poultry research. It rarely causes mortality but effects an immunogenic stimulation sufficient enough to cause reduced and uneven growth that is reflective of a challenging growing environment.

Methods

A mixed microbial challenge was delivered as used litter containing Campylobacter jejuni and coccidial oocysts to 120 male Ross 308 broiler chicks, randomly divided into two groups: control and challenged. On day 12, 15, 18 and 22 (pre- and 3, 6 and 10 days post-addition of the used litter) the proximal jejunum was recovered from 6 replicates per group and differentially abundant proteins identified between groups and over time using 2D DiGE.

Results

The abundance of cytoskeletal proteins of the chicken small intestinal proteome, particularly actin and actin associated proteins, increased over time in both challenged and control birds. Villin-1, an actin associated anti-apoptotic protein, was reduced in abundance in the challenged birds indicating that many of the changes in cytoskeletal protein abundance in the challenged birds were as a result of an increased rate of apoptosis. A number of heat shock proteins decreased in abundance over time in the intestine and this was more pronounced in the challenged birds.

Conclusions

The small intestinal proteome sampled from 12 to 22 days of age showed considerable developmental change, comparable to other species indicating that many of the changes in protein abundance in the small intestine are conserved among vertebrates. Identifying and distinguishing the changes in proteins abundance and molecular pathways that occur as a result of normal growth from those that occur as a result of a challenging microbial environment is important in this major food producing animal.

     

Diversity and prevalence of Arcobacter spp. in broiler chickens

Ninety-nine strains of Arcobacter spp., isolated from 10 chicken carcasses purchased from a supermarket and 15 chicken carcasses collected from a poultry abattoir, were speciated using a variety of phenotypic identification methods. All were tested using API Campy test strips and the 16-test (Preston) identification scheme developed for campylobacters. Fifty strains were selected for examination using a more comprehensive probabilistic identification scheme, and the identity of representative strains confirmed by protein profiling using SDS-PAGE. All 25 carcasses yielded Arcobacter butzleri . Three supermarket and 10 abattoir carcasses also carried A. cryaerophilus , and two abattoir carcasses carried A. skirrowii . The API Campy scheme proved unsatisfactory for identifying these strains: only 20 of 99 strains were accurately identified, all of which were A. cryaerophilus , the only Arcobacter sp. included in the database. Moreover, 76 of 99 strains were misidentified. The 16-test scheme identified all the arcobacter strains as A. cryaerophilus, since neither A. butzleri nor A. skirrowii had been described when the scheme was developed. The computer-assisted probabilistic scheme succeeded in identifying all but one strain, the identity of which was clarified by the use of SDS-PAGE. To our knowledge this is the first time that arcobacters other than A. butzleri have been reported in poultry meat or any other food of animal origin. Their high prevalence in poultry products may be of significance to public health.     

Bacteriophage therapy to reduce salmonella colonization of broiler chickens

Acute enteric infections caused by salmonellas remain a major public health burden worldwide. Poultry, particularly chickens, are known to be the main reservoir for this zoonotic pathogen. Although some progress has been made in reducing Salmonella colonization of broiler chickens by using biosecurity and antimicrobials, it still remains a considerable problem. The use of host-specific bacteriophages as a biocontrol is one possible intervention by which Salmonella colonization could be reduced. A total of 232 Salmonella bacteriophages were isolated from poultry farms, abattoirs, and wastewater in 2004 and 2005. Three phages exhibiting the broadest host ranges against Salmonella enterica serotypes Enteritidis, Hadar, and Typhimurium were characterized further by determining their morphology and lytic activity in vitro. These phages were then administered in antacid suspension to birds experimentally colonized with specific Salmonella host strains. The first phage reduced S. enterica serotype Enteritidis cecal colonization by > or = 4.2 log10 CFU within 24 h compared with controls. Administration of the second phage reduced S. enterica serotype Typhimurium by > or = 2.19 log10 CFU within 24 h. The third bacteriophage was ineffective at reducing S. enterica serotype Hadar colonization. Bacteriophage resistance occurred at a frequency commensurate with the titer of phage being administered, with larger phage titers resulting in a greater proportion of resistant salmonellas. The selection of appropriate bacteriophages and optimization of both the timing and method of phage delivery are key factors in the successful phage-mediated control of salmonellas in broiler chickens.     

Assessment of age-related glucose oxidation rates of broiler chickens by using stable isotopes

During their relatively short commercial lifespan of six weeks, broiler chickens undergo very pronounced age- or body weight-related changes in metabolic rate and body composition. The present study was aimed to assess the age-related changes in glucose oxidation rate of broiler chickens by using 13C-labeled glucose. The methodology for this breath test needed to be established first. Broiler chickens aged from two to six weeks were placed in open-circuit respiration cells and received a single intubation of U-13C6-glucose, followed by breath sampling for 4 hours and mass spectrometric analysis of 13C: 12C ratio in the exhaled air. Simultaneously, CO2 concentration in the respiration cell air was continuously monitored in order to calculate the cumulative percentage dose recovery (CPDR). With respect to the methodology, an oral dose of 2 mg U-13C6-glucose per kg body weight while maintaining a CO2 in the concentration of 0.4 to 0.5% was considered to be optimal. The three-parameter Gompertz curve fitted the CPDR values very well. Pronounced age-related changes in exogenous glucose oxidation rates in rapidly growing meat-type chickens were assessed. Young broiler chickens spend only a relatively low percentage of ingested glucose for immediate oxidation. In contrast, broiler chickens approaching the age of maximal absolute growth rate oxidize a greater proportion of the recently ingested glucose relative to the non-oxidative disposal pathways. This shift in the exogenous partitioning is discussed in relation to age-dependent changes in glucose turnover, lipid oxidation and deposition and metabolic heat production.     

Bone response of broiler chickens (Gallus gallus domesticus) induced by corticosterone

Experiments were conducted with chickens exposed to corticosterone (CORT), with the aim of determining its effects on bone characteristics. At 7 d of age, the experimental birds were injected daily with CORT (4 mg/kg of body mass) for 1 week. CORT administration significantly decreased the body weight while increasing relative liver weight of the chickens and the bone parameters were also decreased. Histology and immunohistochemistry of type X collagen revealed that CORT reduced the lengths of proliferative and prehypertrophic zone in growth plate and the number of positive chondrocytes in the prehypertrophic zone. In conclusion exposure to CORT depressed the growth performance and retarded the longitudinal growth of the long bones by inhibiting the proliferation and differentiation of chondrocytes in growth plate in broilers.     

Campylobacter infection of broiler chickens in a free-range environment

Campylobacter jejuni is the most common cause of bacterial gastroenteritis worldwide, with contaminated chicken meat considered to represent a major source of human infection. Biosecurity measures can reduce C. jejuni shedding rates of housed chickens, but the increasing popularity of free-range and organic meat raises the question of whether the welfare benefits of extensive production are compatible with food safety. The widespread assumption that the free-range environment contaminates extensively reared chickens has not been rigorously tested. A year-long survey of 64 free-range broiler flocks reared on two sites in Oxfordshire, UK, combining high-resolution genotyping with behavioural and environmental observations revealed: (i) no evidence of colonization of succeeding flocks by the C. jejuni genotypes shed by preceding flocks, (ii) a high degree of similarity between C. jejuni genotypes from both farm sites, (iii) no association of ranging behaviour with likelihood of Campylobacter shedding, and (iv) higher genetic differentiation between C. jejuni populations from chickens and wild birds on the same farm than between the chicken samples, human disease isolates from the same region and national samples of C. jejuni from chicken meat.