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AIMS: To assess the potential of essential oils and structurally related synthetic food additives in inhibiting the growth of Clostridium perfringens for the control of necrotic enteritis in chickens. METHODS AND RESULTS: The antimicrobial activity of essential oils/compounds was measured by determining the inhibition of bacterial growth. Thirty-three of 66 oils/compounds exhibited > or =80% inhibition. Seven with the highest potency were further studied. The oils/compounds had MIC(95) values between 167 and 425 microg ml(-1). Most of them were tolerant to low pH (2.0) and exhibited minor or no inhibition of Lactobacillus isolates from the chicken intestine. When mixed with chicken ileal digesta, the oils/compounds retained their efficacy against C. perfringens, but had little effect on the total number of lactobacilli and anaerobic bacteria in the digesta. CONCLUSIONS: Some essential oils/compounds demonstrated good potential in controlling C. perfringens. SIGNIFICANCE AND IMPACT OF THE STUDY: This study has identified candidates of essential oils/compounds for in vivo studies for the control of necrotic enteritis in chickens.

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AIMS: Determining the effects of zinc bacitracin, bird age and access to range on bacterial microbiota in the ileum and caeca of broilers. METHODS AND RESULTS: 16S rRNA gene-based polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) profiling, DNA sequencing and real-time quantitative PCR techniques were used. The richness of both ileal and caecal microbiota increased with chicken age. The microbiota from those birds of the same age demonstrated relatively similar PCR-DGGE profiles and tended to form closely related clusters in the relatedness analyses. Dietary treatment with bacitracin (50 mg kg(-1)) and access to range did not change the richness but altered the composition of the microbiota. The impact of bacitracin was particularly obvious in 3-day-old chicks. Lactobacilli were abundant in the caecal microbiota of 3-day-old chicks regardless of the dietary treatment with bacitracin. The access to range enriched Bifidobacterium in both the ileum and caeca. CONCLUSIONS: Bacitracin, bird age and access to range all influenced bacterial microbiota in the ileum and caeca of broilers, with bird age having the greatest apparent effect. SIGNIFICANCE AND IMPACT OF THE STUDY: Providing useful information for the development of antibiotic replacement therapy for poultry production.

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The microaerophilic nature of Campylobacter jejuni has complicated its recovery from human and animal sources. In this study, enhancement of the growth and aerotolerance of C. jejuni ATCC 35921 in nutrient broth no. 2 (NB2) was investigated. The efficiency of recovery of C. jejuni in NB2 containing FBP (0.025% [each] ferrous sulfate, sodium metabisulfite, and sodium pyruvate), 5% laked horse blood, hemin, Oxyrase, or activated charcoal in an aerobic atmosphere was compared with that obtained under microaerophilic incubation. The shortest lag time (lamda) for cells grown aerobically was observed with NB2 supplemented with FBP, 5% laked horse blood, 0.01 g/liter of hemin, or 0.15 U/ml of Oxyrase. The efficacy of these media to resuscitate C. jejuni cells in late exponential phase, as well as cells subjected to stress induced by cold, heat, starvation, or acid, was determined in aerobic or microaerobic atmospheres. The h of cells grown aerobically in NB2 containing both FBP and blood was similar to that obtained in the same medium incubated in a microaerobic environment (P > 0.05). However, the X was longer during aerobic growth when low numbers of cells (approximately 1 log CFU/ml) in late exponential phase were used as the initial inoculum. The best recovery of stressed C. jejuni was observed in NB2 supplemented with FBP and blood and incubated aerobically. Enrichment in media incorporating FBP and 5% laked horse blood is a simple, convenient, and time-saving method to replace microaerophilic incubation methods for the resuscitation of C. jejuni.

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(1) Genetic parameters for the sudden death syndrome (SDS) were estimated in meat-type chickens. Data were collected over 11 generations of selection for body weight within two distinct breeds (Cornish and White Rock). (2) The animal model was used exclusively with linear methods (LM) to estimate genetic parameters. Heritability (h2) of SDS on the liability scale was 0.30 +/- 0.002 and 0.25 +/- 0.002 in the Cornish and White Rock breeds, respectively. (3) A positive genetic correlation (r(g)) with ascites (AS) was determined (approximately 0.3 +/- 0.006). However, it was not possible to estimate the rg of SDS with body weight because of the low prevalence of the defect trait studied (1.8% in the Cornish and 1-5% in the White Rock). (4) Heritability of SDS calculated using male records only was 0.45 +/- 0.009 and 0.35 +/- 0.009, and r(g) with body weight was 0.30 +/- 0.010 and 0.27 +/- 0.009, in the Cornish and White Rock breeds, respectively. (5) In conclusion, the heart defect investigated was heritable with a positive genetic correlation with AS and body weight.

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Genetic parameters for the ascites syndrome (AS) were estimated for meat-type chickens. Data had been collected over 11 generations of selection for body weight and other traits within two distinct breeds (Cornish and White Rock). Linear methods (LM) were used to estimate genetic parameters and also to analyze a binary measure of survival. Survival analyses (SA) were also conducted to estimate the effects of various factors influencing the incidence of AS by evaluating the number of days that the birds survived. The animal model was used exclusively with linear methods. Heritabilities (h2) on the liability scale were 0.12 +/- 0.02 and 0.22 +/- 0.01 in the White Rock and Cornish breeds, respectively; however, the genetic correlation (r(g)) with body weight was not possible to estimate due to the low prevalence of the defect trait studied (1.5% in the Cornish and 1.1% in the White Rock). Because males are more prone to AS, the h2 using the male records only were 0.22 +/- 0.017 and 0.41 +/- 0.009, and the r(g) were 0.35 +/- 0.007 and 0.22 +/- 0.009 in the dam and sire populations, respectively. In conclusion, the heart defect investigated was heritable and had a positive genetic correlation with body weight.

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Adaptation to changes in extracellular salinity is a critical event for cell survival. Genome-wide DNA chip analysis has been used to analyze the transcriptional response of yeast cells to saline stress. About 7% of the genes encoded in the yeast genome are induced more than 5-fold after a mild and brief saline shock (0.4 m NaCl, 10 min). Interestingly, most responsive genes showed a very transient expression pattern, as mRNA levels dramatically declined after 20 min in the presence of stress. A quite similar set of genes increased expression in cells subjected to higher saline concentrations (0.8 m NaCl), although in this case the response was delayed. Therefore, our data show that cells respond to saline stress by inducing the expression of a very large number of genes and suggest that stress adaptation requires regulation of many cellular aspects. The transcriptional induction of most genes that are strongly responsive to salt stress was highly or fully dependent on the presence of the stress-activated mitogen-activated protein kinase Hog1, indicating that the Hog1-mediated signaling pathway plays a key role in global gene regulation under saline stress conditions.

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Restriction fragment length polymorphism analyses were used to examine endogenous viral genes (ev genes or ALVE genes) of the avian leukosis viral (ALV) family in semi-congenic lines of meat chickens. The Generation 6 lines examined in this study were semi-congenic in that each contained birds with either zero or with one ALVE gene in hemizygous state plus some solitary long terminal repeat (LTR) elements. Using four restriction enzymes on chicken genomic DNA and two probes, one representing the entire ALV retroviral genome and one with only a small part plus the LTR, four ALVE genes were characterized. Each seemed to be complete with no detectable deletions. None appeared to be similar to known ALVE genes of White Leghorns, whereas two of the four may be the same as ALVE genes reported by others in White Plymouth Rock chickens.

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Swabs of crop contents of 635 broiler chickens were obtained from 9 Ontario and 12 Quebec processing plants and cultured for Salmonella to determine prevalence in broiler crops. Serotypes of positive cultures were determined to evaluate the serotype profile. The overall prevalence of contamination was low (4.3%). Prevalence was higher in broilers sampled in Quebec (5.8%) than in those sampled in Ontario (2.2%). In Quebec, there were differences in prevalence among the groups of broilers sampled at the various plants. These differences were believed to be attributable to differences in Salmonella prevalence among groups of flocks delivered to the plants due to the limited exposure of the chickens to the plant. The serotype profile of Salmonella isolated from the crops of broilers in this study was similar in several respects to profiles obtained from other surveys of Canadian broiler flocks using either environmental samples or cloacal swabs. Similarities included: predominance of Salmonella hadar and Salmonella heidelberg; several other common serotypes at a low prevalence; little Salmonella enteritidis isolated in other studies, and no S. enteritidis isolated in this study. Results of this field survey of Salmonella in crops of broilers are similar to those of Canadian studies of other internal and environmental sites of broilers. The similarity indicates that monitoring of Salmonella environments of flocks of live broiler chickens should define profiles of Salmonella contamination of the carcasses and would also aid in determination of Salmonella contamination status of broiler flocks. Such information would assist efforts to reduce Salmonella contamination in broiler chickens.

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The purpose of this study was to determine the relationship between genetic selection for growth traits and tissue expression of the chicken growth hormone receptor (cGHR) gene. Two different populations of broiler chickens were studied. One population consisted of strain (S) 80, selected for 14 generations for high 9-week body weight (BW), and its progenitor, S90 (a 1950's strain). The second population consisted of S21, selected for 10 generations for high 4-week BW and low abdominal fat, and its progenitor S20 (a 1970's strain). Tissue (liver, fat, breast and leg muscle) and blood samples were collected from six birds/strain at 2-week intervals between 1 and 11 weeks of age. An RNase protection assay was developed to measure mRNA levels of full-length cGHR (3.2 and 4.3 kb) transcripts and chicken glyceraldehyde 3-phosphate dehydrogenase (for normalization) in total RNA prepared from tissue. Analysis of the area-under-curve (AUC) was used for strain comparisons of certain developmental profiles (BW, plasma hormones and tissue cGHR mRNA). The BW AUC showed that the growth rates are different (P < 0.05) among the four strains (S21 > S20 > S80 > S90). Both slow-growing strains (S90 and S80) had a higher (P < 0.05) plasma GH AUC than the two fast-growing strains (S20 and S21). The plasma T3 AUC was highest (P < 0.05) in S90 due to maintenance of higher T3 levels after 3 weeks of age. At 11 weeks of age, hepatic and plasma GH-binding activities were positively related to growth rate (S21 > S20 > S80 > S90). However, the developmental increase in cGHR mRNA in liver and fat was similar among these different populations of growth-selected broiler chickens. Steady-state levels of cGHR mRNA increased in a developmental manner in the liver (5-fold at 9 weeks of age) and abdominal fat (4.5-fold at 11 weeks of age) of all strains. In contrast, there was no developmental increase or strain difference in cGHR mRNA levels in breast and leg muscle. There is a discrepancy between GH-binding activity in liver and plasma, which is different among strains, and steady-state levels of tissue cGHR mRNA which are similar among strains. These observations suggest that the cGHR is under translational or post-translational regulation which would determine the amount of cGHR protein available for GH binding.

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Vermicompost (VC) was produced by earthworms fed with fresh chicken faeces, and was earth-like in appearance and odour. In three experiments, VC was sprinkled on the first feed of newly-hatched broiler chicks. Treated and control groups were challenged on day 6 by the addition of seeder chicks that had been inoculated orally with Salmonella typhimurium. Chickens were killed at intervals during a 6-week period and were tested for colonization of the pathogen in the crop, caecum and internal organs. The VC-treated groups were significantly more resistant (p < 0.01) to colonization by S. typhimurium than the untreated controls. In one experiment, the VC treatment appeared to have conferred complete protection against colonization of both the crop and caecum. Colonization of the crop was increased transiently by addition to the diet of 2.5% fructooligosaccharide, xylo-oligosaccharide or transgalactosylated oligosaccharide. The influence of these carbohydrates on colonization of the caecum was variable.

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Salmonella typhimurium colonization, pH, and density of ceca were measured in 3-, 5-, and 6-wk-old broilers fed either a control ration or rations with added fructooligosaccharides (FOS) or lactose derivatives (LD). The purpose was to compare dietary crude FOS from Jerusalem artichokes with refined FOS and two LD for ability to reduce Salmonella colonization as determined by semi-quantitative procedures. Chicks were challenged commencing at 5 d, by exposure to chicks orally infected with S. typhimurium. With the exception of chicks fed crude FOS, the high prevalence of Salmonella infection at 3 wk declined as chicks aged. At 6 wk, broilers fed crude FOS had higher Salmonella counts than all other broiler groups, whereas broilers fed refined FOS had lower infections than control broilers. The decline of Salmonella infection of broilers fed either refined FOS or LD ceased after dietary additives were discontinued at 5 wk of age. At 6 wk, infection rates of the latter groups were at least as high as those of control broilers. Both FOS and LD reduced cecal pH and density. Broilers fed the control ration had higher pH at 5 and 6 wk and higher cecal densities at 3 and 5 wk than those of broilers fed rations containing 5% (wt/wt) carbohydrates. Treatment differences for cecal pH and density disappeared within 1 wk of withdrawal of carbohydrates from 5-wk-old broilers. No consistent effect of cecal pH and density on Salmonella infection was observed. Density was dependent on cecal volume, and at 6 wk of age, on broiler size.

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To assess the value of DNA fingerprints for the prediction of heterosis in chickens, retrospective analyses of data from three crossbreeding experiments and DNA fingerprints (DEP) of parental strains were conducted using two minisatellite and one middle-repetitive DNA probes. DEP bands were assessed on pooled DNA samples of 10-15 individuals per parental genetic group. The number of DEP bands evaluated in the experiments ranged from 81 to 139. The probes varied in their predictive value, but predictability of heterosis generally increased with multiple probes. Highly significant correlations (0.68-0.87) between band sharing ratios (SH) and heterosis were found in 25 crosses of White Leghorns in the first egg production cycle for age at sexual maturity, egg production, and mature body weight: traits with heterosis of 10% or more of the means. Regressions on SH explained 78.4% of the variation in heterosis in age at sexual maturity, 60.2% in egg production and 46.4% in mature body weight. For "broiler" traits with heterosis of < 1%, none of the correlations, based on 13 crosses, were significant. It was concluded that multilocus probe DFP of pooled DNA samples show promise as predictors of heterosis.

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An in vivo antisense strategy was used to examine the involvement of G-protein subunits in supraspinal (intracerebroventricular; i.c.v.) alpha2-adrenoceptor-mediated antinociception. Mice that were injected with 33-mer antisense oligodeoxyribonucleotides (6 nmol) or vehicle were tested (tail-flick) with an agonist (clonidine, guanfacine or BH-T 920) administered i.c.v. 18 - 24 h later. Gi3alpha antisense treatment attenuated BH-T 920 and clonidine-induced antinociception. Gi2alpha antisense produced differential effects on the three agonists. Gi1alpha and G(s)alpha antisense treatment had no significant effect. Together with the previous demonstration that i.c.v. mu-opioid antinociception is mediated via Gi2alpha, the present results suggest that different receptors may mediate antinociception via different G-protein subunits and, hence, that specific subunits might offer novel targets for drug discovery.

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Restriction fragment length polymorphism analysis was conducted for a set of eight different meat chicken-derived endogenous viral genes (ev genes) of the avian leukosis viral (ALV) family. Each viral element was first isolated into a separate single-element line by selective breeding. Genomic DNA from the founder male for each semi-congenic, single-element line was digested with each of four restriction enzymes, and the resulting Southern blots were each hybridized with up to four probes representing different portions of the ALV retroviral genome. Among the eight elements, there was one that represents the broiler equivalent of locus ev3 of White Leghorn chickens. A second broiler element showed a SacI-specific junction fragment similar to that of ev8. The remainder appeared to be different from any of the 21 ev genes previously described for White Leghorn chickens. Four of the eight elements examined were essentially complete, but the rest have sustained internal deletions.

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The molecular architecture of the sex-linked late-feathering region of the chicken genome is still poorly defined. Current evidence points to a strong association between the presence of the endogenous viral element ev21 and the late-feathering phenotype. However, analysis at the molecular level has demonstrated that this is not a simple case of insertional mutagenesis. Instead, the structure of the region of the chicken genome containing the feathering locus is complex and variable between and within lines of chickens. Significant clues to the molecular structure of this genomic region can be obtained by analyzing rare and revertant genotypes. However, searching for rare genotypes can only be carried out effectively using quick screen methodology. This paper describes a quick, polymerase chain reaction-based test for ev21 that facilities the search for rare genotypes.

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Feed consumption and feed efficiency data obtained over nine generations from three selected strains of broilers were adjusted for differences in either initial or initial and final test BW (at 28 and 42 d, respectively). Genetic and environmental variables were estimated for each strain using an animal model with restricted maximum likelihood procedures to avoid bias due to reduced genetic variation caused by selection. Variance component and heritability estimates before and after adjustment of these traits for differences in test weights were evaluated. Adjustments of feed consumption data for either initial test BW or both initial and final test BW reduced additive genetic and environmental variation. Reductions were relatively greater for additive genetic than for environmental variance, thereby reducing the heritability of the adjusted trait. Adjustment of feed efficiency for initial test BW altered additive genetic variation little and reduced environmental variation slightly; however, correction for both test BW reduced variation of both components. Heritability of adjusted feed efficiency was 40% larger (.33 to .35 vs .21 to .28) than for uncorrected feed efficiency, but phenotypic variation of the adjusted feed efficiency was reduced 42% (5.82 to 6.13 vs 10.30 to 11.35). Heritabilities of feed consumption and efficiency were essentially the same (.34 and .36) after adjustment for both test weights. Changes in genetic and environmental variation and heritability due to adjustments were similar for the three strains.

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In vivo nuclear magnetic resonance (NMR) imaging and spectroscopy techniques were used to monitor changes in P- and H-containing molecules in embryos of two broiler strains (30 and 31) differing genetically in fat content and ranging in age from 0 to 20 days of incubation. Chemical analysis showed that Strain 30 has more carcass fat than Strain 31 at market age (7 wk). Proton (1H) and 31P spectra were obtained on four eggs per strain at Days 0, 2, 4, 6, 8, 11, 12, 14, 16, 17, 19, and 20 of incubation. Fat:water, phosphomonoester (PME):phosphodiester (PDE), and adenosine triphosphate (ATP):PDE ratios were calculated. Chicks were hatched, grown to market weight (2,000 g for females and 2,300 g for males at 7 wk), and the whole intact carcasses were analyzed for crude fat. Hydrogen-1 NMR spectroscopy studies of incubated eggs indicated no significant difference (P > .05) in the fat:water ratio between the two strains. The difference in the PME:PDE ratios between the two strains as obtained by 31P-NMR spectroscopy over all days of incubation analyzed was not significant (P > .05); however, there was a significant difference in this ratio between the two strains at Days 4, 6, and 11. Up to Day 16, Strain 30 had a slightly, but not significantly (P > .05), higher ATP:PDE ratio as shown on 31P-NMR spectra, whereas after Day 17 the ATP:PDE ratio was significantly higher (P < .01) for Strain 31. Strain 31 birds had a significantly lower (P < .05) crude fat content. There was a significant difference (P < .05) in 7-wk carcass fat content between sexes, males having significantly (P < .01) less fat than females, which was correlated with a significantly higher (P < .01) ATP:PDE ratio in male embryos. It might be possible to use ATP:PDE ratios obtained during embryonic development by 31P-NMR to select strains of birds for low fat content at market weight and to distinguish between sexes during late embryonic development.

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The present study was designed to document the complexity of endogenous viral (ev) genes and seek evidence for their association with production traits in selected and control strains of meat-type chickens. Three populations were studied, each consisting of a control strain and one to three strains selected for various production traits. The ev genes were revealed by digesting genomic DNA with restriction enzymes and detecting DNA fragments on Southern blots using radioactive probes from nucleotide sequences of the avian leukosis virus genome. A total of 31 polymorphic ev loci were identified in these populations from a SacI digest, with an average of 7.3 ev genes per bird. There were no significant differences in ev genes per bird between strains within populations or between selected and control strains overall. Thirty of 62 comparisons in the three populations indicated ev gene frequency differences (P less than .05). Within populations, 13 of 93 comparisons of ev gene frequencies between control and selected strains and 8 of 62 between three selected strains of a sire population showed such differences (P less than .05). Selection for body weight and feed efficiency had been observed to reduce gene frequencies of the slow-feathering gene, which usually contains the ev21 locus; however, these effects were not detected (.05 less than P less than .06) between strains of the dam population in the current study. Such differences suggested possible associations between ev genes and production traits in meat-type chickens.

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In an effort to eliminate exogenous avian leukosis virus (ALV) from a susceptible population, albumen of one egg per hen from each of four generations was tested by ELISA for group-specific antigen (gsa) of leukosis/sarcoma viruses. From 1510 to 2099 hens were tested in each generation. Hens were not used as breeders if optical density readings for gsa were 0.4 or greater. Despite this procedure, there was no appeciable change in the occurrence of gsa in eggs from one generation to the next and in the sire population the percentage of hens with levels of 0.4 or greater was 7.1 and 8.7 in the first and fourth generations tested, respectively. There were no consistent differences in antigen levels in any of the eight strains that comprised the two populations. For the most part, antigen detected in eggs resulted from expression of endogenous viral genes since there was a low incidence of exogenous infection. Six of 234 hens of the second generation tested from the sire population were positive for ALV in serum and three of 161 were positive for antibody to subgroup A virus; no antibody to subgroup B was detected. Five of the six hens that tested positive for ALV were from the same strain and had been reared together. With one exception these hens would have been eliminated as breeders based on tests for antigen in eggs. In the fourth generation tested, no virus or subgroup A antibody was found in serum from approximately 250 hens from the two populations.

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Feather pulp from White Leghorns that had been inoculated with the RAV-1 strain of avian leukosis virus (ALV) was tested for antibody by ELISA. In an ELISA on three feather pulps and serum from each of four inoculated chickens, there was a highly significant correlation between antibody levels in feather pulp (1 pulp per 800 mul of buffer) and a 1:500 dilution of serum. Also based on ELISA, there were highly significant correlations between levels of maternal and actively acquired antibody to Newcastle disease virus in feather pulp and serum collected from meat-type birds at intervals between 10 and 108 days of age.