Prediction of heterosis from DNA fingerprints in chickens.

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.

Assessment of inbreeding by DNA fingerprinting: development of a calibration curve using defined strains of chickens.

By analyzing DNA fingerprints of chickens from seven well-defined genetic groups, a calibration curve was established relating the degree of inbreeding with the average band frequency, allelic frequency and band sharing. The probe used was bacteriophage M13 DNA and digestion of the genomic DNA was carried out with the MspI restriction enzyme. The analysis also provided an estimate of the average allelic frequency at a hypervariable locus and the average mutation frequency per locus and generation. The values of 0.24 and 1.7 X 10(-3), respectively, are similar to the estimates for humans using other probes and hybridization protocols. It is suggested that the calibration curve established can be used for determining inbreeding not only in chickens, but also in other species.

Identification of markers associated with quantitative trait loci in chickens by DNA fingerprinting.

Three approaches for identifying VNTR alleles associated with quantitative traits in chickens are described. One approach is based on the comparison of well-defined selected and non-selected control strains. The second approach is based on analyzing chickens within a breeding population ranked according to specific traits and the third approach involves segregation analysis. In this latter approach a large number of offspring of a single male segregating for a quantitative trait are produced and tested for trait association of the male DNA fingerprinting bands. In all cases pooled DNA samples of birds, rather than individual samples, are analyzed and band intensity is assumed to reflect the relative frequency of an allele. Examples from the literature and from our laboratory indicate that these methods permit the identification of DNA fingerprinting bands associated with quantitative traits. After developing locus-specific probes for these bands it should ultimately be possible to detect and map quantitative trait loci.

Combined effects of glycerol concentration, cooling velocity, and osmolality of skim milk diluents on cryopreservation of ram spermatozoa.

The interaction of glycerol concentration from 0 to 16% and cooling velocity from 1 to 100 degrees C/min on freeze-thaw survival of ram spermatozoa was studied using a diluent based on 15% skim milk (450 mOs/kg water). Optimal spermatozoa survival (percentage motility and rating) was obtained with 4 to 6% glycerol and freezing rates of 10 to 100 degrees C/min. Similar results were obtained with 8% glycerol at freezing rates of 5 to 30 degrees C/min. Although the ram spermatozoa tolerated several cooling velocities at each glycerol concentration, increasing the concentration of glycerol resulted in a downshift in the range of optimal cooling velocities. Glycerol concentrations above 8% were toxic and contributed greatly to the progressive decrease in spermatozoa survival. Comparison of the 15% skim milk diluent (450 mOs/kg water) with a 19% skim milk diluent (600 mOs/kg water) showed that optimal cryosurvival was obtained with 4 to 6% glycerol and freezing rates of 10 to 100 degrees C/min with both diluents.

Evaluation of a new diluent and different processing procedures for cryopreservation of ram semen.

Ram semen was processed for freezing after initial dilution with a modified Tris-fructose diluent. Two aliquots were processed by cooling gradually to 5 degrees C, further dilution, equilibration and freezing in 0.5 ml straws either in pressurized liquid nitrogen (LN(2)) vapor (Method A) or on a block of dry ice (Method B). A third aliquot was cooled rapidly to 16 degrees C and then slowly to 5 degrees C, diluted further, equilibrated and frozen in straws in pressurized LN(2) vapor (Method C). The second dilution was carried out using a new diluent based on dextran-lactose. The diluted semen was equilibrated for 2 h before freezing. Semen was evaluated by artificial insemination (AI). The fertility of ewes bred by a double insemination with frozen-thawed semen processed by Methods A, B and C was 73% (n = 33), 67% (n = 30) and 80% (n = 30), respectively. In comparison, the fertility of ewes inseminated with fresh semen was 93% (n = 31). These preliminary data indicate an acceptable fertility can be achieved by AI with frozen-thawed semen processed using improved procedures.

Genotypic and phenotypic parameters of spur incidence and length in White Leghorn hens.

Spurs were measured on two groups of hens: at 532 days of age in six selected and three control strains (Expt. 1); and at 483 days of age in a full 3 X 3 diallel of three of the selected strains as well as two control strains and two commercial stocks (Expt. 2). There were no significant differences in length between spurs on right and left legs. Strain differences for spur incidence were large and significant, but those for length generally were not. Pooled strain estimates of heritability for spur incidence were .21 for both left and right spurs, and .40 (left) and .44 (right) for spur length (Expt. 1). General combining ability was large and highly significant for spur incidence only, and specific combining ability was not significant for either incidence or length of spurs (Expt. 2). Heterosis was not significant for spur incidence or length. Reciprocal differences were significant only for spur incidence of the crosses of strains 4 and 8. Spur length was negatively correlated phenotypically with egg production, egg weight, specific gravity, and Haugh units. Genotypically, spur length was positively correlated with part-record egg production to 273 days and negatively correlated with 450-day egg weight and specific gravity.

Genetic effects of aging on egg production traits in the first laying cycle of White Leghorn strains and strain crosses.

Three White Leghorn strains, their two-way crosses, and two commercial lines were used to evaluate the effects of aging on mean heterosis, reciprocal, additive, Z-chromosome, and heterotic effects and their variances in two egg production traits during the first laying cycle. Egg number of the survivors (EPF) and egg number including mortality and morbidity (EPM) were evaluated from hens housed one per cage in a randomized block design. For analyses, egg number was divided into 12 periods of 28 d each. Synchronization of the records was achieved by starting recording at age at first egg. The mean heterosis was significant over time and increased with age for both traits (P<0.05). Reciprocal effects were not significant across periods but increased in magnitude with age for EPF. The pattern of age changes in additive, Z-chromosome, and heterotic effects varied among strains, indicating genotypic differences in response to aging. These differences increased, on average, with age. The additive, heterotic, environmental, and phenotypic variances increased with age for both traits. A decline with physiological age was observed in heritabilities, which was due to a faster increase in environmental variance than to an increase in additive variance. Genetic variance of egg production increased with age, indicating that improvement of lifetime performance of layers is possible. This improvement could be achieved by selecting animals at older ages or by favoring individuals with better DNA repair or those who had more genes turned on or off during the course of aging.

Embryonic adenosine triphosphate:phosphodiesters ratios obtained with in vivo nuclear magnetic resonance spectroscopy (phosphorus-31): a new technique for selecting leaner broiler chickens.

Phosphorus-containing compounds of 18-d-old embryos from two broiler chicken lines, differing in body fat content, were measured using in vivo 31P nuclear magnetic resonance (NMR) spectroscopy. Subsequently, the same birds were slaughtered at 8 wk of age and the whole body was analyzed for body fat content. The birds of the fat line had lower (P = .002) embryonic adenosine triphosphate (ATP):phosphodiester (PDE) ratios, higher (P = .002) body fat content when adjusted to common BW, and higher (P = .047) dry matter content than the lean line. No differences (P > .05) were detected for BW between the two lines. Females of the two lines had lower (P = .002) ATP:PDE ratios, lower (P = .001) BW, higher (P = .003) adjusted fat contents, and higher (P = .003) dry matter content than the males. No interaction (P > .05) between sexes and lines was detected for any variables. Regression equations indicating a linear negative relationship (Ybody fat content = 369.05 - 407.27 XATP:PDE + .1295 XBW, R2 = .62 to .78; or Ybody fat % = 30.57 - 19.4 XATP:PDE, R2 = .49 to .71) between embryonic ATP:PDE ratios and body fat content were developed.

The lethal nature of "wiry" mutation in the domestic fowl.

A study of the effect of normal environmental conditions on newly hatched chicks with the feather mutant described as "wiry" (Fiser et al., 1973) was carried out. The results indicated that the abnormality should be classified as a potential facultative lethal expressed after hatching.

Relationship between activity at a young age and feed efficiency in chickens.

Feed conversions of three Ottawa strains of Leghorns were measured from hatch to 27 days and bird activity at 1, 2, and 4 weeks of age was assessed. Males were more active than females. Chicks of the more active strain consumed more feed and had poorer conversion than those from the least active strain. Ranking of the strains on feed conversion to 27 days was similar to their ranking on feed consumed per egg mass produced in an earlier study.

Effects of comb dubbing on the performance of laying stocks.

Three studies were conducted with birds dubbed at hatch vs. dubbed and dewattled at 118 days or 255 to 260 days of age or with normal hens. In the first, involving 5928 pullets of four strains housed 1 per cage, dubbing and dewattling at 255 to 260 days caused small adverse effects on egg numbers as compared with nondubbed controls. Body size was reduced, and both egg weight (EW) and shell strength were increased slightly. In the second study, involving 8180 pullets of nine strains housed 1 per cage, dubbing and dewattling at 118 days slightly increased survivor egg production (SEP) and laying house mortality, and reduced age at first egg, 240-day EW, and 450-day Haugh units (HU), as compared with dubbing only at hatch. There were significant strain by dubbing treatment interactions for hen-day rate of lay (HDR) and SEP. In the third study, involving 1824 pullets of three strain crosses housed 2 and 3 birds per cage (three different cage sizes), there were no significant differences between birds dubbed at hatch and those not dubbed. Variation in age at 50% production, SEP, and HDR was reduced for the dubbed groups. These studies show that the older the birds are when dubbed, the greater the negative effects of dubbing. Hens dubbed at hatch exhibit no effects with the early dubbing or adverse effects in the laying house.

The effect of sodium pentobarbital on the preservation of fertilizing capacity of turkey spermatozoa.

The effect of sodium pentobarbital in modified glycine citrate extender on the preservation of fertilizing capacity of turkey spermatozoa was tested. Extended semen samples were stored for 6 or 24 hr at 4 C before insemination. Although there was an indication that the addition of 120 mg of sodium pentobarbital per 1 liter of extender was beneficial at 24 hr storage, the difference was not statistically significant.

Genetic effects of aging on fitness and nonfitness traits in laying hens housed three per cage.

Three White Leghorn strains, their two-way crosses, and two commercial lines were used to study the effects of aging on several parameters related to performance of fitness and nonfitness traits during the first laying cycle of hens housed three per cage. Egg number of the survivors (EPF) and egg number including mortality and morbidity (EPM) were divided into 12 periods of 28 d each, starting at age at sexual maturity. Egg weight (EW), specific gravity (SG), and albumen height (AH) were measured at 240, 350, and 450 d of age. Mean heterosis was significant over time, except for AH, increasing in magnitude with age for EPF, EPM, EW, and AH. Reciprocal effects were more important for egg quality than for egg production traits and were influenced by age. The age-related changes in additive, Z-chromosome, and heterotic effects varied among strains, showing that strains differ in their genetic schemes in response to aging. Nonadditive, environmental, and phenotypic variances increased with age for all traits. Additive variance increased with age for EPF, EW, and AH. Z-chromosome variance increased with age for EW and AH. Heritabilities decreased with age, except for EPF and AH. On average, genetic variance increased with advancing age. Improvement in lifetime performance may be obtained by selecting birds at older ages. As the relative increase with age in additive variance was larger for egg production than for egg quality traits, selection for the latter could be performed at early stages.

Genetic effects of aging on egg quality traits in the first laying cycle of White Leghorn strains and strain crosses.

Three White Leghorn strains, their two-way crosses, and two commercial lines were used to evaluate the effects of aging on heterosis (H), reciprocal effects, and additive (A), Z-chromosome (Z), and heterotic effects and their variances on egg quality traits during the first laying cycle. Egg weight (EW), specific gravity (SG), Haugh unit (HU), and albumen height (AH) were measured at 240, 350, and 450 d of age from hens housed one per cage in a randomized block design. The mean heterosis was significant over time only for EW. For EW, heterosis increased in magnitude with age. The mean heterosis for both HU and AH was also influenced by age. Reciprocal effects were significant, on average, across periods for all traits and were influenced by age. The age-related changes in additive, Z-chromosome, and heterotic effects varied significantly among strains, indicating differences by genetic group in response to aging for egg quality traits. The heterotic, environmental, and phenotypic variances increased with age for all traits, except for AH. The additive and Z-chromosome variances did not always increase with age. Their age trend varied, depending on the trait. Heritabilities decreased with advancing age, suggesting that selection to improve lifetime performance of egg quality traits can be done early in the cycle.

The effects of geographical area, rearing method, caging density, and lymphoid leukosis infection on adult performance in egg stocks of chickens.

Birds were brooded reared, and laying house-tested at two widely separated sites: Animal Research Centre (ARC) and North Central Region Poultry Breeding Laboratory (NCRPBL). Two strain and a commercial stock were included only at NCRPBL. At ARC, all birds were brooded, reared, and laying house-tested in cages. At NCRPBL, half of the chicks were brooded and reared on floor and half in cages, and all hens were laying tested at one or two birds per cage. Most hens housed one per cage were tested for lymphoid leukosis virus (LLV) infection by the complement fixation test for group specific antigen in egg albumen. When all hens tested at NCRPBL were considered, the method of brooding and rearing affected only egg weight and body weight in favor of the hens reared in floor pens. The commercial stock and the strain crosses performed better than the control strains. Despite very similar genetic backgrounds, there were large differences between control strains 10 and NCR. Hens housed one per cage had lower mortality and higher egg production than those housed two per cage. Average performance of birds at ARC was somewhat better than that at NCRPBL, but there were genotype-site interactions for mortality, egg production, and sexual maturity. The frequency of shedders in the crosses (17.2%) was the same as that of the control strains (20.8%), but the frequency of shedders in the commercial stock (4.5%) was much lower. The method of brooding and rearing had no effect on the frequency of shedders. The LLV infection was associated with significant reductions in egg production and egg quality. For example, the difference between test-positive and test-negative hens was 25 eggs per hen housed and 16 eggs per surviving hen.

Influence of selection for egg production and Marek's disease resistance on the incidence of endogenous viral genes in White Leghorns.

The influence of selection on the frequencies of endogenous viral (ev) genes related to the avian leukosis virus was studied in two genetically distinct sets of White Leghorn strains. Each set consisted of four strains: an unselected control strain, two strains selected for egg production traits, and a strain selected for Marek's disease (MD) resistance as well as egg production traits. Eight different ev genes were observed in Set I and seven in Set II, four being common to both sets. Selection for egg production traits resulted in significant changes of the frequency of four ev genes in both sets. In Set I, increased frequencies were observed for ev-4, ev-7, and ev-8; a decreased frequency for ev-9 was observed. The ev-9 gene expresses the viral envelope protein, whereas the others are transcriptionally silent, with the possible exception of ev-7. In Set II, increased frequencies were observed for the transcriptionally silent ev-8 and for ev-15, a gene which consists of a solitary long terminal repeat. Decreased frequencies were observed for ev-18, which codes for infectious endogenous virus, and for a second ev gene of unknown phenotype. In the resistance-selected strains the frequencies of the ev genes were intermediate between those of the control strains and the strains selected for egg production traits with the exception of ev-6, which expresses the viral envelope protein, and ev-3, which expresses internal viral proteins as well as the envelope protein.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo nuclear magnetic resonance spectroscopy of chicken embryos from two broiler strains of varying fat content.

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.

Effects of changes in photoperiod on freezability of ram spermatozoa.

The effect of photoperiod on freezability of ram spermatozoa was evaluated in ejaculates collected over 52 weekly periods from two groups of rams housed in windowless rooms maintained under either a natural light regimen corresponding to latitude 45 degrees N or its reverse. The survival of spermatozoa after freezing of 0.5-ml straws at 15 degrees C/min, storage in liquid nitrogen, and thawing in a water bath at 39 degrees C was evaluated as freeze-thaw motility percentage and rating and as a cryosurvival percentage. Freeze-thaw motility percentage was highest during the decreasing photoperiod, regardless of season. Motility percentage after freezing was positively correlated with motility percentage before freezing (r = 0.40) and ejaculate osmolality (r = 0.41), and negatively correlated with percentage of abnormal spermatozoa (r = 0.46). Cryosurvival was significantly lower during the winter and spring seasons for semen collected from rams maintained under the natural light regimen. No significant differences in cryosurvival over the year were observed in semen collected from rams maintained under the reverse light regimen. Cryosurvival was positively correlated with ejaculate osmolality. The vigor of frozen-thawed spermatozoa, assessed as motility rating, was significantly lower during the increasing photoperiod for rams exposed to the natural light regimen. However, the motility rating of spermatozoa collected from rams under the reverse light did not differ significantly.

The effects of rapid cooling (cold shock) of ram semen, photoperiod, and egg yolk in diluents on the survival of spermatozoa before and after freezing.

The effects of rapid cooling of semen (cold shock) from 30 degrees C to various temperatures above 0 degrees C on survival of ram spermatozoa suspended in diluents with or without egg yolk were assessed before and after freezing. Rapid cooling of extended semen from 30 to 15 degrees C had little or no effect on spermatozoa survival before or after freezing. Rapid cooling of extended semen from 30 degrees C to 10, 5, or 0 degrees C was accompanied by a progressive decrease in percentage of motile spermatozoa and percentage of intact acrosomes before freezing and a decrease in percentage of motile spermatozoa and after freezing. The ability of spermatozoa motile after cold shock to survive freezing and thawing, evaluated as cryosurvival, was not significantly (P greater than 0.05) affected by the temperature to which semen was cooled. The addition of egg yolk to the initial extender had a beneficial effect on percentage of motile spermatozoa particularly after rapid cooling of semen to 10 and 5 degrees C. Although egg yolk had little effect before freezing on semen rapidly cooled to temperatures above 15 degrees C and therefore not actually cold shocked, it substantially improved the subsequent survival of spermatozoa after freezing and thawing. Percentage of motile spermatozoa after cooling and after freezing was generally higher when the semen was collected during a decreasing photoperiod than during an increasing photoperiod.

Combined effect of glycerol concentration and cooling velocity on motility and acrosomal integrity of boar spermatozoa frozen in 0.5 ml straws.

The interaction of glycerol concentrations of 0-10% and cooling rates from 1 to 1,500 degrees C/min with boar spermatozoa motility and acrosomal integrity (proportion of spermatozoa with normal apical ridge) was studied after thawing 0.5 ml straws at a constant rate. While increasing the glycerol concentration from 0 to 4% progressively improved motility, the percentage of spermatozoa with a normal apical ridge gradually decreased. The magnitudes of the respective changes depended on cooling rate. A peak value of 48.1% and rating 3.8 were obtained in semen protected with 4% glycerol, frozen at 30 degrees C/min. Increasing the glycerol levels above 6% resulted in a gradual decrease in motility. The proportion of spermatozoa with normal apical ridge was highest in semen protected with 0-1% glycerol after cooling at 30 degrees C/min (64.4% and 66.1%, respectively), but at these glycerol concentrations the percentage of motile spermatozoa was low. At the 30 degrees C/min cooling rate, the decline in the proportion of cells with normal apical ridge due to increasing the glycerol levels to 3 and 4% was relatively slow (57.3% and 49.4%, respectively). Cooling at 1 degrees C/min was detrimental to acrosomal integrity, which decreased with increasing glycerol concentration, in contrast to increasing motility, which even at its maximum, remained low. The direct plunging of straws into liquid nitrogen (1,500 degrees C/min) resulted in damaged acrosomes in all spermatozoa with the total loss of motility. Balancing motility and acrosomal integrity, freezing boar semen protected with 3% glycerol by cooling at 30 degrees C/min resulted in optimal survival for boar semen frozen in 0.5 ml French straws.