The control of lymphoid leukosis in a flock White Plymouth Rock chickens

Summary

Lymphoid leukosis (LL) was successfully controlled in a commercial basic breeding line of White Plymouth Rock chickens. The control method has been developed for breeder flocks and consists of three elements: - In the flock under study, homogenates of embryos from all eggs collected during a number of I4‐day periods are tested for the presence of LL viruses.

- Only eggs from hens that have been shown not to shed virus in their eggs are used for the production of progeny. The offspring are reared in isolation during the first two months of life, at which time the age‐related resistance against tumour formation by LL viruses appears to be sufficiently developed.

- The chickens are subsequently inoculated intramuscularly with LL viruses of subgroups A and B transferred to a conventional chicken house.

The vaccination raises a solid immunity to horizontal LL virus exposure and, due to the age‐related resistance, tumour formation does not follow.

No excretion of LL viruses could be detected in three generations of White Plymouth Rock chickens to which the three elements of the control procedure were applied. Clinical disease was not observed in any of the chickens under notice.     

On epizootiology and control of lymphoid leukosis in chickens

Sera and organ extracts from ten different commercial stocks of layer chickens were examined for the presence of lymphoid leukosis (LL) viruses. Virus was recovered from 40.8% of the cockerels between three and six weeks of age. Their female hatch mates were examined at the age of 20 months. A mean of 11.3% of these laying hens was positive in the NP activation test. Lymphoid leukosis was successfully controlled in three inbred strains of White Leghorn chickens and in a commercial White Plymouth Rock line. All flocks were kept in a filtered air positive pressure (FAPP) house during the first two months of life and thereafter transferred to a conventional environment. The control method is based on three elements:

• —from an infected flock, hens are selected in whose eggs no avian lymphoid leukosis viruses can be detected by examination of pooled extracts of groups of embryos;

• —only eggs from hens that are shown not to shed congenitally virus in their eggs are used for the production of progeny. The offspring are reared in isolation until two months of age at which time the age-related resistance against tumour formation appears to be sufficiently developed;

• —the chickens are subsequently intramuscularly inoculated with lymphoid leukosis viruses of subgroups A and B and transferred to a conventional chicken house. The inoculated birds become persistently viremic and resist horizontal virus exposure and intramuscular challenge infections.

Horizontal virus transmission was observed to take place when virus-free non-vaccinated chickens were reared in isolation for two months and then exposed under field conditions.Efficiency of virus recovery was considerably improved when washed buffy coat cells were cocultivated with chick embryo fibroblasts or explant cultures were prepared from various tissues before testing with the NP activation test.     

饲粮中添加丁酸梭菌和丁酸钠对产蛋鸡生产性能和蛋品质的影响

试验旨在研究在产蛋鸡饲粮中添加丁酸梭菌和丁酸钠对不同品种产蛋鸡产蛋率、鸡蛋雀斑、暗斑以及蛋品质的影响。选择270日龄狼山鸡、芦花鸡、北京油鸡各320只,随机分成2组,每组8个重复,每个重复20只鸡,对照组（CON）饲喂基础饲粮,试验组（EXP）饲粮中添加100 mg/kg丁酸梭菌＋500 mg/kg丁酸钠。预饲期3 d,试验期为5周。试验结果表明,与对照组相比,①试验组狼山鸡、芦花鸡、北京油鸡产蛋率分别提高12.5%、12.0%和24.9%（P<0.01）;②试验组狼山鸡鸡蛋雀斑3级率下降34.2%（P<0.05）,芦花鸡和北京油鸡鸡蛋各级雀斑率均无显著差异（P>0.05）,狼山鸡、芦花鸡、北京油鸡鸡蛋各级暗斑率均无显著差异（P>0.05）;③试验组狼山鸡蛋重降低1.7%（P<0.05）,蛋形指数增加2.3%（P<0.05）;芦花鸡蛋重增加1.5%（P<0.05）;北京油鸡蛋白高度增加13.9%（P<0.05）,哈氏单位增加4.7%（P<0.05）。综上,在产蛋鸡基础饲粮中添加一定量的丁酸梭菌和丁酸钠,可以提高狼山鸡、芦花鸡、北京油鸡产蛋率,对改善狼山鸡鸡蛋的雀斑也有一定的作用,且有助于提高芦花鸡、北京油鸡的蛋品质。     

鸡lmbr1基因外显子16的SNP检测和单倍型分析

外显子16是鸡lmbr1基因最大的外显子,本研究进行了该外显子在丝羽乌骨鸡和白洛克肉鸡间的SNP检测和单倍型分析。研究表明,鸡lmbr1外显子16的PCR-SSCP基因型在两个品种间的分布存在明显差异,测序从24个个体中检测到4个变异位点,其中T32C变异在两个品种间存在明显差异,丝羽乌骨鸡均含有32T(纯合的TT或杂合的TC),白洛克肉鸡在T32C位点都为纯合的CC;单倍型分析从24个个体中检测到5种单倍型,丝羽乌骨鸡和白洛克肉鸡的单倍型存在明显的差异,hap1和hap2是丝羽乌骨鸡的特异性单倍型,hap5是白洛克肉鸡的特异性单倍型,hap3和hap4主要存在于白洛克肉鸡中,在丝羽乌骨鸡中的比例极少。     

Ovum formation and multiple ovulation in lines of white Plymouth Rocks and their crosses

The production of double-yolked eggs and the duration of the rapid growth phase of yolks were measured in parental lines of White Plymouth Rock pullets and their crosses over 30 d, commencing with the day of first egg. Significant differences were found between mating combinations in the incidence of multiple-yolked eggs, but not in the period of rapid yolk growth. Heterosis and recombination effects for multiple ovulation were respectively -9% and -66% of the mean, while the corresponding values for the period required for rapid yolk growth were -1% and -5%. Three double-yolked eggs were observed containing yolks which differed by 3 d in their periods of rapid growth. Hypotheses are were presented for the origin of double-yolked eggs.     

8周龄中外鸡种心肌、肝脏组织差异表达基因的初步分析

本文利用mRNA差异显示技术对8周龄中国地方品种丝毛乌骨鸡、引进品种白莱航蛋鸡、白洛克肉鸡进行基因差异表达分析。结果表明：三个品种鸡的心肌、肝脏组织分别检测到18和20条差异表达的基因片段。其中，心肌组织中的差异表达片段在白莱航蛋鸡与白洛克肉鸡之间最多，肝脏组织中的差异表达片段在丝毛乌骨鸡与白洛克肉鸡间最多。实验结果为寻找产蛋、产肉及肉质性状相关的基因提供了遗传学基础信息。     

Quantitative genotyping by amplifying the polymorphic sequences of Pre-Melanosomal Protein (PMEL17) gene using real-time polymerase chain reaction in chickens

1. This study was conducted to develop a quantitative genotyping system of chimaeric chickens by real-time PCR. 2. The polymorphisms in exons 7 and 11 of PMEL17 gene, which is one of the major genes affecting plumage colour, were identified from White Leghorn, Barred Plymouth Rock and Rhode Island Red chickens. 3. Quantitative genotyping was successfully performed by real-time PCR using polymorphic sequence-specific TaqMan Probes. 4. This methodology can support future research of germline chimaeric chickens as well as the application of germ cell transfer technique.     

Genetic control of cellular infection by subgroups A and C RNA tumour viruses in guinea fowl

An investigation was carried out in guinea fowl to determine their susceptibility to infection by Rous sarcoma viruses of subgroups A and C. A standard dose of each subgroup virus was inoculated into 14-day-old embryos via the chorioallantoic membrane (CAM). On the 10th day after inoculation, 50% of the embryonic chorioallantoic membranes were harvested to assess their infection status (CAM(+) or (–)), while the rest were allowed to hatch. The hatchabilities of the embryos inoculated with subgroups A and C were about 50% and 57%, respectively. The relative sensitivities of guinea fowl to infection by viruses of subgroups A and C were observed to be 0.220 and 0.003, respectively, as compared to chickens (1.00). Mortality due to subgroup A virus-induced liver tumours (LT) was 54% and four phenotypic subclasses, namely CAM(+) LT(+), CAM(+) LT(–), CAM(–) LT(+) and CAM(–) LT(–), were observed in guinea fowl as in chickens. However, a higher incidence (31%) of conversely associated phenotypes, i.e. CAM(+) LT(–) and CAM(–) LT(+), were observed in guinea fowl. Mortality caused by subgroup A virus-induced liver tumours was first observed in inoculated guinea fowl keets during the 3rd week after hatching, and 93% of the mortality occurred within 6 weeks. The peak mortality occurred in the 4th week after hatching. The target organs for transformation were considered to be the liver and spleen because of the equal incidence of tumours in these organs. Males and females were equally likely to die from liver tumours. There was also a considerable reduction in the hatchability of guinea fowl embryos from eggs inoculated with either viral subgroup, as reported in chickens.Abbreviations BS Bryan standard - CAM chorioallantoic membrane - LL lymphoid leukosis - LT liver tumour - PCV pock count range - RSV Rous sarcoma virus - tva tumour virus (subgroup A) - tvc tumour virus (subgroup C)     

Field trial in commercial broilers with a multivalent in ovo vaccine comprising a mixture of live viral vaccines against Marek's disease,infectious bursal disease,Newcastle disease,and fowl pox

A multivalent in ovo vaccine (MIV) was tested for safety and efficacy in a commercial broiler complex. The MIV comprised five replicating live viruses including serotypes 1, 2, and 3 of Marek's disease virus (MDV), an intermediate infectious bursal disease virus (IBDV) and a recombinant fowl poxvirus (FPV) vector vaccine containing HN and F genes of Newcastle disease virus (NDV). The performance of MIV-vaccinated broilers was compared with that of hatchmates that received turkey herpesvirus (HVT) alone (routinely used in ovo vaccine in the broiler complex). The chickens that hatched from the MIV-injected and HVT-injected eggs were raised under commercial conditions in six barns. Barn 1 housed 17,853 MIV-vaccinated chickens and each of the barns 2-6 housed 18,472-22,798 HVT-vaccinated chickens. The HVT-vaccinated chickens were given infectious bronchitis virus (IBV) and NDV vaccines at hatch and at 2 wk of age. The MIV-vaccinated chickens received IBV vaccine at hatch and IBV + NDV at 2 wk of age. The relative values of hatchability of eggs, livability and weight gain of chickens, and condemnation rates at processing were comparable between the MIV and the HVT groups (P > 0.05). Chickens from the MIV- and the HVT-vaccinated groups were challenged with virulent viruses under laboratory conditions. The resistance of vaccinated chickens against Marek's disease could not be assessed because of high natural resistance of unvaccinated commercial broilers to virulent MDV. The relative resistances of the MIV- and the HVT-vaccinated groups, respectively, against other virulent viruses were as follows: IBDV, 100% for both groups; NDV, 81% vs. 19%; FPV, 86% vs. 0%. The successful use of MIV under field conditions expands the usefulness of the in ovo technology for poultry.     

Genetics of hyperpigmentation associated with the Fibromelanosis gene (Fm) and analysis of growth and meat quality traits in crosses of native Indian Kadaknath chickens and non-indigenous breeds

1. The study investigated the extent of hyperpigmentation (a trait fixed in native Indian Kadaknath chickens), bodyweight, carcase quality and leanness at 12 weeks of age in F(1) and back-crosses of Kadaknath with White Leghorn, White Plymouth Rock and Aseel Peela chickens. 2. The objective of the study was to determine if hyperpigmentation was affected by the major gene Fibromelanosis (Fm) and to evaluate the effects of different proportions of Kadaknath genes on growth and carcase quality. 3. The pigmentation pattern of skin indicated that Fm behaved as the primary locus affecting dermal-hyperpigmentation and that the sex-linked Id locus produced an epistatic effect. 4. The results suggested that variable allelic forms of Id were acting in different crosses, which resulted in variation in melanosis of the host. However, no conclusive pattern for shank pigmentation could be explained through genotyping of the Id and Fm loci. 5. Analysis of quantitative traits indicated the positive impact of a Kadaknath genomic proportion of 50% or more on meat texture and carcase leanness. Improvement in leanness occurred in White Rock crosses but not in White Leghorn and Aseel Peela crosses. 6. Thigh-meat texture was influenced more by enhanced Kadaknath genomic proportions than the breast-meat. It was concluded that introgression of Kadaknath genomic proportion beyond 50% in a cross with meat-type chickens, irrespective of the impact Fm, brought improvement in meat quality whereas no such advantage was obtained for growth traits. 7. The beneficial impact of the Kadaknath genome on meat quality calls for further studies to identify causative genes for their selective use to improve meat quality in Kadaknath crossbred chickens.     

Characterization of infectious laryngotracheitis viruses, antigenic comparison by kinetics of neutralization and immunization studies.

Five isolates of infectious laryngotracheitis virus were compared by pock formation on the chorioallantoic membrane of embryonated eggs, plaque size in chicken embryo kidney tissue culture, and antigenic relationship using reciprocal kinetics of neutralization. The A4557-5 strain of infectious laryngotracheitis virus, which causes mild respiratory disease, produced pocks with a zone of edema on the chorioallantoic membrane. A virulent virus (Virus 1), isolated from an outbreak of severe disease characterized by a diphtheritic laryngotracheitis, produced the largest plaques in chicken embryo kidney cell culture. Other virulent viruses (Viruses 2, 3 and V154) did not have unique growth characteristics when grown on the chorioallantoic membrane or in chicken embryo kidney cell culture. All viruses were closely related antigenically as shown by kinetics of neutralization but viruses 2 and 3 were not homogeneous with the other three viruses when neutralized by anti-V154 chicken serum. Following aerosol infection, chickens infected with the A4557-5 virus were immune to challenge with virulent V154 virus. However, in comparison to SA-2 virus, this virus was a less effective immunizing agent when administered by the vent or drinking water methods.     

Characterization of two microsatellites in chicken monoamine oxidase A

We surveyed two polymorphic microsatellites in a candidate gene for fear response, monoamine oxidase A ( MAOA ), in chicken. Two hundred and eighty chickens from five breeds (Nagoya, Mikawa, White Leghorn, White Plymouth Rock, and Rhode Island Red) were investigated. A thymine (T) repeat and an adenine (A) repeat were found on MAOA intron 4 ( CMin4T ) and intron 9 ( CMin9A ), respectively. Nine alleles ( 127-bp to 166-bp ) in CMin4T and six alleles ( 184-bp to 198-bp ) in CMin9A were detected. A 128-bp allele in CMin4T was observed in the Nagoya breed only, implying a highly useful marker for discriminating the Nagoya breed from other breeds. In addition, the Mikawa breed had the fixed 127-bp and 198-bp alleles in CMin4T and CMin9A , respectively. The Nagoya breed chickens show densely cowardly behavior, but the Mikawa breed chickens do not show the same behavior. The current results may indicate that MAOA is an informative candidate gene for breed difference.     

Limited susceptibility and lack of systemic infection by an H3N2 swine influenza virus in intranasally inoculated chickens

Chickens were intranasally inoculated with the swine influenza virus (SIV) A/swine/NC/307408/04 (H3N2) (NC/04 SIV) to determine the infectivity of a North American SIV for chickens, as well as the possibility of chicken meat serving as a transmission vehicle for SIV. White leghorn (WL) layer-type chickens were used for initial pathotyping and infectivity tests, and a more comprehensive intranasal pathogenesis study was done with white Plymouth rock (WPR) broiler-type chickens. None of the NC/04 SIV-inoculated WL or WPR chickens displayed clinical signs. Serologic tests showed that the virus was able to infect both intranasally inoculated WL and WPR chickens, but the antibody titers were low, suggesting inefficient replication. Some of the NC/04 SIV-inoculated WL chickens shed low levels of virus, mostly from the alimentary tract, but viral shedding was not detected in NC/04 SIV-inoculated WPR chickens. The comprehensive pathogenesis study demonstrated that the virus did not cause systemic infections in WPR chickens, and feeding breast and thigh meat from the NC/04 SIV-inoculated WPR to WL chickens did not transmit NC/04 SIV.     

Cellular defense of the avian respiratory tract: paucity of free-residing macrophages in the normal chicken

Avian respiratory macrophages (ARM) were obtained from lungs and air sacs of 122 White Plymouth Rock chickens, ranging from 376 to 3800 g in weight. Procedures involved lavaging through the surgically prepared trachea with either a 15-g cannula or French #8 pediatric urinary catheters. Factors, in different combinations, investigated for their effects on the ARM yield, were: lavage fluids (0.85% physiologic saline, 0.1 M phosphate-buffered saline, Ca-Mg-free Hanks' solution, Eagle's minimum essential medium); additives (10 U heparin/ml, 0.1% EDTA, 12 mM lidocaine); lavage repetitions (from 3 to 10); fluid temperature (room and 41 C); and lavage time (fluid retention up to 35 min). None of the lavage methods emerged clearly as the best, with phosphate-buffered saline and 0.85% physiologic saline alone as good as when combined with additives. Although 10 lavages yielded more ARM, it appeared that the majority of ARM washed off into the early lavages. Chickens from a line selected for large body size had more ARM than those from a line selected for small body weight. Regardless of genetic line, however, the chickens yielded a very low number of ARM compared with mammalian species of the same or smaller weight. Most of the birds yielded only 200,000 to 300,000 ARM, with minimum yields being less than 10,000, the maximum being 2 million ARM. Either these results point to a deficiency in the defense system of the chicken's respiratory tract against bacteria, mycoplasma, fungi, and viruses, or mechanisms other than macrophages are primary in resistance to pathogens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Embryo vaccination against Marek's disease with serotypes 1, 2 and 3 vaccines administered singly or in combination

Marek's disease virus (MDV) vaccines of serotypes 1 and 2 administered in 18-day-old embryonated eggs induced better protection against post-hatch challenge at 3 days with virulent MDV than vaccines given at hatch. Embryonal vaccination with a polyvalent vaccine containing equal quantities of serotypes 1 and 2 of MDV and serotype 3 virus (turkey herpesvirus, HVT) was also significantly more effective than post-hatch vaccination. These and earlier results indicate that protective efficacy of single or combined Marek's disease vaccine serotypes against post-hatch challenge at 3 days can be substantially improved if the vaccines are injected into 18-day embryos rather than at hatch. Injection of vaccines of serotypes 1 or 2 into embryonated eggs or hatched chicks did not cause detectable gross or microscopic lesions in chickens. Vaccine viruses of serotypes 1 and 2 could be isolated from spleen cells of chickens 1 week post-vaccination, and the titer of recoverable viruses was higher in chickens that received the vaccines at the 18th day of embryonation than in chickens vaccinated at hatch. Although embryo vaccination with HVT usually provided better protection than post-hatch vaccination against early post-hatch challenge with variant pathotypes of MDV, the protection was poor regardless of vaccination protocol. If challenge with variant pathotypes of MDV was delayed until embryonally or post-hatch HVT-vaccinated chickens were 21 days of age, protection of chickens by HVT was not enhanced. Thus, resistance induced by embryonal vaccination with HVT was qualitatively similar to that induced by post-hatch vaccination with this virus.     

Allele‐specific polymerase chain reaction typing and sequencing of mitochondrial D‐loop region in broiler chickens in Japan

This study aimed to comprehend a feature of single nucleotide polymorphism (SNP) in mitochondrial DNA (mtDNA) mainly of general broiler chickens in Japan. We typed two SNP sites (199C/T and 792A/G) of the D‐loop region in mtDNA by allele‐specific PCR (AS‐PCR) in 359 broiler (182 chunky and 177 cobb) and 506 layer (233 White Leghorn, 140 Barred Plymouth Rock and 133 Rhode Island Red) chickens. The SNP of 199C or 792A by AS‐PCR was observed in the chunky and cobb chickens, and not in the layers. The haplotype 199T/792G was observed in a part of cobb and all layers. By the result of AS‐PCR haplotyping and the broiler brands, the D‐loop region was sequenced in 44 broiler chickens (20 chunky and 24 cobb) and compared with the layers’ sequence data. Among the broiler and layer chickens, 21 SNP sites (including one insertion) and 11 sequence haplotypes were observed. Haplotype variation or correspondence was observed in and between the broiler brands. This study provides important information to establish a chicken meat traceability system by SNP haplotyping of mtDNA in Japan.     

鸡lmbr1部分内含子的克隆及其单核苷酸多态性

以丝羽乌骨鸡和白洛克肉鸡的基因组DNA为模板,采用PCR产物直接测序的方法进行鸡lmbr1基因部分内含子的克隆、单核苷酸多态性检测。结果表明,试验克隆了鸡lmbr1基因的10～14及16内含子序列,共计4 610 kb,从丝羽乌骨鸡和白洛克肉鸡中检测到67个单核苷酸多态位点（SNPs）。将提交序列与NCBI上公布的红色原鸡基因组序列比较,在这6个内含子内发现40个变异位点,其中20个为新的SNPs。基因组结构分析表明,鸡lmbr1基因约60 kb,为人类lmbr1基因的28%,内含子外显子的剪接方式符合GT-AG法则。     

Allele‐specific PCR typing and sequencing of the mitochondrial D‐loop region in four layer breeds

This study aimed to investigate the ability of single nucleotide polymorphism (SNP) haplotypes in chicken mtDNA for presumption of the origins of chicken meat. We typed five SNPs of the D‐loop region in mtDNA by allele‐specific PCR (AS‐PCR) in 556 hens, that is 233 White Leghorn (WL), 50 Dekalb‐TX35 (D‐TX), 140 Barred Plymouth Rock (BPR) and 133 Rhode Island Red (RIR) kept in the National Institute of Livestock and Grassland Science (NILGS, Tsukuba, Japan). Five haplotypes were observed among those chickens by AS‐PCR. WL, D‐TX, BPR and RIR displayed three, two, one and four SNP haplotypes, respectively. By a combination of the haplotypes by AS‐PCR and the breeds, these chickens were classified into 10 groups. After the D‐loop was sequenced in two chickens from every group (20 individuals), 15 SNP sites (including one insertion) and eight sequence haplotypes were observed. In conclusion, haplotype variation was observed in and among the layer breeds of the NILGS. This study demonstrates that SNP haplotypes in mtDNA should be appropriate for the presumption of the origins of chicken meat.     

Field trials with a bivalent vaccine (HVT and SB-1) against Marek's disease

White leghorn chickens on five farms were given a bivalent Marek's disease (MD) vaccine consisting of turkey herpesvirus (HVT) and SB-1 (a nononcogenic MD virus); other chickens received only HVT. The farms had histories of "vaccination failures," presumably owing to an exceptionally virulent challenge MD virus. The bivalent vaccine uniformly protected chickens better than HVT alone between 12 and 16-20 weeks of age, when serious MD losses occurred. During that period, total mortality in groups given both viruses ranged from 0.39 to 1.26% (mean 0.86%), whereas that in HVT-vaccinated groups not exposed to SB-1 varied from 1.92 to 7.44% (mean 3.43%). Chickens in pens or rows with close contact to those given bivalent vaccine also had low MD mortality rates (0.46-1.06%, mean 0.77%), probably from the spread of SB-1.     

The pathogenicity of three Australian fowl plague viruses for chickens,turkeys and ducks

The pathogenicity of three Australian fowl plague viruses, FPV-1, FPV-2, FPV-3, isolated during a natural outbreak of the disease varied for chickens, turkeys and ducks. FPV-1 and FPV-2 were pathogenic for chickens and turkeys, but not for ducks. However, these viruses were not highly pathogenic as they failed to cause illness or death in all birds that became infected. FPV-3 was non-pathogenic for the three species tested.The viruses spread from infected to in-contact birds, and more readily to ducks than to chickens or turkeys. All chickens and turkeys infected with the fowl plague viruses developed specific serum haemagglutination-inhibiting antibody which persisted for up to 85 days after infection. The titre of this antibody wan ed in six of 16 ducks over an 85-day period and two ducks failed to produce detectable specific HaI antibody despite being infected with the virus.