Molecular characterization of MHC class II region in guinea fowl

1. The MHC class II gene was amplified, cloned and sequenced in guinea fowl. 2. The NumeMHC II sequence of 754 nucleotides included complete exon 1 (91 nt), exon 2 (270 nt), exon 3 (282 nt) and exon 4 (110 nt). 3. The size of β(1) and β(2), domains were 89 and 93 amino acids, respectively in guinea fowl. 4. High amino acid variability (38·2%) was observed within guinea fowl in β(1) domain, while in β(2) domain, amino acid variability (6·3%) was low. 5. Among poultry species, the percent amino acid identity between guinea fowl and chicken, quail, pheasant and duck was 38·8, 42·2, 44·4 and 58·8 in β(1) domain; and 13·8, 17·0, 13·8 and 27·6 in β(2) domain, respectively. 6. Sequence alignment with mammalian and avian MHC showed that many of the conserved features of MHC class II glycoprotein was conserved in guinea fowl. 7. Within-species genetic distances (Poisson correction) based on cumulative amino acid variability in β(1) domain and β(2) domains was 0·141 in guinea fowl. 8. Guinea fowl showed low and similar genetic distances with all the poultry species (0·255-0·268) except duck (0·456). 9. Guinea fowl made separate branch within the major cluster having chicken, quail and pheasant, showing equal distance from these poultry species, whereas duck MHC II clustered separately.     

Evaluation of interspecific hybrids of the chicken, guinea fowl, and Japanese quail for innate resistance to coccidia

Experimental chicken/guinea fowl hybrids, guinea fowl, and chickens were orally inoculated with Eimeria acervulina or E. tenella, which are specific for chickens, or with E. grenieri, which is specific for guinea fowl. No intact oocysts were found in feces within 24 hr of inoculation, suggesting that excystation occurred in the normal and abnormal hosts. No oocysts were found in the feces of hybrids during a 9-day postinoculation period. The guinea fowl passed oocysts of guinea fowl coccidia (E. grenieri) but not those of chicken coccidia, and the chickens passed oocysts of chicken coccidia (E. acervulina and E. tenella) but not those of guinea fowl coccidia. Some asexual development (schizogony) occurred in hybrids inoculated with E. tenella, but sexual development (gametogony) did not. In contrast, quail/chicken hybrids became infected with oocysts of chicken coccidia (E. acervulina, E. tenella, and E. maxima) and quail coccidia (E. bateri) and passed a few oocysts during the normal patent period; control chickens and quails became heavily infected with oocysts of chicken and quail coccidia, respectively.     

Effect of enzyme supplementation on the metabolisable energy content of solvent-extracted rapeseed and sunflower seed meals for chicken, guinea fowl and quail

(1) The nitrogen-corrected apparent metabolisable energy (AME(N)) content of solvent-extracted rapeseed and sunflower seed (un-decorticated) meals in relation to species (chicken, guinea fowl and quail) and dietary addition of feed enzymes (0 or 0.5 g/kg diet) was evaluated by a diet replacement method in a 3 x 2 factorial design. (2) The metabolism trial was conducted at two substitution levels (200 and 400 g/kg diet) of each meal with or without supplementation of commercial enzyme preparation in 6 individuals or 6 groups of cockerels, guinea fowls and quails. (3) The experimental diets were fed for a period of 12 d followed by a 3-d collection period during which total feed consumed and droppings output were quantitatively recorded. (4) The AME(N) values of rapeseed meal for cockerels, guinea fowls and quails were 8.4, 8.7 and 8.8 MJ/kg, respectively, while the corresponding values for sunflower seed meal were 6.1, 6.1 and 6.2 MJ/kg dry matter, without enzyme supplementation. (5) The AME(N) value of rapeseed meal did not improve with enzyme supplementation. However, AME(N) values of sunflower seed meal significantly increased with enzyme supplementation, from 6.1 to 6.5 MJ/kg dry matter. (6) Since AME(N) values of rapeseed meal and sunflower seed meal were similar in chicken, guinea fowl and quail, values reported for chicken could, therefore, be used for guinea fowl and Japanese quail.     

Passage of duck hepatitis virus in cell cultures derived from avian embryos of different species

An egg-attenuated strain of duck hepatitis virus was successfully passaged through cell cultures of avian embryos derived from goose, turkey, guinea fowl, Japanese quail, pheasant and chicken. Two field strains of the virus were passaged in a more limited range of species.     

Mapping of expressed sequence tag markers with a cDNA-amplified fragment length polymorphism method in Japanese quail (Coturnix japonica)

In our previous study, a Kobe‐NIBS Japanese quail (KNQ) linkage map was constructed mainly using amplified fragment length polymorphism (AFLP) markers. In order to compare chicken and quail chromosomes, we developed expressed sequence tag (EST) markers derived from cDNA‐AFLP fragments and localized these markers on the linkage map. Using a total of 128 AFLP primer combinations, 24 polymorphic bands were obtained between a neurofilament‐deficient mutant quail line male and a muscular disorder quail line female, which were the parents of the KNQ resource family. Nine of the 24 markers were mapped by linkage analysis. These markers were mapped to seven linkage groups, namely 1, 2, 3, 6, 8, 15 and 42. A subsequent homology search using chicken genome sequences strongly suggests that these linkage groups correspond with chicken chromosomes 1, 2, 3, 5, 15, 23 and 26.     

Some avian and mammalian hosts of Amblyomma hebraeum and Amblyomma marmoreum (Acari: Ixodidae)

Large numbers of birds, wild mammals and domestic stock from a variety of localities within the Republic of South Africa were examined for infestation with the ixodid ticks Amblyomma hebraeum and Amblyomma marmoreum. Every warthog (Phacochoerus aethiopicus), Burchell's zebra (Equus burchelli), impala (Aepyceros melampus) and kudu (Tragelaphus strepsiceros) from the Kruger National Park in the north-eastern Transvaal Lowveld was infested with A. hebraeum. In the eastern Cape Province every helmeted guinea fowl (Numida meleagris), scrub hare (Lepus saxatilis) and kudu from the Andries Vosloo Kudu Reserve; all but 1 of the 22 domestic cattle examined on the farm "Bucklands"; and all Angora goats plus nearly all Boer goats examined on the farm "Brakhill" were infested with this tick. Most animals examined appeared to be good hosts of the immature stages, and the larger the host species the greater the chances of it harbouring large numbers of adult ticks. The largest animals examined, such as eland, buffalo, giraffe and rhinoceros, harboured very large numbers of adult A. hebraeum. No adult A. marmoreum was recovered from any host. However, 50% or more of helmeted guinea fowl and kudu from the Andries Vosloo Kudu Reserve; helmeted guinea fowl, scrub hares and eland (Taurotragus oryx) from the Mountain Zebra National Park; helmeted guinea fowl, kudu, domestic sheep, goats and cattle on the farm "Bucklands", and caracal (Felis caracal) from the Cradock and Southwell areas of the eastern Cape Province were infested with immature A. marmoreum. In the Bontebok National Park in the south-western Cape Province more than 35% of scrub hares, vaal ribbok (Pelea capreolus) and bontebok (Damaliscus dorcas dorcas) were infested with immature ticks.     

Developmental stages of the blue‐breasted quail (Coturnix chinensis)

Chickens and Japanese quail (Coturnix japonica) have traditionally been the primary avian models in developmental biology research. Recently, the blue‐breasted quail (Coturnix chinesis), the smallest species in the order Galliformes, has been proposed as an excellent candidate model in avian developmental studies owing to its precocious and prolific properties. However, data on the embryonic development of blue‐breasted quail are scarce. Here, we developed a normal developmental series for the blue‐breasted quail based on developmental features. The blue‐breasted quail embryos take 17 days to reach the hatching period at 37.7°C. We documented specific periods of incubation in which significant development occurred, and created a 39‐stage developmental series. The developmental series for the blue‐breasted quail was almost identical to that for chickens and Japanese quail in the earlier stages of development (stages 1–16). Our staging series is especially useful at later stages of development (stages 34–39) of blue‐breasted quail embryos as a major criterion of staging in this phase of development was the weight of embryos and the length of third toes.     

Comparative apparent metabolisable energy values of high, medium and low tannin varieties of sorghum in cockerel, guinea fowl and quail

1. Nitrogen-corrected apparent metabolisable energy values (AMEN) of three varieties of sorghum (white-low tannin, brown-medium tannin and red-high tannin) were measured in three species of poultry (cockerel, guinea fowl and Japanese quail) by a practical diet replacement (total collection) method. 2. Each variety of sorghum was tested at two concentrations (200 and 400 g/kg of reference diet) in 6 replications with one cockerel or guinea fowl or two quails per replication. The duration of the trial included a 10 d preliminary feeding period (on conventional grower diet) followed by a 12 d adaptation period (on reference and test diets) and a 3 d balance period (with recording of feed intake and excreta output). 3. The calculated AMEN values of different sorghum varieties were: white--12.9, 12.8 and 12.7; brown--12.7, 12.3 and 12.6; and red--11.4, 11.1 and 11.6 MJ/kg for cockerels, guinea fowls and quails, respectively. The mean AMEN value of red sorghum (11.3 MJ/kg) was significantly lower than those of brown (12.5 MJ/kg) or white sorghum (12.8 MJ/kg). A negative correlation was observed between tannin concentration and AMEN. 4. There was no significant difference in the AMEN values of white, brown and red sorghum varieties to the different poultry species. AMEN values of sorghum for the cockerel could, therefore, be used in practical feed formulation for guinea fowl and quail.     

Complete nucleotide sequence of mitochondrial DNA in White Leghorn and White Plymouth Rock chickens

Among the chicken breeds, White Leghorn (WL) and White Plymouth Rock (WR) are major breeds and have different history in their establishments. Whole mitochondrial DNA of the breeds were sequenced in order to elucidate the genetic relationship between the breeds. The lengths of the two WL and two WR mitochondrial DNA were found to be 16 788 and 16 785 base pairs, respectively. When the DNA sequences were compared, the similarity was found to be 99.96% (six nucleotide differences). In addition, the present study conformed the existence of an extra nucleotide ‘C’ in NADH dehydrogenase subunit 3 (ND3) of the chicken mitochondrial DNA, which has been consistently observed in Galliformes.     

珍珠鸡源IBDV的分离鉴定及其卵黄抗体的制备

从一群发病的珍珠鸡中分离到1株病毒,经鸡胚接种、琼脂扩散试验、PCR快速检测试验和病毒基因序列的测定,证实了所分离的病毒为IBDV。将该病毒制成灭活疫苗,接种开产的蛋鸡制备卵黄抗体。结果显示该卵黄抗体经免疫扩散试验测定抗体效价可达1∶256,符合无菌标准,对雏鸡也没有任何不良反应。临床试验表明:该卵黄抗体对该养殖场珍珠鸡的传染性法氏囊病的治疗有效率达91%、预防有效率达100%。     

Characterization and expression of non‐polymorphic liver expressed antimicrobial peptide 2: LEAP‐2 in the Japanese quail,Coturnix japonica

Liver‐expressed antimicrobial peptide 2 (LEAP‐2) is a cationic peptide that plays an important role in innate immunity for host defense. The aim of this study was to characterize the LEAP‐2 gene in the Japanese quail (Coturnix japonica). Japanese quail LEAP‐2 (CjLEAP‐2) was identified from the Japanese quail draft genome database by a local BLAST analysis using chicken LEAP‐2 (GgLEAP‐2). The exon‐intron structure of CjLEAP‐2, analyzed from three quails, is composed of three exons, as is the chicken LEAP‐2 homolog (GgLEAP‐2). An analysis of the coding sequence revealed that CjLEAP‐2 is 231 bp long, like GgLEAP‐2, and 93% identical to GgLEAP‐2 at the nucleic acid level. The predicted amino acid sequence of CjLEAP‐2 contained the liver‐expressed antimicrobial peptide 2‐precursor domain and four cysteine residues characteristic of the LEAP‐2 protein. The amino acid sequence of the mature peptide of CjLEAP‐2 was 100% identical to that of GgLEAP‐2. We confirmed that CjLEAP‐2 was transcribed in at least seven tissues, including the digestive system. Additionally, the mature peptide region of CjLEAP‐2 exhibited no polymorphisms in 99 quails from six strains. Taken together, these findings indicate that CjLEAP‐2 is non‐polymorphic and therefore, it likely plays an important role in the innate immunity of quail as it does in chicken.     

珍珠鸡蛋与海兰褐鸡蛋蛋品质及营养成分比较分析

珍珠鸡是从国外引进,经人工驯养育成的一个特色品种,其蛋用性能获得了显著进展,为加快蛋产品的开发力度,增加其附加价值,对珍珠鸡蛋和海兰褐鸡蛋蛋品质及营养成分进行比较分析。试验随机抽取新鲜珍珠鸡蛋、海兰褐鸡蛋各100枚,测定其蛋外、内在品质及主要营养成分,采用Duncan新复极差法、皮尔逊相关系数进行比较分析。研究表明,在外部品质方面,海兰褐鸡蛋蛋重高于珍珠鸡蛋15.54g（35.61%）,有极显著差异（P〈0.01）;珍珠鸡蛋蛋壳厚度比海兰褐鸡蛋的蛋壳厚0.14mm（40%）,差异极显著（P〈0.01）;由于蛋重的差异,两种鸡蛋蛋黄比例、蛋白比例、蛋壳比例也均有极显著差异（P〈0.01）。在内在品质方面,两种鸡蛋蛋清pH、蛋黄高度、蛋黄系数均无明显差异;珍珠鸡蛋浓蛋白高度比海兰褐鸡蛋的低3.02mm,有极显著差异（P〈0.01）;浓蛋白系数比海兰褐的高0.87,有显著差异（P〈0.05）;珍珠鸡蛋蛋黄颜色比海兰褐鸡蛋的颜色大一个罗氏比色值,有显著差异（P〈0.05）。在营养成分上,以g/枚为单位,珍珠鸡蛋的主要营养成分除胆固醇高于海兰褐鸡蛋外,其他均低于海兰褐鸡蛋。以g/100g为单位,珍珠鸡蛋水分含量明显低于海兰褐鸡蛋,相差12.62（20.21%）,差异极显著（P〈0.01）,脂肪含量也低于海兰褐鸡蛋,差异显著（P〈0.05）,相差1.34（4.4%）,其它营养成分均高于海兰褐鸡蛋。珍珠鸡蛋是较海兰褐鸡蛋小、壳厚、水分含量低、营养丰富的浓缩型蛋,为珍珠鸡蛋的蛋品开发创造了条件。     

Organic matrix composition and ultrastructure of eggshell: a comparative study.

1. The avian eggshell is a biomineralised composite ceramic consisting of calcium carbonate embedded in an organic matrix. Matrix components are supposed to be involved in the control of mineralisation, crystallographic texture and biomechanical properties of eggshell. 2. The structure and eggshell matrix composition of various domesticated bird species were compared to gain insight into the universality of the eggshell mineralisation process. 3. The SDS-PAGE profiles of soluble eggshell matrix were specific within groups of birds (a: laying hen, breeder hen, quail, pheasant and possibly turkey; b: guinea fowl; c: duck and goose) but some of the protein bands were common to all groups. 4. Analogies between species were confirmed by Western blotting using hen protein antibodies. Ovocleidin-17 (OC-17) and ovalbumin were revealed in all species (except quail for OC-17). Lysozyme was present only in hen eggshell. Another egg white protein: ovotransferrin showed a positive signal in hens, turkey and quail. Osteopontin was observed in laying and breeder hens and quail. 5. Different proteoglycans were localised to discrete regions within the eggshell. Dermatan sulphate was observed within the matrix of the calcified shell of all species except quail which contained chondroitin-6-sulfate. Keratan sulphate was observed in mammillary bodies of breeder and laying hen, quail, pheasant and turkey while chondroitin sulphate was also present in guinea fowl and duck. 6. The general structural organisation of the different avian eggshells was similar but specific differences were observed in the ultrastructure of the mammillary layer. Species of the same taxonomic family could be grouped according to their structural analogies: breeder hen, turkey and pheasant resembled that of the domestic fowl. Guinea fowl was unique. Goose and duck were quite similar with large and confluent mammillary bodies. 7. Some matrix components are therefore common to eggshells of various species but more information is needed to relate differences in matrix composition between taxonomic groups with differences in ultrastructure.     

利用SRAP标记分析鸡、鹌鹑及其属间杂交种的群体遗传结构

为了研究鸡、鹌鹑及其属间杂交种基因组的遗传变异程度及其关系,试验采用相关系列扩增多态性(SRAP)标记对鸡、鹌鹑及其属间杂交种进行了DNA多态性分析。结果表明:选用的7对引物组合共产生31个多态性条带,平均每个引物组合产生4.4个多态性条带,每个引物组合产生多态性条带的比例为22.3%;从聚类分析图可知,母本(鹌鹑)和属间杂交种的相似系数远高于属间杂交种与父本(鸡);生理生态学观察结果也显示属间杂交种和母本在外形、习性上更为相似。     

Egg-laying pattern of the semi-domesticated helmeted guinea fowl (Numida meleagris galeata)

1. Studies on the egg laying pattern of the semi-domesticated helmeted guinea fowl showed that the birds laid eggs between 06.00 and 20.00 h local time (05.00 to 19.00 h GMT). 2. More (67.9%) were laid in the evenings (15.00 to 20.00 h) than at any other period. 3. There were two distinct periods in the reproductive cycle: a breeding season which began in April and ended in September, and a resting or non-breeding period between October and March. 4. Sequence length was predominantly of 4 eggs and July was the month of peak egg production.     

Genetic studies on primary antibody response to sheep erythrocytes in guinea fowl

1. The primary antibody response to sheep erythrocytes was determined by haemagglutination test in guinea fowl. The effects of various genetic and non‐genetic factors on immune response to sheep RBCs in guinea fowl were also estimated.

2. The immune response to sheep RBCs was normally distributed in guinea fowl with mean titre at 1.534 ± 0.014.

3. In guinea fowl, effects on titre values of sire and variety (feather colour) were significant whereas sex and sex × variety interaction effects were non‐significant.

4. The estimate of heritability for immune response to sheep RBCs in guinea fowl was 0.35 ±0.17.     

Identification of novel genetic markers and evaluation of genetic structure in a population of Japanese crested ibis

Japanese population of the Japanese crested ibis Nipponia nippon was founded by five individuals gifted from the People's Republic of China. In order to exactly evaluate genetic structure, we first performed development of novel genetic makers using 89 microsatellite primer pairs of related species for cross‐amplification. Of these, only three primer pairs were useful for the genetic markers. Additionally, we sequenced allelic PCR products of these three markers together with 10 markers previously identified. Most markers showed typical microsatellite repeat units, but two markers were not simple microsatellites. Moreover, over half of the markers did not have the same repeat units as those of the original species. These results suggested that development of novel genetic markers in this population by cross‐amplification is not efficient, partly because of low genetic diversity. Furthermore, the cluster analysis by STRUCTURE program using 17 markers showed that the five founders were divided into two clusters. However, the genetic relationships among the founders indicated by the clustering seemed to be questionable, because the analysis relied largely on a small number of triallelic markers, in spite of the addition of the three useful markers. Therefore, more efficient methods for identifying large numbers of single nucleotide polymorphisms are desirable.     

Aspects of the functional morphology of the ductus epididymidis in domestic anseriform and galliform birds

The structure of the ductus epididymidis, and its capacity to take up luminal particulate material (India ink) were studied, using scanning and transmission electron microscopy, in the domestic fowl, turkey and Japanese quail (Order Galliformes) and drake (Order Anseriformes). The epithelium appeared contractile and stable structurally, especially in the drake in which, among other cytoskeletal features, abundant perinuclear intermediate filaments radiated peripherally in the principal, nonciliated (Type III) cell. The Golgi complex was well developed, except in the turkey in which it was extremely inconspicuous. Sparsely granulated profiles were the main component of the elaborate endoplasmic reticulum, which was best developed in the drake and least in the turkey. The cell in the turkey was uniquely laden with large aggregates of lipid droplets. Generally, characteristics of a cell active in the synthesis and merocrine secretion of proteinaceous material were evident. There was no evidence of apocrine-like secretion in well-fixed tissues. The cell lacked the capacity to take up luminal India ink particles, and displayed very poorly developed endocytic structural features, which are also probably only selectively and minimally absorptive. Ultrastructural features varied between and within the orders studied, necessitating further studies.