Characterization of the cathelicidin cluster in the Japanese quail (Coturnix japonica)

The Japanese quail has several advantages as a low‐fat meat bird with high immunity against diseases. Cathelicidins (CATHs) are antimicrobial peptides that play an important role in innate immunity. The aim of this study was to characterize the CATH cluster in the Japanese quail (Coturnix japonica). The Japanese quail CATH (CjCATH) cluster, contains four CATH genes, as in the chicken. The coding sequences of CjCATHs exhibited >85.3% identity to chicken CATHs. The predicted amino acid sequences of the four CjCATH genes contained the cathelin‐like domain characteristic of CATH proteins. Polymorphisms were detected in the open reading frames (ORFs) of all CjCATH sequences. Two amino acid substitutions were observed in the antimicrobial region of the mature peptide of CjCATH2, and predicted to influence peptide function. CjCATH1 is expressed in lung, heart, bone marrow and bursa of Fabricius (BF). CjCATH2 is expressed in bone marrow. CjCATH3 is expressed in lung, heart, bone marrow, BF, tongue and duodenum. CjCATHB1 is expressed in bone marrow and BF. This study is the first to characterize CATH genes in the Japanese quail, and identifies novel antimicrobial peptide sequences belonging to the cathelicidin family, which may play a role in immunity in this species.     

Analysis of 46-kDa protein in the perivitelline membrane of Japanese quail (Coturnix japonica)

The extracellular matrix surrounding the oocyte before ovulation is called the perivitelline membrane (PL) in avian species. The PL is constructed with two major glycoproteins, ZPC and ZP1, which are synthesized in the ovarian granulosa cells and the liver, respectively. Although the properties of the major components in the PL have been examined, knowledge about the nature of its minor constituents is lacking. In this study we focused on PL protein, which migrates at 46‐kDa in the gel of SDS‐PAGE. N‐terminal sequence analysis demonstrated that the 46‐kDa protein is the C‐terminal fragment of ZP1. Analysis of lysylendopeptidase digests or cyanogens bromide‐degraded fragments of ZP1 confirmed this postulate. Western blot analysis using antiserum against 46‐kDa protein indicated the absence of 46‐kDa protein in the serum. Moreover, small immunoreactive bands, thought to be cleaved fragments of ZP1, were detected in the PL lysate by western blot analysis using antiserum against the N‐terminal peptide of ZP1. These results indicated that the N‐terminal proteolytic processing of ZP1 might take place after the arrival of ZP1 at the ovary, and the resulting product, 46‐kDa protein, is incorporated into the PL.     

Effects of probiotic Bacillus cereus toyoi on abdominal fat accumulation in the Japanese quail (Coturnix japonica)

Effects of a commercial probiotic, Bacillus cereus toyoi on abdominal fat accumulation in the Japanese quail fed either a commercial control diet (crude protein (CP), 23.5%; metabolizable energy (ME), 11.7MJ/kg) or a high‐energy diet (CP, 23.0%; ME, 13.8MJ/kg) were investigated. Four‐week‐old male birds (n = 108) were divided into four diet groups: control diet, control with probiotic, high‐energy diet and high‐energy diet with probiotic. At 8 weeks and 12 weeks of age (4 and 8 weeks of probiotic supplementation period, respectively), abdominal fat, muscles and the liver were weighed. The weight of total fat and protein in the muscles and liver were also determined. A rectal temperature was recorded weekly. Bodyweight and feed intake in both diets were not affected by probiotic supplementation. At 8 weeks of age, birds fed the control diet with probiotic had significantly less abdominal fat than those fed without the probiotic, and a similar tendency was seen for both diet groups at 12 weeks of age. The decrease in abdominal fat coincided with an increase in rectal temperature. These results suggest that greater metabolizable energy was consumed through elevation of heat production by supplementation of B. cereus toyoi, with hardly any energy stored as body fat. Probiotic supplementation appears to induce greater muscle weight, higher protein and lower lipid content in muscles by 8 weeks of age, and lower lipid content in muscles and greater liver weight by 12 weeks of age, in the Japanese quail.     

Primary analysis of DNA polymorphisms in the TRIM region (MHC subregion) of the Japanese quail,Coturnix japonica

Based on sequences of two cosmid clones from Japanese quail (Coturnix japonica, Coja), we confirmed that the syntenic cluster, GNB2L1～BTN1～BTN2, is located in the quail TRIM subregion of the quail major histocompatibility complex (MHC Coja) region. These cosmids also included four CjBG loci and one CjLEC locus; therefore, the quail TRIM subregion was thought to be adjacent to the BG/LEC subregion. We then identified three polymorphic markers – CjHEP21, CjTRIM39.2 and CjBTN2 – in the TRIM subregion that may be useful for the functional analysis of the MHC‐Coja region. We examined MHC‐Coja sequences from 321 individual quails sampled from 11 inbred strains, and we found eight alleles for each of the three genes – CjHEP21, CjTRIM39.2 and CjBTN2. These polymorphisms represent the first avian DNA markers in the TRIM subregion. Additionally, we discovered a quail‐specific VNTR (variable number of long tandem repeats, 133–137 bp) in intron 7 of CjBTN2. We identified 25 haplotypes in the sample of 321 quail; these haplotypes comprised combinations of all 24 alleles of the three polymorphic genes. We suggest that there are two recombination hotspots, one between each pair of adjacent loci. All strains, except AMRP, contained multiple haplotypes; the AMRP strain contained a single, apparently fixed haplotype.     

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.     

A Growth Performance and Nonlinear Growth Curve Functions of Large- and Normal-Sized Japanese Quail (Coturnix japonica)

This study aimed to evaluate the differences between the growth patterns of large- and normal-sized Japanese quail strains and their F₁ progeny, by fitting their growth parameter values to five nonlinear regression growth models (Weibull, Logistic, Gompertz, Richards, and Brody). The Richards model presented the best fit for both sexes of the large-sized quail strain, whereas the Gompertz model presented the best fit for both sexes of the normal-sized quail strain, based on goodness-of-fit criteria (higher adjusted R² and lower Akaike and Bayesian information criteria). Both sexes of F₁ birds derived from the cross between normal-sized females and large-sized males were best fitted by the Richards model. In contrast, growth parameters of the F₁ birds derived from the cross between large-sized females and normal-sized males were best fitted to the Gompertz model. The data could be fitted nearly as well to the Weibull and Logistic models as to the Richards and Gompertz models. The Brody model presented the poorest fit for the growth parameter values. The results indicated that the Richards and Gompertz models could best describe the growth characteristics of both large- and normal-sized quails. Moreover, the observed growth pattern of the F₁ birds was likely inherited from the male parental strain. To the best of our knowledge, this is the first study comparing the growth curves of the reciprocal F₁ generations with their parental strains in quails.     

Histological and histochemical studies on the oviduct microcirculation of the laying Japanese quail (Coturnix japonica)

The health of oviduct circulation is very important for normal egg production. The present work describes the histology and histochemistry of vessels of different types on the oviduct of quail. Arteria, venae and lymphatics of the oviduct were described clearly. In addition, special vascular elements were observed presenting circular, longitudinal and oblique smooth muscle fibres on the blood vessel wall. Glomus cells were observed at typical glomus vessels, atypical glomus vessels and on the wall of some of arteria and arteriolae. Also, spirally oriented blood vessels were observed. Dark brown pigment was detected in the wall of arteria, venae, lymphatics and the special types of blood vessels.     

Hypocholesterolemic effect of karaya saponin in Japanese laying quails (Coturnix coturnix japonica)

The effect of karaya saponin supplementation on the serum and egg yolk cholesterol and fatty acid composition in egg yolk were investigated in Japanese quails. A total of 80 Japanese quails aged 5 weeks were equally divided into four groups of 20. Four levels (0, 25, 50 and 75 mg/kg feed) of karaya saponin were included in the basal diet and experiment was lasted for 6 weeks. The cholesterol fractions in the egg yolk and serum were measured by enzymatic assay, and the fatty acid composition in egg yolk was determined by gas chromatography. The results revealed that the supplementation of 75 mg/kg karaya saponin significantly reduced (p < 0.05) cholesterol and triglycerides concentration in serum and egg yolk. High-density lipoprotein-cholesterol was increased, low-density lipoprotein-cholesterol and the atherogenic index were decreased (p < 0.05) by the dietary supplementations. Hepatic cholesterol was reduced (p < 0.05) by the 25 mg/kg karaya saponin. A higher degree of yolk colour was improved (p < 0.05) when 75 mg/kg saponin was supplemented in the diet. The concentration of polyunsaturated fatty acids (PUFA) in egg yolk was increased (p < 0.05) in a dose-dependent manner in quails fed the supplemented diet than the control diet. The ratio of PUFA to saturated fatty acids in egg yolk was improved (p < 0.05) by 75 mg/kg feed karaya saponin-supplemented diet. Therefore, the dietary supplementation of 75 mg/kg karaya saponin may be a feasible means of producing quail eggs with lower cholesterol and higher PUFA content for health conscious consumers.     

CXCL14-like immunoreactivity in somatostatin-producing cells of the Japanese quail (Coturnix japonica) pancreas

This study examines chemokine CXCL14-like peptide distribution in the Japanese quail (Coturnix japonica) pancreas using a specific anti-human CXCL14 antibody. CXCL14-immunoreactive cells were observed in the pancreatic islet peripheral region. The staining was abolished after pre-absorbing the antibody with recombinant human CXCL14. CXCL14-immunoreactive cells were immuno-positive for somatostatin, but not glucagon and insulin. CXCL14 secreted from somatostatin-producing cells might participate in insulin secretion modulation together with somatostatin. In addition, CXCL14 might participate in glucose homeostasis in co-operation with somatostatin and growth hormone.     

Influence of animal age on body concentrations of minerals in Japanese quail (Coturnix japonica)

Mathematical modelling of the relationships between mineral inputs and outputs would enable the prediction of mineral requirements of poultry under a wide range of conditions. To establish the feasibility of possible modelling of mineral requirements, the current study aimed to describe the individual mineral concentrations of whole bodies of quail over the life cycle from hatching to 70 days of age. Quail were reared indoors without any restrictions that could limit growth. Sampling of birds (n = 6–18) was carried out at 0, 3, 7, 14, 21, 35, 49 and 70 days after hatching. Freeze‐dried samples of whole bodies (digestive contents removed) were analysed for ash, and macrominerals (calcium, magnesium, phosphorus, potassium, sodium) and microminerals (copper, iron, manganese, nickel, selenium, zinc). Ash concentration followed a curvilinear trend, with a maximum of 101.7 g/kg dry matter at 32.77 days. Individual mineral concentrations, expressed as a proportion of ash, were fluctuating over time, with the most prominent changes at 3 days and again at either 14 or 21 days. Dissimilar patterns in individual mineral concentrations resulted that ratios between minerals followed inconsistent patterns over time. Although mineral contents in absolute quantities can be described through modelling over the entire life cycle of the bird, it can be concluded that variable concentrations of individual minerals could complicate further model development.     

Basic characterization of avian β‐defensin genes in the Japanese quail,Coturnix japonica

In this study, we identified a cluster of 14 avian β‐defensins (AvBD; approximately 66 kbp) in the Japanese quail, Coturnix japonica. Except for AvBD12 (CjAvBD12) and ‐13, the CjAvBDs coding sequences exhibited greater than 78.0% similarity to the respective orthologous chicken AvBD genes (GgAvBD). The putative amino acid sequence encoded by each CjAvBD contained six cysteine residues and the GXC (X1‐2) motif considered essential for the β‐defensin family. Each CjAvBDs also formed a sub‐group with the respective orthologous genes of various bird species in a phylogenetic tree analysis. Synteny between the CjAvBD cluster and GgAvBD cluster was confirmed. The CjAvBD cluster was mapped on the long‐arm end of chromosome 3 by linkage analysis based on single nucleotide polymorphisms (SNPs) of CjAvBD1 and CjAvBD12 (approximately 46kbp), as well as GgAvBD cluster. We also confirmed that CjAvBD1, ‐4, ‐5, ‐9, and ‐10 are transcribed in 20 tissues, including immune and digestive tissues. However, our experimental data indicated that the CjAvBD cluster lacks the AvBD3 and ‐7 loci, whereas the CjAvBD101α, ‐101β, and ‐101θ loci arose from gene duplication of the AvBD6 orthologous locus in the CjAvBD cluster after differentiation between Coturnix ‐ Gallus.     

Intestinal mucosa develops in a sex-dependent manner in Japanese quail (Coturnix japonica) fed Saccharomyces cerevisiae

ABSTRACT

1. The aim of study was to investigate whether the impact of the yeast Saccharomyces cerevisiae on the histological structure of the intestine, innervation of the small intestine wall, and basal biochemical serum parameters in Japanese quail was sex dependent.

2. One-day-old healthy male and female Japanese quail were fed either a basal diet containing no yeast (control group) or the basal diet plus 1.5% (15 g/kg of diet) of yeast (S. cerevisiae inactivated by drying). Samples from the duodenum and jejunum were taken from each bird at the age of 42 days. Blood samples were collected at this age and the concentrations of glucose, total protein, creatinine, uric acid, lipid profile (total cholesterol, low density lipoproteins (LDL), high density lipoproteins (HDL) and triacylglycerols (TG)), alanine aminotransferase (ALAT), aspartate aminotransferase (AspAT), lactate dehydrogenase (LDH), amylase (AMY), calcium, phosphorus and iron were determined.

3. Female quail fed diets supplemented with yeast had significantly lower total cholesterol and amylase activity than the control females. The concentration of HDL was higher in the male quail than in the females, irrespective of the treatment. An opposite effect was observed in LDL. The diet treatments influenced the activity of AspAT, which was significantly less in the male quail fed diets with 1.5% yeast.

4. Supplementation with S. cerevisiae increased the myenteron, submucosa and mucosa thickness, villus length and thickness and size of absorptive surface, while the number of villi and enterocytes were decreased in the duodenum in males. Female quail showed an increased absorptive surface in the jejunum. The Meissner (submucosal) plexuses were influenced by the feeding and sex to a greater extent than the Auerbach plexus (in the muscularis propria).

5. The results demonstrated that S. cerevisiae (1.5%) in the diet caused significant positive effects in Japanese quail, exerting an effect on the morphology of the small intestine in a sex-dependent manner.     

The effect of different high‐fat diets on erythrocyte osmotic fragility,growth performance and serum lipid concentrations in male,Japanese quail (Coturnix coturnix japonica)

Poultry diets are formulated with additional animal fat or vegetable oils to improve growth rate and feed conversion efficiency. High‐fat diet feeding in rats and fish has been shown to result in alterations in the phospholipid composition and cholesterol content of the erythrocyte membrane, in turn affecting erythrocyte osmotic fragility. In contrast, the few studies performed using high‐fat diet feeding in avian species show no changes in erythrocyte osmotic fragility. This study made use of the Japanese quail as no data exists on investigation of this species with respect to high‐fat diet feeding and erythrocyte osmotic fragility. Fifty‐seven male quail were randomly divided into six groups and fed either a standard diet (commercial poultry feed) or one of five high‐fat diets (commercial poultry feed with 22% of either coconut oil, lard, palm oil, soya bean oil or sunflower oil on a weight/weight basis) for 12 weeks. All birds on the high‐fat diets were significantly heavier (p < 0.05) after the 12‐week feeding period, than when commencing the dietary intervention. Serum triglyceride concentrations of birds in all high‐fat diet groups were significantly lower (p < 0.05) than birds in the standard diet group, whereas only birds in the palm oil group had significantly lower (p < 0.05) serum cholesterol concentrations compared to the standard diet group. Fragiligrams of erythrocytes from birds in the various dietary groups were similar. High‐fat diet feeding with different types of additional fat did not affect the osmotic fragility of the quail erythrocytes. Feeding quail high‐energy diets of varying degrees of fatty acid saturation was well tolerated and did not seem to affect the overall health status of the birds. Resistance of avian erythrocytes to modification by excess dietary fat may be a general characteristic of avian erythrocytes.     

Haematology of the Japanese quail (Coturnix coturnix japonica)

Blood samples from 30 2‐week‐old unsexed chicks, 25 adult males, 30 non‐laying hens and 19 laying hens were examined to characterise haematological norms in Japanese quail (Coturnix coturnix japonica) under various physiological conditions such as age, sex and laying. Young quail had significantly lower erythrocyte counts, packed cell volumes, haemoglobin content, mean corpuscular haemoglobin concentrations, numbers of thrombocyte::, percentages of lymphocytes and plasma protein levels than did the adult males and the non‐laying hens. The total leucocyte count of the young birds was not significantly different from that of the males and just different from that of the non‐laying hens. The young birds also showed a higher mean corpuscular volume, mean corpuscular haemoglobin and percentages of heterophils and monocytes in comparison with the adult males and the non‐laying hens. A comparison between males and females showed that males had a higher erythrocyte count, packed cell volume and haemoglobin content and a lower concentration of plasma proteins than the adult females. Excepting for an elevated relative count of eosinophils, no influence attributable to laying was observed.     

Cellulitis in Japanese quail (Coturnix coturnix japonica)

A case of cellulitis was observed in Japanese quail (Coturnix coturnix japonica) reared for commercial meat production. This condition in Japanese quail has not been reported in the literature. This incident was the first, and to date only, occurrence of cellulitis in this processing plant. The cellulitis lesions were localized to the subcutis overlying the breast and inner thigh. Carcasses of processed birds and live birds from the affected farm were presented to the Poultry Diagnostic and Research Center, University of Georgia. Escherichia coli was cultured from the lesion. The affected live birds displayed lameness and had osteomyelitis. Pasteurella multocida serotype 3,4 was cultured from the liver and bone marrow of affected birds. Approximately 4.61% of the processed carcasses from the flock were condemned because of cellulitis. This represented a 10fold increase from the typical condemnation rate. Further investigation revealed birds were placed in higher than normal density; therefore, we theorize that the concurrent pasteurellosis and increased placement density resulted in the cellulitis condition.     

A simple PCR method for sexing Japanese quail Coturnix japonica at hatching

1. The quail is a potentially important avian model for molecular studies; a major drawback is the inability to sex visually before 3 weeks of age. Molecular sexing is therefore an absolute requirement when animals are sampled before that age.

2. A low-cost method using common laboratory equipment based on Allele-Specific Multiplex-Polymerase Chain Reaction was developed to undertake reliable molecular identification of the sex of Coturnix japonica directly at hatching.

3. This simple method works with down feathers collected from behind the neck of the newly hatched quail and includes internal controls during the PCR to limit risks of error. Males and females can be discriminated on the basis of the presence of one or two amplicons, respectively.