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.     

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.     

Changes of histochemical profiles of myofibers in pectoralis and supracoracoideus fasciculi induced by breeding for large or small body size in Japanese quails

The histochemical profiles of myofibers in Musculus pectoralis (PT) and M. supracoracoideus (SC) fasciculi were compared among Japanese quail strains with large, normal and small body sizes. In male and female adults, both the PT and SC muscles had attained a 2.5–2.7-fold weight gain in the large strain and conversely a 0.43–0.50-fold change in the small strain relative to those of the normal size. The muscles were composed of fasciculi with a central cluster of type IIA fibers surrounded by a peripheral layer of type IIB fibers. In the large strain, the cross sectional area (CSA) of the fasciculus and CSA of the fibers in each type were significantly enlarged compared with those in the normal size, with the exception of the fasciculus in the deep region of the male PT muscle. The hypertrophied type IIA fibers in the large strain showed considerable variation in nicotinamide adenine dinucleotide dehydrogenase activity, some of which might represent a transitional form into type IIB fibers. In the small strain, the fasciculus CSA did not significantly differ from that of the normal size except for the PT surface region of the male. However, fiber atrophy was observed in type IIB fibers of the PT surface region in both sexes, and type IIA fibers of the PT deep region and SC muscle in the small strain male quails. The relative fiber type composition of a fasciculus in each region showed only a slight change across the strains. These results indicate that breast muscle hypertrophy in the large strain could be based mainly on fasciculus and fiber hypertrophy, but muscle atrophy in the small strain is not induced by fasciculus and fiber atrophy.     

Reproductive responses and nitric oxide activity in Japanese quail, Coturnix coturnix japonica, are altered by specific phase relationships of neural (serotonergic and dopaminergic) oscillations and pineal function

The present study investigates the effects of temporal synergism of 5-hydroxytryptophan (5-HTP) and l-dihydroxyphenylalanine (l-DOPA) and pineal function (pinealectomy and melatonin administration) on gonadal response and nitric oxide (NO) activity in Japanese quail. Three-week-old male quails were divided into 9 groups, as follows: (I) control (treated with normal saline); (II) 8-h and (III) 12-h (receiving l-DOPA after the administration of 5-HTP [5 mg/100 g body weight] at the interval of 8 and 12 h, respectively, over a period of 13 days); (IV) SO (sham operated); (V) Px (pinealectomized); (VI) Veh (vehicle treated control); (VII) Mel (melatonin 25 μg/100 g body weight for 45 days); (VIII) 12-h + Veh; and (IX) 12-h + Mel (same as in (III) but further treated with the vehicle or melatonin, respectively, for 32 days). Body weight and cloacal gland volume were recorded weekly, whereas spermatogenesis, plasma testosterone concentration, and total nitrite and nitrate concentration in plasma, hypothalamus, and testes were measured at the termination of the study. Results indicate that compared to the controls, 8-h, Mel, and 12-h + Mel treatments suppressed whereas 12-h and Px stimulated the reproductive system. Further, a significant increase in plasma testosterone, total nitrite, and nitrate in the 12-h and Px groups and a decrease in these concentrations in 8-h, Mel and 12-h + Mel quail were observed. It is evident that 5-HTP and l-DOPA given at the interval of 8 h, as well as melatonin, are potent inhibitors of gonadal development, and the gonado stimulatory effect of the 12-h temporal relationship of the neurotransmitter precursors is masked by the inhibitory effect of melatonin. It is concluded that modulation of the daily phase relationship of neural oscillations and pineal function alters gonadal function and NO activity. Further, NO exhibits a parallel relationship with gonadal function in Japanese quail; this relationship appears to be mediated through NO activity, suggesting a causal relationship, although the mechanism between the 2 systems remains a topic of discussion.     

Histopathologic effect of in ovo exposure to methotrexate at early embryonic stage on optic tectum of Japanese quail (Coturnix japonica)

The optic tectum of Japanese quail embryos with in ovo exposure to methotrexate 100 ng/g egg on embryonic day 4 was examined from 3 to 24 hour after treatment. At 9 hour after methotrexate exposure, several apoptotic neuroepithelial cells appeared in the ventricular zone of the optic tectum; these increased in number and were diffusely distributed throughout all layers of the ventricular zone of the optic tectum at 12 hour. At 24 hour, neuroepithelial cells in the ventricular zone of the optic tectum were eliminated and showed sparse cell density. Throughout the experimental period, proliferation of neuroepithelial cells in the ventricular zone of the optic tectum of methotrexate-treated embryos was inhibited. These results suggest that neuroepithelial cells in the ventricular zone of the optic tectum in Japanese quail embryos can be affected by folic acid antimetabolites, methotrexate, at an early embryonic stage.     

The effects of transportation stress on Japanese quail (Coturnix Coturnix japonica) fed corn‐based diet in comparison with wheat‐based diet supplemented with xylanase and phytase

Harvesting, handling and transporting quails to the slaughterhouses, other farms and laboratories might covertly reduce their welfare. The aim of this study was to evaluate the effects of two major sources of energy in poultry nutrition on reducing transportation stress in Japanese quail (Coturnix Coturnix japonica). Male quails (n = 60) were divided into two groups. The first group was fed corn‐based diet, and the second was fed wheat‐based diet supplemented with xylanase and phytase. At the end of the experiment (day 35), quails were subjected to 80 km of transportation. Immediately on arrival and after 24 h, heterophil counts, lymphocyte counts and H:L ratios were measured. On arrival, H counts were lower, L counts were higher, and H:L ratios were lower for corn‐fed group. After 24 h, wheat‐fed group showed lower increment of H counts, greater increment of L counts and also decrement of H:L ratios rather than corn‐fed group which showed increment of H:L ratios. However, these ratios were still lower in corn‐fed group. Results indicate that corn‐based diets can help Japanese quail to better resist transportation stress, although it seems that feeding wheat‐based diets supplemented with xylanase and phytase could have positive effects for coping better with stress after journeys.     

Basic characterization of 90 kDa heat shock protein genes HSP90AA1, HSP90AB1, HSP90B1 and TRAP1 expressed in Japanese quail (Coturnix japonica)

In the current study, we describe four novel members of the 90 kDa heat shock protein (HSP90) family expressed in Japanese quail, Coturnix japonica. The coding regions of the genes, CjHSP90AA1, CjHSP90AB1, CjHSP90B1 and CjTRAP1, exhibited more than 94% similarity to their related genes in chicken. The putative proteins encoded by these quail genes contained motifs considered essential for HSP90 gene function. In addition, the predicted proteins were more similar to HSP90AA1, HSP90AB1, HSP90B1 and TRAP1 proteins expressed in vertebrates than they were to other members of the HSP90 family. Exon numbers of CjHSP90AA1 (11), CjHSP90AB1 (12) or CjTRAP1 (18) are the same as the chicken and mammalian orthologs. Furthermore, gene order in the regions surrounding CjHSP90AB1 and CjTRAP1 has been preserved, providing evidence that the genomic regions were orthologous to HSP90‐containing regions in the chicken genome. The promoter regions of the genes also contained conserved motifs identified in related genes of chicken. However, the nucleotide sequences of the 5′‐flanking region of these genes were highly polymorphic. We also found that CjHSP90AA1 exhibited a robust response to heat shock treatment. Taken together, the data suggest that CjHSP90AA1, CjHSP90AB1, CjHSP90B1 and CjTRAP1 encode orthologs of HSP90AA1, HSP90AB1, HSP90B1 and TRAP1, respectively.     

Changes in the predicted function of the rumen bacterial community of Japanese Black beef cattle during the fattening stages according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses

We investigated changes in the predicted functions of the rumen bacterial community in Japanese Black beef cattle during fattening. Nine cattle were fed a high-concentrate diet during the early, middle, and late fattening stages consecutively (10–14, 15–22, and 23–30 months of age, respectively). The rumen fluid and solid samples collected at each stage were subjected to sequencing analyses. The sequencing results were clustered and classified into operational taxonomic units (OTUs). Representative sequences and a raw counting table for each OTU were submitted to the Piphillin website. The predicted functions were revealed by the Kyoto Encyclopedia of Genes and Genomes database as the ratio of the total sequence. In the early stage, “Biosynthesis of secondary metabolites” was significantly higher in the fluid fraction than in the solid fraction. “Two-component system” in the middle stage was significantly lower and “Purine metabolism” in the late stage was significantly higher in the fluid fraction than those in the solid fraction. The fluid fraction was significantly correlated with acetic acid, propionic acid, and bacterial metabolism, such as “Biosynthesis of secondary metabolites” and “Sugar metabolism.” Moreover, the solid fraction was correlated with “Purine metabolism” and “Biosynthesis of secondary metabolism”. These results suggest that the rumen bacterial community in Japanese Black beef cattle adapts to changes in rumen conditions by altering their functions in response to a long-term high-grain diet.     

A novel multiplex PCR assay to detect and distinguish between different types of Paenibacillus larvae and Melissococcus plutonius,and a survey of foulbrood pathogen contamination in Japanese honey

Paenibacillus larvae and Melissococcus plutonius are the causative agents of American and European foulbroods of honey bees, respectively. Since their virulence and resistance to disinfectants differ depending on the genotypes/phenotypes of the strains, the discrimination of strain types is important for the effective control of these diseases. Methods to detect and differentiate pathogens in honey are useful for surveying the contamination status of beehives/apiaries. In the present study, we selected a sequence (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"FI763267","term_id":"224923825"}}FI763267) as the specific target for enterobacterial repetitive intergenic consensus (ERIC) II-type P. larvae strains for the first time and developed a novel multiplex PCR assay that precisely distinguishes between the major types of foulbrood pathogens (ERIC I and II P. larvae and typical and atypical M. plutonius) in one reaction. In addition, we found that commercially available kits designed for DNA extraction from Mycobacterium in feces efficiently extracted DNA from foulbrood pathogens in honey. Using the multiplex PCR assay and DNA extraction kits, all the targeted types of P. larvae and M. plutonius were detected in honey spiked with the pathogens at a concentration of 100 bacterial cells/strain/ml. Moreover, 94% of the Japanese honey samples examined in the present study were contaminated with one or more types of the foulbrood pathogens. These results indicate that the newly developed methods are useful for detecting foulbrood pathogens in honey. The epidemiological information obtained by these methods will contribute to the effective control of foulbroods in apiaries.