Expression of gonadotropin and sex steroid hormone receptor mRNA in the utero-vaginal junction containing sperm storage tubules of oviduct during sexual maturation in Japanese quail.

Sex steroid hormones play an important role in reproductive tissue development of avian species. However, their role in Japanese quail is yet to be established. To understand the physiological role of hormones involved in the development of sperm storage tubules (SSTs) in quail, we investigated expression profiles of gonadotropin (LH-R and FSH-R) and sex steroid hormone (PR-R, ER-α and ER-ß) receptors in the uterovaginal junction (UVJ) containing SSTs before and during sexual maturation i.e. four to eight weeks. Every week four birds were sacrificed to collect blood and UVJ for sex steroid hormone (progesterone and estrogen) estimation and gene expression profiling of sex steroid hormone (PR-R, ER-α and ER-ß) and gonadotropin receptors (LH-R and FSH-R) using qRT-PCR. Receptor expression results showed that the expression of sex steroid receptor (PR-R, ER-α and ER-ß) genes were upregulated significantly (P < .05) in SSTs with the advancement of age. The expression of gonadotropin receptors (LH-R and FSH-R) was only high at week 5 and 6 respectively. Serum hormone analysis indicated a significant (P < .05) rise in estradiol till 7th week and progesterone from 7th week onwards. These results suggest that the gonadotropin and sex steroid hormone receptors may have the role in the development and maintenance of UVJ that contains predominantly SSTs during sexual maturation.

Genome survey sequencing and mining of genome-wide microsatellite markers in yellow-billed babbler (Turdoides affinis).

Turdoides affinis is a species of group dwelling old world passerine of family Leiothrichidae. Unavailability of genome-wide sequence and species-specific molecular markers have hindered comprehensive understanding of cooperative breeding behaviour in T. affinis. Therefore, we generated genome-wide microsatellite markers through whole genome short read sequencing of T. affinis. A total of 68.8 gigabytes of paired-end raw data were sequenced containing 195,067,054 reads. Total sequenced reads spanned a coverage of 17X with genome size of 1.18 Gb. A large number of microsatellite markers (265,297) were mined in the T. affinis genome using Krait, and 50 most informative markers were identified and validated further. In-silico PCR results validated 47 markers. Of these 47 markers, five were randomly selected and validated in-vitro in twelve individuals of T. affinis. Genotyping data on these five loci estimated observed heterozygosity (H0) and expected heterozygosity (He) ratios between 0.333 - 0.833 and 0.851-0.906, respectively. Effective allele size ranged from 6.698 to 10.667, inbreeding coefficient of the population ranged from 0.080 to 0.631 and null allele frequency was calculated at 0.055 to 0.303. Polymorphic information content of all the five loci varied between 0.850 and 0.906. Probabilities of exclusion and identity across 5 loci was estimated to be 0.95 and 0.0036, respectively. All the loci showed significant adherence to Hardy-Weinberg equilibrium. The microsatellite markers reported in this study will facilitate future population genetics studies on T. affinis and other congeneric species.

Isolation of E. coli from foam and effects of fluoroquinolones on E. coli and foam production in male Japanese quail.

Sexually active male Japanese quail (Coturnix coutrnix Japonica) produce a foamy substance from their cloacal gland. It was postulated that bacteria played an important role in production of foam. The primary objective of this study was to isolate and identify bacteria present in the cloacal foam. The secondary objective was to evaluate the effect of fluoroquinolone treatment on bacterial counts and foam production. Healthy adult Japanese quail were maintained in individual cages under uniform husbandry conditions and allocated arbitrarily into three groups (each group consisted of 12 male and 12 female birds). Foam was collected from the cloacal gland of male birds of each group separately into sterile petri dishes and was cultured to isolate and identify bacteria and to determine their sensitivity to various antibiotics. Escherichia coli bacteria, sensitive to various antibacterials (including the fluoroquinolones ciprofloxacin and pefloxacin), were isolated. In the second part of the study, male quails of Group I (control) received I mL vehicle (normal saline 0.9% (w/v) NaCl) daily (via the intraperitoneal route) for 12 days. Male birds from groups II and III were treated intraperitoneally with ciprofloxacin or pefloxacin at the rate of 10 mg and 12 mg per/kg body weight respectively, for 12 days. In antibiotic-treated birds, there was a gradual reduction in foam production during treatment. At the end of treatment, the cloacal gland area was smaller (P < 0.05) in pefloxacin-treated birds compared to the other groups. Furthermore, a trend towards decreasing body weight and fertilizing ability was noted in the same group. A drastic reduction in bacterial counts of foam was recorded only in fluoroquinolone-treated groups during treatment period. After cessation of treatment, all end points were increasing back to pre-treatment levels. In conclusion, E. coli were present in the foam of the cloacal gland of Japanese quail and may have a role in foam production.

Effects of simulated hypo- and hyper-reproductive conditions on the characteristics of circadian rhythm in hypothalamic concentration of serotonin and dopamine and in plasma levels of thyroxine, triiodothyronine, and testosterone in Japanese quail, Coturnix coturnix japonica.

In this study, hypo- and hyper-reproductive conditions, as measured by concentrations of plasma testosterone in male Japanese quail held on long days LD 16:8, were experimentally simulated with injections of 5-hydroxytryptophan (5-HTP) and L-dihydroxyphenylalanine, (L-DOPA) with 8 h and 12 h phase angle differences between them in intact and melatonin-treated birds. The effects of these treatments were assessed on the characteristics of the circadian rhythm in the hypothalamic concentration of serotonin (5-HT), dopamine (DA), and plasma levels of thyroxine (T(4)), triiodothyronine (T(3))(,) and testosterone (T). These rhythms were also studied in sham-operated (SO), pinealectomized (Px), vehicle- (Veh), and melatonin (Mel)-treated birds. On the basis of the circadian mesors of the testosterone rhythms, three distinct categories could be identified: category A (i.e., normal breeding concentrations of plasma testosterone), which includes control, sham-operated, and vehicle-treated groups; category A(+) (i.e., concentrations of plasma testosterone higher than that found in normal breeding quail), which includes 12 h, 12 h+vehicle-treated, and Px quails; and category A(-) (concentrations of plasma testosterone lower than that found in normal breeding quail), which includes 8 h, melatonin-, and 12 h+melatonin-treated groups. It is evident that in normal and hypergonadal conditions (i.e., birds belonging to categories A and A(+)) the circadian rhythm in hypothalamic serotonin maintained a positive phase angle of about 16 h. In contrast, birds of category A(-) (i.e., in a hypogonadal condition) exhibited a negative phase angle of about 2 h. The present results clearly suggest that the internal phase relationship between the circadian rhythms in hypothalamic serotonin and dopamine might play a crucial role in strategizing and conferring a particular reproductive status to the birds. The role of circadian mechanisms involving circulating thyroid hormones in conferring reproductive status is completely ruled out, as no definite internal phase angle between these two hormonal rhythms was witnessed vis-a-vis different treatment groups. The testosterone peaks always occurred at the same time irrespective of breeding status of the bird, but with significant variation in its amplitude (high in hypergonadal and low in hypogonadal condition). It is suggested that administration of 5-HTP and L-DOPA at specific time interval and variation in pineal functions that modulate reproductive responses also alter the circadian pattern (acrophase and amplitude) of hypothalamic serotonin and dopamine, maintaining a specific phase relation between these cycles and breeding status. These findings strengthen our previous reports that a specific circadian phase relation of serotonergic and dopaminergic oscillations regulates reproduction. The present study strongly supports interdependence and specific relation of the two systems (gonadal activity and circadian pattern/phase relation of neural oscillation) in both natural and experimentally simulated conditions.