Chapter Eight - The Study of Career Decisions: Oystercatchers as Social Prisoners

Progesterone concentration and lambing rate of Karakul ewes treated with prostaglandin and GnRH combined with the ram effect during breeding and non-breeding seasons

The aim of this study was to elucidate the regulatory role of androgen in the follicular development of wild female ground squirrels. Immunohistochemical staining of FSHR, LHR, P450c17, P450arom, androgen receptor (AR), estrogen receptors (ERa and ERb) were executed in ovaries of female ground squirrels from both breeding and nonbreeding seasons. In addition, total ovarian proteins were extracted from the ovaries of squirrels from breeding and nonbreeding seasons, and Western blot analysis were performed in order to probe for FSHR, LHR, P450c17, P450arom, AR, ERa and ERb. The results of immunohistochemical staining and Western blotting of P450c17 showed that there was no significant difference between the breeding and nonbreeding seasons. It was found that granulosa cells expressed P450arom during the breeding season. In contrast, there was no positive staining of P450arom in the nonbreeding season. There was no significant difference in immunoreactivity of AR between the breeding and nonbreeding seasons. However, the immunoreactivities of ERa and ERb were both significantly reduced in the nonbreeding season compared to the breeding season. The positive stains of FSHR and LHR were found in the granulosa cells and theca cells of the ovaries of the breeding and nonbreeding seasons. In addition, the Western blotting results of FSHR and LHR showed a significant reduction in the nonbreeding season compared with the breeding season. These findings suggested that androgen might be predominantly converted into estrogen in order to regulate the follicular development via binding of estrogen receptors during the breeding season, whereas androgen might predominantly directly bind androgen receptor to regulate the follicular development during the nonbreeding season in the ovaries of wild female ground squirrels.

The current study was undertaken to elucidate the effect of ovarian status in breeding and non-breeding season on the ovarian biometry, oocyte yield and oocyte quality of Baladi goats. Ovaries were collected by slicing from slaughter houses and classified with or without CLs during breeding (September-December) and non-breeding (March-July) seasons. Ovaries were weighed and measured, while oocytes were recoverded, yielded and categorized with or without CLs in breeding or non-breeding seasons. Results showed that ovarian weight and biometry (length, width and thickness) were higher in breeding than in non-breeding season, but the differences were significantly only for width. Number of follicles and oocytes/ovary (P<0.001) as well as number/ovary and proportion of oocytes at compact (P<0.0001) and denuded (P<0.05) stage were higher in breeding season than in non-breeding one. Number of degenerated oocytes/ovary was not affected significantly by season, but its proportion was lower (P<0.001) in breeding than in non-breading season. Number/ovary and proportion of partial denuded oocytes and proportion of denuded oocyte were not affected significantly by breeding season. Weight and biometry of ovaries was higher on ovaries bearing CL (CL+) than in non-bearing ones (CL-). Only ovarian width was higher (P<0.001) by 38% in CL+ than in CL- group. Ovaries bearing CL had higher (P<0.05) total follicles and oocyte yield/ovary (P<0.01) as well as oocyte recovery rate (P<0.05) than CL- ovaries. Number of compact, denuded and partial denuded oocytes/ovary was not affected by CL bearing. Number of compact oocytes tended to be greater on ovaries without than with CL. Number of degenerated oocytes/ovary was higher (P<0.05) on CL+ ovaries. Proportion of all oocyte categories was not affected by bearing CL. Finally, the effect of interaction between breeding season and bearing CL on all parameters studied was not significant. In conclusion, the goat ovaries without CL in breeding season yielded better oocyte quality than in non-breeding season, in term of COCs proportion. During non-breeding season, goats oocytes were available to be harvested from slaughtered goat does with acceptable yield and quality.

Influence of season and nutritional status on the direct effects of leptin, orexin-A and ghrelin on luteinizing hormone and growth hormone secretion in the ovine pituitary explant model

Buffaloes are tendentially short-day breeders, and seasonality is one of the main factors affecting the feasibility of ovum pickup and in vitro embryo production technology in this species. An improvement of oocyte developmental competence during decreasing daylight months was previously reported in Italian Mediterranean buffalo (Di Francesco et al. 2011 Anim. Reprod. Sci. 123, 48-53). The aim of this work was to evaluate whether season also affects embryo quality and cryotolerance. Abattoir-derived buffalo cumulus-oocyte complexes were collected during the breeding season, characterised by decreasing daylight length (n=349 over 6 replicates), and the non-breeding season, characterised by increasing daylight length (n=770 over 12 replicates). Buffalo cumulus-oocyte complexes were in vitro matured, fertilized, and cultured according to standard procedures (Di Francesco et al. 2011 Anim. Reprod. Sci. 123, 48-53). The embryos obtained by the end of culture (i.e. on Day 7 post-IVF) were scored for quality and developmental stage, and the percentages of total transferable embryos (tight morulae and blastocysts) were recorded. Embryos (n=107 and 110 in the breeding and non-breeding seasons, respectively) were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide, and 0.5M sucrose (Boccia et al. 2013 Ital. J. Anim. Sci. 12, 492-496). Warming was carried out by plunging the cryotop strip into a 0.25M sucrose solution and transferring the embryos into 0.15M sucrose for 5min. Embryos were then washed and cultured in SOF for 24h to evaluate post-culture viability. The resistance to cryopreservation was evaluated by assessing the survival rate, on the basis of morphological criteria, and development rate (i.e. the percentage of embryos that resumed their development and reached a more advanced developmental stage) after 24h post-warming culture. Data were analysed by Student’s t-test. Both cleavage (82.8±4.3v. 73.1±1.7 in the breeding and non-breeding seasons, respectively; P&amp;lt;0.05) and blastocyst (32.9±3.5v. 18.3±1.7 in the breeding and non-breeding seasons, respectively; P&amp;lt;0.01) rates increased during the breeding season, confirming previous observations. Due to the different efficiency, a higher number of replicates was required during the non-breeding season to obtain an equal number of embryos. In addition, a seasonal effect was recorded on embryo quality, indicated by poorer cryotolerance of in vitro-produced buffalo embryos during the non-breeding season. Indeed, both survival (94.6±2.7% and 74.0±5.5% in the breeding and non-breeding seasons, respectively; P&amp;lt;0.01) and development (67.3±7.6% and 40.0±7.2% in the breeding and non-breeding seasons, respectively; P&amp;lt;0.01) rates of vitrified blastocysts decreased after 24h post-warming culture in the non-breeding season. These findings suggest that the reduced developmental competence of buffalo oocytes during the non-breeding season may also lead to lower blastocyst quality. This is in contrast to the evidence in cattle that embryo quality is mainly determined by culture conditions, whereas blastocyst production depends on oocyte quality.

The purpose of this study was to investigate seasonal changes of spermatogenesis and the cellular localization of P450c17 and P450arom in wild male ground squirrels during the breeding and non-breeding seasons. The testicular weight, testicular size and score count of spermatogenesis from April to September were measured, and histological and immunohistochemical observations of testicular tissues were performed in wild male ground squirrels. In addition, total proteins were extracted from testicular tissue in the breeding and non-breeding seasons and were used for Western blotting analysis for P450c17 and P450arom. There were marked variations in testicular weight, testicular size and score count of spermatogenesis from the breeding season (April) to the non-breeding season (September). Histologically, spermatogonia, primary spermatocytes, secondary spermatocytes and spermatozoa were identified in the breeding season (April). Immunolocalization of P450c17 was detected in Leydig cells and spermatozoa during the breeding season and was only found in Leydig cells during the non-breeding season. The positive signals of P450c17 by Western blotting were both observed in the breeding and non-breeding seasons. Immunolocalization of P450arom was observed in Leydig cells, Sertoli cells and all types of spermatogenic cells including mature-phase spermatozoa in the breeding season, while immunoreactivity for P450arom was not present in the testis of the non-breeding season. With P450arom antibody, a band was also only detected in the breeding season by Western blotting. These results suggest that the seasonal changes in testicular weight and size are correlated with spermatogenesis and immunolocalization of P450c17 and P450arom, and androgen and estrogen may play an important role in the spermatogenesis and testicular recrudescence and regression process.

Comparison of follicular dynamics and hormone profiles in Boer goats examined during the breeding and non-breeding seasons in the tropics of Queensland, Australia

The changes in testicular and pituitary functions in response to human chorionic gonadotropin (hCG) in the breeding and non-breeding season were investigated and compared in the Thoroughbred stallion. Five mature Thoroughbred stallions ranging in ages from 7 to 21 years were injected in May (breeding season) and October (non-breeding season). All animals received an intramuscular injection of 5,000 IU hCG in the experiments. Peripheral blood samples were collected in heparinized tubes from the jugular vein for hormonal assays just before the injection (Day 0) and at a daily interval for five days following the injections (Days 1, 2, 3, 4 and 5). Basal levels of immunoreactive (ir) inhibin, testosterone, estradiol-17β, FSH and LH were higher in the breeding season than in the non-breeding season. There were significant differences in estradiol-17β, FSH and LH between the two seasons. Ir-inhibin exhibited significant increases on Day 2 after the hCG treatment in the non-breeding season, though there was no change in the breeding season. Plasma levels of testosterone showed remarkable increases after the hCG injection in both seasons. The peak levels of plasma testosterone were observed on Days 2 and 3 in the breeding and non-breeding seasons. Plasma estradiol-17β was significantly higher than the basal level on Day 3 in the non-breeding season, whereas there was no change in the breeding season. Plasma levels of FSH declined from Day 1 to Day 3, then recovered to the basal level (Day 0) on Day 4 after hCG treatment in the breeding season, whereas there was no change in the non-breeding season. Circulating LH showed a significant decrease on Day 3 compared to the based level in the breeding season, but no significant change in the non-breeding season after a treatment of hCG. A significant negative correlation was observed between testosterone and FSH in the breeding season. In conclusion, hCG treatment stimulates secretion of testicular hormones and these testicular hormones temporarily suppress secretion of gonadotropins from the pituitary gland. These results also suggested that Leydig cells of thoroughbred stallion testes have LH receptors in the non-breeding season, the same as in the breeding season.

Contemporary landscape change driven by socio-economic forces and advances in agricultural technology do not favor northern bobwhites (Colinus virginianus) in the Midwestern United States. The challenge of bobwhite conservation is to provide sufficient quantities of habitat with optimal configurations in proportions that satisfy needs throughout the year on private-owned working lands. We radio-tracked 58 coveys and 98 individual birds throughout the year in 2009-10 and 2010-11 on privately owned farmland in southwestern Ohio. We investigated temporal changes in usable space from use-availability data. Estimated proportions of usable space based on analyses of habitat selection across four study sites were 0.06-0.12 during the non-breeding season compared to 0.10-10.30 during the breeding season. We also modeled probabilities that radio-marked coveys or individual birds used points within individual cover types as a function of distance to other cover types. Locations of radio-marked coveys and individual birds within focal cover types were closer to other cover types compared to random points during breeding (mean = 44 m) and non-breeding (mean = 58 m) seasons. Probability of use within focal cover types declined with distance to other cover types, typically falling below 50% when distances exceeded 9-242 m. Locations of radio-marked coveys were concentrated near edges within used cover types, while locations of individual birds were more dispersed during the breeding season. Estimated proportions of usable space based on distances between locations of radio-marked bobwhites and nearest cover types were higher (0.30-0.53) than estimates based on habitat selection ratios, and were similar between non-breeding and breeding seasons. Potentially usable sites were not fully occupied in either season, but there was no relationship between crude covey densities and proportions of usable space estimated from habitat selection analyses during the breeding and non-breeding seasons across study sites and years (r2 < 0.166, P > 0.316). With distance to cover type estimates of usable space (years combined), two study sites with the highest proportions of usable space (0.43-0.53) also had the largest crude covey densities (0.0065-0.153 coveys/ha) compared to the other two sites with lower proportions of usable space (0.30-0.32) and smaller covey densities (0.004 – 0.006 coveys/ha). Habitat enhancement should focus on providing protective cover near food for coveys during the non-breeding season and protective cover near nesting and brood-rearing habitat during the breeding season. Conserving Midwestern bobwhite populations requires innovative practices that can be implemented on private lands as economic incentives change for farm operators. Improving protective cover along habitat edges can increase usable space by improving cover type juxtaposition (e.g. food near cover) while minimizing impact on farming practices.

Pituitary, gonadal and adrenal activity were compared in free-living, adult African buffalo bulls during the breeding and nonbreeding seasons. Frequent blood samples were collected for 2 h from anaesthetized bulls treated intravenously with saline, gonadotrophin-releasing hormone (GnRH, 200 micrograms), human chorionic gonadotrophin (hCG, 10,000 i.u.) or adrenocorticotrophic hormone (ACTH, 1.5 mg). Electroejaculates also were collected from anaesthetized bulls during the breeding and nonbreeding seasons. Pretreatment testosterone concentrations among bulls varied more during the breeding (0.17-23.0 ng/ml) than the nonbreeding (0.15-2.21 ng/ml) season. The variation within the breeding season was attributed to 8 of 25 bulls producing higher (P less than 0.05) serum testosterone (High-T; 16.28 +/- 2.03 ng/ml) and testicular LH receptor (1.53 +/- 0.22 fmol/mg testis) concentrations compared with their seasonal counterparts (Low-T; 0.95 +/- 0.26 ng/ml; 0.38 +/- 0.04 fmol/mg) or with all bulls during the nonbreeding season (0.90 +/- 0.27 ng/ml; 0.31 +/- 0.04 fmol/mg). The magnitude of GnRH- and hCG-induced increases in serum testosterone was similar (P greater than 0.05) between Low-T bulls and bulls during the nonbreeding season. In the High-T animals treated with GnRH or hCG, serum testosterone did not increase, suggesting that secretion was already maximal. Peak serum LH concentrations after GnRH were greater (P less than 0.05) in bulls during the nonbreeding than the breeding season; FSH responses were similar (P greater than 0.05). ACTH treatment did not increase serum cortisol concentrations above the 2-fold increase measured in bulls treated with saline, hCG and GnRH (P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Seasonal expression of androgen receptor in scented gland of muskrat (Ondatra zibethicus)

Differences in DNA methylation and mRNA levels between the breeding and non-breeding seasons of in vitro produced IVF and SCNT sheep embryos

As a vital male accessory reproductive gonad, the prostate requires vascular endothelial growth factors for promoting its growth and development. In this study, we investigated the localizations and expressions of vascular endothelial growth factor (VEGF) and its receptors including VEGF-receptor1 (VEFGR1) and VEGF-receptor2 (VEGFR2) in the prostate of the wild ground squirrels during the breeding and the non-breeding seasons. The values of total prostate weight and volume in the breeding season were higher than those in the non-breeding season. Histological observations showed that the exocrine lumens of the prostate expanded in the breeding season and contracted in the non-breeding season. The mRNA expression levels of VEGF and VEGFR2 in the prostate were higher in the breeding season than those in the non-breeding season, but the mRNA expression level of VEGFR1 had no significant change between the breeding and non-breeding seasons. Immunohistochemical results revealed that VEGF, VEGFR1 and VEGFR2 were presented in epithelial and stromal cells during the breeding and non-breeding seasons. In addition, the microvessels of the prostate were widely distributed and the number of microvessels increased obviously in the breeding season, while decreased sharply in the non-breeding season. These results suggested that expression levels of VEGF and VEGFR2 might be correlated with seasonal changes in morphology and functions of the prostate, and VEGF might serve as pivotal regulators to affect seasonal changes in the prostate functions of the wild male ground squirrels via an autocrine/paracrine pathway.

The seasonal profile of proliferation and apoptosis in the prostate gland of the wild ground squirrel (Spermophilus dauricus)

Food preference of Sri Lanka Yellow-eared Bulbul (Pycnonotus penicillatus) in Breeding and non-breeding seasons was studied at the montane cloud forests of Horton Plains National Park, situated in the highland plateau of the central highlands, from September 2015 to July 2017 in three consecutive days per month. P. penicillatus is an endemic near threatened species. They have two breeding seasons; February-May and August-October. Direct foraging observations and faecal sample analysis methods were used to collect data. Four mist nets (3x6 m2/25 mm mesh size) were allocated from 0600 to 1800 h in mist netting days. Droppings of the bird was later scraped individually into collecting bottles with 70% alcohol and stored. Some droppings were collected in the field. These samples were taken to the laboratory for further analysis. The samples were first divided into two main parts as high plant matter samples and high insect matter samples by visually observing the faecal matter. ‘High plant matter’ faecal samples were directly analysed. High insect matter samples were analysed after using standard protocols. The seeds of the plants were identified by the keys and, insect parts were identified to order level by using insect identification guides. Within the study period there were 1,903 field observations and 50 faecal sample analysis were performed. In the breeding season 87.9% of the feeding observations were consisted with 18 species of plants and remain 12.1% was consisted with animal parts from one Pheretima species, thirteen species of insects, including three species from Order Diptera, three species from Order Lepidoptera, two species from Order Coleoptera, three species from Order Hymenoptera, one species from Order Blattodea and one species from Order Hemiptera. In the non-breeding season 98.1% of the feeding observations were consisted with 15 species of plants and other 1.9% was completed with four species of animal parts, including one species from Order Diptera, one species from Order Lepidoptera and two species from Order Coleoptera. Present study revealed that P. penicillatus preferred more animal food items in the breeding season, possibly to secure the extra energy requirements during their breeding season. These studies will be important for conservation of P. penicillatus, by using species-specific conservation plans in the future.Keywords: Sri Lanka Yellow-eared Bulbul, Endemic birds, Horton Plains National Park, Food preference, Breeding and non-breeding seasons