Native Thai Chicken Research Articles

Supplementation of sperm cooling medium with Eurycoma longifolia extract enhances native Thai chicken sperm quality and fertility potential.

Cooled semen storage methods result in oxidative stress generated by an imbalance between oxidation rates, specifically reactive oxygen species production, and sperm cell antioxidants, leading to degradation of semen quality. We aimed to investigate the impact of adding Eurycoma longifolia (EL) extract as an antioxidant supplement in semen storage medium (IGGKPh semen extender) on semen quality and fertility potential. EL extract at concentrations of 5, 10, 15, and 20 mg/mL was assessed for its antioxidant capacity in IGGKPh semen extender. Our findings revealed that the total phenolic content in the EL extract did not vary significantly across the various concentrations and temperatures tested. However, incubation at 5°C was found to be the most effective temperature for increasing the EL extract antioxidant capacity as assessed via the 2,2-diphenyl-1-picrylhydrazyl inhibition assay in a dose-dependent manner. Supplementation of the IGGKPh semen extender with 15 mg/mL EL extract was found to enhance semen quality during cold storage for up to 48 h (p < 0.05), as evidenced by decreased malondialdehyde levels in cooled semen (p < 0.05). However, antioxidant enzyme activities showed no significant differences among the various experimental groups (p > 0.05). The fertility test showed that the 15 mg/mL EL extract group stored for 24 h had a higher percentage than the control group (p < 0.05). However, there was no significant difference in percentage between the two groups at 48 h of storage (p > 0.05). The hatchability showed no significant difference in both 24 and 48-h storage periods (p > 0.05). Our results indicated that supplementing the IGGKPh semen extender with 15 mg/mL EL extract may positively influence semen quality during storage, suggesting potential applications for enhancing semen quality.

Genetic diversity among five native Thai chickens and Khiew-Phalee chickens in lower-northern Thailand using mitochondrial DNA barcodes

Abstract. Siriwadee P, Wirot L, Thanapol P, Wirawan N. 2023. Genetic diversity among five native Thai chickens and Khiew-Phalee chickens in lower-northern Thailand using mitochondrial DNA barcodes. Biodiversitas 24: 1962-1970. Native chickens are an important biological resource in Thailand, with each breed having distinctive features. Khiew-Phalee chickens are a valuable investment in cultural wisdom and an invaluable Thai resource worthy of conservation. It is the royal rooster of Phraya Phichai Dab Hak. Due to a crisis concerning a sharp decline in the size of the Khiew-Phalee native chicken population, the identification of potential biomarkers is important for the determination of different chicken breeds for future planning. This study aimed to investigate the molecular identification of different native Thai chicken breeds based on mitochondrial DNA (mtDNA) barcodes, especially Khiew-Phalee chickens, which is difficult using the morphological examination. Partial sequences of 648 bp mtDNA control region fragments were determined for 53 specimens of six breeds of native Thai chickens (Khiew-Phalee, Thao Thong, Leung Hang Khao, Chee, Pra Dhu Hang Dam and Kai Jae). Genetic diversity was accessed based on the number of polymorphic sites, number of haplotypes, haplotype diversity, nucleotide diversity, and average number of differences. A phylogenetic tree was constructed based on maximum likelihood. The results showed that eighteen haplotypes were identified from 9 polymorphic sites with polymorphism between nucleotides 1074, 1107, 1109, 1177, 1213, 1214, 1268, 1276 and 1305. The haplotype diversity, nucleotide diversity, and the average number of differences for all chickens were 0.917, 0.00504 and 2.69086, respectively. All Khiew-Phalee chickens showed 7 haplotypes with 0.897 haplotype diversity, 0.00696 nucleotide diversity, and an average number of differences at 3.1795 clustered with other breeds in the phylogenetic tree. The findings indicated that the DNA barcoding gene is an effective method to identify the Khiew-Phalee and five other native Thai chicken breeds. It is a better choice for using molecular biology DNA barcoding to identify Thai native chicken breeds to plan for further conservation and development by the Department of Livestock Development of Thailand.

Effects of dried chaya leaf meal inclusion in the diet on growth performance and blood profiles in Thai native chicken (Pradu Hangdum).

The objective of the present study was to evaluate the effects of using dried chaya leaf meal (DCLM) as a protein feed in a diet on growth performance, blood profiles, and carcass quality in a native Thai chicken. Eighty 14-day-old Pradu Hangdum chicks were divided into four groups with four replicates each: control (without DCLM), 10%, 20%, and 30% DCLM inclusion in mash feed. Growth performance was recorded weekly until 98 days of age. Blood profile, carcass quality, and visceral organ weight were measured at 98 days of age. The 10%-30% DCLM inclusion in the diet did not affect feed intake or feed efficiency; however, the body weight gain of chicks decreased linearly with the increase in DCLM inclusion. The heterophils, eosinophils, and monocytes were linearly increased with the increasing DCLM levels in the groups. The serum blood chemistry did not differ among the groups, while the AST in 10% and 20% DCLM was lower than in the control. Increasing the level of DCLM in the chicken diet did not affect carcass quality. The DCLM can be used as a feed ingredient in Thai native chicken feed up to 20%.

Neuropeptide Y and maternal behavior in the female native Thai chicken

Maternal care behaviors in birds include incubation and rearing behaviors. During incubating period, the hens stop laying and eating less due to food restriction as a natural fasting when compared with the rearing hens, resulting in low production of eggs and chicks. Neuropeptide Y (NPY), a neurotransmitter/neuromodulator, is very well known to be involved in food intake regulation in birds and mammals. The objective of this study is to elucidate the association between NPY and maternal behaviors in the female native Thai chicken. The distributions of NPY-immunoreactive (-ir) neurons and fibers in the brain of the incubating (INC), nest-deprived (ND), and replaced-egg-with-chicks (REC) hens at day 6 were determined utilizing immunohistochemistry technique. The results revealed that the distributions of NPY-ir neurons and fibers were observed within the septalis lateralis, nucleus rotundus, and nucleus dorsolateralis anterior thalami, with predominantly located within the the nucleus paraventricularis magnocellularis (PVN). NPY-ir fibers were located throughout the brain and the densest NPY-ir fibers were distributed in a discrete region lying close to the ventriculus tertius (third ventricle) through the hypothalamus. Changes in the number of NPY-ir neurons within the PVN of the INC, ND, and REC hens were compared at different time points (at days 6 and 14). Interestingly, the number of NPY-ir neurons within the PVN was significantly higher (P < 0.05) in the INC hens when compared with those of the ND and REC hens at day 14 but not day 6. In addition, the number of NPY-ir neurons within the PVN of the INC hens was significantly increased (P < 0.05) from day 6 to day 14 but not the ND and REC hens. These results indicated, for the first time, the asscociation between NPY and maternal behaviors in the femle native Thai chicken. Change in the number of NPY-ir neurons within the PVN during the transition from incubating to rearing behavior suggested the possible role of NPY in the regulation of the maternal behaviors in this equatorial species. In addition, the native Thai chicken might be an excellent animal model for the study of this phenomenon.

Distribution and variation of neuropeptide Y in the brain of native Thai chicken

Neuropeptide Y (NPY) plays a pivotal role in food intake and body weight regulation in both birds and mammals. Unlike imported broilers and layers, native Thai chicken, a tropical non-seasonal breeding species, has lower body weight and exhibits strongly maternal behaviors which, in turn, affect feeding behavior during the reproductive cycle. The aim of this study was to investigate the role(s) of NPY that might be associated with the reproductive cycle of female native Thai chickens using immunohistochemistry technique. The distributions of NPY-immunoreactive (-ir) neurons and fibers in the brain of laying and fasted chickens was also elucidated. Changes in body weight and number of NPY-ir neurons in the nucleus paraventricularis magnocellularis (PVN) were compared across reproductive stages. The results revealed that NPY-ir neurons and fibers were distributed throughout the brain with the greatest density located in the PVN. Differences in the number of NPY-ir neurons in the PVN were found across reproductive stages. The numbers were lowest in non-egg laying and egg laying stages and significantly higher during egg-incubating and chick-rearing stages. Changes in body weight were inversely related to the number of NPY-ir neurons across reproductive stages. In addition, food restriction caused an increase in NPY immunoreactivity, confirming the role of NPY in response to food restriction. Taken together, the present findings suggest that the NPYergic system in the PVN plays an important role in the regulation of food intake during the reproductive cycle in this non-seasonal breeding tropical species.

Effect of Temperature on Reproductive Hormones and Egg Quality in the Native Thai Chicken

Environmental influence on the avian reproductive system is well recognised. This study investigates the effects of ambient temperature on egg quality in the native Thai Chicken (Gallus domesticus). Effects on prolactin (PRL) and ovarian steroids in chickens housed under different temperatures were also elucidated. Sixty female chickens were divided into four groups. Groups 1, 2, and 3 were reared under controlled temperatures of 35±2 °C, 31±2 °C, and 27±2 °C, respectively. Group 4 was reared under an uncontrolled natural temperature (approximately 27 °C). Blood samples were collected from each chicken once a week for 11 weeks, and plasma PRL, oestradiol (E), progesterone (P), and testosterone (T) levels were determined utilising enzyme-linked immunosorbent assays. The results revealed that E levels were significantly higher in chickens reared under low ambient temperature (27 °C) when compared with those exposed to 31 °C and 35 °C. No significant differences in PRL, P, and T levels were observed. Egg quality was affected by high ambient temperature. Eggshell thickness of chickens housed at 35 °C was thinner than those of other groups. These results indicate that changes in ambient temperature affect circulating levels of E but not PRL, P, and T. Thermally-stressed environments with high temperatures therefore impact egg quality in the native Thai chicken by reducing eggshell thickness.

The effects of replacing eggs with chicks on mesotocin, dopamine, and prolactin in the native Thai hen

The mesotocinergic (MTergic) and dopaminergic (DAergic) systems have been documented to play pivotal roles in maternal behaviors in native Thai chickens. In native Thai chickens, plasma prolactin (PRL) concentrations are associated with maternal behaviors, which are also controlled by the DAergic system. However, the role of MT in conjunction with the roles of DA and PRL on the neuroendocrine regulation of the transition from incubating to rearing behavior has never been studied. Therefore, the aim of this study was to investigate the association of MT, DA, and PRL during the transition from incubating to rearing behavior in native Thai hens. Using an immunohistochemistry technique, the numbers of MT-immunoreactive (-ir) and tyrosine hydroxylase-ir (TH-ir, a DA marker) neurons were compared between incubating hens (INC; n = 6) and hens for which the incubated eggs were replaced with 3 newly hatched chicks for 3 days after 6, 10, and 14 days of incubation (REC; n = 6). Plasma PRL concentrations were determined by enzyme-linked immunosorbent assay. The results revealed that the numbers of MT-ir neurons within the nucleus supraopticus, pars ventralis (SOv), nucleus preopticus medialis (POM), and nucleus paraventricularis magnocellularis (PVN) increased in the REC hens when compared with those of the INC hens at 3 different time points (at days 9, 13, and 17). On the other hand, the number of TH-ir neurons in the nucleus intramedialis (nI) decreased in the REC13 and REC17 hens when compared with those of the INC hens. However, the number of TH-ir neurons in the nucleus mamillaris lateralis (ML) only decreased in the REC13 hens when compared with the INC13 hens. The decrease in the numbers of TH-ir neurons within the nI and ML is associated with the decrease in the levels of plasma PRL. This study suggests that the presence of either eggs or chicks is the key factor regulating the MTergic system within the SOv, POM, and PVN and the DAergic system within the nI and ML during the transition from incubating to rearing behavior in native Thai chickens. The results further indicate that these two systems play pivotal roles in the transition from incubating to rearing behavior in this equatorial species.

Role of vasoactive intestinal peptide during the transition from incubation behavior to rearing behavior in the female native Thai chicken

It is well documented that vasoactive intestinal peptide (VIP) is an avian prolactin-releasing factor, regulating the reproductive cycle, that initiates and maintains incubation behavior in avian species. Native Thai chicken has strong maternal behaviors. Thus, it is an excellent animal model to study this phenomenon. Changes in the numbers of VIP neurons within the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) are associated with maternal behaviors in the native Thai chicken. The objective of this study was to elucidate the role of VIP during the transition from egg incubation to rearing behavior in this species. The distributions of VIP-immunoreactive (-ir) neurons and fibers within the nucleus septalis lateralis (SL), nucleus anterior medialis hypothalami (AM), regio lateralis hypothalami (LHy), nucleus commissurae pallii (nCPa), nucleus ventromedialis hypothalami (VMN), and IH-IN were compared between incubating (INC) hens and replaced-egg-with-chicks (REC) hens, in which eggs were replaced by 3-day-old chicks after 6, 10, and 14 d of incubation period for 3 days. Using an immunohistochemistry technique, the results revealed that VIP-ir neurons and fibers were found within the SL, AM, LHy, nCPa, VMN, and IH-IN, with the greatest expression observed in the IH-IN in both groups. The number of VIP-ir neurons in the IH-IN was markedly decreased (P <0.05) in the REC hens when compared with those of the INC hens at 3 different time points (at d 9, 13, and 17). However, changes in the number of VIP-ir neurons between the INC and REC hens were not observed within other hypothalamic areas tested. These findings indicate that the presence of eggs and chicks affects the VIPergic system, and VIP neurons in the IH-IN might play a role in the transition from incubating to rearing behavior in the native Thai chicken.

Effects of nest-deprivation on hypothalamic mesotocin in incubating native Thai hens (Gallus domesticus)

Avian mesotocin (MT) is homologous to oxytocin in mammals. Native Thai chickens (Gallus domesticus) strongly express maternal behaviors including incubation and rearing. However, the role of MT during incubation behavior has never been studied. The objective of this study was to determine the physiological function(s) of the MTergic system in incubation behavior in native Thai chickens. The brains were collected from incubating (INC) and nest-deprived (ND) hens at different time points (days 3, 6, 8, 10, 14, 18, and 21; n=6). Immunohistochemistry technique was used to compare the numbers of MT-immunoreactive (-ir) neurons between the INC and ND hens within the Nucleus supraopticus, pars ventralis (SOv), Nucleus preopticus medialis (POM), and Nucleus paraventricularis magnocellularis (PVN). The results revealed that the numbers of MT-ir neurons within the SOv, POM, and PVN remained high during the incubating stage. The number of MT-ir neurons in the SOv was lower than that of the POM and PVN. Disruption of incubation behavior by nest deprivation caused the numbers of MT-ir neurons within the SOv, POM, and PVN to decrease throughout the observation periods. For the first time, this study demonstrates that the MTergic system within the SOv, POM, and PVN may be involved with incubation behavior. In addition, these results further suggest that the MTergic neurons in these nuclei are not only regulated by rearing behavior but also might have a role in the initiation and maintenance of incubation behavior in this tropical species.

Ovarian steroids involvement in maternal care in the native Thai hen (Gallus domesticus)

Maternal care in birds generally includes incubation, brooding and rearing behavior. The neuroendocrine regulation of such maternal behavior is not fully understood for many bird species. The aim of this study was to elucidate whether changes in the circulating levels of ovarian steroids might be associated with brooding behavior in the female native Thai chicken. Changes in the levels of ovarian steroids of rearing hens were compared with those of non-rearing ones. Plasma estradiol, progesterone, and testosterone levels were determined by enzyme-linked immunosorbent assays. The results revealed differences in circulating levels of ovarian steroids between hens rearing their chicks and those deprived of their young after hatching. The non-rearing hens had higher circulating estradiol, progesteron, and tertosterone levels than those of the rearing hens. These differences were significant during the first two weeks after hatching. The results of the present study demonstrate the inhibitory effect of brooding behavior on ovarian steroids in the native Thai chickens. Indeed, disruption of brooding behavior by removing the chicks from the hens increased circulating levels of ovarian steroids and reinitiated reproductive and egg laying activities.

Dopamine and prolactin involvement in the maternal care of chicks in the native Thai hen (Gallus domesticus)

The dopaminergic (DAergic) system plays a pivotal role in incubation behavior via the regulation of prolactin (PRL) secretion in birds, however the role of the DA/PRL system in rearing behavior is poorly understood. The objective of this study was to investigate the relationship between the DA/PRL system and rearing behavior in a gallinaceous bird, the native Thai chicken. Incubating native Thai hens were divided into two groups. In the first group, hens were allowed to care for their chicks (rearing hens; R). In the second group, hens were deprived of their chicks immediately after hatching (non-rearing hens; NR). In both groups, blood samples and brain sections were collected at different time points after the chicks hatched (days 4, 7, 10, 14, 17, 21, 24, and 28; 6 hens/time point/group). In this study, tyrosine hydroxylase (TH) was used as a marker for DAergic neurons. The numbers of TH-immunoreactive (-ir) neurons in the nucleus intramedialis (nI) and in the nucleus mamillaris lateralis (ML), which regulate the vasoactive intestinal peptide (VIP)/PRL system, were determined in R and NR hens utilizing immunohistochemical techniques. Plasma PRL levels were determined by enzyme-linked immunosorbent assays. The results revealed that both the number of TH-ir neurons in the nI and the plasma PRL levels were significantly higher in the R hens compared with the NR hens during the first 14days of chick rearing (P<0.05). However, there was no significant change in the DAergic activity in the ML in either the R or NR groups throughout the 28-day rearing periods. These results suggest that the DA/PRL system is involved in early rearing behavior. The additional decline in DAergic activity and plasma PRL levels during the disruption of rearing behavior further supports their involvement in rearing behavior in this equatorial precocial species.

Mesotocin and maternal care of chicks in native Thai hens (Gallus domesticus)

Oxytocin (OT) is known to induce and regulate maternal behaviors in mammals via the supraoptic nucleus and paraventricular nucleus (PVN), whereas the function of mesotocin (MT; the avian homolog of OT) is poorly understood in birds. To elucidate the association of MT and the regulation of maternal behaviors in birds, we studied changes in the number of MT-immunoreactive (ir) neurons in native Thai chickens using immunohistochemistry. We observed that MT-ir neurons and fibers appeared in discrete regions located close to the third ventricle from the level of the preoptic area through the anterior hypothalamus with an abundance observed in the nucleus supraopticus, pars ventralis (SOv), nucleus preopticus medialis (POM), and PVN. The number of MT-ir neurons was low in the SOv, POM, and PVN of non-laying hens, but it increased gradually when the hens entered the laying stage, and peaked in incubating and rearing hens. We compared the number of MT-ir neurons in the SOv, POM, and PVN of native Thai hens rearing chicks (R) with that of non-rearing chicks (NR). The number of MT-ir neurons was high in the R hens, but low in the NR hens in these nuclei. For the first time, these results indicate that the association between the MT neurons and the presence of chicks might, in part, play a role in the neuroendocrine reorganization to establish and maintain maternal behaviors in native Thai chickens. MTergic activity is likely related to the contribution of rearing behavior in this equatorial precocial species.

The effects of ambient temperature on egg quality of native Thai chickens

The effect of increased ambient temperature on egg quality is well known in commercial layers, whereas in native Thai chicken has not been elucidated. This study investigated the relationship between the ambient temperature and egg quality in the native Thai chicken. Laying native Thai hens were used. Groups 1, 2, and 3, were reared in the close housed under 35 ± 2 °C, 31 ± 2 °C, and 27 ± 2 °C, respectively. Group 4 (control group) was reared in the open housed. Eggs were randomly collected and weighed from each group. Ten samples of eggshell from each group were taken from sharp, blunt, and equatorial parts. Shell thickness was measured using scanning electron microscope. The results revealed that egg weight (g) and eggshell weight (g) were higher in the control group (45.69 ± 0.65 and 4.77 ± 0.08) than those of the treatment groups (27 ± 2 °C, P&lt;0.05; 42.85 ± 0.76 and 4.35 ± 0.08; 31 ± 2 °C, P&gt;;0.05; 44.58 ± 0.52 and 4.73 ± 0.06; 35 ± 2 °C, P&gt;;0.05; 43.95 ± 0.64 and 4.60 ± 0.06). The shell thickness (μm) tended to decrease in the groups that reared under high temperature (35 ± 2 °C; 302.73 ± 6.62, 31 ± 2 °C; 316.13 ± 10.98, 27 ± 2 °C; 317.80 ± 7.75) when compared to the control group (316.20 ± 6.04). These findings provide the evidence that changed ambient temperature had a negative effect on egg weight, eggshell weight, and shell thickness in the native Thai chickens. Supported by The Research and Development Fund, Suranaree University of Technology

Gonadotropin releasing hormone and brooding behavior in the native Thai hen

Changes in the number of hypothalamic gonadotropin releasing hormone-I (GnRH-I) neurons within the Nucleus commissurae pallii (nCPa) were associated with the reproductive cycle of native Thai chickens. In order to further understand the association of GnRH-I in the regulation of brooding behavior in this bird, the native Thai chickens were divided into two groups; chick-rearing (R) and non-chick-rearing (NR) hens. Numbers of visible of GnRH-I-immunoreactive (GnRH-I-ir) neurons in the hypothalamus of R and NR hens were compared utilizing immunohistochemistry. Numbers of visible GnRH-I-ir neurons within the Nucleus anterior medialis hypothalami, Nucleus suprachaiasmaticus, pars medialis, Nucleus septalis lateralis, Nucleus paraventricularis magnocellularis, and Regio lateralis hypothalami areas were observed in both groups, but no differences were seen between R and NR hens. The number of visible GnRH-I neurons in the nCPa was higher (P<0.05) in the NR than in R hens, and increased in NR hens by day 14 after chick removal. These findings suggest, for the first time, an association of the GnRH system with brooding behavior in continuously breeding birds. Furthermore, the expression of brooding behavior of native Thai chickens might be regulated, in part, by GnRH-I neurons in the nCPa.

Plasma leptin concentrations during the reproductive cycle in the native Thai chicken (Gallus domesticus)

Plasma leptin concentrations were investigated during the reproductive cycle in the native Thai chicken. The plasma leptin concentration was high during non-laying (0.69±0.15ng/ml), lowered to a minimum concentration during egg laying (0.07±0.02ng/ml), and gradually increased during egg incubation and rearing of the chicks (0.53±0.22 and 0.74±0.29ng/ml, respectively). However, the differences were not significant. Incubating chickens that were deprived of their nests for 3 weeks showed a significant decrease in plasma leptin concentrations (0.29±0.04ng/ml, P<0.05) compared to those of their corresponding incubating controls (0.77±0.08ng/ml). Similarly, plasma leptin concentration of chickens that were deprived of their chicks for 4 weeks was significantly lower (0.09±0.11ng/ml, P<0.05), when compared to those of chickens that rearing their chicks (0.71±0.18ng/ml). These findings taken together with the results that the low plasma leptin concentrations were observed in chickens having relatively greater ovary and oviduct weights led to the suggestion that circulating leptin concentrations are associated with the reproductive states of the birds, especially the ovarian activity (i.e. ovarian steroid hormone concentrations) in the native Thai chicken, a tropical and continuous breeding species.

Neuroendocrine regulation of rearing behavior in the native Thai hen

Vasoactive intestinal peptide (VIP) is the avian prolactin releasing factor and changes in the concentrations of plasma prolactin (PRL) are found during the avian reproductive cycle. This study investigated the changes in the VIP/PRL system of native Thai hens rearing their young as compared to hens deprived of rearing their chicks. The number of VIP-immunoreactive (VIP-ir) neurons in the Nucleus inferioris hypothalami (IH) and Nucleus infundibuli hypothalami (IN) of hens rearing chicks (R) were compared with those of non-rearing chicks (NR). Plasma PRL levels were determined by enzyme-linked immunosorbent assay. The localization and number of VIP-ir neurons were determined by immunohistochemistry. The numbers of VIP-ir neurons in the IH–IN areas were high in the R hens, whereas the number of VIP-ir neurons decreased in the NR hens as compared to their respective R hens. During the rearing period, changes in the VIP-ir neurons within the IH–IN were correlated with plasma PRL levels. The results of the present study indicate for the first time that the VIP/PRL system plays a role in neuroendocrine reorganization to establish maternal behavior in native Thai chickens. The VIP/PRL system functions not only as a well established key regulator of incubation behavior, but is also involved in the regulation of rearing behavior. It is possible that VIP and the decline in the number of VIP-ir neurons and in turn VIPergic activity and the decrease in PRL levels are related to their contribution to rearing behavior of this non-seasonal breeding, equatorial precocial species.

Distribution and variation in gonadotropin releasing hormone-I (GnRH-I) immunoreactive neurons in the brain of the native Thai chicken during the reproductive cycle

Gonadotropin releasing hormone-I (GnRH-I) is known to regulate the avian reproductive system. We investigated the roles of GnRH-I in the regulation of the reproductive system of the native Thai chicken. The distribution of GnRH-I neurons and changes in GnRH-I-immunoreactive (-ir) neurons throughout the reproductive stages and between incubating and nest-deprived hens were analyzed utilizing immunohistochemical techniques. The results revealed that GnRH-I-ir neurons were distributed in a discrete region lying close to the third ventricle from the level of preoptic area through the anterior hypothalamus, with the greatest abundance found within the nucleus commissurae pallii (nCPa). The number of GnRH-I-ir neurons in the nCPa was highest in laying hens when compared with that in the other reproductive stages. Nest deprivation caused an increase in the number of GnRH-I-ir neurons in the nCPa of nest-deprived hens when compared with incubating hens. These results indicate that GnRH-I expression is correlated with the reproductive state in the native Thai chicken and may be, in part, regulated by it. This study also confirms a pivotal role of GnRH-I in controlling avian reproduction of this non-seasonal breeding, equatorial species.

The dopaminergic system in the brain of the native Thai chicken, Gallusdomesticus: Localization and differential expression across the reproductive cycle

Dopamine (DA) has a pivotal role in avian prolactin (PRL) secretion, acting centrally through D 1 DA receptors to stimulate PRL secretion by operating through vasoactive intestinal peptide (VIP). DA also inhibits PRL secretion by activating D 2 DA receptors at the pituitary level. This study was designed to investigate the distribution of DA neurons in the native Thai chicken, utilizing tyrosine hydroxylase (TH) as a marker for dopaminergic neurons. The differential expression of hypothalamic TH immunoreactive (TH-ir) neurons was also compared across the reproductive cycle. The results revealed that TH-ir neurons and fibers were found throughout the brain of the laying hen and were predominantly located within the diencephalon and mesencephalon. The observed distribution pattern of TH immunoreactivity was consistent with that reported previously in several avian species. However, changes in the number of TH-ir neurons in the nucleus intramedialis (nI) were observed across the reproductive cycle and correlated directly with variations in PRL levels. The population of TH-ir neurons in the nI increased significantly during the egg incubation period, where circulating PRL levels were the greatest. This study indicates, for the first time, that an association exists between DA neurons and the regulation of the reproductive system in the native Thai chicken. There is a paucity of information about the reproductive neuroendocrine regulation of tropical non-seasonally breeding avian species and it is suggested that the differential expression of DA neurons in the nI might play a role in the control of VIP secretion and subsequent PRL release in such birds.

Vasoactive intestinal peptide and its role in continuous and seasonal reproduction in birds

Native Thai chicken, an equatorial species breeds throughout the year, whereas turkeys are seasonal temperate zone breeder whose reproductive cycle is terminated by the onset of photorefractoriness. This study investigated VIPergic activity throughout a reproductive cycle in both species, hypothesizing that the differential expression of vasoactive intestinal peptide (VIP) would provide an insight into the differing reproductive strategies of the two species. Distribution of VIP neurons in the native Thai chicken and a comparison of VIPergic activity in the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) were investigated. VIP immunoreactivity was found throughout the native Thai chicken brain, predominantly located within the IH–IN. The pattern of VIP distribution in the native Thai chicken supports the findings reported in temperate zone species. Unlike the turkey, where there is a dissociation between VIPergic activity and prolactin levels during photorefractoriness, in the native Thai chicken, which do not express photorefractoriness, changes in VIP immunoreactive (VIP-ir) neurons within the IH–IN were directly correlated with prolactin throughout the reproductive cycle. VIPergic activity reached its lowest level after hatching of the chicks in the native Thai chicken, while in the turkey VIPergic activity was lowest only after exposure to a short day photoperiod and the acquisition of photosensitivity. This suggests that VIP neurons in the IH–IN may play a pivotal role in regulating the reproductive cycle and its differential expression following hatching of the young may, in part, account for the difference in reproductive mode between equatorial, continually breeding, non-photoperiodic birds and seasonally breeding, photoperiodic birds.

Occurrence of Germanium

Plasma leptin concentrations were investigated during the reproductive cycle in the native Thai chicken. The plasma leptin concentration was high during non-laying (0.69 ± 0.15 ng/ml), lowered to a minimum concentration during egg laying (0.07 ± 0.02 ng/ml), and gradually increased during egg incubation and rearing of the chicks (0.53 ± 0.22 and 0.74 ± 0.29 ng/ml, respectively). However, the differences were not significant. Incubating chickens that were deprived of their nests for 3 weeks showed a significant decrease in plasma leptin concentrations (0.29 ± 0.04 ng/ml, P < 0.05) compared to those of their corresponding incubating controls (0.77 ± 0.08 ng/ml). Similarly, plasma leptin concentration of chickens that were deprived of their chicks for 4 weeks was significantly lower (0.09 ± 0.11 ng/ml, P < 0.05), when compared to those of chickens that rearing their chicks (0.71 ± 0.18 ng/ml). These findings taken together with the results that the low plasma leptin concentrations were observed in chickens having relatively greater ovary and oviduct weights led to the suggestion that circulating leptin concentrations are associated with the reproductive states of the birds, especially the ovarian activity (i.e. ovarian steroid hormone concentrations) in the native Thai chicken, a tropical and continuous breeding species.