POLIMORPHIC CHARACTERIZATION OF ESR1 AND FOXL2 GENES AND THE ASSOCIATION OF THEIR INTERACTION WITH PRODUCTIVE TRAITS OF BROWN LOCAL IRAQI CHICKENS

The estrogen receptor 1(ESR1) and fork-head box L2 (FOX L2) genes play a pivotal role in regulation of egg formation in poultry. this study aimed to investigate interaction between ESR1 and FOXL2 in the productive performance of brown local Iraqi chickens (BLIC). A total of 104 BLIC represented from the F4th generation of local chicken selected for high egg productions were used. Two novel single nucleotide polymorphism (SNPs), one of them in ESR1 (T57198C) intron 3, and another of FOX L2 (C919T) gene within encoding region were identified through PCR-DNA sequencing. Six haplotypes (H1=TTCC, H2=TTCT, H3=TCCC, H4=TCCT, H5=CCCC, H6=CCCT) were obtained due to interaction between these two SNPs. Chi-square analysis showed no significant in genotypic and allelic frequencies for each SNP which revealed that both genes were agreement with Hardy-Weinberg equilibrium. Association analysis of haplotypes with production traits revealed that individuals have H4 genotype achieved higher body weight at sexual maturity, at 60 weeks of age and egg weight at 45 and 60 weeks of age, whereas, the higher number of eggs were exhibited in individual with H6 from onset egg till 60 weeks of age. The two haplotypes; H4(TCCT) and H6(CCCT) showed better combination than others with respect to production performance. In conclusion, our findings provided new evidence that the two genes (ESR1 and FOXLl2) with special interaction may have potential effects on productive traits of chickens and beneficial effects in laying breeding programs.

Avian eggshell coloration predicts shell-matrix protoporphyrin content

Avian eggshell pigmentation may provide information about a female’s physiological condition, in particular her state of oxidative balance. Previously we found that female house wrens (Troglodytes aedon Vieillot, 1809) with lighter, less-maculated, and redder ground-colored shells were older and produced heavier offspring than females laying darker, browner eggs. The strong pro-oxidant protoporphyrin is responsible for this species’ eggshell pigmentation, so differences in pigmentary coloration may be related to eggshell protoporphyrin content and reflect female oxidative balance and condition during egg-formation. Therefore, we tested the assumption that egg-surface coloration is related to the amount of protoporphyrin in the shell matrix. We analyzed digital photographs of eggs to determine maculation coverage as a measure of the overall ground coloration of the egg and its red-, green-, and blue-channel pixel values. Pigments were then extracted from these same eggs and analyzed using high-performance liquid chromatography. There was a strong, positive relationship between eggshell redness and protoporphyrin content of eggshells, but no relationship between percent maculation and protoporphyrin content. Thus, when older, larger females deposit more protoporphyrin in their eggshells, this may reflect a tolerance for high levels of circulating protoporphyrin or an effective mechanism for off-loading protoporphyrin into the eggshell matrix. Keywords: Eggs, Female quality, HPLC, Eggshell pigmentation, Eggshell speckles

Polimorphic Explore of Esr1, Esr2 and Foxl2 Genes and Interaction Effect of Esri and Foxl2 with Productive Traits of Brown Local Iraqi Chickens

The estrogen receptor 1(ESR1) and estrogen receptor 2 (ESR2) plays a crucial role in the growth and development of follicles and ovulation in chickens. Fork-head box L2 (FOX L2) gene and its protein is active in several tissues, such as the ovaries, it has a pivotal role in ovarian development and function, like regulation of egg formation in poultry. This study aimed to detect the polymorphism of ESR1, ESR2 and, FOXL2 genes and investigate interaction between ESR1 and FOXL2 in the productive performance of brown local Iraqi chickens (BLIC). A total of 104 BLIC, at 17 weeks of age. Four novel single nucleotide polymorphism (SNPs). Every polymorphism a number had been given according to its location within gene, one of them in ESR1 (T57198C) intron 3 with three genotypes: Wild(TT), Heterozygous(TC),and Homozygous(CC), and two SNPs within ESR2 were assessed, one of the identified SNPsT39565C transition located within intron 7 with two alleles(T and C)and three genotypes were found: Wild(TT), Heterozygous(TC),and Homozygous(CC), another SNPG40100A transition within (3 prime UTR), with two alleles (G and A)and three genotypes were found: Wild(GG), Heterozygous(GA)and Homozygous(AA) and the SNP in FOXL2 (C919T) gene with two genotypes: Wild(CC) and Heterozygous(CT) within encoding region were identified through PCR-DNA sequencing. An interaction between the ESR1 and FOXL2 genes has been proposed, and six haplotypes (H1=TTCC, H2=TTCT, H3=TCCC, H4=TCCT, H5=CCCC, H6=CCCT) were obtained due to interaction between these two SNPs. Chi-square analysis showed no significant in genotypic and allelic frequencies for each SNP which revealed those genes were agreement with Hardy-Weinberg equilibrium. Association analysis of haplotypes with production traits revealed that individuals have H4 genotype achieved higher body weight at sexual maturity, at 60 weeks of age and egg weight at 45 and 60 weeks of age, whereas, the higher number of eggs were exhibited to individual with H6 from onset egg till 60 weeks of age. The two genotypes; TCCT and CCCT showed better combination than others with respect to production performance. In conclusion, our findings provided new evidence that the two SNPs (ESR1 and FOXLl2) with special interaction may have potential effects on productive traits of chickens and beneficial effects in laying breeding programs.

Egg productivity of quails of Japanese breed under different phase feeding schemes

The primary cause of injuries and inflammation of the reproductive system in female quails in industrial quail farming is the process of egg formation and its passage through the oviduct. With technological injuries females aged from 42 to 55 days are involved into the risk zone, whose weight of laid eggs ranges from 11,6 to 13,7 g. In previous morphological and histological studies, the following patterns of development of the organs of the reproductive system of quails under conditions of intensive technology have been established. The development of the reproductive organs of female quails of Japanese breed proceeds sequentially in seven periods: the first (1–21 days) is the period of relative rest of the growth of organs, the second (21–35 days) is the period of intensive morphogenesis of the ovaries and oviduct departments, the third (35–45 days) is the period of preparation and the beginning of egg laying, the fourth (45–75 days) is the period of egg laying growth or functional formation of the oviduct departments, the fifth (75–185 days) is the period of intensive folliculogenesis and high secretory activity of the departments of the oviduct or the peak of egg production, the sixth (185–200 days) is the beginning of the involution of the reproductive organs and a decrease in the rate of egg production, the seventh (200 days and older) is the involution of the reproductive organs and the termination of egg laying. Taking into account these patterns of development of the reproductive organs of quails, the feeding scheme of female egg quails has been adjusted, which provides for the division of the productive period into 4 phases: phase 1 is the beginning of egg production (36–45 days), phase 2 is the growth of egg production (46–75 days), phase 3 is the peak of egg production (76–185 days) and phase 4 is the decline of egg production (185–200 days). The proposed scheme of phase feeding of female quails of Japanese breed during the productive period is the advantageous technological technique, since it allows to reduce the injuries of the poultry oviduct and improve the quality indicators of the quail egg, as a result of reducing the weight and size of eggs in the initial phase of egg production and the number of large eggs during the entire productive period. The use of the new scheme of phase feeding of female quails of Japanese breed allows us to get more egg mass by 67,5 kg, due to the increase in the gross harvest of commercial quail eggs by 4,5 % and the livability of the quails by 5,6 %.

Whole blood and blood components from vertebrates differentially affect egg formation in three species of anautogenous mosquitoes

Background Most female mosquitoes are anautogenous and must blood feed on a vertebrate host to produce eggs. Prior studies show that the number of eggs females lay per clutch correlates with the volume of blood ingested and that protein is the most important macronutrient for egg formation. In contrast, how whole blood, blood fractions and specific blood proteins from different vertebrates affect egg formation is less clear. Since egg formation is best understood in Aedes aegypti, we examined how blood and blood components from different vertebrates affect this species and two others: the malaria vector Anopheles gambiae and arbovirus vector Culex quinquefasciatus. Methods Adult female mosquitoes were fed blood, blood fractions and purified major blood proteins from different vertebrate hosts. Markers of reproductive response including ovary ecdysteroidogenesis, yolk deposition into oocytes and number of mature eggs produced were measured. Results Ae. aegypti, An. gambiae and C. quinquefasciatus responded differently to meals of whole blood, plasma or blood cells from human, rat, chicken and turkey hosts. We observed more similarities between the anthropophiles Ae. aegypti and An. gambiae than the ornithophile C. quinquefasciatus. Focusing on Ae. aegypti, the major plasma-derived proteins (serum albumin, fibrinogen and globulins) differentially stimulated egg formation as a function of vertebrate host source. The major blood cell protein, hemoglobin, stimulated yolk deposition when from pigs but not humans, cows or sheep. Serum albumins from different vertebrates also variably affected egg formation. Bovine serum albumin (BSA) stimulated ovary ecdysteroidogenesis, but more weakly induced digestive enzyme activities than whole blood. In contrast, BSA-derived peptides and free amino acids had no stimulatory effects on ecdysteroidogenesis or yolk deposition into oocytes. Conclusions Whole blood, blood fractions and specific blood proteins supported egg formation in three species of anautogenous mosquitoes but specific responses varied with the vertebrate source of the blood components tested.

Haematologic and Biochemical Parameters of Haemogregarine-infected ‎and Non-infected African Hinge-Back Tortoises in Ibadan, Nigeria

This study was conducted to evaluate haematological and biochemical parameters of haemogregarine-infected (h-infected) and non-infected African hinge-back tortoises in Ibadan, Nigeria. Blood samples were collected from 120 tortoises, of which 70 were Kinixys belliana and 50 were K. homeana. Stained thin smears were examined for haemogregarines using light microscope. Haematological and biochemical analyses were carried out following standard procedures. A total of 91(75.83 %) tortoises were positive for haemogregarines. Significantly (P<0.05) lower values of haematocrit (23.92 %), haemoglobin (5.21g/dl) and mean corpuscular haemoglobin concentration (MCHC) (21.78 %) were recorded for h-infected tortoises with haematocrit (33.29 %), haemoglobin (8.31g/dl) and MCHC (24.96 %). Higher values of white blood cells (WBC) (7.26 x 109/L) and lymphocytes (2.71x109/L), were observed in h-infected than non-infected with WBC (5.58 x 109/L) and lymphocytes (2.15x109/L). Higher values of haematocrit and haemaglobin were recorded for K. Homeana. Males had higher haematocrit (27.27 %) and WBC (7.09 x 109/L) than females with haematocrit (24.35 %) and WBC (6.93 x 109/L). Females had higher MCHC, haemoglobin and calcium values than males.The lower values of haematocrit, haemoglobin and MCHC obtained for h-infected tortoises were expected since haemogregarines are usually found intra-erythrocytic in their host thereby destroying affected erythrocytes and causing a decrease in haematocrit value. Higher WBC counts in h-infected tortoises is typical in diseased conditions. The higher level of calcium in female tortoises is due to their reproductive cycle especially vitellogenesis and egg formation. Hypo-proteinaemia recorded in h- infected tortoises was attributed to parasitism. It is concluded that majority of haematological and biochemical analytes showed considerable variations with level of infection status, species and gender.

Extraordinary sperm to egg ratios in seabirds

Following copulation, females of many seabird species spend a prolonged period of time away from the colony, building up reserves for egg formation and incubation. Here, we report that the number of sperm associated with eggs of single-egg clutch seabirds was almost an order of magnitude greater than predicted from the relationship between ovum size and sperm numbers in multi-egg clutch non-seabirds. Sperm numbers were also several times greater than the estimated number necessary for maximal fertilization success. Our results are consistent with 3 unusual features of seabird reproduction: (1) single-egg clutches, (2) prolonged sperm storage, and (3) a lag period between the end of yolk formation and ovulation. We hypothesize that sperm release from storage is under precise temporal control in these species, with high sperm numbers acting as an insurance against infertility in single-egg clutches. If true, the lag period may have evolved to provide sufficient time for sperm to be released simultaneously from storage and accumulate at the site of fertilization prior to ovulation.