Animal reproduction science

Publisher: Elsevier


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    ABSTRACT: A total of 150 does of different breeds were used to evaluate the accuracy of transrectal (TRU) and transabdominal ultrasonography (TAU) for pregnancy diagnosis and fetal number estimation and to study the effect of breed and age of goats on the accuracy of ultrasonography. All does were scanned by TRU at days 17-22, 24-29 and 31-36 and by TAU at days 39-44 and 46-51 after breeding. The sensitivity (Se) of TRU for detecting pregnant does increased significantly (86.1% vs. 97.7%; P< 0.0005) from days 17-22 to days 24-29. There was a significant (P< 0.05) effect of breed of goats on the specificity (Sp) of TAU for diagnosing non-pregnancy between days 39 and 51. In addition, there was a significant (P< 0.01) effect of breed of goats on the Se and Sp of TRU for determination of does carrying multiples and singles, respectively between days 24 and 36. The sensitivity of TAU for diagnosing pregnant does tended to be significantly (P= 0.06) higher in young does (1.5-2 yr) than in older does (>2-9 yr) at days 39-44. The accuracy of TRU for determination of does carrying singles was significantly (P< 0.05) higher in young does than that in older ones at days 24-29. In conclusions, TRU could be accurately used for early pregnancy diagnosis and fetal number estimation at days 24-29 and days 31-36, respectively with accuracy similar to that of TAU at days 39-51. Furthermore, breed and age of does might influence the accuracy of ultrasonography for pregnancy diagnosis and fetal numbers estimation.
    Animal reproduction science 06/2014;
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    ABSTRACT: The molecular mechanisms operating on a seasonal time-scale and regulating functions such as reproduction are poorly understood in animals living in desert environments. Kisspeptin, the product of the KiSS-1 gene, plays a critical role in control of the hypothalamic-pituitary-gonad axis that orchestrates the reproductive system in vertebrates. We report a sequence analysis of KiSS-1 and the pituitary luteinising hormone-beta (LHβ) and follicle-stimulating hormone–beta (FSHβ) in the Libyan jird (Meriones libycus), a seasonal breeding rodent that is sexually active during spring and quiescent in fall. We also assessed gene expression by quantitative real-time reverse transcription-polymerase chain reaction during the non-breeding and breeding seasons. The KiSS-1 cDNA sequence analysis showed high homology between M. libycus and all other rodents (94%) and humans (92%). KiSS-1 expression was higher during the breeding season than that during the non-breeding season. In contrast, LHβ and FSHβ expression levels were higher during the non-breeding season in autumn and varied in an opposite manner with testicular, seminal vesicle weights and plasma testosterone levels. Our results extend the role for KiSS-1 in activating the HPG axis in this desert rodent in its natural biotope by relaying environmental cues as in other seasonal non-desert rodent models.
    Animal reproduction science 06/2014;
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    ABSTRACT: Deleted in azoospermia-like (DAZL) is used as a germ cell marker in several species, including mice, rats, pigs, rhesus monkeys, bulls, and humans. Our objectives with this study were to investigate DAZL expression in stallion germ cells by using immunofluorescence, immunocytochemistry, and western blotting, and to determine the effects of reproductive stage and breeding season on the DAZL-positive cell population in seminiferous tubule cross sections. Testes were obtained during routine castration procedures at a large animal clinic and routine field service castration. The reproductive stage of the stallions was classified as pre-pubertal (<1 yr), pubertal (1–1.5 yr), post-pubertal (2–3 yr), or adult (4–8 yr). Using immunofluorescent staining, we showed that DAZL is localized to the cytoplasm of some, but not all, spermatogonia in pre-pubertal and pubertal horses. In the post-pubertal and adult testes, DAZL immunostaining was observed in spermatogonia proximal to the basement membrane of seminiferous tubules; however, few spermatogonia attached to the basement membrane were not immunolabeled. DAZL immunostaining was also observed in primary spermatocytes, but not in secondary spermatocytes, spermatids, or spermatozoa. DAZL protein was not detected in Leydig, Sertoli, or myoid cells of the testes at any reproductive stage. The immunocytochemistry analysis showed that DAZL immunolabeling was also localized to the cytoplasm of isolated germ cells such as spermatogonia or primary spermatocytes. We conclude that DAZL can be used as a marker of pre-meiotic germ cells in stallions.
    Animal reproduction science 06/2014;
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    ABSTRACT: Avian spermatozoa are characteristically high in polyunsaturated fatty acids which predispose them to lipoperoxidation, thereby diminishing their fertility. As a by-product, well-known for antioxidative properties, dried apple pomace (AP) was fed to aging Ross 308 breeder roosters (n = 80) at the dietary inclusion levels of 0 (AP0), 10 (AP10), 20 (AP20), or 25 (AP25) percent for 14 consecutive weeks. Seminal traits were studied every other week. At the end of the trial, sperm fatty acids profile, seminal plasma total antioxidant capacity (TAC), fertility, hatchability, and hatchling quality were evaluated, using 240 artificially-inseminated young hens. Dietary AP increased sperm motility and livability and decreased the seminal content of thiobarbituric acid reactive species. Dietary treatment and age interacted to positively affect sperm concentration and sperm membrane integrity. Feeding AP was associated with an increase in C20:4(n-6), C22:4(n-6), and total unsaturated fatty acids percentages. Birds in the AP25 and AP20 treatment groups respectively showed 6 and 7% increases in fertility rate. Interestingly, a higher hatchability rate was found for AP20 group, associated with a higher number of sperm penetration holes in the perivitelline membrane and a lower rate of early embryonic mortality. However, hatchling quality was not affected by dietary AP. Overall, these data suggest that AP could remarkably improve several sperm characteristics, seminal TAC, fertility, and hatchability rate in aging breeder roosters. These improvements were also associated with a higher content of total unsaturated FA in the sperm plasma membrane. Future studies are needed to disclose the causal mechanisms involved.
    Animal reproduction science 06/2014;
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    ABSTRACT: The enzyme 20α-hydroxysteroid dehydrogenase (20α-HSD) catalyzes the conversion of progesterone to its inactive form, 20α-hydroxyprogesterone, and this enzyme has an important role in the regulation of luteal function in mammals. It has previously been determined that the 20α-HSD gene is primarily expressed by large luteal cells during the late stage of the estrous cycle. In the present study, the amounts of mRNA were determined in cultured cells of the corpus luteum (CL) cells. The localization of 20α-HSD was also determined in ovaries, placenta, and endometrium during early pregnancy. The amount of 20α-HSD mRNA in cultured luteal cells increased with time and by treatment with the luteolysis agent prostaglandin F2α (PGF2α). Immunofluorescence assays detected increased protein in cultured luteal cells. The 20α-HSD mRNA and protein were presnet in the ovaries, placenta, and endometrium on Days 30, 60, and 90 of pregnancy. In particular, gene expression was much greater in the ovary than in the placenta and endometrium. Immuno-histochemical analysis indicated that bovine 20α-HSD was primarily localized in ovarian large luteal cells, placental cytotrophoblast villus, and glandular epithelial cells of the endometrium during early pregnancy. Furthermore, in situ analyses demonstrated colocalization of 20α-HSD mRNA and protein. Taken together, results of the present study indicate that 20α-HSD mRNA and protein are co-localized in large luteal cells, the placenta, and the endometrium during early pregnancy, suggesting that 20α-HSD regulates mechanisms involved in the maintenance of early pregnancy.
    Animal reproduction science 06/2014;
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    ABSTRACT: Epigenetic instability of donor cells due to long-term in vitro culture may influence the success rate of subsequent somatic cell nuclear transfer (SCNT). Therefore, the present study was designed (1) to investigate the epigenetic changes after prolonged culture in vitro of porcine embryonic germ (EG) cells, including differences in expression levels of both DNA methylation and demethylation-related genes and catalyses of histone modifications, and (2) to assess the efficiency of SCNT using EG cells from different passages. Results showed that genes either associated with DNA demethylation including DNMTs and TET1 or genes related to histone acetylation including HDACs were highly expressed in EG cells at higher passages when compared to EG cells at lower passages. In addition, the expression level of H3K27me3 functional methylase EZH2 increased while no changes were observed on H3K27me3 demethylase JMJD3 in relation to passage number. Moreover, the expression levels of both the H3K4me3 methylase MLL1 and the H3K4me3 demethylase RBP2 were increased at high passages. By using lower passage (numbers 3-5) EG cells as donor cells, the SCNT efficiency was significantly lower compared with use of fetal fibroblast donor cells. However, similar blastocyst rates were achieved when using higher passage (numbers 9-12) EG cells as donor cells. In conclusion, the present study suggests that the epigenetic status of EG cells change with increasing passage numbers, and that higher passage number EG cells are better primed for SCNT.
    Animal reproduction science 06/2014;
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    ABSTRACT: Due to the scarcity of information about the reproduction of bats, it is necessary to perform studies on different species to identify the occurrence of annual variations in their reproductive process. Therefore, the aims of this study were to describe and quantify the intertubular components within the testes of the bat Sturnira lilium and to verify whether seasonality takes place in spermatogenesis of this species. The animals were collected in different seasons and time of collection was grouped into dry and rainy seasons. Testicular fragments were routinely processed for light and transmission electron microscopies and blood samples from each animal were collected for quantification of plasma testosterone. Overall, the Leydig cells (LC) were the main intertubular component (83.2%), with abundant lipid droplets in their cytoplasm. Ultrastructural analysis indicated collagen fibers in the connective tissue and lymphatic spaces, with thin walls, surrounding the seminiferous tubules. The proportion and volume of each intertubular component did not vary significantly between seasons. On average, testosterone concentrations did not vary between rainy and dry seasons (21 ng/ml). The total number of LC in the testis (50.0 × 105) and per gram of testis (11 × 107) did not vary nor did the Leydigosomatic Index (0.03%). Therefore, it is concluded that S. lilium had significant development in intertubular tissues, especially in LC. Most of the variables that were assessed were did not vary with season of the year, which leads to the assumption that S. lilium has a continuous reproductive cycle in southeastern Brazil.
    Animal reproduction science 06/2014;
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    ABSTRACT: Present study was undertaken to characterize the presence of histamine H1R using molecular biology tools and unravel the influx and release mechanism(s) involved in calcium signaling cascades in histamine-induced myometrial contraction in buffaloes. Presence of H1R mRNA transcript and immunoreactive membrane protein in buffalo myometrium was confirmed by RT-PCR and Western blot analysis. Further, histamine produced concentration-dependent (1 nM - 10 μM) contraction in buffalo myometrium with potency of 7.13 ± 0.11. When myometrial strips were pre-incubated either with Ca2+ free solution or with nifedipine, a L-type Ca2+ channel blocker, dose response curve (DRC) of histamine was significantly (P < 0.05) shifted towards right with decline in maximal contraction (Emax). Reduction in Emax of histamine in the presence of nifedipine (55.75 ± 3.10%) was significantly (P < 0.001) greater than in the presence of ruthenium red (93.61 ± 3.43%), a blocker of IP3-gated and RyR-sensitive Ca2+-channels. Moreover, histamine produced only 26.87 ± 1.99% of the maximum contraction in the resence of both nifedipine and CPA (blocker of sarco-endoplasmic reticulum Ca2+-ATPase). Interestingly, following concurrent exposure to U-73122 (a PL-C inhibitor) and nifedipine, the DRC of histamine was significantly (P < 0.05) shifted towards left with increase in maximal contraction (126.30 ± 3.36%). Our findings in buffalo uterus thus suggest that influx of extracellular calcium plays a major role in histamine–induced myometrial contraction while release of intracellular calcium through calcium-release channels of sarcoplasmic reticulum have minor role. Although possible involvement of non-selective cation channels in histamine-induced myometrial contraction cannot be ruled out, but it requires further investigations.
    Animal reproduction science 05/2014;
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    ABSTRACT: The aim of this study was to investigate the clinical relevance of measuring blood concentrations of serum amyloid A (SAA), haptoglobin (Hp) and fibrinogen (Fib) in horse reproductive management, and changes in response to artificial insemination (AI) with frozen-thawed semen. Standardbred mares (n = 18) with different reproductive status (eight healthy mares in first postpartum oestrus, five healthy barren mares and five mares with endometritis) were inseminated with frozen-thawed semen. Endometritis was evaluated during oestrus by bacteriological culture, cytology and presence of ultrasonically visible intrauterine fluid during oestrus. Concentrations of SAA, Hp and Fib were analysed in the blood in every 48 hours during oestrus and until 5, 6 or 7 days after AI. The day of sampling and number of blood samples varied between mares because of length of the oestrus and time of AI. Changes in concentrations of SAA, Hp and Fib were evaluated based on the day of sampling regard to AI and classification of the mares. There were no differences in SAA, Hp and Fib concentrations over time before or after AI or between the groups of mares. The insemination of mares with frozen-thawed semen did not increase the plasma concentrations of SAA, Hp and Fib above clinical threshold concentration and there were no differences between susceptible or healthy mares.
    Animal reproduction science 05/2014;
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    ABSTRACT: Determining the volume of an ejaculate is an important part of processing semen in bovine AI laboratory practice. A multi AI laboratory study was performed to estimate the density of whole bull semen, and whether the use of this value as a standard is suitable for practical use when semen of different breeds is processed at different AI laboratories. The density of whole bull semen had been determined for 90 ejaculates at five AI laboratories (five breeds). The results showed no effect of bull (p = 0.766), breed (p = 0.279) and laboratory (p = 0.183). All duplicate measurements within the same sample were within level of agreement (5%). Using the mean value of 1.053 g/ml as a golden standard for the density of whole bull semen is therefore suitable for use in routine bovine AI laboratory practice.
    Animal reproduction science 05/2014;
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    ABSTRACT: The objective of this study was to evaluate the effect of equine chorionic gonadotropin (eCG) administration associated to fixed–time AI (FTAI) on follicular dynamics, ovulation, corpus luteum (CL) development and serum progesterone concentrations. Multiparous suckled Hereford cows (n = 46) in anestrus with 60 to 75 days postpartum were used. Females received an intravaginal device containing 0.5 g of progesterone during 8 days and 2 mg of estradiol benzoate i.m. at device insertion. At device removal 500 μg of cloprostenol and 0.5 mg of estradiol cypionate were administered i.m., and FTAI was performed 52 to 56 hours later. Cows were divided into two experimental groups to receive 400 IU of eCG i.m. at device removal (n = 23), while control group did not receive eCG (n = 23). Daily ovarian ultrasonography (7.5 MHz transducer) and progesterone concentrations determined by RIA were assayed from device removal until 30 or 14 days after FTAI, respectively. Treatment with eCG increased ovulation rate [65.2% (15/23) vs. 30.4% (7/23); P = 0.018], ovulatory follicle diameter (14.5 ± 0.4 mm vs. 13.1 ± 0.7 mm, mean ± SEM; P = 0.081), CL area from days 6 to 14 after FTAI (344.3 ± 25.1 mm2 vs. 274.2 ± 23.9 mm2; P = 0.045) and mean serum progesterone concentrations from FTAI to 14 days later (3.0 ± 0.2 ng/ml vs. 1.8 ± 0.2 ng/ml; P = 0.001), in comparison with control cows. In conclusion, the addition of eCG to a progesterone and estradiol’ based treatment for FTAI improves ovulation rate and luteal function in anestrous cows. These findings have implications in order to increase pregnancy rates in FTAI treatments in Bos taurus beef cattle.
    Animal reproduction science 05/2014;