Bio-economic and operational feasibility of introducing oestrus synchronization and artificial insemination in simulated smallholder sheep breeding programmes

Low selection intensity due to few selection candidates available at any one time due to thinly spread year-round lambings in villages and prohibitively large nucleus requirements to provide sufficient improved rams to the production tier are the major challenges for designing effective village-based and central nucleus-based breeding programmes, respectively, for smallholder sheep farmers. To tackle these challenges, we used deterministic simulation to design three schemes in village-based programmes introducing hormonal oestrus synchronization (natural oestrus (VNE), single oestrus synchronization (VSE1) and double oestrus synchronization (VSE2)) and three schemes in central nucleus programme introducing artificial insemination (AI) (natural mating with nucleus sizes of 5% (CNM1) and 1% (CNM2) of the total ewe population and natural mating in breeding tier and AI in production tier (CAI)). The schemes were evaluated for their bio-economic and operational feasibility, taking Bonga sheep of Ethiopia as a case study. The selection intensities achieved in VNE, VSE1 and VSE2 were 2.0, 2.3 and 2.4, respectively, for selecting rams for the breeding tier and 0.0, 0.8 and 1.0, respectively, for the production tier. The profits per ewe per year from VNE, VSE1 and VSE2 were Birr 12.2, 21.7 and 24.5, but the profit from VNE for the production tier was zero. CAI generated more genetic gains in the breeding objective (Birr 4.8) than CNM1 (Birr 2.5) and CNM2 (Birr 0.0) in the production tier. However, CAI was less profitable than CNM1 and CNM2. In conclusion, hormonal oestrus synchronization was found to be a feasible technological aide to accelerate genetic progress in village-based programmes. CNM1 and CNM2 could not be recommended as CNM1 requires large nucleus of 10 325 ewes and CNM2 results in zero genetic gain in the production tier. CAI could overcome the challenge in central nucleus programmes, namely unaffordable large nucleus, but the scheme needs to be subsidized by the public sector to be economically feasible for farmers.     

Fertilization environment of the non-copulating marine sculpin, Hemilepidotus gilberti

To clarify the extracellular environment for external fertilization in the non-copulating marine sculpin Hemilepidotus gilberti, sperm motility was measured in NaCl, KCl, mannitol solutions, seawater, and ovarian fluid. Spermatozoa of H. gilberti actively moved in seminal plasma the moment they were removed from the genital papilla. Spermatozoa showed higher motility in NaCl solution at osmolalities between 300–400 mOsmol kg^-1. In KCl and in mannitol solutions, spermatozoa actively moved at osmolalities between 500 and 800 mOsmol kg^-1, and at osmolality 300 mOsmol kg ^-1, respectively. The ovarian fluid was a transparent and viscous gelatinous material, rich in sodium with an osmolality of 340 mOsmol kg^-1. Sperm motility in the ovarian fluid lasted more than 90 min, which was six times longer than in seawater. This sperm motility under conditions isotonic to body fluid is similar to that of copulating marine sculpins rather than to other non-copulating marine fishes. In addition, eggs of H. gilberti could be fertilized in the ovarian fluid. This suggests that external fertilization takes place under physiological conditions similar to the internal conditions of the ovary provided by the ovarian fluid, which isolates the eggs from sea water for several hours after spawning. This manner of fertilization is thought to be one of the evolutionary pre-adaptations allowing copulation among marine sculpins.     

亲本死亡时间对近太湖新银鱼人工授精受精率的影响

银鱼捕起后易死,较难用活的亲本进行人工繁殖,在实际生产中,多采用死的亲本进行人工受精,亲本死亡的时间对卵的受精率影响较大。本文对徐家河水库近太湖新银鱼亲本死亡时间与卵受精率的关系进行了系列研究,亲本死亡后240分钟内,每隔15分钟进行一次人工受精,结果表明,卵的受精率随着亲本死亡时间的延长,其受精率越来越低,240分钟后,受精率趋于0。水温在15℃以下,亲本死亡105分钟后进行人工受精,平均受精率为43%,因此建议水温在15℃以下近太湖新银鱼的人工受精应在亲本死亡后105分钟之内进行。       

大熊猫繁殖生物学初步研究

近年来,在动物园内人工饲养下的大熊猫(Ailuropoda melanoleuc),由于雌雄个体常常不能同时发情,甚至都不发情,因而不能正常产仔。故大熊猫的繁殖问题就成为国内外各动物园所关注的问题之一。 北京动物园大熊猫于1963年自然交配产仔首次成功。1978年以后,国内外动物园又相继进行了上百次的人工授精试验,据报道,真正受孕产仔的仅有中国的北京、上海、成都、杭州以及西班牙的马德里和墨西哥等动物园。可见其受精率很低。     

Porcine sperm zona binding ability as an indicator of fertility

The escalated use of artificial insemination in swine has increased the importance of determining fertility of a semen sample before it is used. Multiple laboratory assays have been developed to assess fertilizing potential but they have yielded inconsistent results. This experiment sought to determine the relationship between in vitro competitive zona binding ability and in vivo fertility based on heterospermic inseminations and paternity testing. The zona pellucida binding ability and fertility of sperm from 15 boars was assessed by comparing sperm from one boar with sperm from other individual boars in a pairwise fashion using four ejaculates. The relationship of zona binding ability to the mean number of piglets sired per litter for each boar as well as historic fertility data (litter size and farrowing rate) was assessed. The in vitro competition assay consisted of labeling sperm from each boar of the pair with a different fluorophore and incubating an equal number of sperm from each boar in the same droplet with porcine oocytes. The competitive assay was highly effective in ranking boars by zona binding ability (R2=0.94). Paternity testing using microsatellite markers was used to determine the mean number of piglets sired per litter for each boar during heterospermic inseminations. The pairwise heterospermic insemination assay was effective in ranking boar fertility (R2=0.59). Using historical data from these boars, average litter size and farrowing rate were correlated (r=0.81, p<0.001). However, zona binding ability was not significantly correlated with historic farrowing rate data or historic average litter size. Boar sperm zona binding ability was also not correlated significantly with the mean number of piglets sired per litter following heterospermic insemination. But the number of piglets sired by each boar was related to a combination of zona binding ability, sperm motility, normal morphology, acrosomal integrity, and the presence of distal droplets (R2=0.70). These results suggest that zona binding ability is not an accurate predictor of fertilizing ability when used alone; however, when coupled with other sperm assessments, fertility may be predicted successfully.     

The influence of cysteine and taurine on microscopic-oxidative stress parameters and fertilizing ability of bull semen following cryopreservation

Oxidative stress significantly damages sperm functions such as motility, functional integrity, endogenous antioxidant enzyme activities and fertility due to lipid peroxidation induced by reactive oxygen species (ROS). The aim of this study was to determine the effects of antioxidants such as taurine and cysteine in Bioxcell^® extender on standard semen parameters, fertilizing ability, lipid peroxidation (LPO) and antioxidant activities comprising reduced glutathione (GSH), glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) after the cryopreservation/thawing of bull semen. Nine ejaculates for each bull were included in the study. Three groups, namely taurine (2 mM), cysteine (2 mM), and control, were designed to analyze the antioxidants in Bioxcell^®. Insemination doses were processed so that each 0.25-ml straw contained 15 × 10⁶ sperm.The addition of cysteine led to higher motility, compared to the other groups (P < 0.001). Cysteine showed a greater protective effect on the percentages of acrosome damage and total abnormalities in comparison to the other groups (P < 0.001). No significant differences were observed in hypo-osmotic swelling test (HOST), following supplementation with antioxidants during the freeze-thawing process. No significant difference was observed in non-return rates among groups. In biochemical assays, the additives did not show effectiveness on the elimination of malondialdehyde (MDA) formation and maintenance of GSH and GSH-Px activities, when compared to controls. CAT activity (35.1 ± 8.1 kU/g) was demonstrated to be significantly higher upon the addition of 2 mM taurine (P < 0.001), while the level of MDA increased, indicating oxidative stress in this group. SOD activity (21.4 ± 2.9 U/g protein) was significantly elevated in the group with cysteine, compared to the other groups (P < 0.001).     

Superovulation and embryo transfer in Holstein cattle using sexed sperm

The use of sexed bull sperm in multiple ovulation and embryo transfer (MOET) programs for Holsteins was evaluated for (1) heifers housed at a commercial embryo transfer (ET) facility (Experiments 1 and 2), and (2) heifers and cows on dairy farms (Experiment 3). In Experiment 1, superstimulated heifers were inseminated with 5 × 10⁶ sexed (X-sorted; n = 5) or unsexed (n = 5) frozen-thawed sperm from one bull at 12 and 24 h after estrus detection. No difference was observed in the rates of transferable embryos (53.4% vs 68.1%), degenerate embryos (24.8% vs 26.6%) and unfertilized ova (21.8% vs 5.3%) between sexed and unsexed sperm, respectively, except for the percent of female transferable embryos diagnosed by embryo sexing (100% vs 49.3%, P < 0.0001). In Experiment 2, donors were inseminated twice with 5 × 10⁶ sexed unfrozen sperm (n = 10) or sexed frozen-thawed sperm (n = 9). Embryo production rates for both treatments were similar to that observed on a commercial ET facility using unsexed sperm. Pregnancy rates for frozen-thawed embryos were similar for sexed and unsexed sperm (70.4% vs 72.4%, respectively). In Experiment 3, 99 flushes were conducted using sexed frozen-thawed sperm from nine bulls but an overall statistical analysis was not completed because the use of bulls was not balanced. However, for one bull with balanced usage, the rate of transferable embryos was higher in heifers than in cows (P < 0.05) inseminated twice with 5 × 10⁶ sperm/dose (10 × 10⁶ total). We concluded that the use of sexed frozen-thawed sperm (≥90% X-sperm biased and 10 × 10⁶ total sperm) may be economically viable for commercial MOET programs in Holstein heifers.     

Implementing artificial insemination as an effective tool for ex situ conservation of endangered avian species

Approximately 503 of the known species of birds are classified as ‘endangered’ or ‘critical’. Captive propagation programs have proven useful in maintaining genetic diversity and restoring wild populations of certain species, including the Peregrine falcon, California condor and Whooping crane. Artificial insemination (AI) has the potential of solving problems inherent to reproductive management of small, closed populations of endangered birds, including dealing with demographic instability, physical and behavioral disabilities, sexual incompatibility, lack of synchrony, and need to maintain gene diversity. In this review, we address the necessary methods and factors that allow AI to be applied effectively to manage rare bird populations. It is clear that semen availability and quality are the greatest limiting factors to implementing consistently successful AI for birds. Behavioral sensitivity to animal handling and the ability to minimize stress in individual birds also are keys to success. Multiple, deep vaginal inseminations can improve fertility, particularly when semen quality is marginal. Laparoscopic methods of semen transfer also have produced fertile eggs. All of these practices leading to successful AI remain dependent on having adequate basic knowledge on female reproductive status, copulatory behavior, endocrine profiles and duration of fertility, especially as related to oviposition. The overall greatest challenge and highest priority is defining these normative traits, which are highly species-specific.     

Effect of natural or vaccine-induced porcine circovirus type 2 (PCV2) immunity on fetal infection after artificial insemination with PCV2 spiked semen

The objectives of this study were to determine if vaccination against porcine circovirus type 2 (PCV2) or previous PCV2 infection of the dam are sufficient to prevent fetal infection when dams are artificially inseminated with PCV2-spiked semen. Nine sows (Sus domestica) were allocated into three groups of three dams each: The PCV2 naïve negative control Group 1 was artificially inseminated with extended PCV2 DNA negative semen during estrus, whereas the extended semen used in the vaccinated Group 2 (PCV2 vaccine was given 8 wk before insemination) and PCV2-exposed Group 3 (infected with PCV2 12 wk before insemination) was spiked with 5 mL of PCV2 inoculum with a titer of 10^4.2 tissue culture infectious dose (TCID₅₀) per milliliter at each breeding. The dams in the vaccinated and PCV2-exposed groups were positive for PCV2 antibody but negative for PCV2 DNA in serum at the time of insemination. Three negative control dams, two vaccinated dams, and three dams with previous PCV2 exposure became pregnant and maintained pregnancy to term. After artificial insemination, viremia was detected in one of three vaccinated dams and in two of three dams with previous PCV2 exposure. At farrowing, PCV2 infection was not detected in any piglets or fetuses expelled from the negative control dams or from dams with previous PCV2 exposure. In litters of the vaccinated dams, 15 of 24 live-born piglets were PCV2 viremic at birth, with 6 of 26 fetuses having detectable PCV2 antigen in tissues. In conclusion, vaccine-induced immunity did not prevent fetal infection in this sow model using semen spiked with PCV2.     

Sex-sorting sperm by flow cytometry in pigs: issues and perspectives

Several hundred thousand offspring of preselected sex of various species have been born since sperm sexing technology based on flow cytometric sorting of X- and Y-chromosome-bearing sperm and DNA was first demonstrated in 1989. The advantages derived from application of sexing technology to commercial dairy cattle production have been demonstrated worldwide. Utilizing sex-sorting technology for pig production systems offers many similar advantages. However, several factors currently limit implementation of sexing technology in pigs. Anatomical and physiological features inherent to the female pig, together with the relatively low sperm output of a flow sorter, are the main limitations to widespread use of this technology in pig production systems. This review analyzes the factors that limit the efficiency of sperm sorting technology for commercial swine production. In addition, this review discusses recent innovations in technical instrumentation and applied reproductive techniques that may help to overcome some of these limitations.