Application of Artificial Insemination in Selective Breeding of Native Beef Cattle in Wenzhou

[Objective]The paper was to improve the quality and growth rate of native yellow cattle in Wenzhou.[Method]Three groups of cattle were subjected to artificial insemination using frozen semen from Wagyu bull.The groups consisted of 20 native yellow cattle,20 Angus catle,and 20 Luxi yellow cattle.The heifers were subsequently evaluated for body size and body weight,and underwent treatment with estrus synchronization,artificial insemination,and early pregnancy diagnosis.[Result]The mean body size and body weight of native yellow cattle were found to be significantly lower than those of Angus cattle and Luxi yellow cattle(P<0.05).The estrus synchronization rate and mating rate of native yellow cattle were both 100%,while the corresponding rates for Angus cattle and Luxi yellow cattle were 90%and 95%,respectively.The ultrasound examination conducted on the 32m day following the mating revealed a distinct image of gestation sac,which,when considered alongside the findings of the rectal examination,may be indicative of pregnancy.The conception rates were 75%,72%,and 74%for native yllow cattle,Angus cattle,and Luxi yellow cattle,respectively.[Conclusion]The reproductive performance of native beef cattle is relatively superior.

Bacteriospermia among smallholder artificial insemination boars in the Philippines and potential associated factors

Objective:To determine the prevalence of bacteriospermia,the bacterial load,and the potential factors associated with bacterial contamination in boar semen collected by local smallholder artificial insemination operators.Methods:Fifteen individual raw semen samples were collected from locally available artificial insemination boars owned by different smallholder boar operators within the 5th district of Leyte,Philippines and were subjected to standard bacteriological culture and identification,including a survey of potentially associated factors.Prevalence and bacterial count were determined accordingly,while boar characteristics and collection practices were clustered following agglomerative hierarchical clustering technique.Results:One hundred percent contamination with a bacterial count of(2.01±0.38)×10^(3) CFU/mL was observed.At least 73.33%of the samples were positive for Bacillus spp.,while other identified isolates included Enterobacter spp.,Staphylococcus spp.,E.coli,Pseudomonas spp.,Citrobacter spp.,and Klebsiella spp.Conclusions:Despite the high prevalence of bacteriospermia,the bacterial count is low.Nevertheless,on-farm practices on boar health and management,semen collection,and sanitation as well as the enhancement of basic protocols to control contamination should be conscientiously considered in smallholder artificial insemination operation.

Study on Key Technique of Artificial Insemination for Silkworm,Bombyx mori

[Objective] The aim of this study was to investigate the efficient technique of artificial insemination for silkworm. [Method] Sperms were extracted from bursa copulatrix of female moths mated for 30 min through extruding and centrifugal method, and then the semen was injected into other virgin moths with trypsinase. [ Result] A high-effective collection technology of spermatids from silkworm was established successfully, 50 μl semen could be collected by only one person in each hour. The survival rate of spermatids was over 80% in vito after collected from bursa copulatrix, while the obtained semen was quite pure and the average fertilization rate of silkworm was 76,5%. [ Conclusion] The establishment of high-effective semen extraction technique of silkworm provides the technical basis for studies on other related techniques for silkworm sperm.

Efficiency of Artificial Insemination in Giant Pandas at the Wolong Breeding Center

The goal of the giant panda ( Ailuropoda melanoleuca ) breeding program is to develop a self sustaining,genetically diverse population.Due to the common problems about sexual incompatibility and a limited number of captive born males that breed naturally,artificial insemination (AI) has become a critical genetic management tool.It is common practice,however,to combine natural mating and AI using semen from non breeding males.From 1998 to 2000 at the Wolong breeding facility,12 of 18 (66.7%) females produced 20 cubs following combined natural mating and AI.The objective of this study was to determine the efficiency of AI without natural breeding.In 1998 and 2000,seven females were anesthetized for transcervical AI on two consecutive days.Ejaculates from six males were collected by electroejaculation,diluted in an egg yolk diluent containing 0% or 4% glycerol and used either fresh or following cold storage at 4℃ (for 24 or 48 h) or cryopreservation using the pellet freezing method.Mean (±SEM) ejaculate traits in six male sperm donors were:ejaculate volume,3.3±0.5 ml;sperm concentration,1,429.8±235.4×10 6/ml;sperm motility,81.7±2.1%;progression (0～5,5=best),3.1±0.1;and normal sperm,79.3±9.2%.For AI (n = 14) in seven females,mean inseminate traits were:spermic volume inseminated,2.4±0.3 ml;sperm motility,73.5±2.9%;progression,2.5±0.1;and total motile sperm inseminated/AI,684.2±118.2×10 6.Four of seven (57.1%) females became pregnant and produced five cubs of which four survived.Mean gestation and litter size was 131.5±9.7 days and 1.3±0.3 cubs/litter,respectively.These results indicate that the efficiency of AI is sufficient for recovering valuable genes from non breeding individuals to enhance genetic diversity in the ex situ population of giant pandas.

Artificial Insemination of Cherry Valley Ducks with Muscovy Ducks

[Objective] To explore artificial insemination technique for production of mule ducks. [Method] Female Cherry Valley ducks were artificially inseminated with semen collected from male Muscovy ducks by massage method and vagina-insemination method to investigate the effects of insemination dose, insemination interval, insemination time and diluents on fertilization rate. [ Result ] The average fertilization rate was only 39.58%, when the female Cherry Valley ducks naturally mated with the male Muscovy ducks. However, it was increased to 74.79% by artificial insemination. The fertilization rate was 75.24% after semen was diluted by PBS buffer, while it was 75.16% after semen was diluted by Lake＇s buffer; however, there was not significant difference between the fertilization rate of the diluted semen and that of fresh semen （74.10%）. After semen was respectively diluted by PBS buffer and Lake＇s buffer and then stored at 5 ℃ for 24 h, the fertilization rate was low, respectively 23.76% and 34133%. [ Conclusion] Artificial insemination technology can reduce insemination dose and increase fertilization rate in production of mule ducks.

Factors Influencing the Superovulation and Artificial Insemination of Sheep

[Objective] This study aimed to find out a method for low-cost and highly efficient sheep superovulation treatment and artificial insemination. [Method] The factors those probably influencing the results of conventional superovulation and insemination, such as combination of FSH hormone and sponge suppository, estrus interval, number of insemination, and ram individuals were analyzed. [Result] The combination of sponge suppository and FSH produced in Beijing exhibited the poorest effect to superovulation, significantly worse than that of other combinations （P0.01）. The FSH produced in Ningbo, combined with sponge suppository or CIDR produced better effect to superovulation. The superovulation effect was better when the interval from the last FSH injection to estrus was 12 h, significantly better than that when the interval was 36 h （P0.01）; and there was no difference in the superovulation results when the interval was 0, 12 and 24 h. The pregnancy rate of two artificial inseminations was significantly higher than that of only one insemination （P0.01）. Rams themselves had significant influence on fertilization results. [Conclusion] The combination of domestic FSH and domestic sponge suppository cost much less and dose not reduce the superovulation results. Better fertilization result can be obtained if the ewes are inseminated twice with the sperm those gave high pregnancy rate.

Lazzaro Spallanzani: pioneer of artificial insemination, multidisciplinary research, and scientific dissemination

Lazzaro Spallanzani(1729-1799)was a father of modern biology,with peculiar traits such as a multidisciplinary approach and penchant for scientific dissemination.Spallanzani consistently contributed to modern reproductive medicine by implementing experimental methods for the first historically successful artificial insemination.Nevertheless,he participated in discoveries pertaining to blood circulation,digestion,and respiration.Widely known in Europe in the eighteenth century,his fame prolonged to the following century,not exclusively through scientific acknowledgments,but even in literature.Nowadays,the figure of Spallanzani experience a kind of neglect and it would appear essential to maintain his work in the light of the history of medicine.

Effect of Uterine Anomalies on Pregnancy Rates and Reproductive Outcomes in Women Undergoing Artificial Insemination by Husband

Objective: Congenital uterine anomalies are common; however, their effects on artificial insemination by husband(AIH) and the period during which AIH is converted to in vitro fertilization(IVF) are unclear. We examined the influence of uterine malformations on reproductive outcomes following AIH and the optimum number of AIH cycles before resorting to IVF-embryo transfer(IVF-ET).Methods: We retrospectively recruited 168 patients with uterine malformations(anomalous group) undergoing AIH between January 2011 and December 2016. Meanwhile, 168 patients with infertility but with normal uteri(normal group) were matched as controls according to age.Results: The clinical pregnancy rate was similar in both groups(12.4% vs. 12.3%, P=0.950); the cancellation(21.6% vs. 4.4%, P< 0.001),early pregnancy loss(35.7% vs. 11.4%, P = 0.032), and preterm birth rates(21.4% vs. 2.9%, P = 0.038) were higher in the anomalous group,resulting in lower term birth(32.1 % vs. 74.3%, P =0.001) and live birth rates(50.0% vs. 77.1 %, P = 0.034). After two AIH cycles, the clinical pregnancy rate was lower(3.6% vs. 23.1%, P = 0.037) among women with uterine anomalies than among those with normal uteri. There was no difference in the pregnancy rates(52.5% vs. 53.7%, P= 0.908) between the two groups of patients with unsuccessful AIH who then underwent IVF-ET.Conclusions: IVF-ET can be performed immediately after two unsuccessful AIH cycles in patients with uterine malformations. In patients undergoing AIH or IVF, uterine malformations increase the risk of adverse obstetric outcomes.

The Impact of Seasonal Variation on Clinical Pregnancy and Live Birth Rates in Assisted Reproductive Technology: A Retrospective Cohort Study in Hainan

Objective:To investigate the influence of season on live birth and clinical pregnancy rates,as well as assisted reproductive technology(ART)outcomes,in the Hainan region.Methods:Patients were categorized into four groups based on the dates of artificial insemination and transplantation:spring,summer,autumn,or winter.The main outcome measures were clinical pregnancy rates and live birth rates.Secondary outcomes included body mass index(BMI),oocyte number,two pronuclei(2PN)cleavage rate,total gonadotropin(Gn)dosage and days,age,2PN fertilization rate,sperm concentration,sperm PR rate,anti-Müllerian hormone(AMH),and endometrial thickness.Outpatient semen quality indicators included sperm PR rate,total sperm count,sperm concentration,and total sperm motility.Results:This retrospective cohort study analyzed 2,016 artificial insemination cycles and 1,783 ovarian retrieval cycles from January 2017 to October 2022,and assessed the semen quality of 6,651 outpatients from May 2017 to October 2022.In artificial insemination cycles,sperm PR rate and clinical pregnancy rate were highest in winter,with a statistically significant difference between groups(P<0.05).Clinical pregnancy rate was influenced by both age and sperm PR rate(P<0.05).In ovarian retrieval cycles,the winter group had significantly higher clinical pregnancy,2PN fertilization,and 2PN cleavage rates than the other groups.The autumn group had higher live birth rates,though not significantly different.Additionally,winter months showed higher total sperm concentration and total sperm number compared to other seasons.Conclusion:Seasonality affected clinical pregnancy and live birth rates in artificial insemination cycles but not in ovarian retrieval cycles in the Hainan region.These findings suggest that while there is no need to choose a specific season for ovarian retrieval cycles,artificial insemination in winter may be preferable for patients.

Study on Artificial Propagation and Embryonic Development of Poyang Lake Catfish

Poyang Lake catfish （Silurus asotus Linnaeus） is native to Poyang Lake Basin. Artificial propagation of Poyang Lake catfish was carried out by using artificial insemination. Under water temperature of 18-26 ℃, embryonic development of Poyang Lake catfish in impounded water was observed and recorded in detail. Embryonic development of Poyang Lake catfish encountered seven stages, and it last- ed for 47.08 h from the fertilized egg stage to the stripping stage. The hatching time of Silurus meridionalis Chen was shortest, followed by that of Poyang Lake catfish, and the hatching time of Pelteobagrus vachelli was longest.

Harnessing male germline epigenomics for the genetic improvement in cattle

Sperm is essential for successful artificial insemination in dairy cattle,and its quality can be influenced by both epi-genetic modification and epigenetic inheritance.The bovine germline differentiation is characterized by epigenetic reprogramming,while intergenerational and transgenerational epigenetic inheritance can influence the offspring’s development through the transmission of epigenetic features to the offspring via the germline.Therefore,the selec-tion of bulls with superior sperm quality for the production and fertility traits requires a better understanding of the epigenetic mechanism and more accurate identifications of epigenetic biomarkers.We have comprehensively reviewed the current progress in the studies of bovine sperm epigenome in terms of both resources and biological discovery in order to provide perspectives on how to harness this valuable information for genetic improvement in the cattle breeding industry.

Assessment of the Quality of Frozen-Thawed Semen or Epididymal Sperm in Three Native Vietnamese Pig Breeds

Our aim was to evaluate the quality of ejaculated and epididymal frozen-thawed pig sperm of endangered Vietnam native pig breeds. Ejaculated sperm was collected from live boars and epididymal sperm was collected from slaughtered boars of the MuongTe, Kieng Sat and Co BinhThuan breeds and frozen in 0.25 ml straws using a protocol established earlier for modern pig breeds. We evaluated the sperm quality after thawing in terms of motility and rates of viable and abnormal spermatozoa. Our results revealed that the sperm motility and rates of viable and abnormal frozen-thawed sperm were >30%, >44%, and <14%, respectively. The origin of sperm had an effect on the production of pig embryos in vitro. In the Co BinhThuan breed, ejaculated sperm generated higher cleavage, blastocyst and hatching rates than did the epididymal sperm (60.11% vs 56.02%, 17.23% vs 14.31%, 3.78% vs 2.34%, respectively, P < 0.05). Although no difference in cleavage rate, blastocyst formation rate and the average number of cells/blastocysts, the hatching blastocyst rate was different between the breeds (P > 0.05). In the Co BinhThuan breed, the rate of pregnancy of ejaculated groups was similar to that of the epididymal group. In conclusion, the ejaculated and epididymal sperm of native Vietnamese pigs were successfully frozen. We succeeded in creating embryos in vitro and pregnant pigs after artificial insemination from frozen-thawed semen in three native Vietnamese pig breeds for the first time. The use of the ejaculated sperm improved the production of native pig embryos in vitro efficiency.

Sperm donation and its application in,China： a 7-year multicenter retrospective study

Sperm donation in China is different from that in other countries due to cultural, social and political factors. This research presents the current status of sperm donation in China's Mainland and highlights some problems. Between January 2003 and December 2009, 19 471 sperm donors were screened totally and 6467 donors （33.2%） were recruited. The primary reasons for non-recruitment were either inadequate semen parameters （55.0%） or positive results for sexually transmitted diseases （7.9%）. There were 327 （1.7%） qualified donors who withdrew from the program because of frustration related to failed semen parameters, participation merely for free medical tests or job transfer. A questionnaire investigating donor intention, as well as other concerns associated with sperm donation, was distributed to 516 potential donors. All potential donors indicated their primary motivation as altruism, while 90.9% mentioned monetary reward as a second motivating factor. Approximately 93.4% of donors expressed some apprehension about the risk of consanguineous mating and the protection of their identity. Over the past 7 years, 488 389 vials of donors＇ semen have been cryopreserved. In 36 438 artificial insemination with donor sperm （AID） cycles, the clinical pregnancy rate was 23.9% and the live birth rate was 16.6%, In 7148 in vitro fertilization cycles, the clinical pregnancy rate was 45.8% and the live birth rate was 35.2%. Human sperm banks have been strictly monitored to ensure that each sperm donor can only impregnate five women nationwide. There is still a large gap between the supply and demand for sperm donation which may be solved by updated guidelines.

Timing of IUI Treatment after hCG Administration 1-48h Affecting Pregnancy Rate

Objective To compare the different time of administration of hCG affecting pregnancy rate of IUI. Methods A total of 189 infertile couples underwent 331 cycles of lUI with husband＇s sperm. They were separated into 3 groups according to the time of hCG administration in IUI： hCG 1-23 h （group A）; hCG 24-36 h （group B）; hCG 37-48 h （group C）. Results There were no statistical differences among 3 groups. None of the other relative factors, such as the female age, the different methods of ovulation and the cause of infertility, showed differences in pregnancy rate among 3 groups. Conclusion IUI can be performed any time after administration of hCG （1-48 h）.

Oxytocin in pig seminal plasma is positively related with in vivo fertility of inseminated sows

Background:Identification of relevant in vivo biomarkers for fertility remains a challenge for the livestock industry.Concentrations of the small peptide hormone oxytocin(OXT),involved in male reproductive function and present in the seminal plasma(SP)of several species could be a robust one.This study characterized concentrations of SPOXT in ejaculates from boars used in artificial insemination(AI)programs aiming to evaluate its relationship with sperm quality variables and in vivo fertility of their liquid-stored AI-semen.Seminal OXT concentrations(ng/mL)were measured in 169 ejaculates from 61 boars of the Duroc,Pietrain,Landrace and Large White breeds using a direct competitive immunoassay test based on AlphaLISA®technology.Ejaculate(ejaculate volume,sperm concentration,total sperm count)and sperm parameters(motility,viability,intracellular generation of reactive oxygen species,plasma membrane fluidity)were assessed at 0 h and 72 h in AI-semen samples stored at 17℃.In vivo fertility included only 18 Large White and Landrace boars whose AI-semen was used to inseminated>100 sows and evaluated both farrowing rate and litter size of 3,167 sows.Results:The results showed that SP-OXT differed between boars and between ejaculates within boar(P<0.05)but not between breeds(Duroc,Pietrain,Landrace and Large White).Ejaculates with higher SP-OXT concentration/mL(hierarchically grouped;P<0.001)had larger volume and came from younger boars(P<0.05).Ejaculates of boars showing positive farrowing rate deviation exhibited higher(P<0.05)SP-OXT concentration/mL than those with negative farrowing rate deviation.Conclusion:The SP concentrations of OXT are boar,ejaculate and age dependent,and positively related with ejaculate volume and farrowing rates of liquid-stored semen AI-doses.

Current and future prospects for nanotechnology in animal production

Nanoparticles have been used as diagnostic and therapeutic agents in the human medical field for quite some time, though their application in veterinary medicine and animal production is still relatively new. Recently,production demands on the livestock industry have been centered around the use of antibiotics as growth promoters due to growing concern over microbial antibiotic resistance. With many countries reporting increased incidences of antibiotic-resistant bacteria, laws and regulations are being updated to end in-feed antibiotic use in the animal production industry. This sets the need for suitable alternatives to be established for inclusion in feed.Many reports have shown evidence that nanoparticles may be good candidates for animal growth promotion and antimicrobials. The current status and advancements of nanotechnological applications in animal production will be the focus of this review and the emerging roles of nanoparticles for nutrient delivery, biocidal agents, and tools in veterinary medicine and reproduction will be discussed. Additionally, influences on meat, egg, and milk quality will be reviewed.

A Comparison on Four Methods for Cryopreserved Spermatozoa

The relative merit of four different methods (method of fine tube, semen pellets, ampule, and test tube)of cryopreserved spermatozoa from 43 male donors with normal fertile ability was evaluated.The mobility and viability in post-thawed 172 samples were examined. The method of fine tube was recommedned to be the method of choice. A discussion on these four methods was given.

Factors Affecting Technology Adoption in Small Community Farmers in Relation to Reproductive Events in Tropical Cattle Raised under Dual Purpose Systems

The impact of technology adoption among farmers in developing countries has been an issue difficult to measure and interpret. Hence, research to improve production has been centered in technology that possible is not adequate for this region. The present review aims to utilize subjects such as Genetics, Reproduction, Nutrition, Management and Animal Health as examples of research endeavors illustrating the scarcely bond existing between research and adoption of technology. A final section is an attempt to illustrate the possible reasons why the transfer of technology is failing to be of any consequence at least under the conditions of developing countries.

Seasonal variation in semen quality of swamp buffalo bulls （Bubalus bubalis） in Thailand

Aim： To test the hypothesis that season affects the semen quality of swamp buffalo （Bubalus bubalis） bulls used for artificial insemination （AI） under tropical conditions in Thailand, as it does in Bos taurus and Bos indicus. Methods： Clinical and andrological examinations, and monitoring of semen production and quality were carried out on five mature, healthy swamp buffalo AI bulls in Thailand from July 2004 to the end of June 2005. Sperm output, motility, morphology and plasma membrane integrity （PMI） were compared between three seasons of the year （rainy, i.e. July-October; winter, i.e. November-February; and summer, i.e. March-June） with distinct ambient temperature and humidity. Results： All bulls were diagnosed as clinically healthy and with good libido throughout the study. Ejaculate volume, pH, sperm concentration, total sperm number and initial sperm motility did not differ between seasons, whereas PMI and the relative proportion of morphologically normal spermatozoa were highest in summer and lowest in winter （P 〈 0.05）. Buffalo age, week of collection and season influenced sperm morphology （P 〈 0.05-0.001）. Among morphological abnormalities, only proportions of tail defects were affected by season, being highest in the rainy season and lowest in summer （P 〈 0.001）. In conclusion, climatic changes did not seem to largely affect semen sperm output or viability. Although the proportions of PMI and tail abnormalities were affected by season, they were always below what is considered unacceptable for AI bull sires. Conclusion： Seasonal changes did not appear to cause deleterious changes in sperm quality in swamp buffalo AI-sires in tropical Thailand.

Metabolomic fingerprinting of pig seminal plasma identifies in vivo fertility biomarkers

Background:Metabolomic approaches,which include the study of low molecular weight molecules,are an emerging-omics technology useful for identification of biomarkers.In this field,nuclear magnetic resonance(NMR)spectroscopy has already been used to uncover(in)fertility biomarkers in the seminal plasma(SP)of several mammalian species.However,NMR studies profiling the porcine SP metabolome to uncover in vivo fertility biomarkers are yet to be carried out.Thus,this study aimed to evaluate the putative relationship between SPmetabolites and in vivo fertility outcomes.To this end,24 entire ejaculates(three ejaculates per boar)were collected from artificial insemination(AI)-boars throughout a year(one ejaculate every 4 months).Immediately after collection,ejaculates were centrifuged to obtain SP-samples,which were stored for subsequent metabolomic analysis by NMR spectroscopy.Fertility outcomes from 1525 inseminations were recorded over a year,including farrowing rate,litter size,stillbirths per litter and the duration of pregnancy.Results:A total of 24 metabolites were identified and quantified in all SP-samples.Receiver operating characteristic(ROC)curve analysis showed that lactate levels in SP had discriminative capacity for farrowing rate(area under the curve[AUC]=0.764)while carnitine(AUC=0.847),hypotaurine(AUC=0.819),sn-glycero-3-phosphocholine(AUC=0.833),glutamate(AUC=0.799)and glucose(AUC=0.750)showed it for litter size.Similarly,citrate(AUC=0.743),creatine(AUC=0.812),phenylalanine(AUC=0.750),tyrosine(AUC=0.753)and malonate(AUC=0.868)levels had discriminative capacity for stillbirths per litter;and malonate(AUC=0.767)and fumarate(AUC=0.868)levels for gestation length.Conclusions:The assessment of selected SP-metabolites in ejaculates through NMR spectroscopy could be considered as a promising non-invasive tool to predict in vivo fertility outcomes in pigs.Moreover,supplementing AI-doses with specific metabolites should also be envisaged as a way to improve their fertility potential.