Effective bovine artificial insemination (AI) training requires balancing technical skill development with animal welfare considerations. Commercial simulators typically replicate Bos taurus anatomy, limiting utility in regions where Bos indicus breeds predominate. This study validates the TrAI4Nel simulator, customized for Nelore cattle AI training. Validation employed a multi-dimensional framework encompassing face, physical, content, construct, and concurrent validity, plus usability and training effectiveness assessments. Of the 85 participants in standardized AI technician courses who were randomly allocated to control (abattoir specimen-based) and experimental (simulator-integrated) groups, 61 provided feedback about TrAI4Nel (19 in the control group and 42 in the experimental group). The simulator was also independently evaluated by 14 AI experts. Trainees rated the simulator highly for anatomical realism and procedural consistency. Compared with abattoir specimens, TrAI4Nel significantly enhanced skill transfer to live animals, particularly cervical pipette navigation and semen deposition identification. The simulator increased trainee confidence, reduced anxiety, and improved perceived preparedness. Performance assessments demonstrated simulator-trained participants achieved significantly higher success rates (78.6%) versus controls (52.6%; p = 0.043), without prolonging completion times. Qualitative feedback emphasized the simulator's pedagogical value in enhancing anatomical comprehension, skill acquisition, and learner autonomy while supporting animal welfare. Training sequence analysis indicated biological specimen exposure before simulator use may optimize learning efficiency. These findings validate TrAI4Nel as an effective, ethically sound tool for Bos indicus AI training. Simulator integration into curricula provides scalable improvement of reproductive management in zebu-dependent regions.

Efficient use of frozen-thawed semen in sheep is limited by low fertility following cervical insemination (CAI), whereas laparoscopic insemination (LAI) offers higher success but requires specialised skills and equipment. The current study aimed to evaluate the fertility outcomes of two artificial insemination techniques, laparoscopic and transcervical, in Lohi sheep using frozen-thawed semen. Lohi ewes (n = 44) were synchronised through progestin-sponges (day 0-day 11), administering PGF2α (d-cloprostenol 75 μg/mL; i.m @ 75 μg/ewe) on day 09 and eCG (i.m @ 300 IU/ewe) on day 11. Synchronised ewes (n = 44) were divided into two groups for artificial insemination (AI) procedures: cervical (CAI; n = 24; sperm concentration = 200 million/dose) and laparoscopic (LAI; n = 20; sperm concentration = 50 million/dose). All ewes were inseminated at 54 h post-sponge removal. Results of the experiment revealed that LAI demonstrated higher pregnancy rates (65%) than CAI (33.3%) (p = 0.05). However, no significant differences were observed in prolificacy or twinning rates between the two techniques (p = 0.11). In conclusion, LAI as compared to CAI is a superior method of insemination for achieving optimum pregnancy rates in estrous synchronised Lohi ewes.

The cryopreservation of Chengde polled goat semen plays a critical role in conserving genetic resources, enhancing the utilization efficiency of superior breeding bucks, and advancing artificial insemination techniques. However, spermatozoa are vulnerable to oxidative stress during the freezing process, which can significantly compromise sperm motility. In this study, pooled ejaculates from multiple bucks were divided into six groups, including a control group cryopreserved with conventional extender and five treatment groups supplemented with sericin at concentrations of 0.2%, 0.4%, 0.6%, 0.8%, and 1.0% (w/v). The results demonstrated that supplementation of the semen cryoprotectant with 0.6% sericin significantly improved post-thaw sperm viability to 65.25% in Chengde hornless goats, while concurrently reducing both the sperm abnormality rate (p < 0.05) and intracellular ROS levels (p < 0.05). Integrated TMT proteomics and LC/MS metabolomics further compared the 0.6% sericin group with the frozen control group and identified 162 differentially expressed proteins and 109 differential metabolites between the sericin supplementation and frozen control groups. Functional analysis revealed the significant enrichment of differential metabolites, such as glutamine, in the alanine, aspartate, and glutamate metabolism pathway, concomitant with the marked upregulation of antioxidant proteins including LRP8, GSTM3, and SIRT2. Thus, 0.6% sericin enhances cryotolerance primarily by improving sperm viability, reducing oxidative damage, and sustaining energy metabolism. These findings indicate that sericin enhances cryotolerance by reducing oxidative damage and supporting metabolic function, providing preliminary molecular insights for improving goat semen cryopreservation.

ABSTRACT Laparoscopy is the preferred technique for artificial insemination (AI) with frozen semen in sheep, given the ovine cervix’s anatomical challenges. This study assessed if and when laparoscopic AI alters stress and inflammation markers. Ewes underwent estrus synchronization and were divided into Control (hormonal protocol only; n=4) and LP (laparoscopic AI 54-60 h post-protocol; n=5) groups. Blood samples were collected at D-1 (pre-fasting), 0 h (during AI), 30 min, 60 min, 24 h, and 48 h post-procedure to measure serum cortisol, total protein (TP), albumin, and paraoxonase 1 (PON1). LP ewes showed elevated cortisol (p=0.01) at 0 h and 48 h compared to Controls. Albumin decreased acutely in LP at 0 h, 30 min, and 60 min. PON1 activity and TP remained unchanged, though data variability suggests larger samples for future validation. In conclusion, laparoscopic AI modulates cortisol and albumin, highlighting the need for strategies to reduce procedure-induced stress and inflammation, which can impair reproductive performance.

This study aims to evaluate the fertility and hatchability of Nunukan chickens responding to the rooster semen collected using the dorso-abdominal massage method and a collection tube by presenting a teaser. A total of 20 male Nunukan chickens were divided into two treatment groups, in healthy condition, aged 12-36 months, body weight 2.5-3.4 kg, and housed individually (35x45x60 cm3); the semen was collected every three days. Semen characteristics evaluated macroscopically included color, odor, consistency, volume, and pH; microscopically included sperm concentration, mass motility, progressive individual motility, total sperm per ejaculate, viability, abnormality, and sperm quality factor (SQF); furthermore, the artificial insemination technique was used to evaluate the effect of sperm quality on fertility and hatchability. Both methods showed no difference in macroscopic parameters; color, odor, consistency, and pH, and microscopic parameters (p&gt;0.05); concentration, viability, abnormality, mass motility, and progressive individual motility. While semen volume, total sperm per ejaculate, and SQF showed significant differences (p&lt;0.05). However, the fertility and hatchability rates are not significantly different (p&gt;0.05). Despite the high quantitative qualities of rooster semen obtained with collection tubes and female teasers, Nunukan chickens' fertility and hatchability did not differ significantly (p&gt;0.05) from other semen collection methods. Keywords: collection method; fertility; hatchability, semen, quality

This experiment was conducted to evaluate the effects of dietary vitamin E supplementation in female rabbits on reproductive performance and economic efficiency, monitored across three consecutive litters using artificial insemination. The experiment was arranged in a completely randomized design with 32 crossbred doe rabbits (New Zealand White × local), aged 8-8.5 months, with an average body weight of 2.693 ±115 g, and they were divided into 4 treatments, each consisting of 8 replicates. The diet of all rabbits in the experiment was the same, contained 21% crude protein (CP) and 10.5 MJ/KgDM of metabolizable energy (ME). The experimental treatments were as follows: Treatment 1 served as the control, without vitamin E supplementation (E0), while the other treatments included dietary supplementation with vitamin E at levels of 40 mg/kg DM (E40), 80 mg/kg DM (E80), and 120 mg/kg DM (E120). Artificial insemination was used for all experimental rabbits. The experiment results showed that the E80 performed better than the others in terms of conception rate, mean weight at birth, milk yield of doe, mean weight at weaning (P&gt;0.05), and produced the highest economic efficiency.

Given the potential of artificial insemination (AI) to enhance goat productivity, this study conducted a strategic assessment of AI adoption among goat farmers in Kamrup district, Assam, covering both rural and urban subdivisions. A total of 120 goat farmers (60 each from Kamrup Rural and Kamrup Metro) were selected using stratified random sampling to ensure diverse representation. Data were collected through a structured, pre-tested interview schedule, with goat farmer perceptions captured under a SWOT framework. Analysis was performed using Mean Percent Score (MPS), Z-score, and percentile ranking to prioritize the top five factors under each SWOT category. Key strength identified included favourable agro-climatic conditions, presence of indigenous breeds, traditional knowledge, herd health benefits, and available institutional support. Major barriers to adoption were limited awareness of AI, cultural resistance, shortage of trained AI technicians, distant AI service centres, and low conception rates. Opportunities emerged in the form of increasing demand for goat meat and milk, prospects for genetic improvement, rising youth interest in livestock-based livelihoods, entrepreneurial potential, and the relevance of climate-resilient breeding programs. Notable threats included poor AI accessibility in remote areas, heightened disease risk, climate-related stress, and complications such as dystocia. Based on the SWOT outcomes, a TOWS matrix was formulated to develop actionable strategies such as mobile AI delivery units, farmer-led awareness campaigns, capacity building, and integration of veterinary and extension services. The study offers practical insights to guide sustainable AI adoption in goat farming across Kamrup, Assam.

In vitro embryo production is critical for accelerating genetic improvement in livestock. Nigerian cattle breeds were not exposed to assisted reproductive technology other than artificial insemination techniques. The efficiency of producing embryos using in vitro technologies rarely exceeds the 40% threshold, indicating that the proportion of oocytes that fail to develop after in vitro fertilization and culture is significant. The aim of this study was to determined oocytes collection and grading. Seventy-five bovine ovaries were collected from Sokoto Modern Abattoir and transported in a thermos container maintained at 37-38oC and processed in the laboratory. The ovaries were categorized based on the presence or absence of corpus luteum. The oocytes were recovered using aspiration technique. The oocytes transferred in minimum essential medium (MEM) and were graded using stereomicroscope based on cumulus investment and the nature of the cytoplasm into four grades: (I-IV). Greater numbers of oocytes were aspirated from ovaries without corpus luteum (CL- ovaries) 156 cumulus oocytes complex than those with corpus luteum (CL+ ovaries) (60 COCs). The CL- ovaries contributed higher normal oocytes (Grades I and II) than that of CL+ ovaries. It is highly recommended that ovaries without corpus luteum should be utilized for in vitro oocytes recovery and possible embryo production.

BackgroundDespite significant advancements in artificial insemination techniques, male reproduction continues to pose a considerable challenge in cattle breeding. Circular RNAs (circRNAs), a class of non-coding RNAs (ncRNAs), play a crucial role in regulating testis growth and spermatogenesis. Therefore, it is essential to comprehend the involvement of circRNAs in bull reproduction for livestock production. However, the identification of differentially expressed circRNAs during testis development and their underlying mechanisms remains largely unknown.ResultsIn this study, RNA-seq was employed to investigate the expression of circRNAs in neonatal and sexually mature bovine testes. We identified 28,065 candidate circRNAs, of which 987 circRNAs showed differential expression between the two stages (P-adjust < 0.05). Notably, circSMC1B was significantly up-regulated in sexually mature testis. Overexpression of circSMC1B promoted the proliferation and apoptosis of bovine male germline stem cells (mGSCs). Further analysis revealed that circSMC1B acts as a molecular sponge for let-7i, while High mobility group AT-hook 1/Nuclear receptor subfamily 6 group A member 1 (HMGA1/NR6 A1) were identified as direct targets of let-7i. Furthermore, circSMC1B levels exhibited a significant positive correlation with HMGA1/NR6A1 mRNA expression in bovine mGSCs, highlighting the critical role of competing endogenous RNA (ceRNA) mechanisms.ConclusionOur research elucidates that circSMC1B, through let-7i binding, promotes bovine mGSCs proliferation and apoptosis by targeting HMGA1/NR6A1. These findings provide valuable resources for studying the functional aspects of circRNAs in testis development and enhance our understanding of the biological function of circSMC1B in promoting bull spermatogenesis.

Beekeeping is one of the oldest and most profitable sectors of agriculture. Since ancient times, humans have bred, propagated, and maintained honey bees (Apis mellifera L.). The favorable soil and climatic conditions, along with the rich flora of Azerbaijan, create an ideal environment for the development of beekeeping. One of the challenges in bee breeding research is that honey bees mate in the air, often at considerable distances from their colonies. This complicates controlled breeding efforts. To address this issue, selection specialists are exploring artificial insemination techniques and the establishment of controlled mating zones. This article examines the development of beekeeping in Azerbaijan, with a particular focus on honey production in the Western region. Additionally, it explores the most effective modern approaches to bee breeding, especially in the context of increasing resistance to adverse environmental factors, which has become a critical priority in contemporary apiculture.

Biodiversity is increasingly threatened by intensive agriculture, environmental pollution, climate change, and habitat loss, and many mammal species, including ruminants, have disappeared or are threatened with extinction. Therefore, reproductive biotechnologies represent an important alternative for the conservation of endangered species, being fixed-time artificial insemination (FTAI) the most widely used method, which has been extrapolated from livestock to wild ruminants. The main benefit of artificial insemination is the maintenance of the genetic diversity of populations through the preservation and use of semen from genetically valuable individuals. Variables, such as hormones and semen condition, can modify the efficiency of FTAI in domestic and wild animals. The aim of this review was to evaluate the different protocols and variations that have been reported in FTAI applied to different species of wild ruminants.

Ethiopia's genetic enhancement efforts have included directly importing exotic cattle from other countries or introducing genes from an external source via artificial insemination (AI) to enhance the breed composition of local cattle. The study aimed to evaluate the status of artificial insemination and identify its constraints in the selected districts of East Arsi Zone, Oromia regional state. The data were collected from 301 farmers and 9 AI technicians (AIT) using semi-structured questionnaires. Five-year secondary data were used from the annual summary of the casebook to evaluate AI status. Data on AI status, satisfaction, breeding method, controlled mating and AI delivery were analyzed using Statistical Analysis Systems (SAS) chi-square procedures. Secondary data were analyzed using General Linear Model SAS methods. The ranking coefficient was analyzed using the R software Plackett–Luce model procedure. The study found that 43.52% of participants were dissatisfied with AI services, while 56.48% were satisfied. Furthermore, 72.43% of respondents indicated an increase in AI services, whereas 13.62% reported a decrease and 13.95% no change. The respondents' satisfaction with AI and AI status differed (P&lt;0.05) among districts, but no difference (P&gt;0.05) between the production systems. The secondary data revealed a gradual increase in AI delivery from 2018 to 2022. The average AI delivery was 2281.5±275.6 per year. While AI services did not differ (P &gt; 0.05) across districts, there was a significant (P&lt; 0.05) variation over time. About 55.48% of the districts' dairy producers used AI for breeding. Most respondents (61.79%) received AI from government administrations, while 36.21% received from government and private and only 1.99% obtained it from private suppliers. Breeding methods varied significantly across production systems (P&lt;0.0001), but controlled mating and AI provision were non-significant. The farmers preferred neighbor bulls with estimated coefficients of 2.24 followed by their bulls (1.05) for breeding purposes. The respondents indicated that conception failure (0.72) and poor conception rates (0.56) were the biggest challenges for AI in the study areas. Transportation (2.89) was the main constraint in delivering AI services identified by AITs. Despite these challenges, there has been an increase in the use of AI in study areas over the past five years. Because AI is the only accessible technology for increasing dairy cow performance in the country, it is vital to address these challenges to increase AI utilization in the study regions. Focusing on semen quality is necessary to achieve a high conception rate per service. Supporting commercial AI businesses could improve farmers' access to services. Key words: artificial insemination, breeding method, controlled mating, satisfaction, status

Graphene oxide and silymarin combination: A novel approach to improving post-cryopreservation quality of ram sperm.

Abstract. Cryopreservation of semen has advanced significantly with the development of artificial insemination techniques, but post-thawed sperm often exhibit reduced viability, membrane integrity, and acrosome integrity compared to fresh sperm, leading to decreased fertilization capacity. Oxidative stress is a major concern during cryopreservation. This study investigated the use of resveratrol (RSV), a potent antioxidant, in the cryopreservation of Mongolian horse semen. Different concentrations of RSV were incorporated into semen cryopreservation extenders, and the morphological and antioxidant indices of post-thawed sperm were assessed to determine the optimal RSV concentration. The study also employed tandem mass tag (TMT) quantitative proteomics technology to explore differential proteins and their pathways. The results showed that sperm quality parameters were positively correlated with RSV concentration within a certain range (10–40 µmol L−1) and were significantly higher than the control group. RSV also enhanced the antioxidant capacity of sperm, with the optimal effect observed at 40 µmol L−1. Proteomics analysis identified 10 differential proteins between the control and optimal RSV concentration groups, with 7 upregulated proteins primarily involved in antioxidant activity and maintaining intracellular redox balance. These findings were further validated through real-time fluorescent quantitative PCR and protein immunoblotting, suggesting that RSV has potential as an effective antioxidant for improving the cryopreservation of Mongolian horse semen.

Community Service was conducted to increase local people’s economy by Poté goat population using Artificial Insemination in Alaskokon, Modung, and Bangkalan. The partner in this Community Service are local Poté goat farmers in Alaskokon village, Modung, Bangkalan. The analysis of this situation shows a problem that the Partner has been facing. Even though the Poté population is not lacking, their rearing is conventional where it is not focused on selling but more on saving for when there is no money (Rojokoyo). To tackle this problem, Artificial Insemination was introduced to the Poté. Moreover, socialisation was done by covering various aspects. Sosialisation was given on caring for pregnant does, semen collection using an artificial vagina, and buck semen dilution using different types of extenders such as coconut water, milk, and egg yolk-citrate. Socialisation of the marketing strategy for goats was also given. Artificial Insemination Technology is a solution for improving the goat population. Goats are fast-bred livestock with a relatively short gestation time and can utilise forage like grasses. Artificial Insemination technique can be administered three times in two years to increase the goat population, thus, the local people’s economy may follow. After the programme was completed, farmers were enthusiastic about collecting goat semen and inseminating livestock.

In the context of modern pig farming, the central role of boars is underscored by large-scale centralized breeding and the widespread application of artificial insemination techniques. However, previous studies and breeding programs have focused mainly on product efficiency traits, such as growth rate, lean meat yield, and litter size, often neglecting boar semen traits. In this study, we estimated the genetic parameters and assessed the genomic prediction accuracy of boar semen traits with phenotypes evaluated from 274,332 ejections in a large population consisting of 2467 Duroc boars. Heritability of sperm morphological abnormality rate (ABN), fresh semen volume (VOL), sperm concentration (DEN), and motility (MOT) were estimated to be 0.43, 0.22, 0.23, and 0.16, respectively. GBLUP achieved a moderate predictive ability of semen traits, with a range of 0.32-0.50. Incorporating gene interactions indicated by the KEGG pathways (biBLUP) significantly improved predictive accuracy over the classical additive model (GBLUP) and epistatic model (RKHS). Moreover, biBLUP showed an improvement from 9.50% to 20.10% among the studied traits compared with GBLUP, with the greatest improvement (0.40 vs. 0.48) observed in sperm morphological abnormality rate. In conclusion, moderate to low heritability was estimated for the Duroc boar semen traits. Genomic prediction was able to achieve moderate accuracy, with a range from 0.32 to 0.56, for the studied traits. Considering gene interactions within KEGG pathways enhanced the predictive ability of boar semen traits.

Objective: To report pregnancy outcomes following intratubal insemination without catheterization using the FemaSeed device in a population of low sperm count couples. Methods: A prospective, single-arm, historical control, pivotal trial (Clinicaltrials.gov NCT04968847) of females aged 19-40 years with male-factor or unexplained infertility and total motile sperm count of 1-20 million undergoing intratubal insemination (ITI) with the FemaSeed device, conducted at twelve academic and private medical centers in the USA. Pregnancy was confirmed at 3 weeks (-7 days) by serum β-hCG and follow-up continued at 7 weeks (±7 days) for safety, including pregnancy test by serum β-hCG and ultrasonography. Results: The pregnancy rate for ITI with FemaSeed per subject was 26.3% (95%CI: 13.4‒43.1%; n=10/38) and 17.5% per cycle (95%LCB: 7.6%, 95%CI: 5.7‒29.4%; n=10/57), which was significantly higher than the performance goal of 7% based on the historical control (one-sided P=0.041). The cumulative pregnancy rates through the first and second ITI cycles were 15.8% (95%CI: 7.4‒31.8%) and 30.7% (95%CI: 16.5‒52.5%), respectively. Safety reports were consistent with intrauterine insemination (IUI), of note ectopic pregnancy and uterine perforation occurred in 0.5% (95%CI: &lt;0.05‒2.5%; n=1/222) and 0.0% of cycles, respectively. Conclusion: Targeted intratubal insemination of washed spermatozoa using the FemaSeed ITI device is a safe artificial insemination technique that demonstrated high effectiveness for couples with male-factor/unexplained infertility associated with low male sperm count. Delivery of washed spermatozoa directly into the utero-tubal ostium and fallopian tube without catheterization likely increases sperm-oocyte interaction, suggestive of improved efficiency over conventional intrauterine insemination particularly for male-factor infertility

Unlocking the potential of propolis: Concentration-dependent effects on Ossimi Ram (Ovis aries) semen quality

Dairy farming plays a significant role in the socio-economic development of the northern region of Iraq, constituting a crucial segment of the rural economy. The primary aim of this study is to investigate the sociodemographic characteristics of dairy farms and management practices in the Northern Region of Iraq. Data were gathered from 280 dairy farmers in the Erbil, Sulaymaniyah, Dohuk, and Halep provinces and analyzed using frequency tables and descriptive statistics. The findings revealed that the average number of cows per farm stood at 28.2, with an estimated average milk yield of 10.1 liters per cow. Additionally, 53.2% of farms in the surveyed area reported the utilization of artificial insemination techniques. Despite these achievements, several challenges confront milk producers in the region, such as a lack of digital skills, the burden of high feeding costs, inadequate feeding, challenges related to hygiene and disease control within dairy herds, uncertainties regarding market access, and limited veterinary services.

This review paper summarizes information on the efficiency, major challenges and economic loss of cattle Artificial Insemination (AI) service in developing countries. Efficiency of AI service can be measured by number of services per conception (NSC), conception rate at first insemination, and calving rate (CR). The optimum recommended NSC for profitable dairy cow ranges from 1.0-2.0. The averages of NSC of the conventional AI (1.73) and fixed time AI (1.78) in the reviewed publications were in the ranges of the recommendations of NSC. The NSC varies with animal breeds, animal factor (body condition score, age and parity), semen factor (handling procedure and quality), inseminator factor (knowledge, skill and experience), production systems, management level provided and AI breeding methods. The NSC and calving rate vary with conventional AI and fixed time AI breeding methods. Likewise, calving rate (CR) is influenced by poor semen quality, poor semen handling procedure, inadequate insemination skill, poor oestrus detection and wrong time of insemination. The average success rate (CR) of conventional AI (35.15%) in the reviewed publications is far below the success rates reported in some developing countries. Nevertheless, widespread use and longtime use of AI service on selected genotype(s) can cause cattle biodiversity loss. Dairy producers incur additional costs when cows fail to conceive at their first AI services. Moreover, challenges of AI service comprised of feed scarcity, animal diseases, climate change, poor AI infrastructure, poor livestock husbandry practices, weak livestock extension systems and incapability of Artificial Insemination Technicians (AITs). This calls designing suitable interventions to improve the efficiency of AI services. Adequate feed and high level of management should be provided to the breedable cows and heifers. AI service centers should be established as per the recommended number of breedable cows and heifers. AI inputs including AITs should be always available at each AI service center, and proper AI service recording should be practiced. Sexed semen should be used to enhance the number of replacement heifers, and capacity building should be provided to AITs and community cattle breeders. Radio, television and printed medias should be used to enhance the awareness of the community cattle breeders. Frequent extension support is essential for the success rate of the AI service.