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Single Layer Centrifugation (SLC) through a low density colloid offers an alternative solution to antibiotic use in boar semen extenders, with lower costs compared to high density colloids. The aim of this study was to explore the reproductive performance of sows when using SLC-prepared semen doses without antibiotics, employing low density Porcicoll to prepare semen doses for artificial insemination in a commercial swine herd in Thailand. Ejaculates were divided into two equal parts to create insemination doses, with each dose containing 3000 × 106 sperm/80 ml for intra-uterine insemination in individual sows. The sows were inseminated twice, with the interval between the two inseminations ranging from 8 to 16 h. The CONTROL group consisted of 206 semen doses treated with antibiotics, prepared for insemination in 103 sows, while the SLC group comprised 194 SLC-prepared semen doses without antibiotics for inseminating 97 sows. Fertility and fecundity traits, including non-return rate, conception rate, farrowing rate, and litter traits (i.e., the total number of piglets born per litter, number of piglets born alive per litter, number of stillborn piglets, and number of mummified fetuses), were compared between groups. Furthermore, data on piglet characteristics, including live-born and stillborn piglets (i.e., the prevalence of stillbirth (yes, no), birth weight, crown-rump length, body mass index (BMI), and ponderal index (PI)), were determined. No significant differences in non-return rate (75.7 % vs. 77.3 %), conception rate (73.8 % vs. 73.2 %), and farrowing rate (71.8 % vs. 73.2 %) were observed between the CONTROL and SLC groups, respectively (P > 0.05). Nevertheless, the total number of piglets born per litter in the SLC group was higher than in the CONTROL group (14.6 ± 0.9 vs. 12.3 ± 0.6, respectively, P = 0.049). Interestingly, the prevalence of stillbirth in the SLC group was lower than in the CONTROL group (6.2 % vs. 11.6 %, respectively, P < 0.001). Moreover, the newborn piglets in the SLC group exhibited higher birth weight and BMI compared to those in the CONTROL group (1.36 ± 0.03 vs. 1.26 ± 0.02 kg, P = 0.005, and 18.3 ± 0.3 vs. 17.3 ± 0.2 kg/m2, P = 0.003). In conclusion, employing sperm doses after SLC through a low density colloid in artificial insemination within a commercial breeding operation did not have a detrimental impact on either fertility or fecundity traits but showed potential benefits in increasing the total number of piglets born per litter. Moreover, improvements were observed in the birth weight and body indexes of piglets, and the percentage of stillbirths was reduced. Our findings introduce new possibilities for antibiotic alternatives in semen extenders to reduce the risk of antimicrobial resistance in the swine industry. Additionally, they provide compelling reproductive outcomes supporting the integration of SLC-prepared semen doses into artificial insemination practices. • Single Layer Centrifugation is an alternative solution to antibiotic use in boar semen extenders. • Farrowing rate did not differ between control and SLC groups. • The total born in the SLC group was higher than in the control group. • The prevalence of stillbirth in the SLC group was lower than in the control group. • Newborn piglets in the SLC group had higher birth weight and BMI than control group. [ABSTRACT FROM AUTHOR]

Melatonin (MLT) has strong antioxidant capacity and can reduce the damage caused by oxidative stress in sperm, but there is still little content in the field we have studied. In this study, we are committed to scientific research on adding melatonin to Belgian blue bull semen diluent for cryopreservation. Different concentrations (0, 0.1, 0.3, 0.5 or 0.7 mg/mL) of MLT were added diluent. Sperm kinetic parameters, enzyme activity, antioxidant gene expression and fertility were analyzed after thawing. The results showed that MLT concentration of 0.3 mg/mL exerted positive effects on post-thaw kinetic parameters. Compared with other groups, 0.3 mg/mL MLT treated sperm acrosome and plasma membrane integrity, catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels significantly increased. Meanwhile, the mRNA expression of antioxidant genes SOD2 , CAT and GPx increased in the 0.3 mg/mL MLT treatment group, and the mRNA expression of apoptosis genes Caspase-3 and Bax were significantly reduced. In addition, in vitro fertilization (IVF) embryo cleavage, blastocyst rate and artificial insemination (AI) pregnancy rate were higher in 0.3 mg/mL MLT. Therefore, MLT showed cryoprotective capacity to the freezing diluent used for Belgian blue bull sperm during the process of freezing-thawing, and the optimal concentration of MLT for the frozen diluent was 0.3 mg/mL. • Optimizing our laboratory's semen diluent formulation by adding MLT. • Belgian blue bulls have good research and production advantages. • Adding 0.3 mg/mL MLT significantly improves bull semen storage efficiency. [ABSTRACT FROM AUTHOR]

Background: Centrifugation is a common procedure to improve the quality of chilled and frozen canine semen by removing debris and seminal plasma and adding semen extenders. The aim of this study was to evaluate the efficacy and influence of a second centrifugation after 48 h of storage at 5 °C on the sperm quality of canine semen. The ejaculates of 45 healthy male dogs, divided into three groups according to body weight, were analyzed for macro- and microparameters such as ejaculate volume, sperm concentration, kinematic parameters, morphology, and integrity of plasma membrane. Samples were analyzed at baseline conditions (T0), after 24 h (T24) and after 48 h (T48) to assess the effects of the different treatments on sperm quality. Results: The results showed a significant effect of a second centrifugation on the improvement of chilled sperm quality compared to the other techniques, especially up to 48 h. Conclusions: Analysis of the data showed that the semen samples centrifuged and then cooled at 5 °C had acceptable semen parameters, especially in terms of motility, with a gradual decrease in serial evaluations after 24 and 48 h. A second centrifugation after 48 h of storage may lead to better semen quality and improve the kinetics of sperm parameters, the percentage of morphologically normal sperm and the percentage of sperm with intact membranes. [ABSTRACT FROM AUTHOR]

Boar semen production plays a pivotal role in modern swine breeding programmes, influencing the genetic progress and overall efficiency of the pork industry. This review explores the current challenges and emerging trends in liquid‐preserved boar semen production, addressing key issues that impact the quality and quantity of boar semen. Advances in new reproductive technologies, boar selection, housing, semen processing, storage and transport, and the need for sustainable practices including the use of artificial intelligence are discussed to provide a comprehensive overview of the field. [ABSTRACT FROM AUTHOR]

Context: The conflicting findings regarding the impact of equilibration time on post-thawed sperm quality underscore the need for further research to evaluate the impact of equilibration time and cooling rate on post-thaw sperm quality of ram semen. Aims: The current study aimed to assess the combined impact of cooling rates and pre-freezing equilibration times on post-thaw sperm quality in Kail ram semen (n = 5). Methods: Semen collection was performed using an artificial vagina at 42°C. The pooled semen was divided into equal aliquots and subjected to either slow cooling (SC, −0.27°C/min) or moderate cooling (MC, −0.36°C/min) rates, transitioning from 37°C to 4°C. Equilibration times of 0, 4, 8, and 12 h were employed before freezing. Key results: Semen samples undergoing the SC rate and equilibrated for 4 h exhibited higher (P < 0.05) percentages of progressive motile (PM), rapid progressive (RP), and medium progressive (MP) sperm compared with the MC rate. However, total motility remained unaffected by the cooling rate (P < 0.05). Semen equilibrated for 4 h demonstrated higher (P < 0.05) percentages of PM and RP sperm, as well as improved kinematics (curvilinear velocity, average path velocity, and straight-line velocity) compared with other equilibration times. Nevertheless, equilibration time had no (P > 0.05) impact on the amplitude of the lateral head displacement for semen samples subjected to the MC rate. Notably, the cooling rate did not affect post-thaw sperm kinematics, plasma membrane integrity, or live-sperm percentage (P > 0.05). Semen samples equilibrated for 4, 8, and 12 h exhibited a higher (P < 0.05) percentage of sperm with intact plasma membrane and viability than did those equilibrated for 0 h. Conclusions: In conclusion, slow cooling rate and a 4 h equilibration period were shown to be optimal for preserving post-thaw sperm quality in Kail rams. Implications: The findings highlighted the combined effect of cooling rate and equilibration time on post-thaw sperm quality for optimising sperm cryopreservation protocols in the context of ram semen. The conflicting results regarding the impact of equilibration time on sperm quality warrant further research to improve the post-thaw sperm quality of ram semen. The findings of the present study indicated that slow cooling and equilibration time of 4 h better-preserved sperm quality. The optimisation of sperm-freezing protocol will be beneficial for the widespread use of artificial insemination in sheep. [ABSTRACT FROM AUTHOR]

Background: Infertility affects 186 million people worldwide, with male factors contributing to 50% of infertility cases. Semen analysis is a key for diagnosing male factor infertility, but sperm parameters can be influenced by ejaculatory abstinence (EA) duration. Shortening or prolonging EA can impact on semen quality and assisted reproductive technology (ART) outcomes, but the optimal EA duration remains unclear, particularly for infertility patients. Objectives: This study conducts a comprehensive meta‐analysis to explore the impact of varying abstinence durations on semen quality and fertility outcomes. Methods: Three English database (PubMed, Embase, and Cochrane Central Register of Controlled Trials) as well as four Chinese database (China National Knowledge Infrastructure, Chinese Scientific Journals database, WanFang database, and Chinese Biomedical Literature database) were searched from 2000 to August 2023. The classical meta‐analysis and "one‐stage" dose–response meta‐analysis were conducted to compare the associations of different abstinence durations (short‐term abstinence vs. long‐term abstinence) on semen quality in healthy adult and different type of infertile patients. Results: There were 85 eligible studies were finally included. The meta‐analysis of volume (mean difference [MD] = −0.95 mL, 95% confidence interval [CI]: −1.16 to −0.74 mL), total sperm count (TSC) (MD = −102.45×106, 95% CI: −117.98×106 to −86.91×106), sperm concentration (SC) (MD = −11.88×106/mL, 95% CI: −18.96×106/mL to −4.80×106/mL), DNA fragmentation index (DFI) (MD = −2.37%, 95% CI: −4.73% to −0.01%) in healthy men showed a significant decrease with different abstinence durations (short‐term abstinence vs. long‐term abstinence). The meta‐analysis of infertile men showed significant decrease in volume in various subgroups (MD range: −0.73 to −1.17 mL) with P < 0.01; TSC (MD = −61.93×106, 95% CI: −88.84×106 to −35.01×106), SC (MD = −5.39×106/mL, 95% CI: −9.97×106 to −0.81×106/mL), DFI (MD = −5.63%, 95% CI: −10.19% to −1.06%) in unexplained infertility subgroup; significant increase in viability (MD = 6.14%, 95% CI: 3.61% to 8.68%) in the unexplained infertility subgroup. The dose–response meta‐analysis showed that TSC in oligozoospermia showed a nonlinear increase (coefficient from 3.38 to −5.76, P from 0.02 to 0.22) and the truncation point was around the 4th to 5th abstinence day. The percentage of progressive motile sperm (PR) in asthenozoospermia showed a significant decrease (coefficient = −2.39, 95% CI: −4.28 to −0.50). For fertility outcomes of different ARTs, only the clinical pregnancy rate (CPR) in the intrauterine insemination (IUI) subgroup showed a significant decrease around the 3rd day (coefficient = 0.85, 95% CI: 0.75 to 0.97). Conclusions: Short‐term abstinence may be associated with limited improvements in semen quality in healthy men but could be more beneficial for infertile men, especially within the first 4 days of abstinence. Caution is urged in making definitive conclusions about the causal relationship between abstinence time and semen quality changes due to potential confounding and interactions. [ABSTRACT FROM AUTHOR]

Due to the pollution from the copper smelter factory in Shahrebabak County, the dust on the plants of this area contains substantial amounts of copper, which sometimes causes copper poisoning in sheep. This study aimed to investigate the correlation of serum concentration of trace mineral elements: copper, molybdenum, and iron with the lambing rate of ewes in Shahrebabak County using laparoscopic artificial insemination. For this purpose, 500 ewes were oestrus synchronized in herds in Shahrebabak County located at various distances from the copper factory. The laparoscopic artificial insemination was performed 52±4 hours after eCG hormone injection but before the laparoscopy, serum sampling was done from 17% of the herd to assess the concentration of trace mineral elements: copper, molybdenum, and iron as well as estrogen. The results showed that the Khatoon-Abad herd with the shortest distance to the copper factory (10 km) had significantly higher levels of copper, molybdenum, and iron compared with other herds. However, there was no relationship between the concentration of copper, molybdenum, and estrogen with the lambing rate. There was a relationship between the amount of iron and the lambing rate, so the chance of fertility increased marginally with the increase in iron. Hence, the present findings indicate that the low distance of the herd from the copper factory results in high levels of copper, molybdenum, and iron in the ewe's serum; however, it has no effect on the lambing rate by laparoscopic artificial insemination. More research is needed to prove this claim. [ABSTRACT FROM AUTHOR]

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Background: Mating control is a crucial aspect of honeybee breeding. Instrumental insemination of queens gives the breeder maximum control over the genetic origin of the involved drones. However, in addition to the drones' descent, the breeder's control also extends over the number of drones to use for inseminations. Thus far, this aspect has largely been ignored in attempts to optimize honeybee breeding schemes. The literature provides some comparisons between single drone inseminations (SDI) and multi drone inseminations (MDI) but it is unclear whether the number of drones used in MDI is a relevant parameter for the optimization of honeybee breeding programs. Methods: By computer simulations, we investigated the effect of the number of drones per inseminated queen in breeding programs that relied on best linear unbiased prediction (BLUP) breeding values. We covered a range of 1 to 50 drones per queen and observed the developments of genetic gain and inbreeding over a period of 20 years. Hereby, we focused on insemination schemes that take the drones for one queen from a single colony. Results: SDI strategies led to 5.46% to 14.19% higher genetic gain than MDI at the cost of 6.1% to 30.2% higher inbreeding rates. The number of drones used in MDI settings had only a negligible impact on the results. There was a slight tendency that more drones lead to lower genetic gain and lower inbreeding rates but whenever more than five drones were used for inseminations, no significant differences could be observed. Conclusion: The opportunities to optimize breeding schemes via the number of drones used in inseminations are very limited. SDI can be a viable strategy in situations where breeders are interested in genetically homogeneous offspring or precise pedigree information. However, such strategies have to account for the fact that the semen from a single drone is insufficient to fill a queen's spermatheca, whence SDI queens will not build full-strength colonies. When deciding for MDI, breeders should focus on collecting enough semen for a succesful insemination, regardless of how many drones they need for this purpose. [ABSTRACT FROM AUTHOR]

Introduction: Targeted single nucleotide polymorphisms (SNPs) have been used in genomic prediction methodologies to enhance the accuracy of associated genetic transmitting abilities in Holstein cows. The objective of this study was to identify and validate SNPs associated with fertility traits impacting early embryo mortality. Methods: The mRNA sequencing data from day 16 normal (n = 9) and embryo mortality (n = 6) conceptuses from lactating multiparous Holstein cows were used to detect SNPs. The selection of specific genes with SNPs as preliminary candidates was based on associations with reproductive and fertility traits. Validation of candidate SNPs and genotype-to-phenotype analyses were conducted in a separate cohort of lactating primiparous Holstein cows (n = 500). After genotyping, candidate SNPs were filtered using a quality control pipeline via PLINK software. Continuous numeric and binary models from reproductive traits were evaluated using the mixed procedure for a generalized linear model-one way ANOVA or logistic regression, respectively. Results: Sixty-nine candidate SNPs were initially identified, but only 23 passed quality control procedures. Ultimately, the study incorporated 466 observations for statistical analysis after excluding animals with missing genotypes or phenotypes. Significant (p <0.05) associations with fertility traits were identified in seven of the 23 SNPs: DSC2 (cows with the A allele were older at first calving); SREBF1 and UBD (cows with the T or G alleles took longer to conceive); DECR1 and FASN (cows with the C allele were less likely to become pregnant at first artificial insemination); SREBF1 and BOLA-DMB (cows with the T allele were less likely to be pregnant at 150 days in milk). It was also determined that two candidate SNPs within the DSC2 gene were tag SNPs. Only DSC2 SNPs had an important allele substitution effect in cows with the G allele, which had a decreased age at first calving by 10 days. Discussion: Candidate SNPs found in this study could be used to develop genetic selection tools to improve fertility traits in dairy production systems. [ABSTRACT FROM AUTHOR]

Simple Summary: Cooling rooster semen to a temperature of 5 °C is a technique utilized to extend the preservation period of sperm. However, rooster sperm is highly susceptible to damage during preservation, which involves numerous sources of oxidative stress from dilution, limiting its viability and fertility potential to less than 24 h under cool storage conditions. Therefore, researchers are continuously working to optimize preservation methods to enhance the viability and longevity of rooster semen. The aim of this study was to investigate the effectiveness of Aloe vera gel, known for its beneficial antioxidant properties, on the quality of cooled semen in Thai native roosters (Pradu Hang Dum) and to evaluate the fertility rate after artificial insemination. The results showed that Aloe vera gel contained various bioactive compounds with antioxidant activity. The supplementation of the extender with 1.0% Aloe vera gel proved to be beneficial, reducing lipid peroxidation and simultaneously improving sperm quality during cold storage at 5 °C for up to 72 h. The cooling of semen storage at 5 °C from a Thai native rooster (Pradu Hang Dum), supplemented with herbs possessing antioxidant properties, provided limited research. This study was conducted to evaluate the efficiency of Aloe vera (AV) gel supplementation at various levels on the quality of cooled semen and subsequent fertility after artificial insemination. Sixty-four chickens had semen pooled, diluted, and supplemented with different levels of AV gel (0% as control, 0.25%, 0.50%, 1.0%, 2.5%, 5.0%, 10%, and 20%), and then stored for 72 h. In Experiment 1, semen quality, malondialdehyde (MDA) levels, and pH values were assessed at 0, 24, 48, and 72 h after storage. Experiment 2 assessed fertility potential using the most effective cooled storage semen from Experiment 1. Results showed a decrease in semen quality with prolonged storage time (p < 0.001). The highest semen quality was observed in the group supplemented with 1.0% AV gel (p < 0.001), whereas the lowest was noted in the 20% AV gel group (p < 0.001). Furthermore, the 1.0% AV gel group exhibited the highest semen quality at 24, 48, and 72 h of storage. The evaluation of fertility and hatchability rates revealed a statistically significant improvement in fertility potential (p < 0.05) in the group supplemented with 1.0% AV gel. In summary, this study represents the first investigation of stored Thai native rooster semen using a semen extender supplemented with Aloe vera gel at 5 °C, demonstrating its efficacy for storage up to 72 h. The addition of 1% AV gel was recommended as an antioxidant supplementation during the semen storage process at 5 °C to enhance semen quality and fertility rates. [ABSTRACT FROM AUTHOR]

Objective: To compare the ovarian reserve and the results of infertility treatment, as well as to investigate the relapse rate in the first year after the assisted reproductive technology (ART) cycle in patients with multiple sclerosis (MS) referred to Royan Institute. Materials and methods: This retrospective study was carried out to evaluate all women diagnosed with MS and referred to Royan Institute for assessment and treatment of possible infertility between 2011 and 2022. The control group consisted of randomly selected healthy women with tubal factor infertility who were referred for treatment during the same time period and matched in terms of age. A comparison was made between groups in terms of ovarian reserve and infertility treatment outcomes. Additionally, patients with MS who met the criteria were monitored via telephone to evaluate the symptoms, disability and relapse rate both pre- and post-ART. Results: Over the course of a decade, the database documented a total of 60 cases diagnosed with MS. Upon examination of the records, it was found that in 27 patients only admission was done without any hormonal assessment or infertility treatment cycle and 5 patients proceeded with the intrauterine insemination cycle. Eventually, 28 women with MS underwent the ART cycle and all of them were treated with interferon beta, glatiramer acetate, or some oral disease modifying therapies. No statistically significant difference in terms of the basal levels of luteinizing hormone, follicle-stimulating hormone and anti-Müllerian hormone was found between the MS and control groups (P > 0.05). Two groups were comparable in terms of menstrual status. The study revealed that both groups exhibited similarities in terms of the controlled ovarian stimulation protocol and duration, the dosage of gonadotropin administered, as well as the ovarian response type, clinical pregnancy rate, and live birth rate (P > 0.05). After follow up, only 2 patients (9.5%) reported relapse of symptoms within one year after ART. Conclusion: The ovarian reserve and ovarian stimulation cycle and pregnancy outcomes following the ART cycle in MS patients were similar to the age-matched control group. The relapse rate of multiple sclerosis did not show a significant increase within a year following the ART cycle. [ABSTRACT FROM AUTHOR]

This study aimed to investigate if washing ram sperm from seminal plasma (SP) could be an effective tool to extend sperm lifespan in medium-term preservation in liquid form to optimize ovine artificial insemination protocols. To this end, in Experiment 1 SP was added to a sperm model without previous contact with this substance (ram epididymal sperm) at the beginning or the end of a 48-hour preservation protocol at 5 °C (n = 13). Sperm motility and kinetic parameters and sperm functionality in terms of sperm viability, apoptosis, mitochondrial activity and reacted acrosomes were assessed after 6 h of storage at 15 °C (standard liquid preservation method) and 24 and 48 h at 5 °C. Extended sperm showed better results after 48 h when stored in the absence than in the presence of SP in most sperm quality parameters. Moreover, the final SP supplementation of this experimental group resulted in the highest sperm motility and kinetic parameters, viability and mitochondrial activity. These results suggested that initial SP deprivation could be beneficial in a medium-term ram sperm preservation protocol in liquid form, as well as a final supplementation. Therefore, we conducted Experiment 2 to evaluate the effect of SP removal from freshly ejaculated ram semen under the same storage conditions as in Experiment 1 (n = 12). Surprisingly, SP withdrawal impaired sperm functionality, leading to increased apoptosis and decreased mitochondrial activity after 24 and 48 h at 5 °C. Conversely, SP supplementation at the end of the preservation protocol of the ejaculate processed as usual had a positive effect on sperm quality and fertility. To summarize, SP absence was beneficial for a medium-term preservation protocol (up to 48 h at 5 °C) of ram epididymal sperm, but the same preservation protocol for ram ejaculated sperm revealed a possible failure of the SP removal method in avoiding the sperm-SP interaction effect. Meanwhile, SP supplementation of ram semen at the end of the preservation protocol increased in vitro sperm quality and fertility after artificial insemination. [ABSTRACT FROM AUTHOR]

Objective Study Design Results Conclusion Recurrent pregnancy loss (RPL) has a multifactorial etiology, with a majority of cases remaining unexplained. To account for these unexplained cases, possible male factors are being explored. Conventional semen analysis lacks a qualitative assessment of sperms and information regarding sperm DNA integrity. Sperm DNA fragmentation (SDF) has diagnostic value in unexplained RPL, and it may account for a number of unexplained cases. Hence, we planned a study to explore and evaluate the impact of sperm DNA fragmentation in couples with unexplained recurrent pregnancy losses.Analytical cross‐sectional study was conducted at a tertiary‐level referral facility in India between August 2021 and July 2023. Participants (n = 70) were divided into two groups—male partners of couples with unexplained RPL (following spontaneous conceptions) (n = 35) and men with at least one previous live birth (spontaneous or following fertility treatments for female factor infertility such as ovulation induction or intrauterine insemination) as controls (n = 35). Neither of the two groups of couples recruited for this study had undergone ART as fertility treatment. Primary outcome assessed was mean DNA fragmentation index (DFI). Secondary outcomes included differences in semen parameters such as sperm concentration, progressive sperm motility and morphology, proportion of men with high (≥30%) and low DFI in the two groups, and the association between various semen parameters and DFI.Univariate logistic regression revealed that sperm DNA fragmentation was higher in men with unexplained RPL (30.0; IQR (interquartile range) 19.0, 46.0) as compared to controls (22.0; IQR 14.0, 30.0) although it was not statistically significant (OR, odds ratio, 1.02; 95% CI 1.0–1.1, p = 0.08). A higher proportion of men with unexplained RPL had DFI ≥30% compared to controls (54.2% vs. 25.7%; OR 3.43 (95% CI 1.2–9.4); p = 0.02). No statistically significant differences were observed in semen volume, sperm concentration, progressive motility, and morphology between the two groups. Sperm DNA fragmentation index also showed a weak but significant inverse relationship with sperm morphology (r = −0.336, p = 0.004).The current study did not show any significant difference in the mean sperm DNA fragmentation levels in male partners of couples with unexplained RPL compared to controls. However, a higher proportion of men with DFI ≥30% were observed in unexplained RPL population when compared to controls. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to evaluate the effectiveness of a random forest (RF) model in predicting clinical pregnancy outcomes from intrauterine insemination (IUI) and identifying significant factors affecting IUI pregnancy in a large Chinese population. Methods: Results: A total of 11 variables, including eight from female (age, body mass index, duration of infertility, prior miscarriage, and spontaneous abortion), hormone levels (anti-Müllerian hormone, follicle-stimulating hormone, luteinizing hormone), and three from male (smoking, semen volume, and sperm concentration), were identified as the significant variables associated with IUI clinical pregnancy in our Chinese dataset. The RF-based prediction model presents an area under the receiver operating characteristic curve (AUC) of 0.716 (95% confidence interval, 0.6914–0.7406), an accuracy rate of 0.6081, a sensitivity rate of 0.7113, and a specificity rate of 0.505. Importance analysis indicated that semen volume was the most vital variable in predicting IUI clinical pregnancy. Conclusions: The machine learning–based IUI clinical pregnancy prediction model showed a promising predictive efficacy that could provide a potent tool to guide selecting targeted infertile couples beneficial from IUI treatment, and also identify which parameters are most relevant in IUI clinical pregnancy. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. The initial ovulatory response during synchronization programs is often low in dairy heifers, largely due to follicular dynamics and hormonal dynamics. Specifically, the progesterone (P4) concentration at the time of the first GnRH treatment in a breeding program can influence the LH response, often resulting in a suboptimal ovulatory response. The objective of this study was to determine the effect of the highest label dose 200 μg (100 μg vs. 200 μg) of GnRH (50 μg of gonadorelin hydrochloride per mL; Factrel, Zoetis Inc. Madison, NJ) at the first GnRH of a 6-d CO-Synch plus P4 device program on ovulatory response and pregnancy per AI (P/AI) in first service in Holstein heifers. A total of 1,308 Holstein heifers were randomly allocated at the beginning of a 6-d CO-Synch program at day 0 to receive either i.m. treatment of 100 μg (2CC, n = 655) or 200 μg (4CC, n = 653) of GnRH. Also, at d 0, heifers received an intravaginal insert with 1.38 g of P4 (Eazi-Breed CIDR Cattle Insert, Zoetis Inc.). On day 6, the insert was removed, and i.m. treatment of 25 mg of PGF 2α (12.5 mg of dinoprost tromethamine/mL; Lutalyse HighCon Injection, Zoetis Inc.) was administered. On d 7, a second i.m. treatment of 25 mg of PGF 2α was given, followed on d 9 by concurrent i.m. treatment of 100 μg of GnRH, and timed AI. A subset of 396 heifers had their ovaries scanned to evaluate ovulatory response, and blood samples were collected to measure the serum concentration of P4 at d 0 and d 6 of the study. The P4 concentrations at d 0 were categorized as low (≤3 ng/mL) or high (>3 ng/mL). The ovulatory response was greater for heifers receiving 4CC than 2CC at d 0 (54.7% vs. 42.8%). The ovulatory response was greater for low P4 than high P4 at d 0 (54.3% vs. 37.8%). However, we did not observe an interaction between treatment and P4 concentrations (low P4 2CC = 48.6% vs. high P4 2CC = 30.0%; low P4 4CC = 60.0% vs. high P4 4CC = 45.5%). The receiver operating characteristic curve analysis indicated that P4 concentrations at d 0 treatment could predict the ovulatory response, although the area under the curve was only 0.6. As expected, heifers that ovulated had increased P/AI (no = 55.6% vs. yes = 67.7%); however, we found no effect of treatment on P/AI (2CC = 63.3% vs. 4CC = 59.6%), and no interactions between treatment and ovulation and treatment and P4 (high vs. low) for pregnancy outcomes. In summary, P4 concentration and increasing the dose of GnRH at d 0 positively affected ovulatory response in Holstein heifers. However, there was no interaction between treatment and P4 on ovulation and no subsequent impact of GnRH dose on P/AI. [ABSTRACT FROM AUTHOR]

The objective of this study was to verify the causes of the lower response of primiparous Bos indicus cows to the ovulation synchronization protocol. Two experiments were performed to evaluate the biochemical profile, oocyte and follicular cell quality (Experiment 1) and pregnancy (Experiment 2). In Experiment 1, suckled primiparous (n = 24) and multiparous cows (n = 24) were submitted to ovum pick up (OPU). On Day 0 (D0), all cows received 2 mg of estradiol benzoate (EB) and an intravaginal progesterone insert (P4). Five days (D5) after the first hormonal administration (EB + P4), all follicles larger than 3 mm were counted on each ovary, and ovum pick-up (OPU) was performed. On day 12 (D12), the intravaginal progesterone insert was removed, and measurement and aspiration of the largest follicle were performed. Blood samples were collected on D5 and D12 to evaluate the concentrations of glucose, cholesterol, NEFA, IGF-1 and insulin. In Experiment 2, suckled primiparous (n = 50) and multiparous (n = 50) cows were subjected to an ovulation synchronization protocol based on E2/P4 (D0: 2 mg EB plus P4 intravaginal insert; D8: 500 μg of cloprostenol, 1 mg cypionate estradiol and 300UI of eCG; D10: artificial insemination). In addition, blood samples were collected on D10 for evaluation of the same hormones and metabolites described in Experiment 1. In all studies, calves remained with the cows during the experimental period. In experiment 1, the number of oocytes grade 1 (P = 0.83), grade 2 (P = 0.23) and grade 3 (P = 0.51), total number of retrieved oocytes (P = 0.14), oocyte quality index (P = 0.93) and total viable oocytes (P = 0.38) did not differ between primiparous and multiparous cows. The number of follicles at the time of follicular aspiration (20.7 ± 1.5 vs. 18.0 ± 1.9; P = 0.05) and the diameter of the largest follicle on D12 (13.5 ± 0.6 vs. 11.4 ± 0.6; P = 0.04) were greater in multiparous cows, and the number of degenerated oocytes was greater in primiparous cows (1.9 ± 0.7 vs. 1.2 ± 0.3; P = 0.05). On D5, the concentrations of IGF-1 (P = 0.47), insulin (P = 0.08), total cholesterol (P = 0.47), NEFA (P = 0.77) and glucose (P = 0.55) in the blood and IGF-1 (P = 0.97) and insulin (P = 0.11) in the follicular fluid did not differ between parity groups. On D12, there was no difference in the concentrations of IGF-1 (P = 0.26), total cholesterol (P = 0.32), NEFAs (P = 0.31) and glucose (P = 0.93) in the blood between primiparous and multiparous cows, however, the serum insulin concentration (P = 0.04) was higher in primiparous cows. There was no correlation between serum and follicular fluid insulin concentrations (r = 0.17; P = 0.31), however, there was a low correlation between serum and follicular fluid IGF-1 concentrations (r = 0.47; P = 0.002). Quantification of transcripts did not differ between parity groups. In experiment 2, concentrations of NEFA (P = 0.12) and insulin (P = 0.16) in the blood and P/AI (P = 0.93) did not differ between parity [60 % (30/50) primiparous vs. 60 % (30/50) multiparous]. In contrast, blood concentrations of IGF-1 (P = 0.0001), total cholesterol (P = 0.005) and glucose (P = 0.01) were greater in primiparous cows. It was concluded that the oocyte quality and expression of the genes evaluated in the granulosa cells were not different between primiparous and multiparous cows. Unexpectedly, the pregnancy rate did not differ between parity. Nevertheless, the blood concentrations of IGF-1, total cholesterol and glucose were greater in primiparous cows. • The concentrations of metabolic markers and hormones were greater in primiparous cows. • Oocyte quality did not differ between primiparous and multiparous cows. • Primiparous cows had a higher number of degenerated oocytes. [ABSTRACT FROM AUTHOR]

Objective: To compare two cancellation policies in controlled ovarian stimulation‐intrauterine insemination (COS‐IUI) cycles to lower the risk of multiple pregnancies (MP). Design: We performed a bicentric retrospective cohort study in two academic medical centers: Angers (group A) and Besançon (group B) University Hospitals. We included 7056 COS‐IUI cycles between 2011 and 2019. In group A, cancellation strategy was based on an algorithm taking into account the woman's age, the serum estradiol level, and the number of follicles of 14 mm or greater on ovulation trigger day. In group B, cancellation strategy was case‐by‐case and physician‐dependent, based on the woman's age, number of follicles of 15 mm or greater, and the previous number of failed COS‐IUI cycles, without any predefined cut‐off. Our main outcome measures were the MP rate (MPR) and the live‐birth rate (LBR). Results: We included 884 clinical pregnancies (790 singletons, 86 twins, and 8 triplets) obtained from 6582 COS‐IUI cycles. MPR was significantly lower in group A compared with group B (8.1% vs 13.3%, P = 0.01), but LBR were comparable (10.8% vs 11.8%, P = 0.19). Multivariate logistic regression found the following to be risk factors for MP: the "cancellation strategy" effect (adjusted odds ratio [aOR] 1.63, 95% confidence interval [CI] 1.02–2.60) and the number of follicles of 14 mm or greater (aOR 1.39, 95% CI 1.16–1.66). Cycle cancellation rate for excessive response was significantly lower in group A compared with group B (1.3% vs 2.4%, P < 0.001). Conclusions: The use of an algorithm based on the woman's age, serum estradiol level and number of follicles of at least 14 mm on trigger day allows the MPR to be reduced without impacting the LBR. Synopsis: A cancellation strategy based on an algorithm allows reduction of the risk of multiple pregnancies without impacting the live birth rate. [ABSTRACT FROM AUTHOR]

Simple Summary: The fertility program Presynch-11-Ovsynch (Presynch-11) improved pregnancies/AI (P/AI) compared with AI following observed standing estrus + timed AI (Estrus + TAI) in first, second, and third+ parities. First parity cows that received AI following an observed standing estrus had greater P/AI compared with second and third+ parity cows. Multiparous cows treated with Presynch-11-Ovsynch had a >60% greater chance of pregnancy compared with cows receiving AI following estrus. Artificial insemination 12 h following observed standing estrus is a standard estimate of the fertility levels of cattle. The main objective of this study was to determine if controlling ovarian development with a fertility program could alter the fertility of lactating dairy cows. Lactating dairy cows (n = 1356) 60–66 days in milk (DIM) were randomly assigned to receive timed AI following Presynch-11/Ovsynch (Presynch-11) or a combination of estrus detection and timed AI using the Presynch-11/Ovsynch program (Estrus + TAI). Cows in standing estrus, following the first two cloprostenol sodium (CLO) injections, in the Estrus + TAI group were artificially inseminated using the AM/PM rule. Cows in this group that were not observed in standing estrus received Ovsynch and TAI beginning 11 d after the second CLO injection. Cows in the Presynch-11 group received two injections of 500 µg CLO 14 d apart but were not observed for estrus. The first GnRH (100 µg) of Ovsynch was administered 11 d following the second CLO injection. All cows in the Presynch-11-Ovsynch group received TAI following Ovsynch. Cows in this treatment were then assigned randomly to receive either CLO or dinoprost 7 d following the first GnRH of Ovsynch. The final GnRH of Ovsynch was administered 56 h later and TAI 16 h after the final GnRH. Pregnancies/AI (P/AI) were greater (p < 0.001) for the Presynch-11 group compared with the Estrus + TAI group (45 vs. 31%). Primiparous cows had greater fertility following observed standing estrus compared with multiparous cows. Days in milk at the first AI were greater (p < 0.01) in the Presynch-11 group vs. the Estrus + TAI group (98 vs. 80) but less variable (p < 0.01). The range of DIM at the first AI was 95 to 101 in the Presynch-11 group and 60 to 101 in the Estrus + TAI group. Within the Presynch-11 group, there were no differences in the rate of luteolysis or P/AI for the prostaglandin type at the final PGF2α of Ovsynch. Multiparous cows treated with Presynch-11 had >60% chance of pregnancy compared with multiparous cows receiving AI following standing estrus. In summary, lactating dairy cows receiving timed AI following the Presynch-11/Ovsynch program had improved fertility compared with a group of cows that received AI following standing estrus or, if not observed in estrus, timed AI. This comparison indicated that controlling ovarian development with GnRH and PGF2α positively impacted the fertility of lactating dairy cows. [ABSTRACT FROM AUTHOR]

Simple Summary: Artificial insemination is a common breeding technique employed in the contemporary livestock industry. However, the efficacy of this technique is contingent upon the quality of the sperm utilized. Several factors influence the quality of sperm during liquid storage. It is, therefore, essential to select an effective physiological regulator in order to maintain sperm quality during this process. This study aimed to evaluate the expression of MTFMT in different motile sperm and to investigate the effects of the addition of amino acids and methionine on sperm quality during liquid storage of boar sperm. The results demonstrated that MTFMT was highly expressed in high-motility sperm. Methionine could act as a substrate for protein translation, affecting mitochondrial function by regulating mitochondrial protein translation and directly influencing boar. Boar sperm quality serves as an important indicator of reproductive efficiency, playing a direct role in enhancing the output of livestock production. It has been demonstrated that mitochondrial protein translation is present in sperm and plays a crucial role in regulating sperm motility, capacitation and in vitro fertilization rate. The present study aimed to determine whether methionine supplementation enhances mitochondrial translation in boar sperm, thereby improving sperm quality. The results showed a significant elevation in the abundance of mitochondrial methionyl-tRNA formyltransferase (MTFMT), a crucial enzyme for mitochondrial protein translation, and mitochondrial DNA-encoded cytochrome c oxidase subunit 1 (COX1) in boar sperm exhibiting high motility. Both amino acids and methionine supplementation significantly enhanced boar sperm motility during storage. Moreover, methionine supplementation mitigates the loss of acrosomal integrity, enhances the expression of COX1, and boosts mitochondrial activity. Furthermore, the positive impact of methionine was negated in the presence of the mitochondrial translation inhibitor chloramphenicol. Together, these findings suggest that boar sperm may utilize methionine as a protein translation substrate to enhance sperm motility by stimulating mitochondrial protein translation. The supplementation of methionine may enhance the quality of boar sperm, thereby providing guidance for the optimization of diluent formulations for liquid storage and the identification of physiological regulators that regulate sperm motility. [ABSTRACT FROM AUTHOR]

STUDY QUESTION What is the relationship between late follicular phase progesterone levels and clinic pregnancy and live birth rates in couples with unexplained infertility undergoing ovarian stimulation with IUI (OS-IUI)? SUMMARY ANSWER Late follicular progesterone levels between 1.0 and <1.5 ng/ml were associated with higher live birth and clinical pregnancy rates while the outcomes in groups with higher progesterone levels did not differ appreciably from the <1.0 ng/ml reference group. WHAT IS KNOWN ALREADY Elevated late follicular progesterone levels have been associated with lower live birth rates after fresh embryo transfer following controlled ovarian stimulation and egg retrieval, but less is known about whether an association exists with outcomes in OS-IUI cycles. Existing studies are few and have been limited to ovarian stimulation with gonadotrophins, but the use of oral agents, such as clomiphene citrate and letrozole, is common with these treatments and has not been well studied. STUDY DESIGN, SIZE, DURATION The study was a prospective cohort analysis of the Assessment of Multiple Intrauterine Gestations from Ovarian Stimulation (AMIGOS) randomized controlled trial. Frozen serum was available for evaluation from 2121 cycles in 828 AMIGOS participants. The primary pregnancy outcome was live birth per cycle, and the secondary pregnancy outcome was clinical pregnancy rate per cycle. PARTICIPANTS/MATERIALS, SETTING, METHODS Couples with unexplained infertility in the AMIGOS trial, for whom female serum from day of trigger with hCG was available in at least one cycle of treatment, were included. Stored frozen serum samples from day of hCG trigger during treatment with OS-IUI were evaluated for serum progesterone level. Progesterone level <1.0 ng/ml was the reference group for comparison with progesterone categorized in increments of 0.5 ng/ml up to ≥3.0 ng/ml. Unadjusted and adjusted risk ratios (RR) and 95% CI were estimated using cluster-weighted generalized estimating equations to estimate modified Poisson regression models with robust standard errors. MAIN RESULTS AND THE ROLE OF CHANCE Compared to the reference group with 110/1363 live births (8.07%), live birth rates were significantly increased in cycles with progesterone 1.0 to <1.5 ng/ml (49/401 live births, 12.22%) in both the unadjusted (RR 1.56, 95% CI 1.14, 2.13) and treatment-adjusted models (RR 1.51, 95% CI 1.10, 2.06). Clinical pregnancy rates were also higher in this group (55/401 clinical pregnancies, 13.72%) compared to reference group with 130/1363 (9.54%) (unadjusted RR 1.46, 95% CI 1.10, 1.94 and adjusted RR 1.42, 95% CI 1.07, 1.89). In cycles with progesterone 1.5 ng/ml and above, there was no evidence of a difference in clinical pregnancy or live birth rates relative to the reference group. This pattern remained when stratified by ovarian stimulation treatment group but was only statistically significant in letrozole cycles. LIMITATIONS, REASONS FOR CAUTION The AMIGOS trial was not designed to answer this clinical question, and with small numbers in some progesterone categories our analyses were underpowered to detect differences between some groups. Inclusion of cycles with progesterone values above 3.0 ng/ml may have included those wherein ovulation had already occurred at the time the IUI was performed. These cycles would be expected to experience a lower success rate but pregnancy may have occurred with intercourse in the same cycle. WIDER IMPLICATIONS OF THE FINDINGS Compared to previous literature focusing primarily on OS-IUI cycles using gonadotrophins, these data include patients using oral agents and therefore may be generalizable to the wider population of infertility patients undergoing IUI treatments. Because live births were significantly higher when progesterone ranged from 1.0 to <1.5 ng/ml, further study is needed to clarify whether this progesterone range may truly represent a prognostic indicator in OS-IUI cycles. STUDY FUNDING/COMPETING INTEREST(S) Oklahoma Shared Clinical and Translational Resources (U54GM104938) National Institute of General Medical Sciences (NIGMS). AMIGOS was funded by the Eunice Kennedy Shriver National Institute of Child Health and Human Development: U10 HD077680, U10 HD39005, U10 HD38992, U10 HD27049, U10 HD38998, U10 HD055942, HD055944, U10 HD055936, and U10HD055925. Research made possible by the funding by American Recovery and Reinvestment Act. Dr Burks has disclosed that she is a member of the Board of Directors of the Pacific Coast Reproductive Society. Dr Hansen has disclosed that he is the recipient of NIH grants unrelated to the present work, and contracts with Ferring International Pharmascience Center US and with May Health unrelated to the present work, as well as consulting fees with May Health also unrelated to the present work. Dr Diamond has disclosed that he is a stockholder and a member of the Board of Directors of Advanced Reproductive Care, Inc. and that he has a patent pending for the administration of progesterone to trigger ovulation. Dr Anderson, Dr Gavrizi, and Dr Peck do not have conflicts of interest to disclose. TRIAL REGISTRATION NUMBER N/A. [ABSTRACT FROM AUTHOR]

The objective of the present study was to investigate the association among the largest follicle (LF), preovulatory estradiol (E2), and predominant vaginal epithelial cell at the completion of hormonal ovarian stimulation for fixed‐time artificial insemination (FTAI) in goats. Thirty‐seven crossbred Boer does received gonadotropin‐releasing hormone (GnRH) and intravaginal progesterone (P4)‐releasing devices (day 0). On day 5, P4 devices were removed and does received prostaglandin F2α and equine chorionic gonadotrophin. On day 7, does received GnRH, and FTAI was undertaken. On day 7, does were divided into three groups, i.e. small‐sized (3–3.9 mm; n = 5), medium‐sized (4–4.9 mm; n = 8), and large‐sized (≥5 mm; n = 24) according to the diameter of the ovarian LF; follicular characteristics (number and diameter) were identified, and blood samples and vaginal smears were collected. The average diameters of total antral follicles and LF and the percentage of superficial cell were greatest in large‐sized LF does (p <.01). The average diameters of total antral follicle (r =.68) and LF (r =.71), number of preovulatory follicle (r =.58), and plasma E2 concentrations (r =.61) were positively correlated with the percentage of superficial cells (p <.01). The likelihood of a pregnancy outcome after the FTAI increased by 13.71 times in does with a greater average diameter of antral follicle, 14.18 times with emergence of a large preovulatory follicle, and 36.83 times with a higher percentage of vaginal superficial cells (p <.01). It was concluded that there is a relationship between the cell types of the vaginal epithelium, the diameters of the largest ovarian follicles, and the concentration of E2 in goats subjected to FTAI protocols. [ABSTRACT FROM AUTHOR]

In this study, we examined the usefulness of a simpler and more feasible method for determining the optimal timing of artificial insemination and the conditions for its success in six female common bottlenose dolphins. Pregnancy was successfully achieved in five dolphins by performing intrauterine insemination, using chilled semen stored for less than 3 days or frozen semen within 24 hr of exhibiting a peak serum estradiol (E2) level of 100 pg/mL or higher or on the day with a serum E2 level of approximately 100 pg/mL, measured with a simple measuring device. We concluded that the determining the optimal timing of intrauterine insemination by measuring serum E2 levels is a simpler and more useful method compared with the conventional approach. [ABSTRACT FROM AUTHOR]

Equine breeding would benefit greatly from reliable biomarkers of stallion or ejaculate fertility. The aim of the study was to investigate how several in vitro sperm characteristics correlate with fertility after artificial insemination, to explore the potential to build a fertility prediction model for stallions. Cooled insemination doses (3–5 per stallion) were obtained from various studs. Sperm membrane integrity, acrosome integrity, chromatin integrity, mitochondrial membrane potential, and reactive oxygen species production were evaluated by flow cytometry 24–30 h after semen collection, and sperm motility was assessed by computer aided sperm analysis. Calcein violet was used to differentiate viable spermatozoa. Per season pregnancy rates for these stallions were available the following year. Positive correlations were found between pregnancy rate and straightness (r = 0.43, p ≤ 0.001), as well as pregnancy rate and the proportion of living hydrogen peroxide positive spermatozoa (r = 0.32, p ≤ 0.05). There were negative correlations between pregnancy rate and amplitude of lateral head displacement (r = -0.26, p ≤ 0.05), and between pregnancy rate and the mean fluorescence of dead superoxide positive spermatozoa (r = -0.46, p < 0.001). Principal component analysis indicated that motility, membrane integrity, DNA fragmentation, and reactive oxygen species production were associated with pregnancy rate. Therefore, a combination of these factors could be used as a biomarker of fertility when assessing ejaculates. However, data from more individuals would be required to construct a model for fertility prediction. [ABSTRACT FROM AUTHOR]

Background: In the field of male infertility, when sperm is normal/subnormal, a few "add-on" routine tests can complete the basic semen examination. Objectives: The aim of this study was to develop and evaluate a faster, simplified motile sperm organelle morphology examination (MSOME) technique for selected infertile patients with apparently normal/subnormal sperm and, in their background: failure of two or three intrauterine insemination (IUI) cycles, repeatedly fragmented embryos, embryonic development to blastocyst-stage failures, repeated miscarriages, a long period of infertility or 2 or more IVF attempts without pregnancy. Our test results were correlated with IUI, conventional in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected sperm injection (IMSI) outcomes. Materials and methods: We validated an adapted version of the MSOME analysis called the pre-IMSI test (PIT), based on vacuole evaluation alone. 248 infertile patients from our assisted reproductive technology (ART) Center were retrospectively selected and split into three PIT score subgroups (patients with ≤8% (score I), 9 to 15% (score II) and ≥16% normal spermatozoa (score III)) based on the correlation between PIT results and each ART technique outcome. The choice of one or another of these ART techniques had been made according to the usual clinico-biological criteria. Results: Clinical outcomes for each of the three PIT subgroups were compared individually for the different ART techniques. For ICSI, the effect of the PIT score subgroup was significant for clinical pregnancies (p = 0.0054) and presented a trend for live births (p = 0.0614). Miscarriage rates of IVF attempts were statistically different depending on the PIT score (p = 0.0348). Furthermore, the odds ratios of clinical pregnancy rates were significantly different according to PIT score subgroup when comparing ICSI vs. IMSI or IVF vs. ICSI attempts. Discussion: IMSI appears to be recommended when sperm belongs to PIT score I, ICSI when it belongs to PIT score II and IVF or IUI when sperm is of PIT score III quality in selected infertile couples. The lack of statistical power in these PIT subgroups means that we must remain cautious in interpreting results. Conclusion: Our results support the interest of this simplified test for certain couples with normal/subnormal sperm to help choose the most efficient ART technique, even as first-line treatment. [ABSTRACT FROM AUTHOR]

BACKGROUND: The impact of the endometrium’s structure and spiral artery blood flow parameters on the pregnancy rate of artificial insemination by husband remains unclear. This study identified the independent factors and constructed a prediction model with good clinical application efficacy after calibration of other confounding factors. OBJECTIVE: To construct and validate a clinical pregnancy prediction model for artificial insemination by husband based on endometrial structure and uterine spiral artery blood flow parameters. METHODS: A retrospective analysis was conducted on 1 299 patients who underwent artificial insemination by husband treatment at Changzhou Maternal and Child Health Hospital from January 2017 to January 2021. The non-pregnancy group consisted of 1 182 patients, while the pregnancy group included 117 patients. Out of these patients, 93 cases were successfully matched between the pregnancy and non-pregnancy groups using a 1:1 propensity score matching method. Single-factor and multi-factor analyses were used to screen the endometrial structure and uterine spiral artery blood flow parameters to determine their influence on artificial insemination by husband outcomes. The optimal cutoff value was established for each independent influencing factor through receiver operating curve analysis and their risk trend affecting artificial insemination by husband pregnancy outcomes was analyzed using a restricted cubic spline. The clinical efficacy of this combined forecast model was tested by using clinical decision curve and clinical influence curve methods. RESULTS AND CONCLUSION: (1) There was no statistical significance in non-endometrial factors between the pregnancy group and the non-pregnancy group, and the data had a good balance by propensity score matching (P > 0.05). (2) Single-factor analysis identified several subendometrial parameters as significant influencing factors of artificial insemination by husband pregnancy outcomes, including vascularization index, flow index, vascular flow index, resistance index, pulsatility index, maximum systolic velocity/end-diastolic velocity, thickness of average junction zone and maximum junction zone from the basal endometrium to the outer myometrium inner layer (P < 0.05). (3) Multivariate logistic regression analysis revealed that thickness of average junction zone, pulsatility index, and vascular flow index were independent influencing factors of pregnancy outcomes of artificial insemination by husband, vascular flow index > thickness of average junction zone > pulsatility index. (4) Receiver operating characteristic curve analysis indicated that the area under receiver operating characteristic curve of vascular flow index was 0.704 (0.629, 0.779), and the optimal cutoff value was 6.26; the area under receiver operating characteristic curve of thickness of average junction zone was 0.660 (0.582, 0.739), and the optimal cutoff value was 6.38; the area under receiver operating characteristic curve of pulsatility index was 0.642 (0.563, 0.721), and the optimal cutoff value was 1.18. (5) The restricted cubic spline analysis revealed that artificial insemination by husband pregnancy outcomes were significantly positively affected when the vascular flow index was > 6.24 or the thickness of average junction zone was ≤6.55 mm, while a negative risk was associated with pulsatility index > 1.27. (6) The clinical decision curve and clinical influence curve analyses exhibited that the combined prediction model had the maximum clinical net benefit at the threshold probability value of 0.17-0.93, and the ratio of loss to benefit was consistently less than 1 in the threshold probability range, indicating that the model had good clinical efficacy. (7) It is concluded that after adjusting for other confounding factors outside of the endometrium using propensity score matching and multifactorial logistic regression, the thickness of average junction zone, pulsatility index and vascular flow index were independent factors that influenced pregnancy outcomes of artificial insemination by husband. Through determining their optimal cutoff values and assessing their risk trends, it was confirmed that the combined prediction model had good predictive value and clinical efficacy. [ABSTRACT FROM AUTHOR]

Background: Most sows will experience negative energy balance during lactation resulting in impaired follicular development. This study aimed to treat 28-day lactating sows with altrenogest (ALT) to suppress follicle enlargement during lactation, and to assess the estrus and reproductive performance post-weaning. Methods: In this study, we conducted two trials. In trial 1, we monitored the follicular development of lactating sows including 10 primiparous sows and 10 multiparous sows during the whole lactation to confirm the ALT administration time. In trial 2, a total of 42 primiparous and 111 multiparous sows were allocated to three treatments: Ctrl (control group, n = 51): no treatment; TAI (timed artificial insemination group, n = 51): sows were injected with equine chorionic gonadotropin (eCG) after weaning 24 h and gonadotropin-releasing hormone (GnRH) when they expressed estrus; and AT-TAI (ALT treatment-timed artificial insemination group, n = 51): base on the process of TAI group, the sows were fed with 20 mg ALT per day before weaning 10 days. All sows were artificially inseminated twice at 12 h and 36 h after estrus. The follicle size changes and serum hormone levels were explored in this process. Results: Although the follicle size of multiparous sows was larger than primiparous sows during the whole lactation (P < 0.05), similar change trends of follicle size were observed in primiparous and multiparous sows. Meanwhile, the FSH, LH and E2 levels of multiparous sows were higher than primiparous sows. The ALT treatment significantly inhibits the increase in follicle size (P < 0.05) and reduces the serum levels of FSH, LH and E2 (P > 0.05). Additionally, ALT treatment increases estrus concentration and the preovulatory follicle size (P < 0.05), meanwhile, it delays the weaning-to-estrus interval (WEI, P < 0.001). However, the estrus rate, pregnancy rate, total pigs born and born alive did not differ between treatments (P > 0.05). Conclusions: There were significant differences in the size of follicles in the lactation between primiparous and multiparous sows. ALT treatment during the last ten days of lactation concentrated estrus expression leading to higher work efficiency of breeder in batch production, however, with no improvement in reproductive performance. [ABSTRACT FROM AUTHOR]

Objective: To determine whether endometrial thickness (EMT) differs between i) clomiphene citrate (CC) and gonadotropin (Gn) utilizing patients as their own controls, and ii) patients who conceived with CC and those who did not. Furthermore, to investigate the association between late-follicular EMT and pregnancy outcomes, in CC and Gn cycles. Methods: Retrospective study. Three sets of analyses were conducted separately for the purpose of this study. In analysis 1, we included all cycles from women who initially underwent CC/IUI (CC1, n=1252), followed by Gn/IUI (Gn1, n=1307), to compare EMT differences between CC/IUI and Gn/IUI, utilizing women as their own controls. In analysis 2, we included all CC/IUI cycles (CC2, n=686) from women who eventually conceived with CC during the same study period, to evaluate EMT differences between patients who conceived with CC (CC2) and those who did not (CC1). In analysis 3, pregnancy outcomes among different EMT quartiles were evaluated in CC/IUI and Gn/IUI cycles, separately, to investigate the potential association between EMT and pregnancy outcomes. Results: In analysis 1, when CC1 was compared to Gn1 cycles, EMT was noted to be significantly thinner [Median (IQR): 6.8 (5.5-8.0) vs. 8.3 (7.0-10.0) mm, p<0.001]. Within-patient, CC1 compared to Gn1 EMT was on average 1.7mm thinner. Generalized linear mixed models, adjusted for confounders, revealed similar results (coefficient: 1.69, 95% CI: 1.52-1.85, CC1 as ref.). In analysis 2, CC1 was compared to CC2 EMT, the former being thinner both before [Median (IQR): 6.8 (5.5-8.0) vs. 7.2 (6.0-8.9) mm, p<0.001] and after adjustment (coefficient: 0.59, 95%CI: 0.34-0.85, CC1 as ref.). In analysis 3, clinical pregnancy rates (CPRs) and ongoing pregnancy rates (OPRs) improved as EMT quartiles increased (Q1 to Q4) among CC cycles (p<0.001, p<0.001, respectively), while no such trend was observed among Gn cycles (p=0.94, p=0.68, respectively). Generalized estimating equations models, adjusted for confounders, suggested that EMT was positively associated with CPR and OPR in CC cycles, but not in Gn cycles. Conclusions: Within-patient, CC generally resulted in thinner EMT compared to Gn. Thinner endometrium was associated with decreased OPR in CC cycles, while no such association was detected in Gn cycles. [ABSTRACT FROM AUTHOR]

Introduction Material and Methods Results Conclusions Intrauterine insemination (IUI) is one of the most widespread fertility treatments. However, IUI protocols vary significantly amongst fertility clinics. Various add‐on interventions have been proposed to boost success rates. These are mostly chosen arbitrarily or empirically. The aim of this systematic review and meta‐analysis is to assess the effectiveness and safety of add‐on interventions to the standard IUI protocol and to provide evidence‐based recommendations on techniques used to optimize the clinical outcomes of IUI treatment.Systematic review and meta‐analyses were performed in accordance with PRISMA guidelines. A computerized literature search was performed from database inception to May 2023. Randomized controlled trials (RCTs) were included reporting on couples/single women undergoing IUI with any protocol for any indication using partner's or donor sperm. A meta‐analysis based on random effects was performed for each outcome and add‐on. Three authors independently assessed the trials for quality and risk of bias and overall certainty of evidence. Uncertainties were resolved through consensus. Primary outcomes were ongoing pregnancy rate (OPR) or live birth rate (LBR) per cycle/per woman randomized. Registration number PROSPERO: CRD42022300857.Sixty‐six RCTs were included in the analysis (16 305 participants across 20 countries). Vaginal progesterone as luteal phase support in stimulated cycles was found to significantly increase LBR/OPR (RR 1.37, 95% CI 1.09–1.72, I2 = 4.9%) (moderate/low certainty of the evidence). Endometrial scratch prior/during stimulated IUI cycles may increase LBR/OPR (RR 1.44, 95% CI 1.03–2.01, I2 = 1.8%), but evidence is very uncertain. Results from two studies suggest that follicular phase ovarian stimulation increases LBR/OPR (RR 1.39, 95% CI 1.00–1.94, I2 = 0%) (low certainty of evidence). No significant difference was seen for the primary outcome for the other studied interventions.The findings of this systematic review and meta‐analysis suggest that vaginal luteal phase progesterone support probably improves LBR/OPR in stimulated IUI treatments. In view of moderate/low certainty of the evidence more research is needed for solid conclusions. Further research is also recommended for the use of endometrial scratch and ovarian stimulation. Future studies should report on results according to subfertility background as it is possible that different add‐ons could benefit specific patient groups. [ABSTRACT FROM AUTHOR]

Background: Most commerce of equine seminal doses is carried out using commercial extenders under refrigeration at 5°C. Objectives: To determine if 10 mm pyruvate in a 67 mm glucose extender and storage at 22°C could be the basis of an alternative storage method to cooling to 5°C. Material and methods: Stallion ejaculates were extendedin: INRA96 (67 mm glucose, non‐pyruvate control), modified Tyrode's (67 mm glucose–10 mm pyruvate), supplemented with 0, 10, 50, and 100 μM itaconate. As itaconate was vehiculated in DMSO, a control vehicle was also included. Sperm motility, viability, mitochondrial membrane potential, and production of reactive oxygen species were measured after collection and again after 48 and 96 h of storage at 22°C. To disclose molecular metabolic changes, spermatozoa were incubated up to 3 h in modified Tyrode's 67 mm glucose–10 mm pyruvate and modified Tyrode's 67 mm glucose, and metabolic analysis conducted. Results: After 96 h of storage aliquots stored in the control, INRA96 had a very poor total motility of 5.6% ± 2.3%, while in the 67 mm glucose–10 mm pyruvate/10 μm itaconate extender, total motility was 34.7% ± 3.8% (p = 0.0066). After 96 h, viability was better in most pyruvate‐based media, and the mitochondrial membrane potential in spermatozoa extended in INRA96 was relatively lower (p < 0.0001). Metabolomics revealed that in the spermatozoa incubated in the high pyruvate media, there was an increase in the relative amounts of NAD+, pyruvate, lactate, and ATP. Discussion and conclusions: Aliquots stored in a 67 mm glucose–10 mm pyruvate‐based medium supplemented with 10 μM itaconate, maintained a 35% total motility after 96 h of storage at 22°C, which is considered the minimum acceptable motility for commercialization. Improvements may be related to the conversion of pyruvate to lactate and regeneration of NAD+. [ABSTRACT FROM AUTHOR]

Research question: Can an optimal LH threshold algorithm accurately predict timing of ovulation for natural cycle–intrauterine insemination (NC-IUI)? Design: A retrospective cohort study (2018–2022) including 2467 natural cycles. Ovulation timing for these cycles was determined using a previously developed AI model. Two LH thresholds, low and high, were determined in the LH algorithm. Being below the low threshold meant that ovulation is likely to occur in ≥ 4 days, suggesting another daily blood test. Between the two thresholds meant that ovulation was likely in 2–3 days, suggesting IUI the next day. Above the high threshold meant that ovulation will likely occur tomorrow, suggesting performing IUI on the same day. Results: The optimal LH model with a high threshold of 40 mIU/ml and a low threshold of 11 mIU/ml succeeded in correctly predicting timing for IUI (day − 1, − 2 relative to ovulation) in 75.4% (95%CI 75.3–75.4). In 23.1% (95%CI 23.0–23.2), the algorithm predicted "error," suggesting performing insemination when in fact it would have been performed on a non-optimal day (0 or − 3). A previously described 3-hormone-based (LH, estradiol, progesterone) AI model performed significantly better in all parameters (93.6% success rate, 4.3 "error" rate). Conclusions: An LH threshold model, representing common practice, evaluating all possible high and low LH threshold combinations, was successful in accurately scheduling timing for IUI in only 75% of cases. Integrating all three hormones as performed in the AI model may have an advantage in accurately predicting the optimal time for IUI, over the use of LH only. [ABSTRACT FROM AUTHOR]

The present study compared 2 strategies to initiate a progesterone (P4)-based timed artificial insemination (TAI) protocol for lactating dairy cows: only GnRH or estradiol benzoate (EB) plus GnRH (EB+GnRH). Lactating Holstein cows (n = 487; 184 primiparous and 303 multiparous) from 2 commercial dairy herds were used for their second or greater services postpartum. Each week, cows that were nonpregnant at the pregnancy diagnosis 32 d after a previous AI were randomly assigned to 1 of 2 experimental groups that differed only in the strategy to initiate (d 0) the TAI protocol. On d 0, every cow received a 2.0-g P4 implant; in the EB+GnRH group, cows were treated with 2.0 mg i.m. of EB and 16.8 µg i.m. of the GnRH analog buserelin acetate, whereas in the GnRH group, cows received only 16.8 µg i.m. of GnRH. On d 7 after the initial treatment, 0.530 mg i.m. of cloprostenol sodium (PGF) was administered in all cows, followed by a second dose on d 8, concomitant with 1.0 mg i.m. of estradiol cypionate and P4 implant withdrawal. The TAI was performed on d 10 (48 h after P4 device withdrawal) in both experimental groups. Only conventional Holstein semen was used throughout the study. The percentage of cows with corpus luteum (CL) on d 0 (73%) and overall ovulation rate after d 0 (54%) did not differ between groups. The CL regression between d 0 and the first PGF treatment was greater in the EB+GnRH group than the GnRH group (42% vs. 31%). Consequently, the proportion of cows with CL at PGF was greater when only GnRH was used on d 0 compared with EB+GnRH (86% vs. 82%), and the mean number of CL at PGF was greater (1.23 vs. 1.11). The expression of estrus near TAI was greater in GnRH group (84% vs. 77%), and cows showing estrus had greater (44% vs. 10%) pregnancy per AI (P/AI) on d 32 for both treatments. We found no effect of the presence of CL on d 0 or at PGF, nor of ovulation after d 0 or CL regression between d 0 and d 7 on fertility. However, fertility was critically impaired when cows did not have CL at both times, d 0 and at PGF treatment. We did not observe any interaction between treatment and other variables, and the P/AI was similar in cows receiving EB+GnRH or only GnRH on d 0 (37.8% vs. 36.6%). In summary, although there was no detectable difference in P/AI between treatments, this study demonstrated potential negative physiological outcomes caused by EB treatment on d 0 (greater incidence of luteolysis after d 0 and fewer cows with CL at PGF treatment). Overall, we found no benefit of adding EB at the initiation of a P4-based TAI protocol on fertility compared with using GnRH alone, despite differences in ovarian dynamics and expression of estrus. [Display omitted] [ABSTRACT FROM AUTHOR]

Background and Aim: The use of frozen goat semen for artificial insemination frequently results in a decline in sperm quality following thawing, which can be attributed to cold shock from cryopreservation, reduced motility, and possible DNA damage. Freezing may compromise mRNA stability due to the presence of free radicals. Despite strong post-thaw motility and no visible DNA fragmentation, sperm can still exhibit altered gene expression patterns. To reduce the damaging impact of free radicals during cryopreservation, antioxidants are typically added to the freezing medium. This study assessed the impact of adding coenzyme Q10 (CoQ10) to frozen sperm diluent on the ATP5F1A and CPT2 gene expression, sperm motility, and viability post-thawing. Materials and Methods: CoQ10 was added to sperm at six different concentrations: 0 mg/dL (P0), 6.25 mg/dL (P1), 12.5 mg/dL (P2), 25 mg/dL (P3), 50 mg/dL (P4), and 100 mg/dL (P5). The Statistical Package for the Social Sciences (SPSS) software version 22 was used to conduct comparative tests using one-way analysis of variance followed by Duncan's test for motility and viability and Kruskal-Wallis test followed by pairwise comparison test for membrane integrity and gene expression. Results: The addition of CoQ10 to semen diluent has a notable impact on the post-thawed quality of sperm. The most significant outcomes were observed with a 25 mg/dL dosage (P3) for cell viability, membrane integrity, and ATP5F1A gene expression, and with a 50 mg/dL dosage (P4) for sperm motility, membrane integrity, and CPT2 gene expression. Conclusion: Incorporating CoQ10 into frozen semen diluent improves gene expression and prevents deterioration of the cell quality of thawed goat spermatozoa. While the study demonstrates the benefits of CoQ10, the precise molecular mechanisms through which CoQ10 enhances gene expression and cell quality were not fully elucidated. Further investigation is needed to understand these mechanisms in detail. Comparative studies with other antioxidants and cryoprotectants can help establish the relative efficacy of CoQ10 and potentially develop more effective combinations. [ABSTRACT FROM AUTHOR]

Artificial insemination (AI) plays a critical role in facilitating rapid genetic and production gains within the sheep industry. However, variable rates of AI success remain a concern for the industry and a barrier to adoption. Furthermore, the degree to which female factors influence the success of intrauterine laparoscopic AI rather than natural mating remains unknown. As such, this study investigates the effect of several factors collected during the time of AI, on the success of intrauterine laparoscopic AI. Data was generously donated by artificial breeding companies and stud breeders during routine commercial AI operations. AI data was collected over 3 breeding seasons during commercial AI programs (N = 24 programs) using Merino ewes (N = 24,700). Sire ID (N = 253), time of AI following progesterone removal (approx. 43–59 h post removal), uterine tone and intra-abdominal fat (both scored 1–5) as well as age of the ewe were all recorded at the time of AI. Transcutaneous ultrasound subsequently determined pregnancy rate approximately 55 days post-AI. A multivariate regression analysis was performed and revealed pregnancy success to increase when semen was inseminated into a ewe with a uterine tone score of 4 or 5 (P < 0.001). The remaining factors fell short of significance within the multivariate model. An interclass coefficient variation matrix was also used to determine the proportion of variation contributed to AI success by random factors allocated in the model; site, sire, AI date and breeding season (45.99 %, 29.94 %, 15.15 % and 8.92 %, respectively). These results highlight the influence of uterine tone on ewe fertility following laparoscopic AI, but also that program location and the sire used can further modify this influence on pregnancy rate. These factors must now be considered in combination with semen factors per individual sire used during AI to ascertain the contribution of several factors to the success of laparoscopic AI in Australia. • Uterine tone scores of 4 and 5 greatly increases the chance of successful pregnancy. • Program environment and the sire used for insemination can further modify pregnancy results. • Assessing uterine tone at AI can indicate oestrus synchronisation, ovulation onset and optimal timing of semen deposition. • Reducing pregnancy variation will enhance the uptake of AI, accelerating genetic and production gains in the sheep industry. [ABSTRACT FROM AUTHOR]

Simple Summary: Improving the reproductive performance of boars is a major goal in pig-breeding programmes. Semen traits are significantly associated with the reproductive performance of boars, and thus, identifying single nucleotide polymorphisms (SNPs) and the genes affecting semen traits are important. Genome-wide association study have been widely used to detect significant SNPs associated with economically important traits in pigs. The results of our study can be used in genomic selection models to facilitate future breeding programs aimed at improving reproductive traits in boars. Artificial insemination plays a crucial role in pig production, particularly in enhancing the genetic potential of elite boars. To accelerate genetic progress for semen traits in pigs, it is vital to understand and identify the underlying genetic markers associated with desirable traits. Herein, we genotyped 1238 Landrace boars with GeneSeek Porcine SNP50 K Bead chip and conducted genome-wide association studies to identify genetic regions and candidate genes associated with 12 semen traits. Our study identified 38 SNPs associated with the analyzed 12 semen traits. Furthermore, we identified several promising candidate genes, including HIBADH, DLG1, MED1, APAF1, MGST3, MTG2, and ZP4. These candidate genes have the potential function to facilitate the breeding of boars with improved semen traits. By further investigating and understanding the roles of these genes, we can develop more effective breeding strategies that contribute to the overall enhancement of pig production. The results of our study provide valuable insights for the pig-breeding industry and support ongoing research efforts to optimize genetic selection for superior semen traits. [ABSTRACT FROM AUTHOR]

To study the primary objective of clinical pregnancy (CP) rate per ovarian stimulation with intrauterine insemination (OS-IUI) treatment cycle in patients with repetitive cycles up to a maximum of 8 cycles. Retrospective cohort. Large fertility clinic. A total of 37,565 consecutive OS-IUI cycles from 18,509 patients were included in this study. Those with anovulatory diagnoses, tubal factor infertility, male factor infertility, using donor sperm, canceled cycles, and those with missing data for either baseline characteristics or outcome were excluded. The CP rate was analyzed using generalized estimating equations and controlled for age, stimulation protocol, and body mass index. Clinical pregnancy was defined as intrauterine gestation with fetal heartbeat visible on ultrasound. A total of 37,565 consecutive OS-IUI cycles from 2002 through 2019 at a private practice facility were evaluated. All cycles met inclusion criteria and were used in generalized estimating equation modeling. Patients aged <35 years comprised 47.6% of the cohort. After adjustment for confounders, the mean predicted probability of CP for cycles one to 8 was 15.7% per cycle. The mean predicted probability of CP in aggregated data from cycles 2 to 4 was only 1.7% lower compared with cycle 1 as the referent (16.7% vs. 15.0%, 95% confidence interval [CI] 2nd: 0.88 {0.82, 0.95}, 3rd: 0.86 {0.79, 0.93}, 4th: 0.88 {0.79, 0.98}). However, the 15.0% mean predicted probability of CP for the second through the fourth cycle was concordant with the mean for all included cycles (15.7%). The mean predicted probability of CP of cycles 5 to 8 was not significantly different compared with the referent (16.7% vs. 16.1%, 95% CI 5th: 0.97 [0.85, 1.11], 6th: 0.93 [0.79, 1.10], 7th: 1.01 [0.81, 1.26], 8th: 1.01 [0.76, 1.34]). The modeling of consecutive cycles suggested that the adjusted cumulative predicted probability of CP from OS-IUI continues to increase with each of the 8 successive cycles. Clinical pregnancy rates are satisfactory in up to 8 consecutive OS-IUI treatment cycles. These data are useful for counseling, especially in those patients for whom in vitro fertilization is not financially or ethically feasible. [ABSTRACT FROM AUTHOR]

This study was designed to provide data on the incidence rates of congenital anomalies and their distribution by organs and systems in calves, lambs and kids brought to Van Yuzuncu Yil University, Faculty of Vetrinary Medicine, Surgery Clinic between 2017-2023. The animal material of the study consisted of 1104 ruminants, including 886 calves, 183 lambs and 35 kids, aged 0-6 months. In addition to clinical examination, anomalies were diagnosed by using direct and indirect radiographic examinations when necessary. In some cases, a definitive diagnosis was made by performing experimental laparotomy based on clinical and radiological examinations. 341 out of 1104 ruminants, including 234 calves, 96 lambs and 11 kids, suffered from congenital anomalies. Of the ruminants with congenital anomalies, 208 (61.00%) were male, 132 (38.70%) female and 1 (0.30%) hermaphrodite. Abdominal wall anomalies were determined with the highest number of 112 (32.9%) cases. These were followed by musculoskeletal system anomalies in 83 (24.4%) cases and gastrointestinal system anomalies in 56 (16.5%) cases. Head region anomalies occurred in 41 cases (12.1%), urinary system anomalies in 23 cases (6.8%) and CNS anomalies in 9 cases (2.6%). Multiple anomalies were recorded in 16 (4.7%) cases. In conclusion, congenital anomalies are frequently seen as pathologies in ruminants in our country, especially in our region. Avoiding inbreeding, making artificial insemination widespread, improving the care and nutrition of the dam during pregnancy, correcting adverse environmental conditions and eliminating stress factors, avoiding over-the-counter medication misuse during pregnancy and most importantly informing field veterinarians and animal owners about the subject will contribute to the prevention of congenital malformations and losses in livestock. [ABSTRACT FROM AUTHOR]

The use of assisted reproductive technologies to improve productivity in livestock industry is becoming popular. The study aimed to determine the utilisation of artificial insemination (AI) and to find out the factors associated with adoption of AI by beef farmers in selected areas of Southern and Ghanzi districts. The data were collected through a survey where a questionnaire was administered to beef farmers. Random sampling was used to interview 54 beef farmers in Southern district and 40 beef farmers in Ghanzi district. The data were analysed using Frequency procedure in Statistical Package for Social Sciences (SPSS). The association between the nominal variables was tested using Pearson Chi-square in SPSS. There is significant (P < 0.05) association between AI utilization by beef farmers and these socio-economic factors; district where farming is done, age of the farmer, education level, occupation of the farmer, purpose of rearing cattle (commercial or subsistence) and land (communal or ranches). Gender of the farmer is not associated with AI utilization. [ABSTRACT FROM AUTHOR]

The purpose of this study was to determine the impact of an antimicrobial peptide, BiF2_5K7K, on semen quality and bacterial contamination in boar semen doses used for artificial insemination. A key factor affecting semen quality and farm production is bacterial contamination in semen doses. Using antibiotics in a semen extender seems to be the best solution for minimizing bacterial growth during semen preservation. However, concern regarding antibiotic-resistant microorganisms has grown globally. As a result, antimicrobial peptides have emerged as interesting alternative antimicrobial agents to replace the current antibiotics used in semen extenders. BiF2_5K7K is an antimicrobial peptide that can inhibit Gram-negative and Gram-positive bacteria isolated from boar semen and sow vaginal discharge. In this study, ten fresh boar semen samples were collected and diluted with one of two types of semen extender: with (positive control) or without (negative control) an antibiotic (i.e., gentamicin). The semen extender without an antibiotic contained antimicrobial peptide BiF2_5K7K at different concentrations (15.625, 31.25, 62.5, and 125 µg/mL). The samples were stored at 18 °C until use. Semen quality parameters were assessed on days 0, 1, 3, and 5, and the total bacterial count was also evaluated at 0, 24, 36, 48, and 72 h after storage. A fertility test on a pig farm was also performed via sow insemination with a commercial extender plus BiF2_5K7K at a concentration of 31.25 µg/mL. No significant difference was found in terms of semen quality on days 0 or 1. On days 3 and 5, the total motility, progressive motility, and viability remained normal in the 15.625 and 31.25 µg/mL groups. However, the sperm parameters decreased starting on day 3 for the 125 µg/mL group and on day 5 for the 62.5 µg/mL group. For total bacterial count at 0, 24, 36, 48, and 72 h, the lowest bacterial count was found in the positive control group, and the highest bacterial count was found in the negative control group compared with the other groups. Comparing antimicrobial peptide groups from 0 to 48 h, the lowest bacterial count was found in the 125 µg/mL group, and the highest bacterial count was found in the 15.625 µg/mL group. For the fertility test, artificial insemination was conducted by using a commercial extender plus BiF2_5K7K at a concentration of 31.25 µg/mL. The results showed a superior pregnancy rate, farrowing rate, and total number of piglets born compared with artificial insemination conducted using a commercial extender plus antibiotic. In conclusion, BiF2_5K7K can inhibit bacterial growth in extended boar semen for 24 h, and thereafter, the bacterial count slightly increases. However, the increase in the number of bacterial counts from days 0 to 3 had no negative effect on sperm quality in the positive control, 15.625, or 31.25 µg/mL groups. This indicates that BiF2_5K7K might be an antimicrobial peptide candidate with potential for use as an alternative antimicrobial agent to replace the conventional antibiotic used in boar semen extenders. [ABSTRACT FROM AUTHOR]

The infertility of cows and heifers has been the most pressing problem in the livestock sector. The genitals and mammary gland have an increased functional load associated with pregnancy, childbirth, and lactation. The purpose of this study was to determine the prevalence of symptoms of chronic irreversible infertility in Ayrshire cows culled as a result of multiple artificial inseminations. The study employed analytical, structural-comparative, and statistical methods, as well as a modified differential palpation diagnosis of the clinical state of the reproductive organs in the ovary+oviduct area with lateral localisation was consistently applied in two stages. The findings of the study showed that cows with in vivo diagnosed symptoms of chronic adherent salpingitis and ovosalpingitis (52.17%), i.e., irreversible form of infertility, had significantly higher (+28.05%; P < 0.01) milk production and better lactation compared to those without these pathologies. Furthermore, in the group of cows culled due to numerous ineffective inseminations, related gonadopathies were detected: from 4.35% to 10.87% of ovarian hypotrophy, 4.35% of gonadal sclerosis symptoms and 17.39% of cystic follicular degeneration cases. In 73.91% of females, the presence of sexual cyclicity with unchanged ovarian function in the follicular phase and in 84.78% - in the luteal phase of the cycle was established. The use of differential palpation diagnostics in production conditions allows in vivo predicting the tendency to loss of genetic resources (the ability of cow ovaries to maintain generative and secretory functions) in highly productive cows due to chronic inflammatory gynecological pathologies, which will contribute to the improvement of drug and biotechnological schemes of therapeutic measures for the prevention of infertility in the dairy herd. [ABSTRACT FROM AUTHOR]

Simple Summary: Outbreaks of chicken anemia virus (CAV) had occurred in specific-pathogen-free (SPF) chicken flocks. Removing seropositive chickens and adjacent seronegative chickens from the SPF facility and disinfecting housing with chemical agents did not halt outbreaks. Incidences decreased with an increase in chicks hatched from CAV-seropositive hens as maternal antibodies protected these chicks from CAV infection. We isolated eggs from CAV-seropositive hens through artificial insemination using CAV-free semen from roosters. The eggs were transferred to a new SPF facility and used to produce CAV-free progeny. To date, the colony raised at the new facility has been CAV-free for longer than two years. A specific-pathogen-free (SPF) chicken colony was maintained with successive groups a month apart in age. The absence of specific pathogens, including chicken anemia virus (CAV), was confirmed through periodic serological tests for each group. However, some groups became CAV seropositive. The procedures of removing seropositive and the adjacent seronegative chickens followed with chemically disinfecting the housing did not halt CAV outbreaks. The full genome sequence of the CAV strain that appeared was closely related to low-virulence isolates in China. The outbreaks of CAV decreased with an increase in the seropositive chicken population, indicating that the progeny is protected from CAV infection by maternal anti-CAV antibodies. The persistence of CAV in erythroid and lymphoid tissues or reproductive tissues from CAV seropositive chickens was examined in chickens of various ages using polymerase chain reaction (PCR). Since a low persistence of CAV was observed in the colony, we isolated eggs from CAV seropositive hens through artificial insemination using semen collected from roosters and confirmed as CAV-free by PCR. Fertilized eggs were transferred to a new SPF facility and used for generating CAV-free progeny. To date, chickens reared in the new facility have been CAV-free for longer than two years. Redirection of eggs from seropositive hens was an effective means of eliminating CAV from chickens. [ABSTRACT FROM AUTHOR]

The rapid spread of African swine fever virus (ASFV), causing severe and often lethal disease in domestic pigs and Eurasian wild boar, continues to be a threat to pig populations and dependent industries. Despite scientific achievements that have deepened our understanding of ASFV pathogenesis, alternative transmission routes for ASFV remain to be elucidated. We previously demonstrated the efficient transmission of ASFV from infected boars to naïve recipient gilts via artificial insemination, thereby highlighting the importance of surveillance of boar semen prior to its shipment. Since the accurate and reliable detection of even low amounts of ASFV in boar semen is key to disease prevention and control, we established a suitable diagnostic workflow to efficiently detect the ASFV genome in boar semen. Here, we assessed the sensitivity of various routine nucleic acid extraction kits as well as qPCR protocols in detecting the ASFV genome in the blood and semen of infected boars. The feasibility of the respective kits and methods for future use in boar studs was also considered. Variability in sensitivity mostly concerned samples with low to very low amounts of the ASFV genome. Ultimately, we defined a well-suited workflow for precisely detecting the ASFV genome in boar semen as early as 2 days post ASFV infection. [ABSTRACT FROM AUTHOR]

Poor sperm quality in cooled-shipped semen has been related to subpar fertility in horses. Therefore, this study aimed to evaluate the ability of post-cooling sperm processing to improve sperm parameters of cooled-stored stallion semen for artificial insemination. For all experiments, ejaculates were collected, processed, and diluted in skimmed milk-based (SM) medium and stored at 5 °C/24h. In all experiments an aliquot of unprocessed cooled semen was used as a control. In the first experiment (Exp 1.), cooled-stored semen from 16 stallions (n = 32) was processed by SpermFilter or centrifugation (600× g /10min) and resuspended in an egg yolk-based freezing medium containing permeating cryoprotectants (EY-C) for cryopreservation. Sperm recovery and motility parameters were immediately assessed after sperm resuspension in both groups and compared with unprocessed (Unp) samples. In Exp 2., cooled semen samples from six stallions (n = 18) were processed using SpermFilter and resuspended in SM or EY-C. Motility parameters and plasma membrane integrity were assessed in all groups (Unp, SM, and EY-C). In Exp 3, cooled semen from four stallions (n = 20) was processed by SpermFilter, resuspended in SM, EY-C, or egg yolk-based medium without cryoprotectants (EY-nC); and submitted to a thermoresistance test (37 °C/3h). Motility parameters, plasma membrane integrity and stability, mitochondrial membrane potential, mitochondrial superoxide generation, and DNA fragmentation index were evaluated in all groups. Finally, in Exp 4, 39 estrous cycles of 11 mares were inseminated with unprocessed (n = 6) cooled-stored semen or semen cooled at 5 °C/24h and then processed by SpermFilter and resuspended in SM (n = 5), EY-C (n = 11), EY-nC (n = 11), or centrifuged and resuspended in EY-C (n = 6). Overall, semen processing and resuspension in EY mediums (EY-C and EY-nC) improved sperm parameters compared with those of unprocessed semen (P < 0.05). Centrifugation (91 ± 5 %) recovered more sperm than SpermFilter (84 ± 9 %; P < 0.05). Sperm resuspended in EY-nC maintained better sperm parameters throughout the thermoresistance test than those in the other groups (P < 0.05). The fertility rates were similar between all groups (P > 0.05). In conclusion, processing and resuspension in EY medium can improve sperm parameters in post-cooled-stored stallion semen. • Extender exchange may enhance sperm quality of cooled-transported stallion semen. • SpermFilter and centrifugation yield similar sperm parameters of cooled-shipped processed semen. • Cryoprotectants are detrimental for long-term sperm parameters of cooled-shipped stallion semen. [ABSTRACT FROM AUTHOR]

This research aimed to increase the reproduction of dairy cows with artificial insemination (AI) in Filial Friesian Holstein cows using sexed semen. This research was conducted in Pandesari Village, Pujon District, Malang Regency, East Java. The 114 Filial Friesian Holstein cows were used in this research and divided into three Treatments: T1: 38 cows were inseminated using unsexed semen, T2: 38 cows were inseminated using albumin sedimentation sexed semen, and T3: 38 cows were inseminated using Percoll density gradient centrifugation (PDGC) sexed semen. The material was selected by purposive sampling with a minimum body condition score (BCS) specification of 2.5 (scale 1-5); the material had normal reproductive organs and showed signs of heat/estrus. The parameters of this study are the percentage of non-return rate (NRR) 1, NRR 2, and conception rate (CR). The data obtained were analyzed using descriptive analysis. The differences in NRR and CR between the unsexed sperm, sexed sperm with albumin sedimentation, and sexed sperm with PDGC were analyzed with the chi-square test and were considered significant at p < 0.05. The chi-square test was carried out to compare the observed values with the expected values. The results showed that the success of artificial insemination was greater by using albumin-sedimented sexed semen compared to unsexed semen or PDGC-sexed semen, with NRR values of 1 (95%), NRR values of 2 (87%) and CR values of 63%. The conception rate of artificial insemination using albumin-sedimented sexed semen was 63% greater than that of artificial insemination using unsexed semen and PDGC-sexed semen, which obtained the same value of 47%.

As the beef industry moves towards efficient animal production to improve sustainability in agriculture, new production and management approaches are emerging. Among the many facets of the beef industry, cow–calf operations have the most opportunity for efficiency improvement, including improvements in fertility. This project accounts for measures and methods of (1) twinning reproductive technologies and (2) twin calf perinatal care and pre-weaning rearing. The overall objective was to produce twin calves using two reproductive technologies—embryo transfer and artificial insemination. The subobjectives were to determine accuracy of twin pregnancies embryo/fetal losses using ultrasonography, evaluate parturition and dystocia, and determine the effects of different twin-raising methods on neonatal behavior and growth. A fixed-time artificial insemination (FTAI) protocol was applied to 77 multiparous Angus-cross cows from a commercial beef herd in northcentral South Dakota during the summer of 2019. Cows were assigned to two different treatments groups: only artificially inseminated (AI) or received an embryo transfer following artificial insemination (ET + AI). They were estrous-synchronized, artificially inseminated (AI) with black Angus semen at day 0, and received and embryo transfer (ET) at day 7. Ultrasound examination detected 56% pregnancy risk for both groups, with sensitivity, specificity, and accuracy of 75%, 100%, and 90.5%, respectively, for bilateral twin detection. Calves were born during spring 2020. Twin calves (n = 34) and singleton calves (n = 11) were assigned to one of three raising methods: (1) twin born and twin raised (TT; n = 16), (2) twin born and single raised (TS; n = 18), and (3) single born and single raised (S; n = 11). Neonatal nursing behavior and birth weights were recorded, and adjusted day 200 and day 280 were calculated measures of vitality and growth. Blood samples were collected at age 24 h for colostrum intake measures (total serum protein, IgG1, and IgM). Twin calves were born 20% (p < 0.05) lighter in body weight than singletons; however, weights did not differ at day 280 between TT and S calves. TS calves had the shortest average latency to stand, but immunoglobulin concentrations did not differ among treatments. At weaning, cows that had birthed and raised twins produced more kilograms of live weight per pregnancy than cows birthing and raising singletons. Using ET + AI proved to increase twinning rate, and growth was maintained when raising both twins with their dam.