Effects of GnRH administration on pregnancy rate and litter size in ewes subjected to a fixed time insemination program during the breeding season.

Background and Aim: Conventional egg-yolk and milk-based extenders are widely used for semen cryopreservation but pose biosafety concerns and compositional variability that compromise standardization. Liposome technology offers a biosecure, uniform alternative. This study aimed to develop soy lecithin-derived nanoliposomes (NLs) using an ultrasonic-based process and to evaluate their efficacy as a Tris-based extender for Ongole-grade bull semen cryopreservation. Materials and Methods: Soy lecithin NLs were prepared through probe ultrasonication (15–45 min) and ultracentrifugation, followed by physicochemical characterization using particle size analysis, scanning electron microscopy (SEM), and high-resolution transmission electron microscopy. Tris-NL (TNL) extenders were formulated at concentrations of 5–25 mg/mL and compared with a Tris–egg-yolk (TEY, 20%) control. Fresh semen from five Ongole-grade bulls was evaluated for motility, viability, and morphology before and after freezing. Kinematic parameters were assessed through computer-assisted semen analysis, plasma membrane integrity by hypoosmotic swelling test, acrosome integrity using fluorescein isothiocyanate-conjugated peanut agglutinin/propidium iodide staining, and DNA fragmentation by Halomax-sperm chromatin dispersion assay. Results: Optimized sonication (45 min) produced stable NLs (mean diameter 76 nm, zeta potential −43.2 mV) with uniform spherical morphology. Among the tested formulations, TNL 5–10 mg/mL showed significantly higher (p < 0.05) post-equilibration motility (up to 98%), progressive motility, and kinematic parameters (velocity curved line, velocity average path, and velocity straight line) than TEY. Post-thaw evaluations demonstrated improved sperm viability (≈66%), reduced abnormalities (<7%), enhanced plasma-membrane and acrosomal integrity, and lower DNA fragmentation (~1.2%) in the 5–10 mg/mL groups. SEM confirmed smoother sperm surfaces with minimal cryo-damage compared with TEY. Conclusion: Soy lecithin-derived NLs at 5–10 mg/mL serve as an effective and biosecure substitute for egg yolk in Tris extenders, enhancing motility, viability, and structural integrity of Ongole-grade bull spermatozoa. This locally developed, plant-based nanotechnology supports biosafety, import substitution, and sustainability of artificial insemination programs in Indonesia. Keywords: computer-assisted semen analysis, cryopreservation, lecithin, nanoliposome, Ongole-grade bull, semen extender, sperm integrity.

This study evaluated the impact of pre-freezing semen dilution rate and dimethyl acetamide (DMA) concentration on the post-thaw motility and fertility of cryopreserved rooster sperm. Rooster ejaculates were diluted with a standard EK extender to achieve low (LSC; 1 × 10⁹ sperm/mL) and high (HSC; 2 × 10⁹ sperm/mL) sperm concentrations. Each dilution group was further treated with three DMA concentrations (3%, 6%, or 9%) before cryopreservation. Post-thaw sperm motility traits were obtained by computer-assisted sperm analysis (CASA), and fertility features were evaluated through artificial insemination in hens. The current results showed that HSC significantly improved total motility (TM), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF), but reduced linearity (LIN) and straightness (STR) compared to LSC. DMA concentration had a quadratic effect on motility, with 6% yielding the highest progressive motility (PM), straight line velocity (VSL), and BCF. Fertility outcomes revealed that HSC resulted in higher fertilization rates, while neither DMA concentrations nor their interaction with dilution rates exerted significant effects on fertility traits. VCL, ALH, and BCF showed positive correlations with pipping-chicks rates, whereas STR, LIN, and WOB displayed negative correlations. These findings underscore the critical interplay between dilution rate and cryoprotectant concentration and provide practical guidance for developing more reliable cryopreservation protocols that can be applied under field conditions to enhance fertility management in poultry production.

Background. Cryopreservation of sperm from cattle is an effective method of breeding work. To increase the efficiency of cryopreservation, the improvement of synthetic diluents for freezing sperm is required. Special attention is paid to the search for new cryoprotectors capable of increasing sperm survival and preserving their fertility during cryopreservation. Purpose. Evaluation of the effectiveness of artificial insemination of cows with sperm containing molecular hydrogen. Materials and methods. For the experiment, 2 groups of animals were selected, 50 heads each. The cows of the control group were inseminated with sperm frozen in a BioXcell medium, and the animals of the experimental group with sperm frozen in a BioXcell medium with molecular hydrogen. Insemination of cows was carried out by a group method by synchronizing sexual hunting according to the "Ovsinh" scheme. The effectiveness of insemination was evaluated on the basis of rectal ultrasound examination data after insemination. A comparison was made of the effectiveness of artificial insemination of cows with sperm cryopreserved in an improved medium containing molecular hydrogen with the indicators of fertilization obtained using sperm frozen in an environment without molecular hydrogen. Results. Fertilization of cows with sperm containing molecular hydrogen increased by 11.5% after 3 months compared with the use of cryopreserved sperm without molecular hydrogen. Conclusion. The results of the experiments indicate that the use of molecular hydrogen in the composition of the medium for diluting the seed makes it possible to increase the fertilizing ability of bull sperm and increase the fertilization rates of cows during artificial insemination.

Background: Geographic location is one of the key factors influencing the reproductive performance of cattle. Environmental variables such as temperature, humidity, and rainfall directly affect the reproductive efficiency of cows. Purpose: This study aimed to compare the reproductive performance of beef cattle raised in highland and lowland areas. Method: A total of 60 productive female cows were surveyed, comprising 30 cows from the lowland area of Borobudur (230–240 m above sea level) and 30 from the highland area of Sawangan (450–1400 m above sea level). Data on calving interval and service per conception were obtained through field surveys of farmers maintaining artificial insemination records. The data were analyzed using a one-way ANOVA test. Results: Reproductive performance was assessed based on calving interval and service per conception. Cows raised in the highland area of Sawangan demonstrated superior reproductive performance, with a calving interval of 429.66 ± 145.16 days and a service per conception rate of 1.5 ± 0.73, compared to 516.33 ± 168.72 days and 2.5 ± 1.61 in the lowland area of Borobudur. These findings suggest that higher altitudes with cooler and more stable climatic conditions contribute to improved reproductive efficiency. Conclusion: The results indicate that altitude has a significant influence on cattle reproductive performance. Cows raised in highland areas exhibited shorter calving intervals and lower service per conception rates compared to those in lowland areas. Cooler temperatures and lower humidity in highlands likely enhance reproductive outcomes. Farmers in lowland regions may improve reproductive efficiency by implementing cooling systems or adjusting breeding schedules. Further studies are recommended to identify additional environmental and management factors influencing cattle reproduction across different agroecological zones.

This study analyzes the level of satisfaction of broiler farmers with the performance of Artificial Insemination (IB) officers in Central Lombok Regency. A quantitative survey method with a descriptive approach was used to collect data from 99 farmers spread across four sub-districts. Service per Conception (S/C), Conception Rate (CR), and Non Return Rate (NRR) parameters were used to assess the technical performance of IB officers. The results of the study show that the level of satisfaction varies between sub-districts, with the performance of IB officers in Jonggat and Pringgarata Districts being more optimal than other regions. This analysis provides recommendations in the form of increasing the technical capacity of officers, as well as increasing education for farmers to support the success of IB programs in the region.

This study aimed to evaluate the effect of platelet-rich plasma (PRP) on sperm motility in men with asthenozoospermia and assess its potential use in intrauterine insemination (IUI). A prospective randomized controlled pilot study was conducted with 40 men diagnosed with asthenozoospermia. Semen samples were divided into two equal parts: one treated with autologous, non-activated PRP and the other serving as control. Sperm motility was assessed at 1, 3, 6, and 24h using a Makler chamber. A two-step centrifugation method was used for PRP preparation, and all samples underwent identical swim-up protocols. Data were analyzed using two-way repeated measures ANOVA. PRP-treated samples showed significantly higher motility than controls at all time points (p < 0.001). PRP also slowed the decline in motility over time. At 24h, motility was 50.5% in the PRP group versus 21.1% in controls. Statistical analysis confirmed normal data distribution. PRP significantly enhances and preserves sperm motility during in vitro incubation. These findings suggest that PRP may serve as a supportive option in sperm preparation for IUI, potentially reducing the need for more invasive techniques. Further studies are warranted to evaluate clinical outcomes.

Semen cryopreservation is an essential technique in artificial insemination (AI) and assisted reproductive technology (ART). The addition of antioxidants to the freezing medium is a promising strategy to reduce cryo-induced damage to sperm cells. While sperm freezing has been extensively studied, this research aimed to improve the post-thaw durability and functionality of sperm. A total of 110 semen samples 55 from fertile and 55 from infertile individuals were collected and processed. After centrifugation, each sample was mixed with a cryoprotectant medium at a 1:1 (v/v) ratio. Three different cryoprotectant formulations (S1, S2, and S3), each containing varying concentrations of protective agents, were evaluated. The samples were then frozen in liquid nitrogen at -196°C. Post-thaw analyses included assessments of sperm count, motility, vitality, morphology, DNA fragmentation, and reactive oxygen species (ROS) levels representing a major strength of the study due to the comprehensive evaluation of sperm quality. The results showed significant differences in DNA fragmentation between fresh and cryopreserved samples. Among the tested formulations, S3 supplemented with citric acid anhydrous and taurine produced the best outcomes. It significantly improved sperm motility and vitality, while effectively reducing oxidative stress and cryodamage. Statistical analysis using one-way ANOVA and Tukey's HSD test revealed significant differences in sperm morphology between fresh and post-thaw S3-treated samples (p < 0.05), with a 95% confidence level. A limitation of the study was the long interval between the use of fresh control samples and the analysis of cryopreserved samples, which may influence comparative accuracy. Despite this limitation, the findings suggest that the S3 formulation is highly effective in preserving sperm quality. These results support its potential integration into fertility preservation protocols in ART. Furthermore, the outcomes may guide clinical practice and support the development of national policies aimed at improving access to advanced cryopreservation techniques as part of comprehensive reproductive healthcare services.

Changes in maternal nutrition in the periconceptional period can have long term effects on the offspring. The aim of the current study was to determine the influence of feeding 15 g of rumen-protected methionine (RP-Met) during the periconceptional period on female progeny during the post-weaning phase until puberty. A total of 114 beef cows were fed a roughage-based diet and randomized to receive corn gluten feed supplemented with 15 g of RP-Met (Smartamine M, Adisseo) or not (CON) from day -7 to +7 relative to artificial insemination with female sex-sorted semen. Female calves (n = 32) remained with their dams until weaning. A month after weaning, calves were weighed every 14 d, and withers height (WH), body length (BL), and heart girth (HG) were measured during the post-weaning phase. At 420 ± 13 d of age, a 56-d performance test was conducted, and initial body weight (IBW), final body weight (FBW), dry matter intake (DMI), average daily gain (ADG), and residual feed intake (RFI) were measured. At the end of the performance test, the 12th rib back fat thickness (BF), ribeye area (REA), intramuscular fat (IMF), and rump fat thickness (RF) were determined via ultrasound. Data from the post-weaning phase, performance test, and body composition were analyzed as a RCBD using generalized linear mixed models through the MIXED procedure of SAS, with body measurements analyzed as repeated measures. A treatment-by-day interaction (P = 0.003) was observed for HG, with larger HG for RPM from day 308 onwards. Final BW was greater for RPM heifers (CON = 436.13 ± 1.43 kg; RPM = 445.63 ± 1.52 kg; P < 0.01). Also, RPM heifers had a greater ADG than CON heifers (CON = 1.40 ± 0.04 kg; RPM = 1.65 ± 0.04 kg; P < 0.01). No treatment difference (P > 0.05) was observed for DMI (CON = 12.06 ± 0.19 kg/d; RPM = 12.14 ± 0.18 kg/d) or RFI (CON = -0.12 ± 0.15; RPM = 0.11 ± 0.14). There was also no effect of treatment (P > 0.05) on REA or IMF. RF was less for RPM (CON = 1.16 ± 0.07 cm; RPM = 0.91 ± 0.06 cm), and there was a tendency (P = 0.07) for lesser BF in RPM. In conclusion, supplementing dams with RP-Met enhanced FBW of their heifer progeny during the performance test; furthermore, a treatment-by-day interaction tendency was observed for HG. RPM heifers also had reduced RF and a tendency for reduced BF.

Our bone health as an adult is defined by patterns of development in early life, with perturbed growth during fetal and neonatal periods predisposing individuals to poor bone health in adulthood. Studies have identified poor maternal diet during pregnancy as a critical factor in shaping offspring bone development, with significant impacts on adult bone structure and health. However, the association between a father's diet and the bone health of his offspring remains poorly defined. To address this knowledge gap, we fed male C57BL/6 mice either a control normal protein diet (NPD; 18% protein) or an isocaloric low-protein diet (LPD; 9% protein) for a minimum of 8 wk. Using these males, we generated offspring through artificial insemination, in combination with vasectomized male mating. Using this approach, we derived offspring from either NPD or LPD sperm but in the presence of NPD or LPD seminal plasma. Using micro-computed tomography and synchrotron X-ray diffraction, we observed significant changes in offspring femur morphology and hydroxyapatite crystallographic parameters from just 3 wk of age in offspring derived from LPD sperm or seminal plasma. We also observed that differential femur morphology and hydroxyapatite crystallographic parameters were maintained into adulthood and into a second generation. Analysis of paternal sperm identified a down regulation of 26 osteogenic genes associated with extracellular matrix levels and maintenance, transcription and growth factors, and bone ossification. These observations indicate that poor paternal diet at the time of conception affects offspring bone development and morphology in an age and generation specific manner.

Population was of primary concern in the mid-twentieth century and Australia’s declining birth rate in the 1930s and 40s brought forth concerns about the reproductive nation. One of the key professions which engaged with ideas of a reproductive Australia was medicine, particularly gynaecologists, obstetricians, and surgeons. Their medical discourse in the mid-twentieth century when exploring infertility exhibited a tension between the burgeoning medical understanding of infertility within marriages - shared between the husband and wife - and the social understanding of reproductive responsibility resting on the woman. Through new approaches to diagnosis and therapeutic technologies, such as artificial insemination, this tension around responsibility was elucidated in the medical discourse. Even when incorporating or exclusively assessing the man’s fertility status there was an underlying presumption of, if not female fault, female responsibility. As the majority of practitioners were gynaecologists, obstetricians and surgeons, the continued focus on women could be justified through their particular medical specialisation which exclusively examined women. Despite the increased knowledge of male fault, often incurable male fault, in infertile marriages the very nature of examination, diagnosis and treatment was grounded in ideas of female reproductive responsibility.

BACKGROUND Fetal growth restriction (FGR), or intrauterine growth restriction, is more common in twin pregnancies. Pravastatin enhances nitric oxide synthesis and placental vasodilation, and L-arginine, a precursor to nitric oxide, reduces pulmonary artery resistance. There is evidence that combined pravastatin and L-arginine can improve blood flow to the placenta, prolong pregnancy, and prevent preeclampsia in complicated pregnancies, including monochorionic twin pregnancies. CASE REPORT A 35-year-old patient, gravida 1, with dichorionic twin gestation conceived via intrauterine insemination presented at 16 weeks of gestation with severe FGR of both fetuses. At 21 weeks and 5 days of gestation, umbilical artery Doppler studies demonstrated absent end-diastolic flow for twin B. No anatomic fetal anomalies were detected. Treatment with pravastatin 40 mg and L-arginine 1.5 g daily was initiated at that point and continued until delivery. The pregnancy was prolonged until 32 weeks and 1 day of gestation, with inpatient monitoring when preeclampsia with severe features developed and delivery was recommended. She underwent a primary classical cesarean delivery. Both fetuses were severely growth-restricted, less than the 1st percentile for gestation. Both twins survived without major morbidity. CONCLUSIONS This report presents the case of a 35-year-old woman with a dichorionic twin pregnancy with FGR diagnosed at 16 weeks of gestation, managed with pravastatin and L-arginine, resulting in twin delivery at 32 weeks. This report supports findings from recent studies that combined pravastatin and L-arginine can improve fetal development and survival in FGR within monochorionic twin pregnancies. Adequately powered and randomized trials are needed to confirm this finding.

This study aimed to investigate the effect of maternal taurine (Tau) supplementation during gestation on the development of brown adipose tissue (BAT) and the thermogenesis of offspring lambs. Sixty ewes were randomly divided into three groups and daily supplemented with 0 g (control group), 2 g (low Tau), and 4 g (high Tau) during the gestation period after artificial insemination. The body surface temperature of newborn lambs was measured daily. At 15 days of age, lambs were slaughtered for sample collection. Compared to the control group, lambs in the high-Tau group exhibited higher body temperatures (p < 0.05). Maternal Tau-treated lambs had higher expression of UCP1 (p < 0.05), a greater number of multilocular brown adipocytes, and increased mitochondrial count in the BAT (p < 0.05). Additionally, there were higher copies of mitochondrial DNA and expression of mitochondrial proteins in the maternal Tau-treated lambs (p < 0.05). Enhanced AMPK and ULK1 activation (p < 0.05) were also observed in the BAT of 15-day-old lambs. Further analysis of the effects of Tau on BAT stromal vascular fraction (SVF) cells revealed that Tau treatment upregulated UCP1 during the brown adipogenic differentiation stages (p < 0.05). Consistently, Tau treatment upregulated mitochondrial biogenesis genes and increased the number of mitochondria (p < 0.05). Tau treatment significantly upregulated PGC1a and SIRT3, as well as activated AMPK and ULK1 (p < 0.05). Mitophagy is initiated, but the mitochondrial morphology remains stable. In conclusion, maternal Tau supplementation during gestation promotes brown adipogenesis and enhances the thermogenic activity of offspring lambs by activating the AMPK-PGC1a signaling pathway.

The article explores the current state, dynamics, and development prospects of the reproductive rights movement in European countries. Particular attention is paid to the forms of struggle by women and men for access to artificial insemination, abortion, and surrogacy.The contribution of public, medical, and feminist organizations to the advancement of these rights is examined, as well as the role of state policy, legislative initiatives, and international support. Examples are provided from France, Germany, Poland, Sweden, Denmark, theNetherlands, Austria, Finland, and also Ukraine. It is shown that under the conditions of the armed conflict in Ukraine, there arises a need to preserve reproductive material in case of injury or death of one of the partners, which has led to new legislative changes. The text includes a table comparing national approaches, charts of protest movement activity, and a flowchart ofjudicial protection of reproductive rights.

Objective: To compare pregnancy outcomes between patients undergoing combined hysteroscopy and hysterosalpingo-contrast sonography (HyCoSy) versus hysteroscopy alone prior to intrauterine insemination, and to evaluate the safety and clinical value of the combined procedure in the diagnosis and treatment of infertility. Methods: A retrospective analysis was conducted on clinical data from 385 patients who underwent hysteroscopy at Peking University Third Hospital between October 1, 2020 and September 30, 2022, and subsequently received their first cycle of artificial insemination with donor sperm (AID) within six months. Pregnancy outcomes were compared between the group receiving combined hysteroscopy with four-dimensional HyCoSy (hysteroscopy+4D-HyCoSy group) and the group receiving hysteroscopy alone (hysteroscopy group). Multivariate logistic regression was used to analyze factors influencing pregnancy outcomes after AID. Results: Among the 385 patients included, 79 achieved clinical pregnancy. The clinical pregnancy rate (24.9%, 53/213) and live birth rate (21.1%, 45/213) in the hysteroscopy+4D-HyCoSy group were significantly higher than those in the hysteroscopy group [15.1% (26/172) and 12.8% (22/172), respectively; all P<0.05]. There was no significant difference in tubal patency between the two groups (P>0.05); however, the time interval from tubal patency assessment to intrauterine insemination was significantly longer in the hysteroscopy group compared to the hysteroscopy+4D-HyCoSy group (median: 4.0 vs 2.0 months; P<0.001). Multivariate analysis showed that double insemination significantly increased clinical pregnancy rate compared to single insemination (OR=2.42, 95%CI: 1.02-5.72; P=0.044). An interval exceeding 6 months between tubal patency assessment and intrauterine insemination was identified as a risk factor for reduced clinical pregnancy (OR=0.35, 95%CI: 0.14-0.92; P=0.047). Additionally, neither the time interval from hysteroscopy to intrauterine insemination nor hysteroscopic findings and pathological diagnoses had significant effects on clinical pregnancy rates (all P>0.05). Conclusions: The combination of hysteroscopy and HyCoSy provides a safe and efficient approach for fertility assessment in infertile patients and improves clinical pregnancy rate and live birth rate in intrauterine insemination cycles. Hysteroscopy is recommended for patients with suspected endometrial or intrauterine abnormalities. If no previous tubal patency assessment has been performed or the last assessment was more than six months prior, combined hysteroscopy and HyCoSy may be considered to enhance the likelihood of clinical pregnancy.

Pig breeding as one of the livestock industries has a decisive impact on the level of food security, independence of the country and ensures the health of the people. When reproducing livestock, artificial insemination is used, the result of which depends to a greater extent on the quality of the semen used. One of the qualitative indicators to anticipate successful fertilization is sperm motility. The purpose of the research was to study the motility of sperm of freshly obtained undiluted, freshly obtained diluted and frozen sperm of breeding boars of Large White, Landrace, Duroc and Pietrain breeds. Sperm motility and movement patterns were determined by CASA (Computer-Assisted Semen Analysis) using Andro-Vision (MiniTube) on a ZeissAxioLab.A1 phase contrast microscope. It was found as a result of research that sperm motility varies significantly depending on the breed of breeding boars and the type of sperm. Thus, native sperm exceeds in total, forward and fast motility diluted sperm by 9.07 abs.%, 35.27 and 56.96 abs. % and frozen sperm by 28.01 abs. %, 43.26 and 66.72 abs.%, respectively. When comparing breeds, the highest total and forward motility of native and diluted sperm was established in boars of Large White breed (91.04 ± 0.25 and 90.69 ± 0.22 %; 83.10 ± 0.79 and 59.22 ± 2.08 %, respectively), the lowest in boars of Duroc breed (84.01 ± 2.64 and 83.10 ± 2.73 %; 75.12 ± 1.20 and 46.95 ± 2.66 %, respectively). The quality characteristics of the semen of breeding boars of Pietrain breed change in terms of motility 5 minutes after thawing and 5 hours after incubation at a temperature of +37°C.. Thus, the total, forward and fast forward motility of sperm after 5 hours of incubation is reduced by 7.83 % abs%, 6.96 and 2.49 abs%, respectively.

In 2025, it is our considered opinion, based on 40 years of experience in full-time andrology and reproductive sciences, that varicocele as a cause of male infertility remains largely unproven. While some meta-analyses suggest a marginal benefit in specific patient subsets, these findings are often debated regarding their clinical significance or methodological limitations. Couples with impaired semen parameters can be effectively treated by intrauterine insemination and, if necessary, by advanced assisted reproductive technologies like intracytoplasmic sperm injection (ICSI). With unproven benefits, an invasive nature, no clear explanation for the actual mechanism of causation, an unpredictable outcome, and the availability of simpler, better alternatives like IUI and ICSI, I feel varicocelectomy is the unkindest cut of all.

Artificial insemination (AI) is a critical tool for genetic improvement and fertility management in goats. This study examined the effects of semen deposition site (uterine body, cervix, vagina) and vaginal mucus type (cloudy, turbid, clear) on pregnancy rate, kidding rate, and average litter size in 300 CIDR-synchronized Alpine does in southern Taiwan. Semen deposited in the uterine body combined with cloudy mucus yielded the highest pregnancy rate (55.9%), while vaginal deposition with clear mucus resulted in the lowest (30.7%). Two-way ANOVA showed significant main effects and interactions for pregnancy rate and average litter size (p < 0.05), but no significant effect on kidding rate. Pregnant does exhibited lower vaginal mucus electrical conductivity, higher pH, and elevated temperature compared to non-pregnant does, suggesting these parameters as potential biomarkers for estrus detection. Findings highlight the importance of precise semen placement and optimal mucus condition for improving AI protocols in goats.

Simple SummaryIntestinal atresia is a congenital disorder in calves where a segment of the intestine is closed or absent, causing digestive obstruction. Affected calves usually show abdominal swelling, lack of fecal output, and reduced appetite within the first days of life. In this study, we found that calves with atresia coli exhibited significantly elevated anion gap and plasma lactate, along with decreased ionised calcium and pH, compared to healthy calves. Anion gap and ionised calcium showed high diagnostic accuracy, suggesting their utility in distinguishing atresia coli from other neonatal conditions such as sepsis or diarrhea. Better understanding and timely management of intestinal atresia can improve animal welfare and reduce economic losses in cattle production.This study aimed to evaluate blood lactate, anion gap, and ionised calcium levels as potential diagnostic biomarkers in calves with atresia coli, and to identify possible predisposing factors such as breed, gender, age, method of conception, number of lactations, and births. The study included twenty-two calves with atresia coli and ten healthy controls, all aged 1–11 days (median, 3 days), brought to Erciyes University Veterinary Faculty from Kayseri and nearby provinces due to non-defecation and abdominal swelling. Prominent clinical findings among the 22 calves with atresia coli included abdominal distension in 90.9%, anorexia in 81.8%, and depressed general posture in 86.4%. Blood gas analysis revealed significantly elevated lactate and anion gap and decreased ionised calcium and pH in atresia coli calves compared to controls (p < 0.05). Anion gap (>14.05 mmol/L) and ionised calcium (<1.205 mmol/L) demonstrated high diagnostic accuracy (AUC: 0.964 and 0.872, respectively), suggesting their potential as supportive biomarkers for early detection of atresia coli. The study data revealed that male gender, artificial insemination, and calves born from the third or subsequent pregnancies are statistically significant risk factors for the development of atresia coli. Atresia coli in calves is characterized by specific clinical signs and significant changes in blood gas parameters, such as elevated lactate and anion gap and reduced ionised calcium and pH. Early detection using these markers can improve diagnosis, and further studies should focus on prevention by addressing these risk factors.

Sperm cryopreservation is a critical technique for improving the efficacy of artificial insemination in sheep breeding programs. This study evaluated the effects of different extenders and packaging methods on post-thaw ram sperm quality, ultrastructure, and potential reproductive performance. Semen was collected from five healthy Ossimi rams and extended using three cryopreservation media: Tris-egg yolk (Tris-EY), Tris supplemented with 1% soybean lecithin (Tris-SBL), or Tris supplemented with 2 mM butylated hydroxytoluene (Tris-BHT). The extended semen was then packaged in either plastic straws or pellets and cryopreserved in liquid nitrogen. Post-thaw sperm quality was assessed by evaluating progressive motility, viability, and membrane integrity. Sperm ultrastructure was examined using transmission electron microscopy (TEM) and reproductive index. A two-way ANOVA analysis was used to study the effects of the extender and the method of packing. Cryopreservation using straws significantly improved post-thaw sperm quality compared to pellets, demonstrating higher progressive motility (p = 0.003), viability (p < 0.0001), and membrane integrity (p < 0.0001). However, neither the packaging method nor the extender type significantly affected the plasma membrane status, acrosome integrity, or mitochondrial, tail, and nuclear damage (p > 0.05). Ultrastructural analysis confirmed that straws, regardless of the extender used (Tris-EY, Tris-SBL, or Tris-BHT), effectively preserved sperm ultrastructure, including the nucleus, head, acrosome, plasma membrane, and mitochondria. The methods of packaged and extender types did not significantly affect the conception rate and litter size in sheep (p > 0.05). However, it is worth noting that Tris-SBL in straws had the highest conception rate (85.7%) and litter size (1.5 per ewe), while Tris-BHT in pellets had the lowest values (71.4% and 1.2, respectively). This study confirmed that plastic straw packaging significantly improved post-thaw sperm quality. While Tris-EY showed higher reproductive performance, further research is needed to establish statistical significance in pregnancy rates and litter size.