Veterinary Obstetrics and Gynecology

Fertility is the most important aspect in the efforts to increase livestock populations. Protamine and various proteins in sperm and seminal plasma are the results of the molecular analysis which can be used as a marker of fertility. Each of the proteins plays an important role in the normal function of sperm, starting from the formation of sperm structure, motility, capacitation, cell protection, acrosome reactions, successful fertilization, egg activation, and embryonic development. Finally, these molecular components can be a marker of fertility and can help to diagnose the cases of infertility/subfertility in livestock in the field.

Aim: This study aimed to investigate the protective effect of Rosmarinus officinalis (L.) essential oil on rooster sperm motility during 4°C short-term storage. Materials and Methods: R. officinalis essential oil was analyzed using gas chromatography coupled to mass spectrometry to identify the active components. 10 of 45-week-old Hubbard commercial broilers were subjected to biweekly semen collections during 3 weeks. At each collection, sperm was pooled and divided into four aliquots and then diluted with Tris extender supplemented with 870, 87, or 8.7 µg/ml of R. officinalis essential oil, identified as treatments R, R5, and R10, respectively. Tris-based extender without any supplementation was considered as a control group. Diluted sperm was then stored at 4°C in the refrigerator and analyzed at 0, 6, 24, and 48 h using a computer-assisted sperm analyzer. Different semen parameters were measured including total motility, progressive motility, gametes velocities (straight line velocity [VSL], curvilinear velocity [VCL], and average path velocity [VAP]), amplitude of the lateral head displacement [ALH], and beat-cross frequency [BCF]. Results: The phytochemical analysis of R. officinalis essential oil revealed the presence of 25 active components including seven major molecules: Camphor (18.88%), camphene (5.17%), 1,8-cineole (7.85%), β-thujene (13.66%), α-thujene (4.87%), chrysanthenone (12.05%), and β-cubenene (7.97%). The results showed a beneficial effect of R. officinalis essential oil on sperm cells motility, particularly when using the lowest concentrations, 8.7 and 87 µg/ml. Progressive motility and gametes velocities (VCL, VSL, and VAP), materializing the quality of gametes motility, showed highly statistically significant values (p<0.01) in 8.7 and 87 µg/ml treatments, especially from 6 h of storage at 4°C. Conversely, the highest concentration (870 µg/ml) showed harmful effects with a total spermicidal activity after 24 h of storage. Conclusion: The current results revealed the positive impact of R. officinalis essential oil on rooster sperm at 4°C short-term storage probably through fighting against oxidative stress and cold shock damages.

Aim: The present study was conducted to evaluate the quality of rooster sperm at 5°C after treatment with a diluent containing coconut water, fructose, and chicken egg yolk and stored the semen sample at 5°C.

Materials and Methods: Ten semen samples from 10 healthy roosters were subjected to four different treatments. For the treatments, 0.2 ml fresh semen with a sperm concentration of 5.2X109 cell/ml was mixed with T0 (no diluent), T1 (0.34 ml coconut water and 6 μl fructose), T2 (0.274 ml coconut water, 0.12 ml egg yolk, and 6 μl fructose), and T3 (0.34 ml egg yolk and 6 μl fructose) solutions. Each treated solution was stored at 5°C and evaluated both macroscopically and microscopically. Macroscopically, semen volume, pH, and sperm concentration were evaluated. The microscopic sperm characteristics examined included total motility (i.e., rapid, medium, or slow), progressive and non-progressive motility, viability, and spermatozoa abnormalities noted at different storage times. The results showed that spermatozoa motility was under 40%.

Results: The results indicated that sperm viability significantly affected (p<0.05). The highest mean value of sperm viability on day 7 of storage was found after treatment with the T2 solution (46.100±0.5677%). Similarly, spermatozoa abnormalities were significantly lower after treatment with the T2 solution (6.680±1.702%).

Conclusion: The addition of a diluent containing coconut water, egg yolk, and fructose helped in the better preservation spermatozoa motility, as well as viability for up to 7 days when the semen samples were stored at 5°C.

(2017) Effect of different concentrations of soybean lecithin and virgin coconut oil in Tris-based extender on the quality of chilled and frozen-thawed bull semen, Veterinary World, 10(6): 672-678. Abstract Aim: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL) and virgin coconut oil (VCO) in Tris-based extender on chilled and frozen-thawed bull semen quality parameters. Materials and Methods: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%). A 20% egg yolk in Tris used as a positive control (C+). The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain), membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test. Results: The results showed that all the quality parameters assessed were significantly (p<0.05) improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+) in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups. Conclusion: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.

Background and Aim: Despite many trials, buffalo embryos have poor cryosurvivability because of their high lipid content. L-carnitine was found to be a lipid-reducing agent when added to oocyte and embryo culture media. The study aimed to determine the most effective concentration of L-carnitine to improve the oocyte developmental competence and cryotolerance of buffalo embryos.

Materials and Methods: In vitro maturation and embryo culture media were supplemented with four concentrations of L-carnitine: 0 (control), 0.25, 0.5, and 1 mM. Good-quality embryos on 7 days were vitrified using mixtures of dimethyl sulfoxide and ethylene glycol at two concentrations (3.5 and 7 M).

Results: The result showed that the cleavage and morula rates were significantly (p<0.05) higher in the 0.5 mM group. Blastocyst rates were significantly (p<0.05) higher at both 0.5 and 1 mM. The rates of viable embryos directly after thawing were significantly (p<0.05) increased in the 0.5 mM group. No significant difference was found in embryos cultured for 24 h after warming among all the groups.

Conclusion: The addition of L-carnitine at a concentration of 0.5 mM to the culture media improves the oocyte developmental competence and cryotolerance of buffalo embryos directly after warming but not after 24 h of culture. Nevertheless, further studies must identify how L-carnitine exerts its beneficial micromechanisms.

Background and Aim: Measuring blood progesterone (P4) concentration has become an essential diagnostic tool in small animal reproductive medicine. Methods enabling precise and rapid on-site measurements are in high demand, especially for the optimization of breeding management in bitches. This study aimed to compare two commercial on-site methods (Speed™ P4, Virbac [M1] and mini VIDAS®, bioMérieux [M2]) and a well-established radioimmunoassay (RIA), which was used as a reference method.

Materials and Methods: Comparative measurements were performed on 52 blood serum samples collected from 45 clinically healthy bitches of different breeds. The dogs had been presented to determine the estrus cycle stage and predict the time of ovulation. Each sample was divided into three aliquots. In aliquot 1, P4 was measured immediately applying M2. Aliquots 2 and 3 were stored at –20°C until analysis was performed using RIA and M1. The consistency of the three methods was investigated by pairwise linear regression analyses.

Results: In RIA, the P4 concentrations ranged between 1.1 and 25.4 ng/mL. Regression analyses revealed highly significant (p<0.0001) positive correlations between the three methods applied (M1 vs. RIA: R=0.94; M2 vs. RIA: R=0.98; and M1 vs. M2: R=0.91).

Conclusion: The results show that the two commercial on-site methods tested exhibit approximately equal, high consistency with the radioimmunological reference method and can, therefore, be used beneficially in a clinical setting. However, biological interpretation of data must be performed in a method-specific manner.

Aim: An experiment was designed to evaluate the role of Vitamin E and glutathione in improving the seminal parameters during hypothermic storage of liquid semen at 4°C for 72 h. Materials and Methods: Thirty-six semen ejaculates were collected by artificial vagina from 6 bucks (Beetal) during the normal reproduction season (September to November) at weekly interval. The samples were centrifuged, and the seminal plasma was removed. The sperm pellet was diluted with Tris-based extender and divided into three groups. Group T1: Control samples without antioxidants, Group T2: Samples supplemented with tocopherol at 3 mM, and Group T3: Samples supplemented with glutathione at 1 mM. The samples were evaluated for progressive motility, percent liveability, percent abnormal spermatozoa, and acrosome integrity after liquid preservation for 0, 24, 48, and 72 h. The level of lipid peroxidation and antioxidant enzymes, namely, glutathione peroxidase (GPx) and superoxide dismutase (SOD) were estimated after liquid preservation for 0 and 72 h. Results: It was observed that, after storage of semen at 4°C up to 72 h, the progressive sperm motility, percent liveability, percent abnormal spermatozoa, and percent intact acrosomes were significantly (p˂0.05) higher in group T2 and T3 samples as compared to control. However, the level of lipid peroxidation in T2 and T3 groups was significantly (p˂0.05) lower after 72 h of incubation at 4°C. Similarly, GPx and SOD values were significantly (p˂0.05) increased in T2 and T3 groups after 72 h of storage at 4°C. Conclusion: Thus, it can be concluded that Vitamin E and glutathione supplementation at 3 mM and 1 mM, respectively, while preserving the semen samples at 4°C helped in maintaining the seminal parameters up to 72 h and protected the spermatozoa from oxidative damage.

Background and Aim: The Javan leopard (Panthera pardus melas Cuvier, 1809) is a subspecies of Panthera pardus spp., spread across the African and Asian regions. Information on reproductive aspects is crucial for wild animals, including the Javan leopard. In this study, we aimed to develop electroejaculator (EE) techniques and evaluate cryopreservation success in Javan leopard semen.

Materials and Methods: The semen of four adult Javan leopards was collected once a week using EE. Placement of the EE probe in the rectum was performed after ultrasound imaging (ultrasonography) to determine the prostate body location. The semen obtained was then evaluated macroscopically and microscopically. Three Javan leopards were used for cryopreservation. The ejaculate was divided into two parts [i.e., one part diluted with AndroMed® (Minitüb, Tiefenbach, Germany) and the other part with Steridyl® (Minitüb)] at a 1:1 ratio immediately after collection and evaluation. The semen was then packed in a 0.25 mL MiniStraw® (Minitüb) then equilibrated at 4°C for 2 h. After equilibration, the straw was then frozen in liquid nitrogen vapor. Frozen semen was then stored in containers until further evaluation.

Results: The results showed that ejaculation response occurred at all levels of stimulation, while erections did not always occur. The fastest ejaculation and erection occurred at the fourth voltage. The macroscopic evaluation showed that the semen volume was 0.80±0.26 mL, cloudy white, pH 7.44±0.14, and with watery semen consistency. The microscopic evaluation showed that the sperm motility was 66.98±0.39%, with sperm viability of 75.6±1.79%. Sperm concentration was 62.17±46.95×106 mL–1 with a total concentration of 42.14±23.51×106 cells. Normal sperm morphology is only 40.72±6.26%.

Conclusion: This study concluded that the development of a semen collection technique using an EE preceded by imaging of the EE probe location using ultrasound was effective for the ejaculation of Javan leopards. The characteristics of the semen of the Javan leopard showed moderate semen volume, sperm motility, and viability. Javan leopard showed low sperm concentration and normal sperm morphology.

Background and Aim: The Barb horse occupies a prominent place in the history, culture, and equestrian traditions of the Maghreb (Algeria, Morocco, and Tunisia). Although many studies on the breed standard and morphometry have been published, there are no studies on its fertility and reproductive potential. Thus, this work aimed to study the fertility of Barb, Arabian Purebred, and Thoroughbred horses in Algeria.

Materials and Methods: A total of 168 stallions and 1202 mares at the Chaouchaoua Stud farm in Tiaret, Algeria, were included in the study. The reproductive performance during 1592 cycles over 10 consecutive mating seasons (2003-2012) was evaluated. Multivariate analysis with logistic regression was used to analyze the reproductive parameters such as the number of cycles operated, number of pregnant mares, pregnancy rate per cycle, seasonal pregnancy rate, and embryonic mortality rate, and to determine the influence of breed, stallion's age, and year of the study on reproduction.

Results: Statistical analysis showed that stallion breed was a significant influencing factor for the number of pregnant barren mares (Odds ratio [OR]=1.72; p=0.03; 95% confidence interval [CI]=1.05, 2.84) seasonal pregnancy rate (OR=1.40; p<0.001; 95% CI=1.29, 1.53). Additionally, the reproductive performance of the Barb stallion was superior to that of the Thoroughbred stallion. With regard to the significant influence of stallion's age of >5 years on the number of pregnant foaling mares and seasonal pregnancy rate, significant interactions were observed between the stallion's breed and age, and the pregnancy rate per cycle: in the Barb breed, the pregnancy rate per cycle increased with the age of the stallion, while in the Thoroughbred, it decreased with age. Moreover, a significant effect of the year of the study on the pregnancy rate per cycle and number of pregnant foaling mares was observed. In contrast, the number of cycles and embryonic mortality rate were not influenced by the breed and age of the stallion, or the year of the study.

Conclusion: The Algerian Barb horse attained a similar level of fertility compared with that of the Arabian Purebred and Thoroughbred stallions depending on its age and reproductive performance.

Bovine somatotropin (bST) is a natural metabolic protein hormone produced by the pituitary gland in all cattle and used to increase milk production in dairy cows. Recombinant bovine somatotropins (rbST), that has several amino acids, have been synthesized using recombinant DNA techniques. rBST is administered subcutaneously at day 60 of a cow’s lactation cycle when milk production normally begins to decrease and repeated every 14 days. Even though bST has the potential to increase the efficiency of milk production, there is no change in milk composition. In the case of rBST, potentially 10-15% more milk can be obtained from each cow. rBST is biologically inactive in humans and its residues in food products have no physiological effect. Concentration of Insulin-growth factor-I (IGF-1) is no significant difference in bovine growth hormone levels in milk from rBGH-treated and untreated cows. Even if there were a much higher level of bovine growth hormone ingested by humans, our digestive system would break down and inactivate the hormone protein. In addition, the bovine growth hormone does not affect human growth hormone receptors and good management measures should be taken as per manufacturer to ensure a high response in milk yield to bST administration. Thus, the use of rbST to improve productivity within the lactating cow herd allows for a reduction in resource use and environmental impact per unit of milk.

A four-years-old pluriparours ewe, of a body weight about 40 kg was presented with a history of restlessness, straining, and appearance of a permanent/complete cervico-vaginal prolapse as a smooth, red, and rounded mass at the rear end of the ewe. There was no history of previous cervico-vaginal prolapse during or after previous pregnancies. Complete procedures of the appropriate successful treatment were described in this study. Briefly, treatment was performed under caudal epidural analgesia; the prolapsed mass was immediately covered with a towel soaked in water, examined for presence of injuries/rupture, raised to evacuate the urinary bladder, replaced by manual fist pressure and by the flat of hands, and finally retention suture was used to prevent the possible reoccurrence. The suture material had been removed before lambing, and the owner was advised to exclude the ewe from further breeding.

Objective
To evaluate the severity of fallopian tubal conditions in patients with previous EP after methotrexate or laparoscopic conservative treatment using the second-look laparoscopy (SLL).

Methods
This was a retrospective clinical study in a university hospital. After reviewing medical records, thirty-eight women with EP were divided into group I (methotrexate therapy) and II (laparoscopic salpingotomy) according to the conservative treatment options they previously received. Then, the secondlook laparoscopy was carried out to evaluate the severity of the fallopian tubal conditions (including integrity, patency and adhesion).

Results
The integrity and patency rates of the treated tubes in group I were 94.12% (16/17) and 52.94% (9/17), higher than those in group II (with 57.69% and 7.69%), respectively. But no difference was found in the adhesion rate of peri homolateral tubes between groups I (41.18%) and II (57.69%).

Conclusion
Our results suggested that conservative medical treatment with MTX by SLL could protect the tubal conditions better than laparoscopic salpingotomy, confirming that methotrexate treatment would be a very good way to preserve the tubes and improve future fertility. The second-look laparoscopy could be recommended to the EP patients with a desire for pregnancy.

Background and Aim: As the number of wild Asian elephants (Elephas maximus) continues to decline, maintaining healthy populations under human care is vital. Male fertility assessment is essential for understanding the reproductive status, which can help to uncover underlying problems and improve the rate of pregnancy success. The objectives of this study in Asian elephants were as follows: (1) To investigate the semen characteristics; (2) to compare the relative seminal vesicle size and semen characteristics; (3) to compare the semen characteristics between good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates; and (4) to investigate the pregnancy success rate after artificial insemination (AI) with combined chilled and frozen semen.

Materials and Methods: In total, 153 ejaculates were collected by manual rectal stimulation from 25 bulls. The volume, pH, sperm concentration, progressive motility, viability, morphology, and membrane integrity were investigated in each ejaculate. Assessment of accessory sex glands was conducted using transrectal ultrasonography to compare the relative seminal vesicle size and semen characteristics, and the bulls were divided into two groups according to the size of the ampulla (<7 or ≥7 cm2). For the comparison of good and poor-motile ejaculates and semen characteristics, the samples were divided into two groups: Good-motile (>60% progressive motility) and poor-motile (<60% progressive motility) ejaculates. Semen ejaculates for AI were collected from three bulls. The estrous cycles of four females were monitored using an enzyme immunoassay. Seven AI attempts were conducted using frozen and/or chilled semen by endoscopic visualization. AI was repeated 1 day before the luteinizing hormone surge, on the day of the surge, and 1 day after the surge. Pregnancy was confirmed by monitoring the serum progesterone profile and the abdomen and mammary glands changes.

Results: From 153 ejaculates, the mean±standard error values of progressive motility, semen volume, sperm concentration, pH, and viability were 40.18%±2.28%, 40.94±3.86 mL, 1,205.58±62.26×106 sperm/mL, 7.50±0.10, and 56.17%±1.96%, respectively. Comparing ampulla size and semen characteristics revealed that the bulls with ampullae of ≥7 cm2 yielded significantly larger volume ejaculates. However, there were no significant differences in sperm motility and concentration. The comparison of semen characteristics between good- and poor-motile ejaculates revealed that the former had significantly higher pH, viability, normal acrosomes, intact membranes, and normal head and tail morphology but often had a significantly lower volume and sperm concentration. From seven AI attempts in four females, one female had a confirmed pregnancy (14.3% pregnancy rate), and delivered a healthy live female baby weighing 128 kg at 21 months and 12 days of gestation. The baby is now 3 years old and in a healthy condition, with normally developing growth and behavior.

Conclusion: The semen characteristics of Asian elephants can be used as the baseline reference for further applications. The ampullae size indicates semen quantity but not quality. Our success in producing an elephant calf from AI using frozen and chilled semen demonstrated that AI can be used as an alternative approach for the breeding management of Asian elephants. However, the semen of Asian elephants is of poor quality, especially in terms of membrane integrity; thus, the improvement in semen quality through intensive and careful management of elephant health and fertility remains a challenge for the future. Furthermore, a sperm bank should be established to develop sperm cryopreservation, which will be invaluable for improving the genetic diversity of the Asian elephant.

Background and Aim: Capacity for sperm production is affected by age, which is related to the morphology of sperm abnormalities and can affect fertility. The aim of this study was to evaluate the relationship between age and concentrations of testosterone and adiponectin with sperm abnormalities in Simmental bulls.

Materials and Methods: The study used 11 bulls, separated into three groups. The first group consisted of five bulls aged 4-5 years, and the second and third groups each consisted of three bulls, aged 6-7 and 8-10 years, respectively. The average sperm motility of the animals ranged from 57.66±2.60% to 70.17±0.22%. Blood samples were obtained from the coccygeal region of the animals. Testosterone and adiponectin analysis was performed using the enzyme-linked immunosorbent assay method. Sperm morphology was evaluated using carbol fuchsin-eosin staining according to the Williams method. Finally, correlations between testosterone and adiponectin concentrations, age, and sperm abnormalities were analyzed using Pearson's correlation analysis.

Results: The findings revealed a significant correlation (p<0.01) between the concentrations of testosterone and adiponectin (–0.538), age (–0.588), and abnormal sperm morphology (–0.912). Moreover, they revealed that the concentration of testosterone in the bulls aged 8-10 years was lower, at 21.89±4.56 ng/mL, compared to that in the bulls aged 4-5 years, at 36.15±1.29 ng/mL, and 6-7 years, at 35.16±5.39 ng/mL. The findings also revealed a positive correlation between adiponectin concentration and age (0.529) and sperm abnormalities (0.506). The increase in testosterone concentration was inversely related to the adiponectin concentration (–0.538). Moreover, the mean amount of abnormal sperm increased with increasing age: 3.82±0.33% in the group aged 4-5 years, and 4.40±0.72% and 10.20±1.97% in the groups aged 6-7 years and 8-10 years, respectively.

Conclusion: The study data indicate that there is a decrease in testosterone concentration, a high adiponectin concentration, and an increase in abnormal sperm with increasing age in bulls.

Background and Aim: Calves have increased morbidity and mortality rates after dystocia. One cause is pain during birth, which reduces their colostrum intake. The administration of nonsteroidal anti-inflammatory drugs (NSAIDs) may break this causality. This study aimed to determine the consequences of a single administration of the NSAID meloxicam to dystocia calves after birth.

Materials and Methods: Fifty Holstein–Friesian calves born with dystocia from four dairy cattle farms were included in this study. The animals were randomized into two groups. The animals in Group I (n=25, treatment group) received subcutaneous 0.5 mg meloxicam/kg body weight 2-8 h after birth. The animals in Group II (n=25, control group) received a control substance (Amynin®, bovine infusion solution, Merial) with the same volume. The newborn calves were clinically examined on the 1st and 10th days of life. The information regarding the days in between was gathered by questioning the farmer.

Results: There was a significant difference (p=0.04) only in calves with thin, mushy fecal consistency on the 10th day in the treatment group compared with the control group. Moreover, meloxicam had no effect on dystocia calves.

Conclusion: Since NSAID administration did not produce a significant clinical effect, its necessity is questionable. Further studies should examine how modifying its application time would have an effect. The primary indicators of well-being, such as pain indicators in the blood, were not measured; however, these should be considered in subsequent studies.

Background and Aim: Reproductive disorders keep the beef cattle population in Jepara Regency, Central Java, Indonesia, from increasing. Ovarian hypofunction and repeat breeding are the most common reproductive disorders, leading to large economic losses for traditional breeders. However, the impact of poor reproductive performance among traditional breeders is not well-known. This study aimed to investigate the reproductive performance of beef cattle with ovarian hypofunction and repeat breeding in Jepara Regency.

Materials and Methods: We determined cattle's reproductive status by rectal examination and anamnesis, assessing reproductive performance in 28 cows with repeat breeding and 27 cows with ovarian hypofunction. The following parameters were measured: Postpartum estrous (PPE), days open (DO), service per conception (S/C), and calving interval (CI). The data came from livestock records from animal recording cards, iSIKHNAS, and estimated births from the insemination date that produced a pregnancy.

Results: In beef cattle diagnosed with ovarian hypofunction, S/C, PPE, DO, and CI were 1.28, 257 days, 265 days, and 18 months, respectively. The length of CI caused by long PPE may be due to improper nutrition and calf weaning delays. In beef cattle with repeat breeding, S/C, PPE, DO, and CI were 4.15, 106 days, 210 days, and 16 months, respectively. The length of CI was caused by long DO due to pregnancy failure at the first estrus.

Conclusion: Hypofunction and repeat breeding reduce the reproductive performance of beef cattle in the Jepara Regency. The cows' health conditions pre- and postpartum can be optimized by providing high-quality feed to enhance reproductive performance.

Background and Aim: Holstein cows and heifers are widely bred in Indonesia by artificial insemination (AI) to increase population and milk production. Sperm fertility is modulated by genetic factors, but the analysis of sperm quality is still based on macro- and microscopic characteristics. This study aimed to analyze both sperm quality and proteins of Holstein bulls at different fertility levels.

Materials and Methods: The frozen semen samples were collected from the Indonesia National AI Center. They were classified based on the reproductive efficiency data and were grouped into high fertile (HF) and low fertile (LF). Sperm qualities were evaluated by microscopic evaluation. The Holstein sperm proteins were extracted using phenylmethanesulfonyl fluoride as a protease inhibitor and the benzidine detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was conducted to analyze the molecular weights (MWs) of the sperm proteins. The data obtained were analyzed by a t-test using the one-factor bull fertility level, and Spearman's correlation analysis was used to identify the correlation between the sperm microscopic evaluation parameters and protein bands.

Results: The sperm motility post-freeze thawing was not significantly different between the HF and LF (p>0.05). The HF level had a higher percentage of viability, intact plasma membrane integrity, and intact acrosomes than the LF (p<0.05). Five protein bands were found in the SDS-PAGE of sperm proteins of Holstein bulls with different concentrations. Sperm proteins with MWs of 17.51 kDa, 14.87 kDa, 33.71 kDa, and 41.97 kDa were abundant in the Holstein bulls with an HF level, while 55 kDa proteins were abundant in the LF level of Holstein bulls. The sperm of Holstein bulls in the HF level contained proteins of about 33.71 kDa that were not detected in the LF.

Conclusion: The sperm protein with a molecular weight of 33.71 kDa was predicted to be a specific protein biomarker that influences bull fertility. Sperm fertilization abilities were also determined by the sperm proteins, the morphology of sperm acrosomes, and the quality of plasma membranes. This method can be used to select bulls with high fertility to increase the population of Holstein bulls.

Background and Aim: Previous findings regarding the effects of farrowing induction on the farrowing characteristics of sows are controversial. This study aimed to investigate the effects of farrowing induction on the following characteristics: (1) Proportion of sows that farrowed during working hours, (2) stillbirth rate, (3) number of stillbirths per farrow, (4) dystocia rate per farrow, (5) dystocia rate (the proportion of farrowings that had at least one dystocia event), (6) number of dystocia events per farrow, (6) farrowing duration, (7) birth interval, and (8) birth weight.

Materials and Methods: Thirty-eight Landrace x Yorkshire sows were randomly allocated into two groups; the control group and the treatment group. In the control group (n = 18), sows farrowed spontaneously. In the treatment group (n = 20), farrowing was induced approximately 2 days earlier than the herd's average length of gestation (7:00 am on day 114) by injecting cloprostenol into the perivulval region. All sows were supervised throughout their farrowing. We recorded the interval between induction and farrowing; total number of births; number of live, stillborn, and mummified piglet births; number of dystocia events; birth interval; farrowing duration; and birth weight. A generalized linear mixed model, a linear mixed-effects model, the Chi-squared test, and Student's t-test were used to compare outcomes between the two groups.

Results: Farrowing induction did not influence the percentage of sows that farrowed during working hours (7 am–5 pm), stillbirth rate, birth weight, and number of dystocia events per farrow. Farrowing induction led to an increase in birth interval, dystocia rate, dystocia per farrow (p < 0.05) and in addition to the percentage of sows that farrowed on the day following induction (60% vs. 27.8%; p < 0.05).

Conclusion: Farrowing induction using a single dose of cloprostenol 2 days before the expected farrowing date can be performed with care to concentrate farrowing into a short interval. This can enhance the optimization of cross-fostering and the practice of an all-in-all-out strategy in the swine breeding industry.

Background and Aim: Indonesia has two National Artificial Insemination centers and 17 Regional Artificial Insemination Centers. The frozen semen production techniques differed between the centers, including the type of diluent and semen dilution technique. The aim of the research was to compare the quality of frozen Limousin bull semen diluted using different techniques. Materials and Methods: Semen was collected from three sexually mature Limousin bulls using an artificial vagina. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen that had &gt;70% motile sperm and &lt;20% sperm abnormality was divided into three tubes and diluted with skim milk-egg yolk (SMEY) using three different dilution techniques: One-step dilution (100% SMEY with 8% glycerol) at room temperature ([RT] 20°C until 25°C) two-step dilution (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol after 1 h stored at 5°C); and three-step dilution ...

Aim: The aim of the present study was to define the onset of puberty in Tazegzawt ram lambs, an Algerian sheep breed in endangered status with a small population in its local area. Materials and Methods: Body growth (body weight and thoracic perimeter), scrotal circumference (SC), penis development stages, and seminal parameters (volume, concentration, and motility) were measured. Data were recorded at fortnightly intervals in 10 animals from 9 to 49 weeks of age. Results: On the basis of seminal analyses, puberty occurred between 29 and 45 weeks of age. At 29 weeks of age, 30% of lambs reached puberty, and at 45 weeks of age, puberty was observed in 100% of the analyzed animals. Body weight appeared as the most determinant factor, and the onset of puberty was observed when animals reached 43.2±6.4 kg body weight with 25.8±3.7 cm of SC. Seminal analyses revealed that all parameters increased regularly from puberty onset except for sperm concentration. The mean semen volume during the study period was 0.48±0.33 mL with 0.84±0.6 mL at 37 weeks of age. Sperm concentration evolved similarly as semen volume; at 29 and 43 weeks of age, the sperm concentration was 942×10 6 and 1904×10 6 spermatozoa/mL, respectively. Kinematic parameters including the percentage of motility, the percentage of progressive motility, and gametes velocities as determined by Computer-Aided Sperm Analyzer showed the highest values at 49 weeks of age. Conclusion: The current results revealed that, in Tazegzawt ram lambs, puberty occurs between 29 and 45 weeks when animals reach 43.2±4.6 kg body weight.

Regulation of pH in spermatozoa is a complex and dynamic process as sperm cells encounter different pH gradients during their journey from testes to the site of fertilization in female genital tract. The precise regulations of pH in sperm cells regulate the sperm functions such as motility, hyperactivity, capacitation, and acrosome reaction. Electrophysiological, pharmacological, and molecular studies have revealed the presence of different ion channels and exchanger systems which regulate intracellular pH in sperm cells as well as regulate sperm functions. Recent studies also have shown the potential involvement of pH in the regulation of fertility competence of sperm cells, and alterations in pH have shown to impede sperm functions. This mini-review discusses the probable mechanisms involved in pH regulation in sperm cells and how pH is involved in regulation of various sperm functions.

The objectives were to investigate the factors (age, sex, breed and season) affecting the occurrence of calf diseases. In this study 497 cases of 0 to 3 months of age from three different farms were investigated. Calf in Savar Dairy farm (n=249), Talbag Dairy Farm (n=85) and BAU Dairy Farm (n=163) were included from April 2011 to March 2012. The occurrence of diseases were classified on the basis of breed (Local and Cross), age (≤ 3 weeks and >3 weeks), sex (Male and Female) and season (Spring, Summer, Rainy and Winter). The overall diseases were diarrhoea (20.3%), anorexia (9.7%), fever (9.1%), naval ill (8.0%), malnutrition (7.7%), indigestion (6.7%), traumatic injury (5.4%), dysentery (5.0%), myiasis (4.6%), pneumonia (3.4%), alopecia (3.4%), atresia ani (3.2%), hernia (2.8%), constipation (2.6%), abscess (2.2%), conjunctivitis (2.2%), lameness (1.8%), FMD (1.2%) and blot (0.6%). The cases of diarrhoea was higher (20.5%) than that of other diseases. The occurrence of diseases was highly significant (p<0.01) in crossbred calves (77.3%) than that of local bred (22.7%). Cases of diseases were higher in male calves (51.7%) than that of female calves (48.3%) but the variation was insignificant (p>0.05). The effect of breeds on diseases were found significantly (p<0.05) higher in ≤ 3 weeks (55.1%) than that of > 3 weeks of age (44.9%). The occurrence of diseases in calves was significantly (p<0.05) higher in rainy season (36.0%) followed by the summer (31.0%), winter (22.1%) and lowest in spring (10.9%). The results demonstrate significant effect of breed, age and season on the occurrence of diseases in calves.

Background and Aim: The Javan leopard (Panthera pardus melas Cuvier, 1809) is a subspecies of Panthera pardus spp., spread across the African and Asian regions. Information on reproductive aspects is crucial for wild animals, including the Javan leopard. In this study, we aimed to develop electroejaculator (EE) techniques and evaluate cryopreservation success in Javan leopard semen. Materials and Methods: The semen of four adult Javan leopards was collected once a week using EE. Placement of the EE probe in the rectum was performed after ultrasound imaging (ultrasonography) to determine the prostate body location. The semen obtained was then evaluated macroscopically and microscopically. Three Javan leopards were used for cryopreservation. The ejaculate was divided into two parts [i.e., one part diluted with AndroMed® (Minitüb, Tiefenbach, Germany) and the other part with Steridyl® (Minitüb)] at a 1:1 ratio immediately after collection and evaluation. The semen was then packed in a ...

Thirty endometritic crossbred dairy cows diagnosed were randomly allotted to three groups with 10 each to evaluate the therapeutic efficacy of some non-antibiotic protocols. Cows in Group I received 30 ml ozonated oil intrauterine (i.u), Group II 500mg of oyster glycogen in 30 ml PBS i.u and Group III 30ml PBS i.u as control. Uterine flushings were collected on the day of estrus before and after treatment (next immediate estrus) and tested for pH, whiteside test, endometrial cytology and determination of common aerobic bacteria. The mean pH of uterine flushings in all the groups before treatment was 7.95. A significant reduction in mean pH was observed following treatment with ozonated oil (7.46) and oyster glycogen (7.31) whereas no significant change in the untreated cows (7.77) was recorded. Aerobic bacteria isolated in the uterus of endometritic cows were E. coli, Staphylococcus sp. Enterobacter sp. and Proteus sp. Following treatment, 60 per cent (6/10) in Group I, 80 per cent (8/10) in Group II and 30 per cent (3/10) in Groups III cows were recovered based on whiteside test and endometrial cytology results. The conception rate was higher (87.50%) in Group II followed by 66.66 per cent in Group I and 33.33 per cent in Group III.

Hymenoplasty is the procedure to restore the torn hymen intactness of which is presumed to be sign of virginity. Usual patient is an unmarried girl who had indulged in premarital sex and is going to be married. Academic literature is devoid of details about hymenoplasty procedures. We have found one report [1] which has just described, in not very descript manner and without photographs, the procedure of hymenoplasty. We have described a procedure for hymenoplasty by using the remnants of torn hymen [2]. However many a times our patients after hymenoplasty complained that though they bled but one or two drops only while they wanted more bleeding during first sexual act. For such patients we developed the technique of creating superhymen in which more bleeding is destined to occur on the eve of first sexual intercourse. Material and method We have done superhymenoplasty in thirty one patients over period of three years. Twenty females were unmarried and eleven were married or divorcee who came for hymenoplasty after which they expected enough bleeding during first sexual act. None of them were aware of two terms and difference between hymenoplasty and superhymenoplasty. Of thirty one females twenty were accompanied by their parents, five accompanied by their boyfriends and fiancée, while others were unaccompanied by anybody. Eleven females were going to be remarried because of divorce or death of their husbands and those were accompanied by parents. Four women were accompanied by their husbands who wanted restoration of hymen of their wives knowing well that they have lost hymen in premarital sexual intercourse. The history was taken and all patients were explained that absence of hymen is not a sign of lost virginity; and same is true for bleeding on first night. But all were determined to get the procedure of superhymenoplasty. They were told that in superhymenoplasty more bleeding is destined. Whether to go for hymenoplasty or superhymenoplasty was their personal decision. Technique The procedure was done in local or general anaesthesia as per wishes of the patients. In 15 patients procedure was done in local anaesthesia and in remaining patients in general anaesthesia. Patients were placed in lithotomy position. Area was prepared and draped. Torn pieces of hymen were incised and incision was continued in remnants up to just below the urethral orifice creating two layers-outer and inner. Inner layer was stitched using chromic catgut 4-0 and outer layer was also stitched using same catgut. When done finally, only small hole was left in hymen just below urethral orifice for menstrual blood and other vaginal secretions to come out. Neosporin ointment was applied over the suture line and

Background and Aim: Lepidium meyenii Walp (Maca) is an herbaceous plant that grows in the Peruvian Andes and it has been widely used as a nutritional supplement and fertility enhancer and has been used in the treatment of a variety of diseases, such as rheumatism, respiratory disorders, and anemia. The most notable feature of Maca is its potent antioxidant capacity, which helps in the scavenging of free radicals and protection of cells from oxidative stress. This study aimed to evaluate the in vitro effect of Maca extract on thawed sperm cells from bulls.

Materials and Methods: Three dilutions of 1, 10, and 100 μg/mL of Maca extract were incubated with frozen–thawed bovine semen and analyzed at 1, 3, and 24 h of exposure time, evaluating the activity of the extract on the DNA, motility, morphology, viability, integrity of the membrane and acrosome of spermatozoa.

Results: The Maca extract improved the studied sperm parameters of motility, acrosome integrity, vitality, and DNA integrity of sperm cells at a concentration of 10 –g/mL, and at 1 –g/mL, an improvement was observed in the morphology and integrity of the membrane. However, the best activity of the Maca extract was observed on the DNA integrity of the sperm, which was effective at the three concentrations evaluated after 24 h of incubation.

Conclusion: The results indicate that L. meyenii can help in maintaining spermatozoa cellular integrity after the frozen–thaw process, especially in the protection against DNA fragmentation. Therefore, Maca would be a feasible supplementation to protect sperm to maintain their fertile ability after thawing.

Background and Aim: Essential oils found frequently in plants are well known for their activities against bacteria, viruses, and fungi, and antioxidant properties. This study aimed to analyze egg yolk replacement by seed oils of Gossypium spp. (cotton), Balanites aegyptiaca (desert date), and Sesamum indicum (sesame) in semen extender, on ram sperm quality chilled at 4°C and frozen-thawed.

Materials and Methods: Ejaculates were collected from adult rams and refrigerated at 4°C in a Tris-based extender containing 1.25%, 2.5%, 5%, and 10% of Gossypium spp., B. aegyptiaca, and S. indicum seed oils, to evaluate which were the two best extenders for comparison with BIOXcell, a commercial extender for deep freezing ram semen.

Results: The data showed that sperm movements analyzed by the CASA system were faster in extenders supplemented with 2.5-5% of cottonseed oil and 1.25-10% of sesame oil, whereas in the extender containing B. aegyptiaca oil, all seminal parameters studied had the worst values. During the sperm-freezing process, 5% of cottonseed oil and 5% sesame seed oil were selected from the first study, with sesame oil reaching the best sperm quality. Thus, sperm motility and velocity were 44.14±13.99%, 24.44±12.6%, and 25.92±11.50%; and 20.26±9.56%, 8.76±6.38%, and 9.42±5.40%, respectively, for sesame oil, cottonseed oil, and BIOXcell.

Conclusion: In summary, 2.5-10% of cottonseed oil and 1.25-10% of sesame seed oil can replace egg yolk in a Tris–egg yolk–based extender. Moreover, a Tris-based extender supplemented with 5% sesame seed oil could be an alternative for deep freezing ram semen, even though these results need to be confirmed with semen collected from rams with appropriate sexual rest.

Background and Aim: Various factors can reduce the quality of semen used for artificial insemination and have an impact on fertility decline, such as poor handling during frozen semen distribution. This study was aimed at assessing the quality of frozen-thawed semen after distribution in the field and its importance in maintaining fertility.

Materials and Methods: The Brahman Cross (BX) breeding program of PT Lembu Jantan Perkasa, Indonesia, was used. This program was preferred due to its adherence to guidelines that limit the effects of extraneous factors that may affect semen quality. Frozen-thawed semen samples from eight bulls with the same production code were analyzed and compared between the production site (artificial insemination [AI] center) and the field (BX breeding program). Total and progressive motility (PM) of sperm were determined using computer-assisted semen analysis. Plasma membrane integrity (PMI) was assessed using hypoosmotic swelling test, sperm viability using Eosin-Nigrosin staining, acrosome integrity using trypan blue-Giemsa staining, morphological abnormalities using William staining, and DNA fragmentation using toluidine blue staining. The fertility rate was determined using the conception rate (%) derived from AI data based on 502 AI services and 478 cows in the BX breeding program. A t-test was used to compare the quality of frozen-thawed semen before and after distribution. The relationship between the qualities of frozen semen after distribution in the field with fertility was analyzed using Pearson correlation.

Results: There was no significant difference (p>0.05) in the quality of frozen-thawed semen (sperm motility, PMI, viability, acrosome integrity, abnormalities, and DNA fragmentation) between the production site (AI center) and after distribution in the field (BX breeding program). The semen met the minimum standards for AI programs. Total motility (r=0.986), PM (r=0.961), sperm viability (r=0.971), PMI (r=0.986), and acrosome integrity (r=0.992) were all positively correlated (p<0.05) with fertility rate; while sperm abnormalities (r=-0.996) and sperm DNA fragmentation (r=0.975) were negatively correlated (p<0.05) with fertility rate.

Conclusion: The study showed that to achieve the maximal and optimal fertility rate in bulls in an AI program, the overall quality of frozen-thawed semen in all aspects is critical. This can be achieved if the handling during distribution and storage, as well as the various factors that may affect the quality of semen in the field, can be controlled properly.

Obstetrics and gynecology, better known by the shorthand OB/GYN, is a branch of medicine devoted to female sexual and reproductive health. Due to the stigma and mystery that often times surrounds the OB/GYN field, men and women alike remain in the dark about all of the various things these physicians do. Here are five facts you may not have known about obstetrics and gynecology.