Veterinary Obstetrics and Gynecology

Obstetrical knowledge and skill is one of the core competencies required at day zero of veterinary graduates. However, in the last three years out of 198 students interviewed in three veterinary colleges in Ethiopia only 6 students have palpated the reproductive organ of a cow 0.02% and none had ever applied ultrasound, showing a huge lag in technological adaption and the non existence of appropriate teaching method. The purpose of this study was to determine the effectiveness of obstetrics simulator training for veterinary students by comparing measures of confidence, anxiety and attitude with and without prior training. In this study it was found that the innovation described here obstetrics and gynecology simulator offers the means by which students can manipulate reproductive organs move and zoom within ultrasound images, sketch, and compare images. It was realized that this solution would provide a more cost-effective and convenient method of teaching to a large numbers of students. Students can be trained to perform invasive procedures prior to examining real animals, which has benefits for animal welfare.

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.

Preeclampsia and HELLP syndrome, both disorders unique to pregnancy, remain a major cause of maternal and neonatal mortality and morbidity worldwide. It is the most common medical disorder complicating pregnancy. In both of these disorders, the liver is a major target with often devastating consequences. The pathogenesis of hepatic damage in cases of severe preeclampsia and HELLP syndrome is not well understood. Diagnosis of HELLP syndrome is heavily relied upon laboratory investigation. In the management, first priority is to assess and stabilize the maternal condition, particularly coagulation abnormalities followed by evaluation of fetal well-being and finally, a decision must be made as to whether or not immediate delivery is indicated. Women with a prior history of HELLP syndrome carry an increased risk of at least 20% (range 16%-52%) risk of recurrence of some form of hypertension, so future pregnancy counseling and follow up is essential. Hypertensive disorders of pregnancy are a dreaded disease for both the mother and the fetus. Preeclampsia and HELLP syndrome, both disorders unique to pregnancy, remain a major cause of maternal  and  neonatal  mortality  and  morbidity  worldwide.  It is the most common medical disorder complicating pregnancy, affecting 7 to 15% of all women. As high as 22% perinatal deaths and 30% of maternal deaths in developed countries are because of preeclampsia. The situation is far by worse in developing countries because of low antenatal turn up, delayed reporting and inefficient management in most of the heath setups [1].

Aim of the work
The purpose of this study was to compare the efficacy and safety of clomiphene citrate (CC) alone versus CC plus N-acetyl cysteine (NAC) and CC plus chromium picolinate (CrP) in management of PCOS.
Patients and Methods
This study, accomplished on 120 women with PCOS and fulfilling Rotterdam criteria 2003. Patients were divided into three equal groups; the First Group (n=40) given out NAC plus CC. The second group (n=40) administered CrP plus CC. The last group (n=40) dispensed CC alone.
Results
The number of follicles (≥18 mm), mean endometrial thickness on the day of hCG administration and number of pregnancies were higher among PCOS cases given NAC+CC group than CrP+CC and CC alone cases. No adverse side-effects and one case of ovarian hyperstimulation syndrome observed in the group receiving CC+NAC.
Conclusion
NAC is a safe and well-tolerated adjuvant to CC. It improves follicular maturation and pregnancy rate in PCOS patients. In addition, chromium picolinate does not improve follicular maturation or pregnancy rate when added to CC in women with PCOS.

(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.

Objective
To explain the pathophysiology, indications, techniques and medicolegal implications of perimortem caesarean section delivery (PMCSD) and introduce criteria based standards for this operation to improve results.

Methods and Search Design
We conducted an Ovid MEDLINE search for the years 1950-2014 with the following key words: maternal mortality, cardiopulmonary resuscitation, maternal collapse, peri-mortem caesarean section, and cardiac arrest. Citations from bibliographies of identified publications were used as a secondary reference list.

Conclusion
A timely performed PMCSD can save the life of a mother and her baby. Awareness of the physiological changes of pregnancy, new advances in resuscitation methods, and proper preparation for a timely surgery, the correct surgical technique, and understanding the medico legal implications are key issues to achieve the goal of maternal and fetal survival. Creation of international and national guidelines and establishment of international and national data base registry with standard improved documentation for these rare events should be considered in order to improve outcomes and provide a more accurate statistically relevant data and results.

The discovery of the role of kisspeptin neurons in the regulation of mammalian reproduction in 2003 was one of the biggest breakthroughs in reproductive endocrinology within the last few decades. Research during the past two decades since the discovery of kisspeptin has been unveiling the mechanism of how the hypothalamic kisspeptin neurons control reproductive functions through regulation of gonadotropin-releasing hormone (GnRH) secretion. This article aims to overview kisspeptin research, including the most recent studies from ours and other research groups, and to discuss the possibility of new strategies to control reproductive functions in mammals. In the first section, we introduce the critical role of kisspeptin neurons in puberty onset and reproductive functions in mammals, including the regulation of two modes of GnRH/gonadotropin secretion, namely pulsatile and surge modes. The next section focuses more on the mechanism of how the kisspeptin neurons in the arcuate nucleus in the hypothalamus precisely controls GnRH pulse using other two neuropeptides, neurokinin B and dynorphin A. The article also discusses the mechanism suppressing reproductive function during lactation and other stress conditions through inhibition of kisspeptin neurons and consequent GnRH/gonadotropin secretion, to provide insights on the possibility of new strategies to improve reproductive performance in mammals including domestic farm animals.

Aim: This study aimed to evaluate the pregnancy rates in hair ewes using an Ovsynch synchronization protocol under a breeding system that combines fixed-time insemination plus a 10-day mating period as an alternative. Materials and Methods: Through an experimental study (n=27), ewes were randomly located into one of three treatments: (1) Pre-synch (n=9): Prostaglandin F2α (PGF2α)+Gonadotropin-releasing hormone (GnRH)+PGF2α+GnRH; (2) Ovsynch (n=9): GnRH+PGF2α+GnRH; and (3) control: Ewes bred by natural mating (NM) (n=9). Ewes were fixed-time inseminated (fixed-time artificial insemination [FTAI]) with fresh semen, collected just before the insemination time through vaginoscopy at 16 h after the second GnRH (gonadorelin) injection. Each experimental group was placed separately during 15 days and, after this time, fertile rams were allowed back with ewes for a 10-day mating period. Control group ewes remained with the rest of the herd suitable for breeding and were bred under NM. Pregnancy diagnosis was performed by ultrasound at 28-, 56-, and 84-day post-breeding to differentiate between FTAI and NM pregnancies. Total (FTAI±NM) pregnancy rates at 56-day post-breeding were used to compared Pre-synch, Ovsynch, and control. For this purpose, two-tailed proportions comparison z-test was used with a 95% confidence level, for testing as the null hypothesis whether two proportions were equal. Results: Pregnancy rates were higher in control ewes (66.4%) than FTAI (46.6%). When pregnancy rates after a 10-day mating period (40%) were added, the final rate (86.6%) was significantly (p<0.05) higher in Ovsynch-based protocols. The pregnancy rate was significantly lower in FTAI ewes compared to FTAI +10-day mating group (p<0.05). The overall pregnancy rate was 88.0, 85.7, and 67.0 (p>0.05) for Pre-synch, Ovsynch, and control ewes, respectively. Conclusion: These results provide evidence on the benefits of combined FTAI protocols for improving the reproductive efficiency of sheep.

India is a developing country with a population of 1.24 billion. Women’s healthcare in India poses two major problems - fertility control and chronic anemia. According to the National Family Health Survey- 3 (NHFS-3) only 49% of the currently married women aged 15-49 yrs use modern contraceptives. Female sterilization is the main type of contraceptive. Two-thirds of women using contraception are sterilized. More than half the women would be sterilized by age of 26 years.

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.

Prolificacy is the ability to reproduce abundantly. Litter size, which is highly dependent on ovulation rate, has a high economic value and it is a fundamental reproductive trait, especially in small ruminants. Considering the low heritability of prolificacy, it is possible to improve the progress of the prolificacy trait selection using DNA markers and molecular biology techniques. In this article, an overview of the various factors affecting prolificacy in small ruminants has been emphasized.
Keywords: goat, growth factors, hormones, prolificacy factors, sheep

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: The follicle-stimulating hormone (FSH) gene is an essential regulator of fertility in livestock. This study aims to provide information on the genetic makeup of Madrasin cattle experiencing hypofunction by the FSH profile and FSH receptors (FSHR) polymorphism.

Materials and Methods: Blood samples were collected from the Bangkalan regency in Indonesia. DNA was isolated and purified following the extraction protocol of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism.

Results: Our results showed that the FSH gene had a band length of 310 bp and produce two alleles (A and B) with restriction enzymes at 250 bp, 230 bp, and 145 bp. Furthermore, the FSHR gene had a band length of 303 bp and produced two homozygous genotypes: GG at bp 239 and CC at bp 188.

Conclusion: Based on these differences, there was no change in allele frequency and genotype between Madura and Madrasin cattle due to crossbreeding with Limousin cattle. Thus, further detailed investigations of Madrasin cattle are required to elucidate the profile of the LH and LHR genes.

How to cite this article: Sheetal SK, Prasad S, Gupta HP (2018) Effect of insulin or insulin-like growth factor-I administration at mid-luteal phase of the estrous cycle during superovulation on hormonal profile of Sahiwal cows, Veterinary World, 11(12): 1736-1741. Abstract Aim: The present study was designed to study the effect of insulin and insulin-like growth factor-1 (IGF-I) administration during luteal phase of the estrous cycle on the hormonal profile of Sahiwal embryo donor cows during superovulation. Materials and Methods: Cows (n=18) were selected and divided into three groups; control (n=6, untreated), T-I (n=6, insulin-treated), and T-II (n=6, IGF-I treated). Insulin and IGF-I were given S/C on 5 th , 6 th , 7 th , and 8 th days of estrous cycle. Superovulatory treatment was started on day 9 th of the estrous cycle. With the sixth dose of follicle-stimulating hormone, prostaglandin was injected to induce superovulatory heat. The superovulated cows were bred, and superovulatory response of each animal was recorded. The embryos were collected non-surgically on the 7 th day of superovulatory estrus. About 15 ml blood without anticoagulant was collected on days 5, 7, 9, 11, 13, 15, 17, 19, and 21 or day of embryo recovery where day 0 of estrous cycle was taken as day of estrus. Serum was separated, centrifuged, and transferred into sterilized serum vials. All samples were stored at −20°C till analysis. Progesterone, insulin, and IGF-I were estimated in blood serum by radioimmunoassay using radioimmunoassay kits. Results and Discussion: The mean concentration of progesterone on days 7 and 11, insulin on days 7 and 9, and IGF-I on days 5, 7, 9, 11, and 15 was significantly higher in insulin-treated T-I groups as compared to untreated control. Conclusion: It may be concluded that exogenous insulin administration during mid-luteal phase may be helpful in follicular and embryonic development through modulation of progesterone, insulin, and IGF-I in indigenous (Bos indicus) Sahiwal embryo donor cows.

Abdominal pregnancy is rare and is usually diagnosed late. It has a higher mortality rate than other ectopic gestations. Delayed diagnosis can lead to catastrophic hemorrhage. This report describes an abdominal ectopic pregnancy, revealed by a massive haemoperitoneum and hemorrhagic shock in a young woman.

Flagellar navigation along the genital tract of male and female in spermatozoa is accomplished through a number of biological, physiological, biochemical, and electrophysiological alterations in spermatozoa. These alterations are highly precise, dynamic, and regulated through a number of ion channels along with their associated pathways. Beating of flagella along with intracellular metabolism of spermatozoa is associated with fluxing of Ca++ as well as release of Ca++ from different sources. Calcium fluxing through the spermatozoa is mediated through sperm-specific calcium channel and also through transient receptor potential (TRP) channels which are diversified multifamily of ion channels which are activated through a number of extracellular agents such as pH, temperature, chemicals, and pathogens. Research has shown the dynamic role of TRP channels in regulating sperm functions such as sperm chemotaxis, rheotaxis, thermotaxis, and eventually fertilization. Diversified forms of TRP and their involvement in regulation of sperm function opens new horizons of understanding of the sperm function and, in specific, issues related to infertility. This mini-review is an attempt to draw some insights into the action of TRP channels in regulating sperm fertility competence through both calcium-dependent and calcium-independent mechanisms.

Elucidating the physiological mechanisms that control reproduction is an obvious strategy for improving the fertility of cattle and developing new agents to control reproductive functions. The present study aimed to identify kisspeptin neurons in the bovine hypothalamus, clarifying that a central mechanism is also present in the cattle brain, as kisspeptin is known to play an important role in the stimulation of GnRH/gonadotropin secretion in other mammals. To characterize kisspeptin neurons in the bovine hypothalamus, the co-localizations of kisspeptin and neurokinin B (NKB) or kisspeptin and dynorphin A (Dyn) were examined. Hypothalamic tissue was collected from Japanese Black or Japanese Black × Holstein crossbred cows during the follicular and luteal phases. Brain sections, including the arcuate nucleus (ARC) and the preoptic area (POA), were dual immunostained with kisspeptin and either NKB or Dyn. In the ARC, both NKB and Dyn were co-localized in kisspeptin neurons during both the follicular and luteal phases, demonstrating the presence of kisspeptin/NKB/Dyn-containing neurons, referred to as KNDy neurons, in cows. In the POA, no co-localization of kisspeptin with either NKB or Dyn was detected. Kisspeptin expression in the follicular phase was higher than that in the luteal phase, suggesting that kisspeptin expression in the POA is positively controlled by estrogen in cows. The kisspeptin neuronal populations in the ARC and POA likely play important roles in regulating the GnRH pulse and surge, respectively, in cows.

Aim: This study aimed to analyze the individual factors influencing the sperm quality of Bali bulls at Baturiti Artificial Insemination (AI) center.

Materials and Methods: Semen that was ejaculated from nine Bali bulls was collected using artificial vaginas (n=5/bull). Semen ejaculates were evaluated immediately after collection to measure the quality of the fresh semen, including semen volume, sperm concentration, progressive motility, membrane integrity (MI), and abnormal morphology. Frozen semen was evaluated for progressive sperm motility, concentration, viability, MI, abnormal morphology, and deoxyribonucleic acid (DNA) fragmentation. Other secondary data, focusing on semen quantity (semen volume and sperm concentration), were also collected from frozen the semen production data of the Baturiti AI center from 2017 to 2019. Data were analyzed statistically using a completely randomized design, and one-way analysis of variance was applied to find differences among individual bulls.

Results: Significant differences (p<0.05) were found among the bulls in semen volume, sperm motility, concentration, and MI of the fresh semen. Significant differences (p<0.05) were also found among the bulls in sperm motility, viability, MI, abnormal morphology, and DNA fragmentation of the frozen semen.

Conclusion: Individual variation in all the tested sperm parameters of the fresh semen of Bali bulls, except sperm viability and abnormalities, was noted. Similarly, individual variation in all the tested sperm parameters in frozen semen, except sperm concentration, was noted. Therefore, individual factors can be used for selecting a superior bull in Bali cattle.

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.

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.

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.

Conventionally, plasma or milk progesterone evaluations are used to determine the reproductive status of female animals. Collection of such samples is often associated with difficulties of animal handling and restraint. Measurable quantities of progesterone metabolites are found in feces of animals. Their concentrations are known to be well correlated to plasma progesterone levels and are, therefore, used as non-invasive samples for assessing reproductive function in a wide range of animal species. Although the analysis of fecal progesterone metabolites has been widely accepted in many laboratories, several factors are known to affect the results from this valuable analytical technique. Some of these factors include storage/ transportation media for fecal samples, type of solvent that is used for extraction of progesterone metabolites from feces, and the type and sensitivity of an assaying technique employed. Although fecal progesterone metabolites analysis is associated with some difficulties, it can effectively be used to monitor reproductive function in a wide range of animal species. This review aims to highlight the usefulness of fecal progesterone metabolite analysis as a non-invasive technique in monitoring reproductive function in animals. The article mainly focuses on the many opportunities and challenges associated with this analytical technique.

Reproduction is fundamental for all living things as it ensures the continued existence of a species and an improved economy in animal husbandry. Reproduction has developed since history, and diverse processes, such as artificial insemination and in vitro fertilization, have been developed. Semen extenders were discovered and developed to protect sperm from harmful factors, such as freeze and osmotic shock, oxidative stress, and cell injury by ice crystals. Semen extenders preserve sperm by stabilizing its properties, including sperm morphology, motility, and viability and membrane, acrosomal, and DNA integrity. Therefore, semen extenders must provide a favorable pH, adenosine triphosphate, anti-cooling and anti-freeze shock, and antioxidant activity to improve semen quality for fertilization. Hence, this review provides precise data on different semen extenders, preservative mechanisms, and essential additives for semen extenders in different animals.

Aim: The aim of this study was to evaluate the effects of the administration of recombinant bovine somatotropin (rbST) at the moment of implementation of the timed artificial insemination protocol, on follicular dynamics and pregnancy rate in crossbred cows.

Materials and Methods: A total of 346 cows were used in two experiments with a factorial 2×2 design. The cycling cows (Tcycling) and the anestrous cows (Tanestrous) were considered as factor 1 and the administration of rbST (TrbST) or not (Tcontrol) as factor 2. The experimental protocol: (1) Tcontrol – day 0 (D0), insertion of a progesterone-release intravaginal device (PRID) plus 2 mg of estradiol benzoate (EB); D8, PRID removal, plus 0.150 mg of prostaglandin F2α, and 400 IU of equine chorionic gonadotropin; D9, 1 mg of EB; and with artificial insemination at day 10; (2) TrbST – similar to Tcontrol plus 500 mg of rbST on D0. In experiment I, ultrasound examinations were performed in all treatments. In experiment II, the cows' pregnancy rate was evaluated. Data were analyzed with 5% probability.

Results: There was no effect of the protocols on cows cyclicity or follicular growth rate (p>0.05). There was no interaction of the effects, administration of rbST, and the cyclicity of cows on the pregnancy rate. The total pregnancy rate observed was 49.0%. The pregnancy rate in cows receiving rbST was lower for anestrous compared with cycling cows (p<0.05).

Conclusion: The administration of rbST did not alter the patterns of follicular dynamics nor the ovulation rate. However, cows in anestrous that received rbST had lower pregnancy rates than cycling cows.

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.

Background and Aim: Protamine (PRM) is the major protein in the sperm nucleus and plays an essential role in its normal function. Moreover, PRM has great potential as a protein marker of semen production and quality. This study aimed to assess the potential of sperm bovine PRM as a protein marker of semen production and quality in bulls at the National Artificial Insemination (AI) Center of Indonesia. Materials and Methods: The semen production capacity of each bull was collected from frozen semen production data at the Singosari AI Center for 6 months, and was then divided into two groups (high and low). A total of 440 frozen semen straws from six Limousin (LIM), six Friesian Holstein (FH), six Peranakan Ongole (PO), and four Aceh bulls aged 4-5 years were used in the study. The frozen semen was used to measure the concentration of PRM1, PRM2, and PRM3 using the enzyme immunoassay method. The frozen semen was also used to assess the quality of the semen, including progressive motil...

Aim: The study aimed to determine both macroscopic and microscopic changes in the reproductive organs of sterile cows.

Materials and Methods: Careful clinical and gynecological examinations (using histological and histochemical methods) of organs of culled sterile cows, such as ovaries, oviducts, the uterus, pars anterior adenohypophysis, thyroid body, and adrenals, were performed.

Results: It was found that 20% of the examined cows in the farms of the Republic of Bashkortostan had pathology of reproductive organs. Ovarian dysfunction was diagnosed in 31% of sterile cows. Histological and histochemical studies revealed that high atresia of all types of ovarian follicles is associated with hypofunction of the ovaries. This was related to stromal vascular dystrophy and was accompanied by atrophy of thecal endocrine elements, resulting in decrease of endocrine and generative function of ovaries.

Conclusion: Essential elements of the ovarian dysfunction pathogenesis are disorders of the functional system "ovary-pituitary-adrenal-thyroid gland" and the abnormality of utero-ovarian relationships, which differ significantly during hypofunction, in case of follicular cysts, and in case of persistent corpora lutea. This difference in abnormalities of utero-ovarian relationships should be considered when developing diagnostic and treatment methods and determining preventive measures.

How to cite this article: Susilowati S, Triana IN, Wurlina W, Arimbi A, Srianto P, Mustofa I (2019) Addition of L-arginine in skim milk extender maintains goat spermatozoa quality in chilled temperature for five days, Veterinary World, 12(11):1784-1789. Abstract Aim: The purpose of this study was to determine the benefits of L-arginine addition in skim milk extender to maintain the quality of goat spermatozoa in chilled storage. Materials and Methods: A total of 18 ejaculates from three healthy goats with weight and age of 45 kg and 4-5 years, respectively, were divided into three groups. The control group contained goat semen diluted in a skim milk extender without L-arginine; Treatment I and Treatment II contained goat semen diluted in a skim milk extender with added L-arginine 4 and 6 mM, respectively. These three groups were chilled at 5°C and evaluated daily for 5 days. Observed variables were viability, motility, intact plasma membrane (IPM), malondialdehyde (MDA) level, necrosis, and apoptosis of spermatozoa. Results: The addition of L-arginine 4 mM was the best treatment in maintaining viability, motility, and IPM and a decreased MDA level, percentage of necrosis, and apoptosis of goat spermatozoa. An ejaculate in this extender can be divided into 37 doses for intracervical insemination in <1 ml volume with 125 million motile spermatozoa. Conclusion: Goat semen retained its quality when kept for 5 days in chilled storage by adding L-arginine in skim milk extender.

Aim: This study aimed to study the electrophoretic properties of seminal plasma and sperm proteins of Black Bengal buck semen and their correlation with in vitro sperm characters and freezability. Materials and Methods: Semen ejaculates from nine Black Bengal bucks were collected by artificial vagina (n=20/buck). Ejaculates were evaluated for in vitro sperm characters and electrophoretic profile of seminal protein. In vitro sperm characters were evaluated immediately after collection, after completion of equilibration period, and after freeze-thawing. For seminal protein studies, seminal plasma proteins were precipitated by ice-cold ethanol method, and sperm proteins were extracted by Triton X detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to assess the molecular weight of seminal proteins. Correlation between in vitro sperm characters and protein bands was determined by Pearson's correlation coefficient, and two-way ANOVA was applied to find the individual buck differences. Results: Significant difference (p<0.01) among the bucks was noticed in the in vitro sperm characters evaluated at all the three stages of semen evaluation such as immediately after collection, after completion of equilibration period, and post-freeze thawing. Progressive loss of sperm motility, membrane integrity, and other in vitro sperm characters were noticed during cryopreservation. A total of ten protein bands in the molecular weight ranging from 17 to 180 kDa were found in the SDS-PAGE of seminal plasma proteins, while nine bands of 17-134 kDa were observed in sperm proteins. Seminal plasma proteins of molecular weight 75, 62-49, 20, and 17 kDa and sperm proteins of 75, 20, and 17 kDa were present in all the nine bucks (100%) screened, and variation among the bucks was noticed for the presence of other proteins. Seminal plasma protein of 180-134 kDa showed a negative correlation with individual motility (−0.716) and functional membrane integrity of sperm cells (−0.724) in post-freeze-thaw analysis and 48 kDa protein had a positive correlation with individual motility (0.649) and functional membrane integrity of sperm cells (0.664) in post-thaw analysis. Sperm proteins of 63 kDa had a negative correlation (−0.616) with sperm concentration in neat semen. Conclusion: Variation among the bucks was noticed in the in vitro sperm characters and semen freezability. Correlation between seminal proteins and in vitro sperm characters and semen freezability had been found which might be useful as a tool to select breeding bucks.

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.

This research article examining the influence of maternal exercise during pregnancy on vascular function in the offspring adds vital knowledge to the area of exercise physiology, fetal programming, and obstetrics research. An abundant amount of research has established the role of maternal behaviors during pregnancy on offspring health. However, much of this research has examined adverse maternal behaviors, such as smoking, drinking, stress, and over and undernutrtion, and their influence on offspring health. Research has shown that these adverse maternal behaviors can influence offspring development in utero and produce offspring that are small or large for gestational age. Changes in growth in utero can lead to greater susceptibility for diseases such as obesity, diabetes, and cardiovascular disease in the offspring later in life. Therefore, the data generated from research examining adverse maternal behaviors has helped health practitioners provide knowledge to pregnant women in order to potentially prevent or decrease the offspring’s risk for developing diseases in adulthood.

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.

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.

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: Oocyte in vitro maturation (IVM) is an appealing approach for several assisted reproductive technologies and dissecting oocyte maturation. Nonetheless, IVM leads to lower developmental competence and usually relies on undefined, serum-containing media. Therefore, biochemical profiling aimed to explore fluctuations in IVM media content during the acquisition of oocyte developmental competence.

Materials and Methods: Bovine cumulus-oocyte complexes (COCs) underwent IVM in TCM199 medium with Earle's salts, supplemented with 2.0 mM L-glutamine, 10% fetal bovine serum, antibiotics, and 0.05 IU/mL porcine follicle-stimulating hormone (FSH+) or vehicle control (CTL) medium for 22 h.

Results: FSH withdrawal (CTL) diminished several processes associated with the acquisition of oocyte developmental competence, such as reduced cumulus cell expansion, diminished estradiol synthesis (FSH+: 116.0±0.0 pg/mL vs. CTL: 97.6±18.0 pg/mL), and lower oocyte nuclear maturation rate (FSH+: 96.47% vs. CTL: 88.76%). Fresh media formulations (i.e., TCM199 with FSH or vehicle) were indistinguishable under biochemical profiling threshold conditions. Biochemical profiling showed similar total protein and lipid concentrations between groups. Further, total sugar concentrations diminished from fresh media to their post-IVM counterparts, albeit in an FSH-independent manner. Glycogen concentrations remained unaltered after IVM within CTL media, albeit were substantially lower after IVM under FSH+ conditions.

Conclusion: FSH mediates the consumption of serum-derived glycogen by bovine COCs during IVM and implies that serum-free media should contain increased glucose concentrations to facilitate the acquisition of oocyte developmental competence.

Background and Aim: Macro- and micro-elements are required to ensure the normal course of biochemical processes in the development of an animal's body. Any excess, deficiency, or imbalance in chemical elements in an animal's body can cause the development of various latent or clinically expressed pathological conditions. Diselementosis in pregnant cows may lead to impaired embryo and fetal development, as well as reduced neonatal viability. The aim of this study was to analyze the content of macroelements and microelements in the blood serum of both calving cows and their calves to evaluate the relationship between indicators of mineral metabolism in the mother and newborn and to establish what role separate chemical elements play in making newborn calves more prone to bronchopneumonia.

Materials and Methods: The content of potassium (K), sodium (Na), iron (Fe), Copper (Cu), Zinc (Zn), Strontium (Sr), arsenic (As), nickel (Ni), cobalt (Co), chromium (Cr), molybdenum (Mo), and selenium (Se) in the blood serum of 33 pregnant cows between 239 and 262 days of gestation and their 33 1-day old calves was determined using the Shimadzu AA6300 (Japan) atomic adsorption spectrophotometer. Calcium (Ca) and magnesium (Mg) content was determined using ion-selective electrodes from the Olympus-400 analyzer (Beckman Coulter, USA). During the 1st month of life, all calves in the sample set had some sort of respiratory diseases and seven of the calves had bronchopneumonia. Retrospectively, the samples of adult and newborn animals were divided into two groups each: Dams I – cows whose calves had uncomplicated bronchitis (n=26); Dams II – cows whose calves got bronchopneumonia (n=7); and Newborns I – calves with uncomplicated bronchitis (n=26); Newborns II – calves with bronchopneumonia (n=7).

Results: The content of Ca, Mg, K, Na, Mo, and Se in dams in both groups of cows was within the reference range; the concentrations of Fe and Ni were higher than the reference range; and the concentrations of Cu, Zn, As, Co, and Cr were lower than the reference range. There were no significant differences in elemental status between the Dams I and Dams II groups. In newborn calves, the concentration of Ca and Mo corresponded to the reference range; the concentrations of Mg, Fe, Co, and Ni in both groups exceeded the reference range; and the concentrations of Cu, Zn, As, Cr, and Se were lower than the reference range. Results highlighted that there was a tendency to decrease concentration of Fe, Mo, and Se and a significant increase in the Ni concentration in calves of the Newborns II group compared with calves of the Newborns I group. It was also found that Zn, Co, Cr, and Mo actively accumulated in the body of newborn animals while the transplacental transfer of Cu, As, and Sr was limited; and transfer of Se and Ni was regulated by concentration ratios in the blood of the mother and the fetus. The excessive concentrations of Ni and Fe in the blood serum of cows and calves and the imbalance in the ratio of elements Fe–Cu–Zn, Fe–Cu–Co negatively affected erythropoiesis, formation of the immune system, and antioxidant status of the fetus and newborn. These changes were considered to be risk factors for the development of bronchopneumonia in calves.

Conclusion: An excess of serum Fe and Ni and deficiency of Cu, Zn, As, Co, and Cr in cows during the gestation period can lead to similar impairments of the mineral status in newborn calves. At the systemic level, dyslementosis in combination with the influence of other adverse factors, can lead to an increased load on the respiratory and hematopoietic systems of calves during postnatal adaptation and can subsequently cause a decrease in the natural resistance of calves and development of bronchopneumonia.

Background and Aim: Dairy cows with uterine torsion often are susceptible to reduced fertility resulting in more costs and effort to restore the economy of those cows. The aim of our study was to examine and evaluate the possible associations between uterine torsion and consequent uterine involution disturbances, on the one hand, and between the degree and duration of uterine torsion with fertility parameters, on the other hand.

Materials and Methods: Within 1.5 years, 115 dairy cows (German Browns, German Holsteins, and German Fleckvieh) that were suffering from uterine torsion were examined to evaluate the incidence of involution disturbances of the uterus and to examine the fertility after calving. Statistical analysis included correlation analyses between the degree and duration of torsion and fertility parameters (days open, days to conception, conception rate and services per conception, and intercalving interval) as well as incidence of involution disturbances.

Results: The study revealed no statistically significant correlation between uterine involution and degree of uterine torsion. However, involution processes were significantly correlated to the time of the expulsion of the fetal membranes. Days to conception and intercalving intervals were significantly influenced by the presence of uterine torsion.

Conclusion: Concerning fertility after uterine torsion, it was shown that reduced fertility is associated with the duration of uterine torsion (p=0.02) and time to drop of fetal membranes (p=0.02) but not with the degree of torsion (p=0.27).

Aim: This study analyzed risk factors influencing the conception rate at the first artificial insemination (CR1) in dairy cows reared in the plain of Mitidja, which is considered a major dairy region in North Algeria. Materials and Methods: A total of 1054 lactations were used in the relational study of fertility conducted using the multivariable logistic regression model using the odds ratio (OR). Results: The breeding season had a specific effect on fertility; the first service was often followed by pregnancy when performed during autumn (AUT) (OR=1.67, p<0.05) and spring (SPR) (OR=1.65, p<0.05). The chances of obtaining conception during the first service increased significantly for a waiting period (WP) (interval between calving and time to first service) of 50-100 days postpartum (DPP) (OR=1.67, p<0.05). Conclusion: From this study, it can be concluded that no specific effect was observed relative to the breed and parity. Furthermore, CR1 significantly increased after summer calving when the first services were performed during SPR and AUT and a WP after 50 DPP.

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 >70% motile sperm and <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 (50% SMEY without glycerol at RT, stored at 5°C; and 50% SMEY with 16% glycerol added twice at 1 h and 1.5 h after being stored at 5°C). The diluted semen was loaded into 0.25 mL mini straws, equilibrated, and frozen using a freezing machine. Sperm motility, viability, membranes, DNA integrity, and concentrations of malondialdehyde (MDA) and aspartate aminotransferase (AST) enzymes were evaluated after thawing.

Results: The results showed that there were no significant differences in sperm motility and DNA integrity between dilutions (p>0.05). However, sperm viability and membrane intactness of one-step dilutions were higher than those of three-step dilutions. The concentrations of MDA and AST enzymes of sperm in one-step dilutions were lower than those of three-step dilutions (p<0.05).

Conclusion: It was concluded that the one-step-dilution technique was better than three-step dilution for cryopreservation of Limousin bull semen.

Background and Aim: Puerperal diseases influence fertility and should be diagnosed as soon as possible. This study aimed to evaluate the applicability of serum concentrations of substance P (SP), vasoactive intestinal polypeptide (VIP), and interleukin (IL)1β in the early diagnosis of uterine involution disturbances.

Materials and Methods: Blood serum samples of 86 dairy cows from six different farms were harvested within the first 20 days after calving from cows with uterine involution disturbances and healthy controls, respectively. Serum concentrations for SP, VIP, and IL-1β were determined using commercially available ELISA test kits. Statistical analyses included timely changes in blood serum levels and group comparisons of healthy cows and cows with uterine disease.

Results: SP concentrations increased significantly within 20 days after calving (p<0.04) with no significant difference observed between the groups. Moreover, no significant differences were found between VIP and log IL-1β.

Conclusion: Results showed that none of the examined serum parameters seems suitable as indicator of uterine involution disorders. Due to the timely changes in serum concentrations of SP after calving, a correlation to diseases might not be precluded. Further research is needed as regards the establishment of normative values concerning this parameter.

Aim: The present study aimed to investigate the effects of different risk factors on stillbirth of piglets born from oxytocin-assisted parturitions. Materials and Methods: Data were collected from a total of 1121 piglets born from 74 Landrace x Yorkshire crossbred sows from a herd. Logistic regression models were used to determine the associations between stillbirth and different risk factors including parity (1, 2, 3-5, and 6-10), gestation length (GL) (112-113, 114-116, and 117-119 days), litter size, birth order (BO), sex, birth interval (BI), cumulative farrowing duration, birth weight (BW), crown rump length, BW deviation, body mass index, ponderal index (PI), and the use of oxytocin during expulsive stage of farrowing. Results: The incidence of stillbirth at litter level and stillbirth rate was 59.5% (44/74) and 8.1% (89/1094), respectively. The final multivariate logistic regression selected BO, BI, PI, GL, and parity as the five most significant risk factors for stillbirth. Increased BO and BI, GL <114 and >116 days, parity 6-10, and low PI increased the stillbirth rate in piglets. Conclusion: Several factors previously determined as risks for stillbirth in exogenous oxytocin-free parturitions also existed in exogenous oxytocin-assisted parturitions. One dose of oxytocin at fairly high BO did not increase stillbirth, whereas two doses of oxytocin were potentially associated with increased values.

Aim: Oxidative stress (OS) is one of the major disruptors of oocyte developmental competence, which appears due to the imbalance between the production and neutralization of reactive oxygen species (ROS).

Materials and Methods: In Experiment 1, buffalo oocytes were in vitro matured, fertilized, and cultured at 38.5°C under 5% CO2 + 20% O2 in standard CO2 incubator (OS) or under 5% O2 + 5% CO2 + 90% N2 (Multi-gas incubator, low O2). In Experiment 2, buffalo cumulus oocytes complexes (COCs) were matured in Basic maturation medium (BMM) composed of TCM199+ 10% FCS+ 10 μg/ml FSH+ 50 μg/ml gentamicin (control group) or in BMM supplemented with 50 μM ascorbic acid (ascorbic acid group) or 3.0 mM glutathione (glutathione group) or 10-5 M melatonin (melatonin group) and cultured at 38.5°C under 20% O2 for 24 h. Matured buffalo oocytes in control, ascorbic acid, or melatonin groups were fertilized and zygotes were cultured for 8 days under the same conditions.

Results: In both experiments, maturation, cleavage, and blastocyst rates were recorded. Results showed that culture of buffalo oocytes under low O2 (5% O2) significantly increased maturation, cleavage, and blastocyst rates (p<0.05). Meanwhile, under 20% O2, addition of 10-5 M melatonin or 50 μM ascorbic acid to in vitro maturation (IVM) medium significantly improved cumulus cell expansion, nuclear maturation rates of buffalo oocytes (p<0.05), and increased cleavage and blastocyst rates (p<0.05).

Conclusion: About 5% O2 is the optimum condition for in vitro production of buffalo embryos, and addition of 10-5 M melatonin to IVM medium for oocytes cultured under 20% O2 could alleviate the adverse effect of high oxygen tension and increased embryo yield.

The aim of this study was to describe the sonographic appearance of abdominal wall at the left flank of laparotomy incision site in 11 mated Ettawah grade does. Brightness-mode ultrasound examination by using transducer with frequency of 5.0-6.0 MHz was conducted to grouping the does based on their pregnancy statuses. The incision site of the abdominal wall at left flank laparotomy was transcutaneous-scanned as long as 8 cm vertically. The sonographic appearance of the laparotomy wall thickness showed that in all groups of does were similar and not different statistically. The thickness of oblique external and oblique internal abdominal muscles was increased in the pregnant does as compared to non-pregnant does (P<0.05). Key words: laparotomy, left flank, ultrasonography, incision site, Ettawah grade does

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 concentration of serum progesterone is commonly used to determine the optimal mating time in bitches, and to diagnose reproductive-related abnormalities. This study aims to compare the serum progesterone results obtained by rapid fluorescence immunochromatography assay (RFICA) with those obtained by chemiluminescent microparticle immunoassay (CMIA) from the same serum samples to develop a standard guideline for optimal breeding time.

Materials and Methods: Serum progesterone levels were measured in 124 bitches using RFICA and CMIA. Simple linear regression and correlation analyses were performed to analyze the data. The percentage difference between the maximum and minimum progesterone values in the same serum sample in the same assay was compared using Wilcoxon's rank-sum test.

Results: The present study showed a strong linear dependence of the results obtained by RFICA on those obtained by CMIA as R2=0.8976, with regression coefficient of 0.9474 and p<0.05, including the regression model was CMIA = (0.9483 × RFICA) - 0.761. Moreover, five critical measurement times during estrous in bitches showed statistically significant differences (p<0.05), except at the fertilizable period, which showed a non-significant difference (p>0.05).

Conclusion: This study demonstrated that it is presumably acceptable to use the RFICA and CMIA methods interchangeably for quality progesterone measurements in serum samples from bitches. However, when considering the use of the RFICA method, it is advisable to carefully interpret the results and follow the interpretation guidelines. Finally, RFICA in the present study provides a reliable and convenient option for veterinarian practitioners to measure canine progesterone levels in-house.

Background and Aim: Hyperglycemia increases advanced glycation end-product (AGE) production, and the activity of receptor for AGE (RAGE) in testis, which leads to testicular histopathological damage and infertility. This research investigated the effect of black soybean tempeh (BST), purple sweet potato (PSP), and its combination on AGE and RAGE expression and spermatozoa quality in streptozotocin (STZ)-induced diabetic rats.

Materials and Methods: The rats were given high-fat diets for 5 weeks, then were injected intraperitoneally with multiple low doses of STZ (30 mg/kg body weight). Diabetes mellitus (DM) rats were divided into seven groups: DM, DM+glibenclamide, DM+BST, DM+PSP, and DM+combination of BST and PSP in ratio 1:3, 2:2, and 3:1 as C1, C2, and C3, respectively. The rats were treated for 30 days. Testicular AGE and RAGE expression and spermatozoa quality were measured.

Results: The combination of BST and PSP significantly decreased AGE and RAGE expression in testicular organs and improved spermatozoa quality compared to the normal group.

Conclusion: The combination of BST and PSP can be used as future alternatives to improve spermatozoa quality in DM patients.

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.

Background and Aim: Cogon grass (Imperata cylindrica L.) (CGG) is a herbal medicine that could be developed into a male antifertility agent. The present study aims to determine the effect of an ethanol extract of CGG roots on mice testicular activity, reproductive hormone levels, and epididymal sperm quality.

Materials and Methods: This study was designed as completely randomized with three different doses, such as an ethanol extract of CGG roots at 0 (control), 90, and 115 mg/kg body weight. In total, 21 male DDY mice strain were treated with the CGG extract (by gavage) for 14 days, followed by an evaluation of reproductive organs, epididymal sperm quality, testis histology, histomorphometry, and reproductive hormone assays. All quantitative data were analyzed by analysis of variance, followed by Tukey's post hoc test at α=0.05.

Results: The results showed that the administration of the CGG root ethanol extract disrupted the testis interstitial area and seminiferous tubules, resulting in decreased epididymal sperm quality as well as serum testosterone levels in a dose-dependent pattern.

Conclusion: Oral administration of a CGG root ethanol extract induced testicular damage, decreased epididymal sperm quality, and impaired testosterone secretion.

Intestinal endometriosis is very rarely manifested by an acute bowel obstruction secondary to ileo-cecal intussusception. We report a case of a 47-year-old female patient with symptoms and signs of an acute small bowel obstruction. CT scan confirmed the intussusception. A right colectomy for suspected malignancy was performed with an ileocolic anastomosis. Histological examination revealed an endometriosis of the ileocecal junction.