Can Panax Ginseng Aqueous Extract Improve Chilled and Cryopreserved Bull Spermatozoa?

This study was to evaluate the influence of Panax ginseng aqueous extract on chilled and frozen-thawed bull sperm quality. Samples of semen were acquired from four bulls through the use of an electro-ejaculator. Extension of the semen was done with tris-egg yolk diluent which was augmented with 0.0, 0.25, 0.5, 1.0, 2.5, 5.0, and 7.5 mg/mL Panax ginseng aqueous extract. Diluted chilled portions of the semen were chilled for 6 days at 5 ̊C whereas the frozen semen was cryopreserved in liquid nitrogen. Results revealed that in chilled and frozen-thawed semen, the control group, T1 and T2 recorded higher percentages in terms of sperm motility and viability in all three groups evaluated compared to others, while the high dose of Panax ginseng aqueous extract in T6 and T5 recorded the lowest percentage. Moreover, the values of sperm morphology for chilled and frozen-thawed semen were not significant among the groups. The results of chromatin stability of the present study showed that T2 and control were higher than for other groups. In conclusion, the low dosage groups (T1, T2 and T3) which were received (0.25 mg/mL, 0.5 mg/mL and 1 mg/mL, respectively) from Panax ginseng aqueous extract were not significant as compared with the control group while high-dosage groups (T4, T5 and T6) which were received (2.5 mg/mL, 5 mg/mL and 7.5 mg/mL, respectively) from Panax ginseng aqueous extract were highly decreased spermatozoa characteristics

Sperm DNA impairment in the bull: causes, influences on reproduction and evaluations

Conventional semen examination involving sperm motility, viability and morphology remains the backbone of assessing the fertility status of a sire. However, there remains instances where these semen parameters appear normal but cases of low conception rates or failure of pregnancy occur. This review highlights the causes of sperm DNA damage and the effectiveness of techniques designed to evaluate the contribution of sperm DNA damage to lowered fertility in bulls. Among the many causes of sperm DNA impairment are imperfect spermatogenesis, faulty apoptosis, reactive oxygen species, in-vitro handling, impact of environment, radiography and the stress of cryopreservation processes. Furthermore, DNA impairment impairs fertilisation, interferes with embryonic development and implantation and blocks blastocyst formation. The most frequently used tests to determine DNA damage are the acridine orange test (AOT) using acridine orange stain with examination under a fluorescence microscope and the sperm chromatin structure assay (SCSA) using the same stain but examined with flow cytometry

Hypo-osmotic swelling test modification to enhance cell membrane integrity evaluation in cryopreserved bull semen

The objective of this study was to enhance the hypo-osmotic swelling test evaluation when it reads under light microscope using 5% formaldehyde-fixed sperm solution buffer (FSSB). Twenty four ejaculates were collected from six crossbred bulls using electro-ejaculator (EE). Tris-egg yolk extender was used to cryopreserve the semen. Concentration, volume, motility, morphology and viability rates of fresh semen were evaluated and samples were cryopreserved in liquid nitrogen. After two weeks of liquid nitrogen treatment (freezing), the motility, morphology and viability rate of the semen were evaluated. In order to carry out a hypo-osmotic swelling test, post-thaw semen was divided into four aliquots based on period of incubation (30 or 60 minutes) adding FSSB to half the samples. The components of FSSB were 5% formaldehyde and 1% eosin-nigrosin stain in PBS. Results showed that F (61.48 ±0.89%) resulted in higher percentage (P0.05) with N60 (60.90±0.70%). In conclusion, adding 10 µl of FSSB after 60 minute of incubation with hypo-osmotic swelling solution (HOSS) will enhance evaluation of the hypo-osmotic swelling test (HOST) under light microscope

Assessment of three different endometrial cytological sampling methods in postpartum beef cows

The aim of this study was to assess three different cytological endometrial sampling methods used to estimate polymorphonuclear leukocytes (PMN) under high power field (HPF) microscopy and to determine subclinical endometritis in postpartum beef cows. Forty beef cows aged 3-7 years were sampled at week three and four after calving by endometrial cytology methods. The cytological sampling methods used included; cotton swab (CS), cytobrush (CB) technique, and low volume flush (LVF), respectively. The mean PMN counts at the third week was higher (p<0.01) (12.2 cells HPF-1 than on the fourth week (4 cells HPF-1). The average PMN counts using CB alone was significantly higher (11.3 cells HPF-1) than CS (7 cells HPF-1) and LVF ( 6 cells HPF-1) methods. Smears from CB had more endometrial cells (58.55 cells HPF-1) at HPF, which was significantly higher (p<0.01) than CS and LVF methods. Both CB and CS methods yielded more intact cells (62.4 % and 61.9 %) (p <0.01) than LVF (52.4 %). The prevalence of subclinical endometritis in the beef cows between 22 and 28 days postpartum using a threshold value of ≥8 % by cytobrush method was 12.5%, which is considered low. In conclusion, CB method was found to be better and effective technique in comparison to other cytological methods used in obtaining endometrial cytology samples

Modification of electro-ejaculation technique to minimise discomfort during semen collection in bulls

The aim of this study was to reduce acute discomfortness experienced in bulls during semen collection by electro ejaculation method. The normal electro ejaculation method of semen collection (Method I) was compared to a modified method involving three stages of graduated electrical stimulation (Method II) in four crossbred bulls. The results showed that intensive muscle spasm, bull struggling and arc back were reduced (p < 0.05), as well as the time of penile protrusion (p = 0.003) and semen emission (p = 0.084) were improved using Method II than Method I. However, the total time taken for semen collection was the same in both methods. Also, there were no significant differences in semen parameters such as sperm volume, motility, morphology, viability, and concentration. In conclusion, gradation of electrical stimulation into three stages (our modified Method II) could help to ease the collection of semen samples from bulls with minimum discomfort signs. Furthermore, the modified method is also recommended to use for other animals, in particular, the wild animals

Effect of Nigella sativa pre-treatment on sub-chronic lead acetate induced hematological and biochemical alterations

Lead acetate (LA) toxicity can occur either by ingestion or inhalation from contaminated surfaces or from the environment. Nigella sativa is a natural product with immense pharmacological properties. In this study, the effects of N. sativa pre-treatment on lead acetate induced hematological and biochemical changes were evaluated. A total of 20 male Sprague Dawley rats were divided into 4 groups with 5 rats each. Group 1 (NC) was the negative control, group 2 was the lead acetate control (PC) and was administered 10 mg/kg/per day of lead acetate (LA) per OS for 30 days, group 3 (T1) was administered 200 mg/kg/daily of Nigella sativa orally for a month and Group 4 (T2) was pre-treated with 200 mg/kg/daily of Nigella sativa orally for one month, followed by administration of 10 mg/kg/daily of lead acetate (LA) orally for another month. At the end of the experiment, whole blood and serum were collected to evaluate the complete blood profile and serum biochemistry. The haemogram showed lower (p 0.05) in the control, T1 and T2 groups. The level of SOD and GSH were lower (p < 0.05) in the PC and T2 groups. In summary, this study showed the prophylactic potential of N. sativa extract in modulating both hematological, biochemical and anti-oxidant enzymes alterations induced by sub-chronic lead acetate administration in rats

Impact of Eurycoma longifolia extract on DNA integrity, lipid peroxidation, and functional parameters in chilled and cryopreserved bull sperm

This study aims to assess the effect of Eurycoma longifolia aqueous extract on chilled and cryopreserved quality of bull sperm. Semen samples were obtained from four Simmental-Brangus. Each sample was divided into two fractions: the first fraction was used for chilling the semen, and the second fraction was used for the freezing process. Both fractions were extended with Tris-egg yolk extender supplemented with 0.0, 0.25, 0.5, 1.0, 2.5, 5.0, and 7.5 mg/ml Eurycoma longifolia aqueous extract. The diluted chilled fraction was chilled at 5 °C for 6 days, whereas the frozen-thawed fraction was frozen in liquid nitrogen. Data revealed that 1 mg/ml E. longifolia aqueous extract yielded significantly (p < .05) higher sperm motility, morphology, viability, and sperm membrane integrity compared with the control group and other treated groups in chilled semen evaluation. For cryopreserved sperm, a significant difference (p < .05) in sperm motility, viability, sperm membrane integrity, DNA integrity, and lipid peroxidation was observed between 5 mg/ml E. longifolia aqueous extract and other treated and control groups. However, no significant difference in the percentage of sperm exhibiting normal sperm morphology was observed among the groups. In conclusion, the addition of 0.25 and 1 mg/ml E. langifolia extract to chilled semen and 5 mg/ml E. longifolia aqueous extract to cryopreserved sperm into Tris-egg yolk extender helps in maintaining superior quality of bull spermatozoa during chilling and freezing

Effects of Panax ginseng and Eurycoma longifolia jack extracts on chilled and frozen-thawed crossbred bull semen collected using modified electro-ejaculation method

Collection of semen from bulls can be done using different methods. Nowadays, electro-ejaculator technique is known as the most popular method to obtain semen samples from wild and domestic males. Chilled semen does not submit to the freeze-thaw procedure and suffers fewer damages, leading to greater capability and an improved capacity to fertilization. Cryopreservation of sperm plays a considerable role in economization of breeding programs in the cattle herd industry, genetic improvement of domestic animals, preservation of endangered species, and is clinically worthy in the controlling of infertility. Hence, the objectives of the present thesis were: to minimize the discomfort signs during semen collection using electro-ejaculator. Secondly, to evaluate the effect of Panax ginseng aqueous extract on the quality of chilled and frozen-thawed bull semen. Thirdly, to evaluate the effect of Tongkat Ali aqueous extract on the quality of chilled and frozen-thawed bull semen. Finally, to assess the synergistic effect of Tongkat Ali and Panax ginseng extracts on the quality of frozen-thawed bull semen. For all experiments, a total of 84 ejaculates were obtained from six crossbred bulls. The normal automatic electro-ejaculation method of semen collection (Method I) was compared to a modified method involving three stages (stage one, two and three) of gradation electrical stimulation (Method II). Discomfort signs, bulls’ response to electro-ejaculator and fresh semen samples were assessed. Tris-egg yolk extender was used to dilute the semen sample. The extender was prepared into two parts, part one (P1) which did not contain glycerol while part two (P2) contained double amount of glycerol (12.8%). Each semen extender was divided into seven groups containing different concentrations either Panax ginseng aqueous extract, Tongkat Ali aqueous extract or combination between them. Chilled and frozen-thawed semen were carried out. Chilled semen groups were placed in test tube and kept in refrigerator at 5 ºC for 6 days. The frozen semen were packaged in (0.25 mL French straws and cryopreserved in liquid nitrogen (-196 ºC). Sperm motility, morphology, viability, membranes integrity, DNA integrity, and lipid peroxidation were carried out to assess the extracts on chilled and frozen-thawed crossbred bulls. The data were analysed as mean ± standard error of the mean and checked for normal distribution. Descriptive statistical analysis of data, independent samples t-test, one or two-way analysis of variance (ANOVA), Fisher’s exact test, and chi square test were used to analyse the data. The results showed that the discomfortness signs were reduced (P ˂ 0.05), as well as using Method II than Method I. The total time taken for semen collection was similar in both methods. Also, there was no significant difference in fresh semen parameters. Panax ginseng aqueous extract did not improve the functional parameters of chilled and frozen-thawed bull semen. Moreover, sperm DNA integrity of frozen-thawed semen was not improved either. The low dosages (0.25 mg/mL) and (0.5 mg/mL) were not significant compared to control group. However, dosages more than (0.5 mg/mL) showed marked decrease in sperm characteristics (P < 0.01). Tongkat Ali aqueous extract significantly improved the chilled and frozen-thawed semen compared to control. The ideal dose of Tongkat Ali in chilled semen was (1 mg/mL) and (5 mg/mL) in frozen-thawed semen. DNA integrity and lipid peroxidation significantly improved in (5 mg/mL) compared to control group of frozen-thawed semen. In addition, different concentrations of combination between PGe and TAe into Tris-egg yolk extender did not improve the frozen-thawed bull semen quality. In conclusion, the discomfort signs prior and during semen collection were reduced by modification of the automatic mode of the electro-ejaculator device. Panax ginseng aqueous extract did not improve the quality of preserved semen. The quality of chilled and frozen-thawed crossbred bull semen was improved by adding Tongkat Ali aqueous extract to the semen diluent. The combination between Panaxginseng and Tongkat Ali extract also did not improve the quality of frozen-thawed bull semen

Semen extenders: An evaluative overview of preservative mechanisms of semen and semen extenders

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

Comparative study between the excision-ligation and autoligation of vas deferens technique for teaser rams preparation

Aim: The present study was designed to demonstrate the autoligation (AL) of vas deferens and the excision-ligation (EL) technique to generate vasectomized rams to reduce the complications, operative time, and price of the vasectomy techniques. Materials and Methods: A total of 12 healthy and mature Iraqi Awassi rams were used, which divided into two groups, six rams for each one. The former group was performed the EL technique while the latter group, the AL of vas deferens technique was used. Results: The results of the present study found that both techniques were same with the reproductive efficient examinations that mean the two techniques had same ability to close the male genital passage for teaser rams preparation. However, the methods were different with the histopathological changes, operation time, prices, and complications, which were minor in the AL of vas deferens compared with the EL technique. Conclusion: The AL technique of vas deferens to prepare teaser animal is recommended over the EL technique due to different aspects such as cost, fewer complications, and active teaser for a long period are the main aspects of AL technique