Effects of different extenders, storage temperatures, and antioxidant supplementation on chilled semen quality: a review.

The successful preservation of ram semen is essential to promote genetic variability, ensure semen transportation, and inseminate multiple ewes. Currently, either animal or plant-based lipoprotein-based extenders are used for semen preservation. Animal product-based extenders include milk and egg yolk, while soybean lecithin is a plant-based extender. Although extenders containing products of animal origin better preserve the quality of chilled semen, the in vivo efficacy after 24 h of storage is still of great concern. Storage temperature is another important and effective factor in preserving sperm quality, whereby different storage temperatures are adopted to enhance the storage life of sperm. Low temperatures (4-5 °C) better preserve sperm quality for a longer duration than high temperatures (15, 20, and 25 °C). Moreover, antioxidant supplementation has a positive impact on sperm quality during liquid storage. The current review summarizes the outcomes of various extenders, different storage temperatures, and antioxidant supplementation on the liquid storage of ram sperm.

Genome Editing and Fatty Liver.

Fatty liver disease is characterized as nonalcoholic fatty liver disease (NAFLD) and alcoholic liver disease (ALD). Fatty liver disease is one of the most common causes of chronic liver disease worldwide among adults and children. It is characterized by excessive fat accumulation in the liver cells. It has a genetically heterogenous background with complex pathogenesis and progressions and is accompanied by significant morbidity, mortality, and healthcare costs. NAFLD's risk factors include metabolic syndrome, abdominal obesity, type 2 diabetes, and atherogenic dyslipidemia. ALD is associated with the excessive consumption of alcohol. Here, we describe the functions of various proteins encoded by gene variants contributing to the pathogenesis of nonalcoholic fatty liver disease and alcoholic fatty liver disease. Advancements in genome engineering technology have generated various in vivo and in vitro fatty liver disease models reflecting the genetic abnormalities contributing toward fatty liver disease. We will discuss currently developed different ALD and NAFLD models using the clustered regularly interspaced short palindromic repeats (CRISPR/Cas9) genome editing tool.Furthermore, we will also discuss the salient features of CRISPR/Cas9 editing technology and Cas9 variants such as prime and base editors to replicate genetic topographies linked specifically to ALD and NAFLD. The advantages and limitations of currently available genome delivery methods necessary for optimal gene editing will also be discussed in this review. This review will provide the essential guidance for appropriate genome editing tool selection and proper gene delivery approaches for the effective development of ALD and NAFLD models, leading to the development of clinical therapeutics for fatty liver disease.

Effects of extender type and storage time on sperm quality parameters of Kail ram semen stored at 5 °C.

The study was designed to evaluate the effect of different extenders and storage times on sperm quality parameters of extended Kail ram semen. Semen was collected from five adult Kail rams using an artificial vagina. Semen samples with >70% total sperm motility were pooled, diluted with Tris (TR), sodium citrate (SC), and skim milk (SM)-based extenders, and stored at 5 °C. Sperm motility and kinematics, viability, and plasma and acrosomal membrane integrity were assessed every 24 hrs for 120 hrs. Sperm longevity was estimated by placing semen in a water bath at 37°C to assess sperm kinematics. Storage time as the main effect had a negative (P < 0.05) impact on sperm quality parameters. The percentages of total motile (TM), progressive motile (PM), rapid, and medium progressive (MP) motile sperm were similar at 24 hrs of storage for the three extenders. However, semen extended in TR had a higher (P < 0.05) percentage of TM, PM, rapid, and MP sperm compared to SM and SC extenders at 48 hrs of storage. The sperm kinematics (VCL, VSL, VAP, and ALH) of progressive sperm were similar for all three extenders up to 72 hrs of storage. In addition, semen extended in TR had a higher (P < 0.05) percentage of sperm with intact plasma and acrosomal membranes at a storage time of 48 hrs. At 37 °C, the percentage of TM and PM sperm was higher (P < 0.05) in the TR extender than in the SM and SC extenders at 60 minutes and beyond. In conclusion, the Kail ram sperm extended in TR and stored at 5 °C maintained better quality after 48 hrs storage than sperm extended in SM and SC extenders. At 37 °C, sperm extended in TR also retained better quality parameters at 60 min and afterward than sperm extended in SM and SC.

Approaches of estrous synchronization in sheep: developments during the last two decades: a review.

The objective of the current review was to summarize the protocols used for estrous synchronization in ewes during the last two decades. Progesterone (P4) is a major hormone used in most protocols. P4 in the form of a controlled internal drug releasing (CIDR) device, medroxyprogesterone acetate (MAP), and fluorogestone acetates (FGA) has been used for estrous synchronization. Also, gonadotropins such as equine chorionic gonadotropin (eCG) and gonadotropin-releasing hormone (GnRH) are often administered at the end of P4-based protocols to improve fertility. Moreover, the administration of prostaglandins (PG) and ram effects have been used for estrus induction and synchronization of ewes. The findings of previous studies indicate that the outcome of administering various synthetics P4 analogues (CIDR, MAP, and FGA) in ewes is comparable in terms of estrous synchronization/induction. The supplementation of P4-based protocols with eCG, however, improves the estrus response and pregnancy rate during breeding and non-breeding season. On the other hand, PG is effective for successful estrous synchronization during the breeding season only. Often, two injections of PG are administered either 11 or 14 days apart along with P4-based protocols to lyse ovine corpus luteum (CL) when it is receptive to PG i.e., 3 days post-ovulation. Alternatively, the "ram effect" has been shown to improve the efficacy of P4-based protocols and can be used as an alternative to eCG in ewes. The current review describes the methods of synchronization and their outcomes during breeding and a non-breeding season in ewes.

Haemoglobinuria And Portal Venous Thrombosis In A Young Male.

Paroxysmal nocturnal haemoglobinuria is a non-malignant stem cell disorder due to acquired somatic mutations in cell surface anchored proteins CD55 and CD59. Both have a compliment inhibitory role and their deficiency leads to intravascular haemolysis. This paper reports a challenging case of a 25 years old male who presented with generalized weakness, exertional dyspnoea and episodic early morning haematuria. Recently, he started developing progressive abdominal distention and dull generalized abdominal pain. Investigations revealed haemoglobin 3.5 g/dl with 10% reticulocytes, total bilirubin 54.5 mg/dl, LDH 3155 U/L, negative Coomb's test and erythroid hyperplasia on bone marrow biopsy. Urine complete exam was significant for haemoglobinuria without red blood cells. Doppler scan of abdomen showed portal vein thrombosis. Loss of expression of CD14, CD16, CD55 and CD59 on leukocytes and erythrocytes was seen on PNH analysis, confirming paroxysmal nocturnal haemoglobinuria. He was managed with blood transfusions and was advised folic acid and bone marrow transplant.

Effects of vitamins C and E in tris citric acid glucose extender on chilled semen quality of Kail ram during different storage times.

Mammalian sperm cells are highly vulnerable to lipid peroxidation by free radicals. Antioxidants such as vitamin E, and vitamin C neutralize the activity of free radicals and protect the sperm from reactive oxygen species. The present study was conducted to investigate the effects of vitamin C, vitamin E, and their combination in a Tris-based extender on the semen quality of Kail Ram. Semen samples from five mature Kail rams were collected in this study. The semen samples were diluted by Tris-glucose-egg yolk. Diluted semen samples were divided into four parts. The first part was added with 1 mg/ml of vitamin C, the second part was added with 1 mg/ml of vitamin E and the third part was added with both vitamin C and E, in combination with a dose of 1 mg/ml. The fourth part was considered as control without any addition. The diluted semen samples were cooled gradually and preserved at 5˚C for three days. Sperms in chilled diluted semen samples were examined for motility, viability, and plasma membrane integrity every 24 h for three days (72 h). Present study results showed significant (P < 0.05) effects of vitamins C, E, and their combination on some parameters such as motility, straightness of average special path, linearity of the curvilinear trajectory, and beat-cross frequency. However, there was no significant (P < 0.05) effect of storage duration and antioxidants (vitamin C, E, and their combination) on viability, sperm plasma membrane integrity, and some CASA parameters. From the present study, it could be concluded that the supplementation of vitamins C, and E, and their combination do not enhance the life span and quality of semen in Kail ram during liquid storage at 5 °C.

Effect of weaning method on body weight, testicular growth, sexual behavior, age of puberty, and testosterone concentration of Beetal bucks.

The current study evaluates the effects of early (8th week) and late (16th week of age) weaning of male goat kids on their body growth, testicular growth, sexual behavior, plasma testosterone concentration, and pubertal age. Early (n = 6) and late (n = 7) weaned Beetal bucks were weekly monitored from 18th to 38th week for their body weight, scrotal circumference, testicular volume, testicular echogenicity (via ultrasonography), sexual activities, and plasma testosterone concentration. In comparison to early-weaned, late-weaned bucks showed a marked increase (p < .05) in body weight (11.4 ± 0.8 vs. 13.7 ± 0.6kg), testicular volume (44.1 ± 7.2 vs. 79.8 ± 18.7cm3 ), scrotal circumference (10.7 ± 0.6 vs. 12.8 ± 0.7cm), and testicular echogenicity (28.3 ± 2.7 vs. 38.3 ± 2.1) from 18th, 28th, 21st, and 24th week onward, respectively. Sexual activities started earlier in late- than early-weaned bucks (22nd vs. 25th week, respectively). Moreover, the sexual behavior index was better (p < .05) after the 34th week in late than early-weaned bucks. The plasma concentration of testosterone (at 39 weeks of age) was relatively more and the onset of puberty was 2-3 weeks earlier (p < .05) in late than early-weaned bucks. In conclusion, age-based early weaning of male kids impairs their testicular growth, sexual behavior, and age at puberty compared to conventional weaning.

Optimal timing of artificial insemination and changes in vaginal mucous characteristics relative to the onset of standing estrus in Beetal goats.

The current study was conducted to determine the optimal timing of artificial insemination and vaginal mucous characteristics relative to the onset of estrus in goats. Does (n = 257) were estrous synchronized using hormonal treatments. Intracervical inseminations with fresh semen were performed at 0, 12, 24 and 36 h after the onset of estrus. Characteristics of vaginal mucus (i.e., color, consistency, and volume) were observed and graded at the time of AI to calculate cumulative mucous score. The vaginal electrical resistance (VER) was recorded (Draminski® detector) at the time of AI. Pregnancy rate was less (P < 0.05) when inseminations occurred at 0 h (28.6%) compared with 12 (58.3%) and 24 h (56.4%) after the onset of estrus; however, pregnancy rate at 36 h (54.5%) did not differ (P> 0.05) compared with inseminations at 0, 12 or 24 h after estrous onset. Relative odds for pregnancy rate at 12, 24 and 36 h were 5.24, 5.20 and 3.29 times greater compared with 0 h. Cumulative mucous score varied (P < 0.05) relative to the onset of estrus and correlated well (P < 0.05) with the color and consistency than volume of the mucus. The VER was less (P < 0.05) at 12 than 0 and 36 h after estrous onset. In conclusion, goats can be inseminated between 12-36 h after the onset of estrus; however, the chances of pregnancy are greater with inseminations at the 12 or 24 h time periods.

Assessment of forest cover and carbon stock changes in sub-tropical pine forest of Azad Jammu & Kashmir (AJK), Pakistan using multi-temporal Landsat satellite data and field inventory.

This study aimed at estimating temporal (1989-2018) change in forest cover, carbon stock and trend in corresponding CO2 emissions/sequestration of a sub-tropical pine forest (STPF) in AJK, Pakistan. Our field inventory estimation shows an average above ground biomass (AAGB) accumulation of 0.145 Kt/ha with average carbon stock (ACS) value of 0.072 Kt/ha. Landsat TM, ETM+ and OLI images of 1989, 1993, 1999, 2005, 2010, 2015 and 2018 were used to extract vegetation fractions through Linear Spectral Mixture Analysis (LSMA) and forest area was calculated for respective years. Based on the forest area and estimated ACS value, the biomass carbon stock with corresponding CO2 emissions/sequestration was worked out for each time and change in forest carbon stock was determined for different time periods from 1989 to 2018. Our analysis shows net increase of 561 ha in forest cover and 40.39 Kt of ACS along with increase in corresponding CO2 sequestrations of 147.83 Kt over the study period. The results based on combination of remote sensing and field inventory provide valuable information and scientific basis to plan and ensure sustainable forest management (SFM) through reforestation, protection and conservation to enhance and maintain adequate forest cover and reduce CO2 emissions.