Deciphering the immune responses in late gestation Sahiwal cows under different microclimate and its carryover effect on progenies.

The current investigation aimed to comprehend the inflammatory and related immune responses in intrauterine calves subjected to heat stress (HS) during late gestation. For this purpose, 48 Sahiwal cows in late gestation were chosen and categorized into four equal groups: naturally heat stressed (NHS), cooling-treated (CLT), spring, and winter, and likewise their neonate calves born in summer (IUHS - intrauterine heat stressed and IUCL - intrauterine cooled), spring, and winter seasons. Environmental parameters were recorded, and the temperature-humidity index (THI) was calculated daily throughout the study period. The average THI values ranged between 84.18 (summer-NHS), 73.88 (summer-CLT), 78.92 (spring), and 64.91 (winter). NHS and spring groups exhibited thermal stress based on THI (> 76.00). Various treatments significantly (P < 0.01) impacted parameters like rectal temperature (RT), respiratory rate (RR), pulse rate (PR), and skin temperature (ST) in Sahiwal cows and their calves during the study, except for heart rate (HR). Blood samples collected during different seasons and from cows housed in a climatic chamber were used to extract plasma. Plasma cortisol, interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), and thiobarbituric acid reactive substances (TBARS) levels were notably higher (P < 0.05) in the NHS compared to the CLT group. Conversely, total antioxidant capacity (TAC) and immunoglobulin G (IgG) levels were higher (P < 0.05) in the CLT and winter groups. IUHS calves exhibited significantly (P < 0.05) lower overall mean plasma TAC and IgG levels but higher inflammatory and oxidative biomarkers, such as IL-6, TNF-α, and TBARS. Additionally, significant impacts on body weight were observed for factors such as interval (P < 0.01) and the interaction between treatment and interval (P < 0.05), exhibiting consistently lower body weight in IUHS calves throughout the study period. These findings suggest that late gestation heat stress may lead to physiological alterations in future calves. Strategies aimed at mitigating heat stress during late gestation should be considered not only for the productivity and well-being of the pregnant dam but also for the development and future performance of the calf.

Optimized addition of nitric oxide compounds in semen extender improves post-thaw seminal attributes of Murrah buffaloes.

Semen dilution and cryopreservation alter the homogeneity of seminal plasma, resulting in a non-physiological redox milieu and consequently poor sperm functionality. Considering the concentration-specific bimodal action of nitric oxide (NO) in the regulation of sperm functions, cryopreservation media supplemented with optimized concentrations can improve the semen attributes. The present study aimed to evaluate the effect of adding an optimized concentration of sodium nitroprusside (SNP) and N-nitro-L-arginine methyl ester (L-NAME) in an extender on in vitro semen quality. An aliquot of semen samples (n = 32) from Murrah buffalo bulls (n = 8) was divided into control (C) and treatment (T-I: SNP in extender at 1 µmol/L; T-II: L-NAME in extender at 10 µmol/L). Fresh semen quality parameters showed no significant difference at 0 h except for the structural integrity in the T-II group. Post-thaw semen quality parameters and sperm kinematics using computer-aided sperm analysis (CASA) revealed significantly higher (p < 0.05) cryoresistance in the treatment groups. Viability, acrosome integrity, and membrane integrity were significantly higher (p < 0.05) in both treatment groups; however, the results were pervasive in T-II. Lower abnormal spermatozoa were observed in both T-I and T-II. SNP supplementation led to a significant rise (p < 0.05) in NO, whereas L-NAME reduced the NO concentration in post-thawed samples, which was directly correlated with different sperm functionality and associated biomarkers viz. total antioxidant capacity (TAC) and thiobarbituric acid reactive substance (TBARS). It was concluded that the cryopreservation media supplemented with SNP and L-NAME at 1 µmol/L and 10 µmol/L, respectively, lower the cryo-damage and improve post-thaw seminal attributes.

Bimodal interplay of reactive oxygen and nitrogen species in physiology and pathophysiology of bovine sperm function.

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) are the mediators of redox activity and are known to perform concentration-specific bimodal roles. At lower concentrations, serves as a molecular messenger and signaling molecule while at higher concentrations induces stress which in turn alters the sperm's functional characteristics. Production of ROS and RNS cannot be prevented entirely and should not be followed as a pragmatic approach as they are involved in numerous sperm physiological functions. When the antioxidants defense armory is meager, excess generation of these species cross the physiological limits and inactivates essential metabolic enzymes and disrupts signal transduction altering normal sperm functions. As per the available literature, oxidants mostly arise as a result of pathological conditions or cryopreservation-induced injury. Dead and debilitated or abnormal spermatozoa and associated leukocytes release free radicals in an excess amount which elicits oxidative and nitrosative stressors that are potentially toxic to cryosurviving sperm. ROS plays a double edge sword effect on sperm function, as regulators of physiological mechanisms at low levels and as toxicants when produced at high concentrations. Recently nitric oxide (NO.) has emerged as a potential regulator of sperm physiology, in addition, found to mediate homeostasis of the seminal plasma microenvironment when semen samples are incubated with optimal concentrations of NO. compounds. The NO. compounds can provide some resistance to future stresses which are not usually harnessed by using the defensive strategy of supplementing antioxidants. Therefore, through the optimized addition of NO. donor and inhibitor in extender, the free radical-induced damage can be avoided without inhibiting their essential physiological effects on fertilization and subsequent embryo development. This article is intended to describe the role of reactive oxidants in the physiology and pathophysiology of spermatozoa and their relationship with various seminal attributes.

Implications of cryopreservation on structural and functional attributes of bovine spermatozoa: An overview.

Sperm cryopreservation is an important adjunct to assisted reproduction techniques (ART) for improving the reproductive efficiency of dairy cattle and buffaloes. Improved understanding of mechanisms and challenges of bovine semen cryopreservation is vital for artificial insemination on a commercial basis. Although cryopreservation of bovine spermatozoa is widely practiced and advanced beyond that of other species, there are still major gaps in the knowledge and technology. Upon cryopreservation, disruption of spermatozoal plasma membrane configuration due to alterations in metabolic pathways, enzymes and antioxidants activity add to lower efficiency with loss of sperm longevity and fertilising ability. Therefore, the effective amalgamation of cryo-variables like ambient temperature, cooling and thawing rates, nucleation temperature, type and concentration of the cryoprotectant, seminal plasma composition, free radicals and antioxidant status are required to optimise cryopreservation. Novel strategies like supplementation of cholesterol-loaded cyclodextrins (CLC), nanovesicles, osteopontin, antioxidants, etc., in an extender and recent techniques like nano-purification and modified packaging have to be optimised to ameliorate the cryodamage. This article is intended to describe the basic facts about the sperm cryopreservation process in bovine and the associated biochemical, biophysical, ultra-structural, molecular and functional alterations.