Seroepidemological investigation of Toxoplasma gondii and Trichinella spp. in pigs reared by tribal communities and small-holder livestock farmers in Northeastern India.

Toxoplasma gondii and Trichinella spp. are critical tissue-dwelling foodborne zoonotic parasites associated with pork consumption and pig rearing. Despite being a major pig-rearing region in the country, Northeastern India has not undergone any investigation regarding the presence of T. gondii and Trichinella spp. in pigs. Therefore, this study aims to determine the seroprevalence of T. gondii and Trichinella spp. and identify associated risk factors in pigs reared by tribal communities and small-holder livestock farmers in the northeastern region of India. In a cross-sectional serological survey, 400 pigs from 400 households across five northeastern states of India underwent testing for the seroprevalence of porcine toxoplasmosis and trichinellosis. Serum samples (80 from each state) were analyzed using commercially available ELISA assays. Data on backyard farm characteristics and various management aspects were collected, and risk factors linked with prevalence were analyzed through univariate and multivariate logistic regression analysis. The findings revealed that the apparent and true prevalence of anti-T. gondii antibodies were 45% (40.12-49.88, 95% CI) and 45.7% (40.7-50.69, 95% CI), respectively. As for anti- Trichinella antibodies, both the apparent and true prevalence were 0.75% (-0.1-1.6, 95% CI). The univariate and multivariate analyses indicated that age above 24 months (OR 7.20, 95% CI 2.45-23.71), exposure to cats (OR = 5.87, 95% CI 2.55-14.05), and farms operating for breeding purposes (OR = 5.60, 95% CI 3.01-11.04) were significant risk factors associated with the seroprevalence of T. gondii. This study marks the initial documentation of the seroprevalence of T. gondii and Trichinella spp. in pigs reared by tribal communities in Northeastern India. The results emphasize the significance of these parasites as foodborne zoonotic threats in the region, potentially posing substantial public health risks, especially within tribal and rural communities. The insights derived from this research could be valuable in formulating targeted preventive and control strategies against T. gondii and Trichinella spp. in pigs, not only in this region but also in areas with similar rearing practices.

First report of Ikeda genotype of Theileria orientalis in Mithun (Bos frontalis) from northeastern hilly region of India.

Oriental theileriosis caused by Theileria orientalis, previously considered a benign disease, is posing a significant threat to the livestock industry across the globe. To elucidate the prevalence of Theileria orientalis in ticks and their host, the Mithun, a comprehensive study was undertaken in the two northeastern states of India, viz. Nagaland and Arunachal Pradesh. A total of 340 of Rhipicephalus microplus ticks and 25 Ambylomma sp. ticks were screened for the presence of Theileria orientalis through PCR. Among the R. microplus ticks examined, 25 of them tested positive for T. orientalis infection whereas none of the Amblyomma ticks was positive. Additionally, a total of 275 blood samples were collected from Mithun from Arunachal and Nagaland and 31 animals were found to be positive for T. orientalis infection. Notably, six positive cases were identified in Porba (Phek district), six in Tening, and one in Bamsiakilwa village (Peren district) of Nagaland. Moreover, out of the 41 animals examined at Medziphema farms, Nagaland, 18 were found to be positive for T. orientalis infection. Moreover, the phylogenetic investigation has unveiled the presence of the highly pathogenic Type 2 (Ikeda) T. orientalis genotype in Mithun, supported by a strong bootstrap value of 100%. This study marks the initial documentation of oriental theileriosis in mithun. It underscores the need for vigilant monitoring and active surveillance of mithun populations in the northeastern states of India. Timely treatment of infected animals is imperative to avert economic losses for the farmers.

Bovine reproductive immunoinfertility: pathogenesis and immunotherapy.

Infertility is one of the primary factors for cattle reproduction in the present scenario. Reproduction-related immunoinfertility mainly involves immunization against the antigens related to reproductive hormones (LHRH, GnRH, Gonadal steroids, PGF2α and oxytocin), spermatozoa, seminal plasma and ovum. Anovulation, delayed ovulation, sperm immobilization, failure of fertilization, prolonged uterine involution, extended calving interval, prolonged post-partum estrus and reduced conception rate could be a result of immunoinfertility that occur due to the blockage of receptor site by antibodies formed against hormones, sperm and ovum. Immunoinfertility can be treated in the animal by giving sexual rest to females, by using various reproductive technologies such as in-vitro fertilization, gamete intra fallopian tube transfer, and intracytoplasmic sperm injection, sperm washing and by treating the animals with immunomodulators such as LPS, Oyster glycogen, etc. This review summarizes the different causes of bovine reproductive immunoinfertility and amelioration strategies to overcome it.

First Seroepidemiological Investigation of Hepatitis E Virus Infection in Backyard Pigs from Northeastern India: Prevalence and Associated Risk Factors.

Hepatitis E virus (HEV) is the leading cause of acute viral hepatitis globally, with zoonotic potential, and pigs are considered the major reservoir. To determine the seroprevalence of HEV infection in pigs reared in backyard conditions in the northeastern region of India, blood samples were collected from 400 pigs from five northeastern states (80 samples from each state) and tested for IgG antibodies against HEV using an ELISA assay. Questionnaires on farm characteristics and management practices were completed, and risk factors associated with HEV were studied using univariate and multivariate analysis. The apparent seroprevalence of HEV infection was 51% (46.1-55.9, 95% CI), with a true prevalence of 52.98% (47.22-58.75, 95% CI). The risk factors significantly associated with higher HEV seropositivity were as follows: lack of disinfection (OR 4.65), feeding swill (restaurant and bakery waste) (OR 2.55), failure to follow the all-in-all-out production system (OR 3.47), and medium holding size (OR 9.83), which refers to mixed rearing of younger and older age groups. This study demonstrates that HEV is widespread among pigs reared in northeastern India. The risk factor analysis conducted in this study provides valuable insights into the prevalence of HEV in the region.

Indigenous chicken production system in different agro-ecology of Indian Himalayan Region: implication on food and economic security.

The indigenous chicken production system (ICPS) has several use values and ecosystem services. In the last few years, ICPS has been recognized for its possible contribution to household food security, income generation, wildlife protection, and bettering the women's lives. This study aimed to collect, for the first time, comprehensive information about ICPS in three different agro-ecologies (tropical, sub-tropical, and sub-temperate) of the Indian Himalayan Region (IHR) and its role in food and economic security of traditional communities. In this study region, ICPS is semi-extensive, providing homegrown feed and temporary night shelter. In sub-temperate agro-ecology, females owned non-significant (p = 0.170) more indigenous chicken flocks than males. Households in sub-temperate agro-ecologies had significantly (p ≤ 0.001) larger flock sizes and tropical livestock units (chicken-TLU). However, the livestock diversity index (LDI) was significantly higher (p ≤ 0.001) in tropical and subtropical agro-ecology. The households in the sub-temperate region highly (p ≤ 0.001) valued indigenous chicken because of its survivability and adaptability. In absolute numbers significant (p ≤ 0.001) higher numbers of adult birds died in past 1 year in sub-temperate agro-ecology. The mortality rate of adult birds in sub-temperate agro-ecology was 9%, and it was 14 and 15% in tropical and sub-tropical agro-ecologies, respectively. In sub-temperate agro-ecology, larger flock size translated into significantly higher (p ≤ 0.001) egg production and subsequently a significant (p ≤ 0.001) higher egg consumption per household per month. In sub-temperate agro-ecology, households' dietary diversity score was significantly (p ≤ 0.001) higher. Similarly, the average annual income from ICPS was significantly higher (p ≤ 0.001) in sub-temperate agro-ecology and accounted for 18% of household income. ICPS' marketing chain was relatively short in the sub-temperate region. In all agro-ecologies, indigenous chicken and egg demand was significantly higher (p ≤ 0.001) in the winter. ICPS litter is used as farmyard manure, enhancing ecological resilience. In all agro-ecologies, the three most frequently cited obstacles to extending the indigenous chicken production system are illnesses, predators, and a lack of chicks availability. ICPS contributes to food and nutritional security, economic stability, and ecological resilience in this hilly and fragile ecosystem. Even though the system is self-sustaining, management and health interventions can increase production and productivity.

Participatory assessment of management and biosecurity practices of smallholder pig farms in North East India.

The present study was aimed at describing the pig production system, farm management, pig movement, and existing biosecurity level of smallholders' pig production system in North East India. A cross-sectional survey of 1,000 pig producers in four districts (two urban and two rural) in core pig-producing regions of India, where ASF occurrence had been reported, was conducted. The mean pig population was significantly (p < 0.05) higher in urban districts. In urban districts, most of the pig houses were isolated but located on the roadside, while in rural districts, commune pig houses along the roadside were more common. The majority of the respondent purchased (91%) or sold (60%) the pigs during the past 12 months. Swill feeding was common in the entire study area. The majority of the respondent (80%) in rural districts were unaware of ASF. Significant pig trade of live pigs and pork products was observed in the urban district. In the case of on-farm biosecurity measures, only 6.9% of respondents had fencing for the pig farm, 99.3% did not have provision for a footbath, and only 17.2% of the respondents restricted visitors' access to the pig farm. The study revealed that the pig production system is dominated by smallholding units with a frequent introduction or exit of pigs along with poor on-farm biosecurity measures. With the current level of farm management and biosecurity practices, smallholder pig farmers are at an increased risk of ASF and other contagious diseases.

Lipid mediated brain disorders: A perspective.

The brain, one of the most resilient organs of the body is highly enriched in lipid content, suggesting the essential role of lipids in brain physiological activities. Lipids constitute an important structural part of the brain and act as a rich source of metabolic energy. Besides, lipids in their bioactive form (known as bioactive lipids) play an essential signaling and regulatory role, facilitating neurogenesis, synaptogenesis, and cell-cell communication. Brain lipid metabolism is thus a tightly regulated process. Any alteration/dysregulation of lipid metabolism greatly impact brain health and activity. Moreover, since central nervous system (CNS) is the most metabolically active system and lacks an efficient antioxidative defence system, it acts as a hub for the production of reactive oxygen species (ROS) and subsequent lipid peroxidation. These peroxidation events are reported during pathological changes such as neuronal tissue injury and inflammation. Present review is a modest attempt to gain insights into the role of dysregulated bioactive lipid levels and lipid oxidation status in the pathogenesis and progression of neurodegenerative disorders. This may open up new avenues exploiting lipids as the therapeutic targets for improving brain health, and treatment of nervous system disorders.

Linseed oil in boar's diet improved in vivo fertility and antioxidant status.

The reactive oxygen species (ROS) which are produced during storage of boar semen are causing oxidative stress and leads to poor fertility. Also, tropical and sub-tropical weather condition adversely impacts the physicomorphological quality and fertility of boar sperm. The aim of this study was to examine the effects of feeding linseed oil to boar on its seminal attributes, sperm kinetics, biomarkers of antioxidant, fatty acid profile of seminal plasma (SP) and sperm and in vivo fertility. Six Hampshire crossbreed boars were fed with 90 ml linseed oil (LIN) whereas six Hampshire crossbreed boars were fed 90 ml canola oil (CON) for 16 weeks. Sperm quality was evaluated (60 ejaculates for each group; a total of 120 ejaculates) for motility, livability, abnormal morphology, acrosomal membrane integrity, hypo-osmotic swelling test (HOST) and sperm kinetic parameters by computer assisted semen analysis (CASA) at 0 h and at 72 h of storage at 17°C. Biomarkers of antioxidant (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC) and malondialdehyde (MDA) were measured in SP and serum. Gas chromatography-mass spectrometry (GC-MS) was used for the estimation of fatty acid composition of SP and sperm. Boars fed with linseed oil had higher semen volume (p < .01) and more total sperm numbers (p < .01). Feeding linseed oil to boar enhanced seminal attributes (p < .05) at 0 h as well as at 72 h of storage. Linseed oil feeding (p < .01) improved biomarkers of antioxidants and significantly (p < .01) lowered the lipid peroxidation in serum and SP. Linseed oil feeding (p < .05) increased the proportion of alpha linolenic (ALA), arachidonic and docosahexaenoic (DHA) fatty acids in SP. The ratio of n-6 to n-3 fatty acids in sperm increased significantly (p < .01) in treatment group. Farrowing rate was significantly (p < .05) higher in treatment group. In conclusion, feeding linseed oil to boar improved the in vivo fertility, enhanced antioxidant capacity and increased the DHA content of SP and sperm.

An evidence of Humanin-like peptide and Humanin mediated cryosurvival of spermatozoa in buffalo bulls.

Buffalo spermatozoa are vulnerable to cryo-injuries due to inherent deficiency of endogenous antioxidants, high polyunsaturated fatty acids (PUFA) content in plasma membrane and low cholesterol/phospholipid (C/P) ratio. Humanin is a potent cytoprotective agent that protects the cells against oxidative stress and apoptosis. The present study was designed to establish the presence of Humanin in buffalo and effect of Humanin supplementation on freezability of buffalo spermatozoa. Indirect immunofluorescence test revealed presence of Humanin in ejaculated and epididymal spermatozoa, and, elongated spermatids and interstitial space in the testicular tissue section. Humanin levels in seminal plasma were significantly and positively correlated with sperm concentration and individual progressive motility (IPM) in good (n = 22; IPM >70%) and poor (n = 10; IPM <50%) quality ejaculates. For supplementation studies, a total of 24 ejaculates (IPM ≥70%) were collected and each ejaculate was then divided into four aliquots. First aliquot was diluted with egg yolk-tris-glycerol (EYTG) extender without Humanin and served as control group (Group I). Rest three aliquots were diluted with extender containing 2 (Group II), 5 (Group III) and 10 µM Humanin (Group IV), respectively. Semen was cryopreserved using standard protocol and evaluated at pre-freeze for lipid peroxidation (LPO) and post-thaw stages for spermatozoa kinematics, LPO, mitochondrial membrane potential (MMP), capacitation, apoptotic status and DNA integrity. The treatment group that showed best results (5 µM) was compared with control group for in vitro fertility assessment by homologous zona binding assay. The LPO levels were lower (p < 0.05) in 5 and 10 µM Humanin supplemented group. The MMP and DNA integrity were higher (p < 0.05) in 5 µM group than other groups. F-pattern was higher (p < 0.05) and B-pattern was lower (p < 0.05) in 5 and 10 µM Humanin supplemented groups. Lower apoptotic and higher viable spermatozoa (p < 0.05) were observed in 5 µM Humanin group. The mean number of spermatozoa bound to zona pellucida was higher (p < 0.05) in 5 µM Humanin treated group than the control group. The study established the presence of Humanin in buffalo spermatozoa and seminal plasma for very first time and concluded that Humanin supplementation at 5 µM concentration improves the freezability and in vitro fertility of buffalo spermatozoa.

Linseed oil in boar's diet during high temperature humidity index (THI) period improves sperm quality characteristics, antioxidant status and fatty acid composition of sperm under hot humid sub-tropical climate.

Environmental heat stress in sub-tropical climates negatively impacts boar semen production and its quality. The present study aimed to examine the heat stress alleviating effects of dietary linseed oil on semen quality and antioxidant status of boar, in the summer and winter seasons in sub-tropical climate. Six Hampshire crossbreed boars were fed with 90 mL linseed oil (treatment) whereas six boars of the same breed were fed 90 mL vegetable oil (control) for sixteen weeks during both season. Sperm quality was assessed for motility, viability, abnormality, acrosomal integrity, and Hypo-osmotic swelling test (HOST). Sperm velocity attributes were assessed by computer-assisted semen analysis (CASA). Antioxidants (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC and nitric oxide; NO) and lipid peroxidation (malondialdehyde; MDA) were measured in seminal plasma and serum. Gas chromatography-mass spectrometry (GC-MS) was used for the estimation of fatty acid composition of seminal plasma and spermatozoa. Feeding linseed oil to the boars significantly (p < 0.05) improved sperm quality at the fresh stage and after 72 h of liquid storage in both season. There was a significant (p < 0.01) effect of treatment and season on semen quality parameters. Significant boar (p < 0.05) effect was recorded on reaction time, semen volume, sperm abnormality, acrosomal integrity and HOST reactive sperm. There was a significant (p < 0.01) effect of treatment and season on the velocity attributes viz. VAP, VSL, VCL, ALH, BCF and STR%. Linseed oil supplementation significantly (p < 0.01) enhanced antioxidant and lowered MDA levels in serum as well as seminal plasma. The concentration of alpha-linolenic (ALA), arachidonic and docosahexaenoic (DHA) fatty acids were significantly (p < 0.01) increased in seminal plasma and sperm after linseed oil supplementation. In conclusion, linseed oil supplementation to boar during high THI months improved the semen quality parameters viz. semen volume, sperm concentration, and progressive motile sperm, along with enhanced antioxidant capacity.

Self-Assembled Biodegradable Core-Shell Nanocomposites of Amphiphilic Retinoic Acid-LMW bPEI Conjugates Exhibit Enhanced Transgene Expression in Hepatocellular Carcinoma Cells With Inherent Anticancer Properties.

Low molecular weight branched polyethylenimines (LMW bPEIs) are almost nontoxic but display poor transfection efficiency due to lack of adequate complexation ability with nucleic acids followed by transportation across the cell membrane. Here, a series of amphiphilic retinoyl-bPEI conjugates (RP-1, RP-2 and RP-3) has been synthesized by allowing the reaction between bPEI (1.8 kDa) and a bioactive and hydrophobic vitamin A metabolite, all-trans-retinoic acid (ATRA), in varying amounts. In aqueous medium, these conjugates self-assembled into core-shell RP nanocomposites with size ranging from ~113-178 nm and zeta potential from ~ +15-35 mV. Evaluation of pDNA complexes of RP nanocomposites revealed that all the complexes exhibited significantly enhanced transfection efficiency without compromising on the cytocompatibility. RP-3/pDNA complex, with the highest content of retinoic acid, exhibited the best transfection efficiency. Further, due to anticancer properties of ATRA, these nanocomposites significantly reduced the viability of cancer cells (HepG2 and MCF-7 cells) without affecting the viability of non-cancerous cells (HEK 293 cells) demonstrating the cell-selective nature of the formulated nanocomposites. The intracellular trafficking and co-localization studies involving RP-3 nanocomposites also showed their higher uptake with intracellular and nuclear accumulation properties. Altogether, the results demonstrate the promising potential of the RP conjugates that can be used in future hepatocellular carcinoma targeted gene delivery applications.

Performance evaluation of the parents of the improved backyard poultry germplasm in sub-tropical condition of North Eastern Hill Region of India.

Productivity of traditional backyard poultry in developing countries can be enhanced by introduction of improved germplasm of backyard chicken which is adaptable to diverse ago-climatic condition. The growth and reproductive performance of parents of improved backyard chicken (Vanaraja and Srinidhi) were evaluated under intensive production system in humid sub-tropical region of North East India. We also evaluated the growth performance and mortality in terminal crosses of above parents until 6 weeks of age. At day 0 and at 25 weeks of age, body weight of Srinidhi male parent was significantly (P<0.05) higher than Vanaraja male parent. During brooding stage, mortality was 15.7% in Srinidhi male parent, 14.5% in Vanaraja male parent, 13.9% in Srinidhi female parent, and 8% in Vanaraja female parent. Hen-day egg production (HDEP) of Vanaraja female parent varied from 0.2 at 25 weeks to 54.9% at 54th weeks of age. In Srinidhi female parent, HDEP reached 60% at 35 weeks and peaked 63.9% at 39 weeks of age. Srinidhi female parent has significantly (P<0.05) lower age at first lay. Average annual egg production was significantly (P<0.05) higher in Srinidhi female parent (202 eggs vs. 137 eggs). Egg weight at 25, 30, 35, 40, 45, and 50 week of age was significantly (P<0.05) higher in Vanaraja female parent. Egg shape index and yolk index at 40 weeks of age were significantly (P<0.05) higher in Srinidhi female parent. Fertility of Vanaraja female parent was significantly (P<0.05) lower in winter. In case of Srinidhi female parent, fertility was significantly higher (P<0.05) in winter as compared to summer. In case of terminal crosses, body weight at 6 weeks of age was significantly (P<0.05) higher in Vanaraja as compared to Srinidhi. Cumulative mortality during brooding phase was 9.2 per cent in Vanaraja and 9.6 per cent in Srinidhi. In conclusion, parents of Vanaraja and Srinidhi are suitable and well-adapted birds in humid sub-tropical condition of North East India. These birds could be introduced in the region on large scale for up-scaling of traditional backyard poultry.

Chol-Dex nanomicelles: Synthesis, characterization and evaluation as efficient drug carriers for colon targeting.

Core-shell structures obtained from amphiphilic molecules on self-assembly in a medium have emerged as an important tool in the area of biomedical sciences. Here, we have synthesized cholesteryl-dextran (Chol-Dex) amphiphiles in sufficiently high yields via conjugation of cholesteryl hemisuccinate to dextran in two different concentrations (5 and 10%). After physicochemical and spectral analysis, the nanomicelles were subjected to size measurements. DLS and TEM confirmed the formation of core-shell type of nanomicelles. Hydrophobic drug-entrapped formulations (Metronidazole and Rifampicin) displayed sustained release behaviour of drugs from them. Sustained release at neutral pH demonstrated usefulness of the non-toxic delivery system for colon specific diseases.

Dietary flaxseed oil improve boar semen quality, antioxidant status and in-vivo fertility in humid sub-tropical region of North East India.

Environmental stress in the form of high temperature humidity index (THI) in tropical and sub-tropical region negatively affects semen quality and fertility of boar. Therefore, the present study was done to evaluate the effect of supplementing flaxseed oil (FLO) to boar's diet on its semen quality, antioxidant status, fatty acid composition of seminal plasma and fertility under sub-tropical climate. For this purpose, six Hampshire crossbreed (50% Hampshire and 50% Gunghroo) boars were divided into two groups i.e control (CON) and treatment (FLO). In FLO and CON group, flaxseed and vegetable oil, respectively, was top dressed at the rate of 3% in basal diets for each boar on daily basis for 16 weeks during monsoon season. A total of 60 ejaculates, comprising 30 ejaculates from each group (ten ejaculates from each boar) were collected. Semen samples were evaluated for sperm quality parameters (SQPs: motility, viability, abnormality, acrosomal integrity and Hypo-osmotic swelling test) and velocity attributes by computer assisted semen analysis (CASA) at fresh and after 72 h of preservation at 17 °C. Antioxidant (glutathione peroxidase; GPx, catalase; CAT, total antioxidant capacity; TAC and malondialdehyde; MDA) were analyzed in seminal plasma and serum. Fatty acid compositions of seminal plasma were analyzed by gas chromatography-mass spectrometry (GC-MS). In-vivo fertility study was also conducted. Reaction time and false mounts were significantly (p < 0.05) reduced in FLO group as compared to CON group. Semen quality parameters were significantly (p < 0.05) improved at fresh stage and after 72 h of liquid storage in FLO group as compared to CON group. Velocity attributes (VAP, VSL, VCL, ALH, BCF and LIN) were significantly (p < 0.05) higher in FLO group. Flaxseed oil supplementation significantly (p < 0.01) enhanced serum GPx and CAT concentration. Serum and seminal plasma MDA concentration decreased significantly (p < 0.01) in FLO group. Similarly, GPx, TAC and CAT were significantly (p < 0.01) elevated in seminal plasma of FLO group. The study revealed that feeding of flaxseed oil altered the fatty acid composition of seminal plasma and significantly (p < 0.05) improved the farrowing rate. In summary, flaxseed oil supplementation improved the semen quality parameters and fertility of boars in sub-tropical climate by improving the antioxidant capacity and altering the fatty acid composition of seminal plasma.

Factors for adoption of artificial insemination technology in pig: evidence from small-scale pig production system.

The average treatment estimation framework was used to estimate the actual and potential adoption rates of artificial insemination technology in pig and their determinants using responses of 390 randomly selected pig farmers. The sample adoption rate of artificial insemination technology in pig is estimated to be 16% while the potential adoption rate is estimated at 47%. Incomplete diffusion or exposure of the technology in the population has lead to significant adoption gap of 31%. Result of the average treatment estimation probit model for determinants of adoption revealed the existence of significant difference in the coefficients and magnitude of marginal effects compared to the classic adoption model. Results showed that variables such as age of household head, piggery farming experience, maximum number of pigs reared in a year during the last 5 years, number of breeding sows, distance to artificial insemination provider centre, access to extension services and participation in training and demonstration programmes on pig were the significant determinants for adoption of artificial insemination technology in small-scale pig production system.

Exogenous cholesterol prevents cryocapacitation-like changes, membrane fluidity, and enhances in vitro fertility in bubaline spermatozoa.

A study was conducted to determine the optimum dosage of the exogenous cholesterol-loaded cyclodextrins (CLC) to get maximum cryoprotection for bubaline spermatozoa. In the present study, 120 × 106 spermatozoa were incubated in 2, 3 and 4 mg of CLC as grouped as Gr II, III and IV, respectively, and sperm progressive motility, intracellular Ca2+ , capacitation status by protein tyrosine phosphorylation (PTP) assay and zona binding per cent (ZBP) and cleavage rate (CR) of the cryopreserved buffalo spermatozoa by in vitro fertility assay were assessed in comparison with an untreated control group (Gr I). Results revealed that there was a significant (p < .05) linear decrease in percentage of sperm population with higher intracellular Ca2+ and percentage of sperm population with medium or high capacitated by PTP in CLC treated from 2 to 3 mg and then increased to 4 mg/120 × 106 spermatozoa whereas sperm progressive motility, percentage of sperm population with low capacitated, ZBP and CR were increased significantly (p < .05) in sperm population treated from 2 to 3 mg CLC and then decreased to 4 mg/120 × 106 spermatozoa. The study has clearly indicated that CLC at 3 mg/120 × 106 spermatozoa has maximum beneficial effects in protection of sperm progressive motility, membrane fluidity (low intracellular Ca2+ ); prevention of cryocapacitation (low capacitation pattern in immunolocalization) and enhancement of in vitro ZBP and CR. Post-thaw motility of the CLC-treated sperm has shown positively significant (p < .05) correlation with sperm population with low intracellular Ca2+ , low capacitated sperm population, ZBP and CR, whereas it was negatively (p < .05) correlated with sperm population with high intracellular Ca2+ , medium or high capacitated sperm. The present study has revealed for the first time that incubation of spermatozoa with CLC of higher dose (>3 mg/120 × 106 spermatozoa) had adverse effects on sperm cryopreservation, although incubation of sperm with 3 mg/120 million prior to processing had minimised the freezing-thawing-associated damages in bubaline species.

A review on antioxidant potential of bioactive heterocycle benzofuran: Natural and synthetic derivatives.

The majority of heterocycle compounds and typically common heterocycle fragments present in most pharmaceuticals currently marketed, alongside with their intrinsic versatility and unique physicochemical properties, have poised them as true cornerstones of medicinal chemistry. In this context, oxygen heterocycles exhibit diverse biological and pharmacological activities due in part to the similarities with many natural and synthetic molecules with known biological activity. Among oxygen containing heterocycles, benzofuran (synthetic and natural isolated) and its derivatives have attracted medicinal chemists and pharmacologists due to their pronounced biological activities and their potential applications as pharmacological agents such as antioxidant, antitumor, antiplatelet, antimalarial, antiinflammatory, antidepressant and anticonvulsant properties. There are also an amazing number of approved benzofuran-containing drugs in the market as well as compounds currently going through different clinical phases or registration statuses. Due to the wide range of biological activities of benzofurans, their structure activity relationships have generated interest among medicinal chemists, and this has culminated in the discovery of several lead molecules in numerous disease conditions. Recently, this scaffold has emerged as a pharmacophore of choice for designing antioxidant drug development as their derivatives have shown excellent results through different mechanism of action. This review focused on the recent development of benzofuran derivatives as antioxidant agents (including natural products) and their antioxidant activities; summarize the structure property, hoping to inspire new and even more creative approaches. Also, this study systematically provides a comprehensive report on current developments in benzofuran-based compounds as antioxidant agents and is also helpful for the researchers working on a substitution pattern around the nucleus, with an aim to help medicinal chemists to develop structure activity relationships (SAR) on these derivatives as antioxidant drugs.

Vitamin E-TPGS stabilized self-assembled tripeptide nanostructures for drug delivery.

Self-assembled peptides and specifically small peptide based nanostructures have been the focus of research in past decade due to their potential biological applications. In this study, we prepared a protected peptide, Boc-Pro-Phe-Gly-OMe, which self-assembled in aqueous solutions leading to the formation of nanostructures and ability to act as a drug carrier. Dynamic light scattering (DLS) measurements showed nanostructures with average size of 119.6 nm containing hydrophobic core, wherein hydrophobic drugs, viz, eosin, aspirin and curcumin, were successfully encapsulated. These encapsulated nanostructures, were further stabilized with Vitamin E-TPGS. In-vitro drug release studies revealed the release of drugs in controlled fashion from the nanostructures. The results advocate the potential of the proposed peptide nanostructures as controlled drug delivery systems and could be used in other biomedical applications.

Sustained delivery of exogenous melatonin influences biomarkers of oxidative stress and total antioxidant capacity in summer-stressed anestrous water buffalo (Bubalus bubalis).

High ambient temperature during summer in tropical and subtropical countries predisposes water buffaloes (Bubalus bubalis) to develop oxidative stress having antigonadotropic and antisteroidogenic actions. Melatonin is a regulator of seasonal reproduction in photoperiodic species and highly effective antioxidant and free radical scavenger. Therefore, a study was designed to evaluate the effect of sustained-release melatonin on biomarkers of oxidative stress i.e., the serum malondialdehyde (MDA) and nitric oxide (NO), and the total antioxidant capacity (TAC). For the study, postpartum buffaloes diagnosed as summer anestrus (absence of overt signs of estrus, concurrent rectal examination, and RIA for serum progesterone) were grouped as treated (single subcutaneous injection of melatonin at 18 mg/50 kg body weight dissolved in sterilized corn oil as vehicle, n = 20) and untreated (subcutaneous sterilized corn oil, n = 8). Blood sampling for estimation of serum TAC and MDA (mmol/L) and NO (µmol/L) was carried out at 4 days of interval from 8 days before treatment till 28 days after treatment or for the ensuing entire cycle length. Results showed serum TAC concentration was higher in the treatment group with a significant (P < 0.05) increasing trend, whereas MDA and NO revealed a significant (P < 0.05) decline. Serum MDA and NO were higher in control compared with those of treatment group. Moreover, buffaloes in the treatment group showed 90% estrus induction with 18.06 ± 1.57 days mean interval from treatment to the onset of estrus. These results report that melatonin has a protective effect by elevating antioxidant status and reducing oxidative stress resulting in the induction of cyclicity in summer-stressed anestrous buffaloes.

Seminal PDC-109 protein vis-à-vis cholesterol content and freezability of buffalo spermatozoa.

Advancements in reproductive technologies have shown seminal plasma (SP) as a nutritive-protective medium for spermatozoa metabolism, function and transport. At the same time quality variables and thus freezability of spermatozoa are influenced by SP proteins originating from male reproductive tract. One such protein, viz. PDC-109 is reported to influence freezability of spermatozoa in cattle. Thus the present investigation was designed to evaluate effect of seminal PDC-109 protein concentration on post-thaw cholesterol content and semen quality variables (SQP) as an indicator of membrane integrity and freezability, respectively of buffalo spermatozoa. Ejaculates (n=42) selected on the basis of mass activity and individual motility were divided into three parts, first part for SP proteins isolation, second for cholesterol estimation and third part was cryo-preserved to evaluate freezability based on post-thaw SQP, viz. individual progressive motility, viability and acrosome integrity of spermatozoa. A total of 28 (66.7%) and 14 (33.3%) ejaculates from four bulls were found as freezable or non-freezable, respectively. Though total seminal plasma protein (TSPP) concentration was found similar in freezable and non-freezable ejaculates, the heparin binding proteins (HBP) content in non-freezable semen was greater (P<0.01) than freezable ejaculates. There was a similar trend for the PDC-109 protein content in respective ejaculates. Cholesterol content of spermatozoa and SQP were greater (P<0.05 and 0.01, respectively) in freezable as compared to non-freezable ejaculates of each bull at post-thaw stage. This study showed that concentrations of HBP and PDC-109 in non-freezable semen might be responsible for greater cryo-damage reflecting in poor freezability of buffalo spermatozoa.