Effect of tyrode albumin lactate pyruvate and modified Krebs Ringers broth media on in vitro capacitation of HD-K75 boar spermatozoa at different period of incubation.

In vitro capacitation allows for a greater understanding of the mechanisms underlying fertilization and the development of improved reproductive techniques for improving fertility rates in porcine. Tyrodes albumin lactate pyruvate (TALP) and modified Krebs Ringers Broth (m-KRB) are two medias that are commonly used in research experiments to induce capacitation in boar spermatozoa (Cañón-Beltrán et al., Theriogenology, 198, 2023 and 231; Oberlender et al., Archivos de Medicina Veterinaria, 44, 2012 and 201; Sahoo et al., International Journal of Biological Macromolecules, 241, 2023 and 124502). Moreover, understanding the morphological and functional changes in boar spermatozoa at different hours of capacitation periods might aid in the development of novel techniques for improving sperm quality and increasing the litter size. This study was carried out to investigate the effect of Tyrode albumin lactate pyruvate and modified Krebs Ringers Broth media on in vitro capacitation of HD-K75 boar spermatozoa at three different periods of incubation. A total of 24 ejaculate from four clinically healthy, 10-12 months aged HD-K75 boars, maintained at ICAR-All India Coordinated Research Project (AICRP) on pig were selected. Semen was collected by 'Simple fist' method using a portable dummy. The semen samples having 200 mL volume, 103 × 106 spermatozoa/ml concentration and 70% initial motility were selected and split into two parts and suspended in TALP and m-KRB media, respectively, and incubated for 5 h at 37°C. Seminal parameters viz. sperm viability, plasma membrane integrity and acrosomal integrity were estimated in the samples at 0, 3 and 5 h of incubation. This study revealed that there was significant variation between media in live acrosome-reacted (p < .05) and HOST-reacted (p < .01) spermatozoa, while between capacitation periods significant (p < .01) variation was observed in hyperactivated spermatozoa, live acrosome-reacted spermatozoa, HOST-reacted spermatozoa, FITC-labelled PSA, extracellular protein and sperm cholesterol. Non-significant variation was observed in total phospholipid. TALP showed overall better consequence on sperm viability, plasma membrane and acrosomal integrity of boar spermatozoa. From this study, it could be concluded that both TALP and m-KRB media were virtuous to induce capacitation in HD-K75 boar spermatozoa. TALP media, however, had a better effect on sperm viability, plasma membrane and acrosomal integrity of boar spermatozoa. Out of the three different periods, 3 h capacitation period resulted in significantly (p < .01) higher incidence of sperm viability, plasma membrane and acrosomal integrity in HD-K75 boar spermatozoa.

Evidence of BVDV in Pigs from North Eastern Part of India- Genetic Profiling and Characterisation.

INTRODUCTION: The work has been attempted to detect and genetically characterise the nature of Bovine Viral Diarrhea Virus (BVDV) isolates from the porcine population of the north east. METHODS AND MATERIAL: The samples have been collected over a two year period and are from areas where there is a mixed and integrated rearing of livestock in close proximity. The isolates were identified, cloned and sequenced using BVD specific genomic primers for two important domains viz., E-2 and 5' UTR. RESULTS: Porcine BVD Sequences were analysed phylogenetically. Divergence in 3 sequences is noted in the 5' UTR region that are forming a clear outlier group while E-2 sequences are coming close to BVDV group but forming a separate cluster.

Porcine circovirus 2 in the North Eastern region of India: Disease prevalence and genetic variation among the isolates from areas of intensive pig rearing.

Porcine Circovirus type-2 (PCV-2) is considered as a major threat to the piggery sector in India. To ascertain the epidemiological status and infection level of PCV2, a pilot study was undertaken to find out the prevalence of PCV2 in swine population by ELISA and PCR in the interior and border areas of Meghalaya which includes the area where accessibility and medical aid is a rare phenomenon. A total of 249 serum samples were collected from October 2014 to February 2016 from three divisions of Meghalaya: Khasi, Jaintia and Garo Hills Divisions. The mean positivity of PCV-2 antibodies in suspected sera was 83.93% whereas 62.25% of the suspected samples respectively were found to contain PCV2 as detected by PCR. Additional 190 tissue samples were collected during necropsy from both symptomatic and asymptomatic animals following reported outbreak in this region, which indicated a mean positivity of 18.94% (36/190); out of which 13 samples were subjected to sequencing to find out the genetic diversity of PCV2 amongst the field isolates. Molecular characterization and phylogenetic analysis of PCV2 isolates based on cap gene depicted genetic diversity among the strains in pig population of Meghalaya as the isolates belonged to PCV2a, PCV2b-1c and PCV2d genotypes; identification of the PCV2d genotype is probably the first report from Meghalaya. Four isolates forming an outlier group in the phylogenetic tree were arising out of natural inter-genotypic recombination between PCV2a and PCV2b. PCV2 being immunosuppressive in nature impairs the host immune response increasing the susceptibility to other co-infections leading to disease severity and high mortality in pig population. This baseline data gives a brief epidemiological status of PCV2 infection and circulating PCV2 genotype in this region which will be useful in the formulation of control and eradication programs in remotes areas of Meghalaya where accessibility is less and vaccination is a rare practice.

Anxiolytic activity of methanol leaf extract of Achyranthes aspera Linn in mice using experimental models of anxiety.

OBJECTIVE: To study the anxiolytic activity of methanol extract of Achyranthes aspera Linn (Amaranthaceae). MATERIALS AND METHODS: Male Swiss albino mice were used. Methanolic extract of Achyranthes aspera (MEAA) was administered in the doses of 100, 300 and 600 mg/kg p.o. Hole board (HB), open field (OF), elevated plus maze (EPM) and light/dark exploration (LDE) tests were used for determination of anxiolytic activity. RESULTS: The methanolic extract of Achyranthes aspera significantly increased the number and duration of head poking in HB test. The extract also significantly increased the time spent and the number of entries in open arm in EPM. In LDE test, the extract produced significant increase in time spent and number of crossings and decreased the duration of immobility in light box. In OFT, the extract showed significant increase in number of rearing, assisted rearing and the squares crossed. CONCLUSION: In the present study, MEAA exhibited anxiolytic activity which might be attributed to its phyto-constituents viz. alkaloid, steroid and triterpenes. Since Achyranthes aspera is ubiquitous and abundantly grown, it could be a fairly economical therapeutic agent for management of anxiety disorders.

Analgesic and anti-nociceptive activity of hydroethanolic extract of Drymaria cordata Willd.

OBJECTIVES: To study the analgesic and anti-nociceptive activity of hydroethanolic extract of Drymaria cordata Willd. MATERIALS AND METHODS: Wistar rats and Swiss albino mice were used for studying analgesic and anti-nociceptive activity of Drymaria cordata hydroethanolic extract (DCHE) at doses 50, 100 and 200 mg/kg p.o. Various models viz. acetic acid induced writhing model (female mice), Eddy's hot plate (mice) and tail flick model (rat) for analgesic study and formalin-induced paw licking model (mice) were used for anti-nociceptive study. RESULTS: In acetic acid induced writhing model, effect of DCHE was better than the standard drug- indomethacin 10 mg/kg (p.o.). In the hot plate model, the maximum effect was observed at 60 min at a dose of 200 mg/kg p.o., which was higher than the standard drug morphine sulfate (1.5 mg/kg i.p.), whereas in the tail flick model, effect was comparable with morphine sulfate. In formalin-induced paw licking model, administration of DCHE completely abolished the early phase at 100 and 200 mg/kg p.o. and in the late phase, the effect of DCHE (200 mg/kg p.o.) was higher than indomethacin (10 mg/kg p.o.). CONCLUSION: DCHE was effective in both non-narcotic and narcotic models of nociception, suggesting its possible action via peripheral and central mechanism. It also abolished the early phase in formalin-induced paw licking model, suggesting complete inactivation of C-fiber at higher dose. The activity can be attributed to the phyto-constituents viz tannins, diterpenes, triterpenes and steroids present in the DCHE extract. In conclusion, DCHE can be developed as a potent analgesic and anti-nociceptive agent in future.

Studies on the anti-inflammatory properties of Plantago erosa leaf extract in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves of Plantago erosa ex Roxb are used traditionally in Northeast India in different illnesses which include wounds, cuts, bruises, insect bites, poison-ivy rashes, minor sores and snakebite, etc. AIM OF THE STUDY: Plantago erosa is one of the commonly used medicinal plants in various inflammatory conditions in this region; however, due to paucity of scientific literature on its anti-inflammatory property, the present study was aimed at evaluating its anti-inflammatory activity in the leaves using in vivo models of inflammation. MATERIALS AND METHODS: Different models like carageenan induced paw edema in rat and mice, formalin induced paw licking in rats and cotton pellet induced granuloma in rats were used for studying the anti-inflammatory activity in methanol extract of Plantago erosa (PEME) leaves. RESULTS: The PEME at the oral doses from 300 to 600 mg/kg showed anti-inflammatory activity in various models. The extract (PEME) reduced carageenan induced paw edema in rat and mice, inhibited the formation of granulomatous tissue in cotton pellet induced granuloma after treatment and also decreased the reaction time in both early and late phases in formalin induced paw licking in rats. CONCLUSION: The study evidently confirmed anti-inflammatory activity of PEME and thus supported the traditional claim. The anti-inflammatory activity could be attributed to the phytoconstituent (flavonoids, alkaloids and steroid) present in the methanol extract of the plant.

Anxiolytic effect of hydroethanolic extract of Drymaria cordata L Willd.

Drymaria cordata hydroethanolic extract (DCHE) at 25, 50 and 100 mg/kg (p.o.) was administered to study anxiolytic effect. Different models for anxiolytic activity viz. Hole board, Open field, Elevated plus maze, Light/dark exploration model were used. In the hole board model, there was dose dependent and significant increase in the numbers of head pokes and the time of head dipping in the treated groups in comparison to the vehicle. In open field test, the number of rearing, assisted rearing and numbers of squares traversed increased significantly. Similarly, in elevated plus maze test, there was significant increase in the time spent and number of entries in open arm as compared to the time spent and number of entries in closed arm in dose dependent manner. In light/dark exploration test, another model for anxiolytic activity, the time spent in lit box, number of crossing and the latency period increased significantly with reduction in time spent in dark box after treatment with DCHE. The presence of phytochemicals viz. triterpenes, diterpenes, steroids and tannins might contribute to its anxiolytic activity.