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ThesisItem Open Access Effect of Paeoniflorin against Testosterone induced Benign Prostatic Hyperplasia and Fibrosis in Wistar Rats(SKUAST Kashmir, 2023) Shahzada Mudasir Rashid; Mir, Manzoor ur RahmanIn men ≥50 years of age benign prostatic hyperplasia is the 5th most prevalent non-cancer related disorder and is reported to affect 6% of population globally. The economic burden of BPH accounts for seventh highest 1-year disease specific medical costs. Benign prostatic hyperplasia (BPH) is a pathologic process that describes proliferative events of prostate gland cells or the voiding inefficiency caused by bladder outlet obstruction due to enlarged prostate. The proliferative events are noticeable in both in stromal and epithelial elements of prostate gland. The development of an alternative medications against BPH based on natural components have become a priority in human and animal therapy. Present study was aimed to evaluate the role of Paeoniflorin-a terpene glycoside against testosterone induced benign prostatic hyperplasia of Wistar rats. We found the paeoniflorin significantly reduced the oxidative damage by increasing the activity of anti-oxidative armory like SOD, CAT, GSH, GPx, and reducing the oxidative marker MDA. A protective role of paeoniflorin against inflammation was established in blocking of NF-κB activation along with the lowering of inflammatory markers, IL-1β, IL-6 and TNF-α in prostatic tissue lysates of rats. Paeoniflorin reduced hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), transforming growth factor β (TGF β1) and confirms its role as anti-angiogenic phytochemical. It increased the epithelial marker E-Cadherin, reduced the activity of mesenchymal marker vimentin and reduced the activity of androgen receptor. Anti-BPH role of paeoniflorin is established in this study by blocking AR–androgen binding. So our study suggests the ameliorating effect of paeoniflorin against BPH of Wistar rats possibly by interfering NF-κB/AR signaling pathway. The whole results confirmed that paeoniflorin has antioxidant, anti-inflammatory, anti-angiogenic and anti-fibrotic properties that could be established as a potential anti-BPH phytocompound for prophylactic or curative use. Western blotting and mass spectrometry-based proteomics will necessities the further validation of research outcomes.ThesisItem Open Access Study on Modulation of Toll Like Receptor-4 Mediated Inflammation in Bovine Mammary Epithelial Cells Using Artemesia absinthium and Curcuma longa(2022) Insha Amin; Mir, Manzoor ur RahmanMastitis is the most prevalent disease of bovines imposing a great economic set back and a public health problem worldwide. Antibiotic resistance in both animals and humans as a result of large use of antibiotics is increasing at an alarming level. To combat this menace, scientists are researching for the development of plant based medicine which have more sidebenefits and less sideeffects in comparison to the synthetic drugs. Ethnoveterinary medicine has been used to treat a number of ailments of animals including mastitis since eons. Artemesia absinthium and Curcuma longa are the two medicinal plant species which are known to possess significant anti-inflammatory, antibacterial, antiparsitic and antioxidative properties. In the current study, we evaluated hexanic and ethanolic extracts of two plants viz A. absinthium and C. longa for their antiinflammatory and antioxidative role in LPS induced inflammation in Buffalo Mammary Epithelial Cell (BuMECs). We found that pretreatment with the extracts of both the plants downregulated the expression of proinflammatory cytokine, TNFα via TLR4/NFκB mediated signaling pathway. However, IL-6 was downregulated in only hexanic C. longa pretreated group. On the other hand, the extracts decreased the Reactive Oxygen Species (ROS) levels as detected by DCFDA assay. Significant upregulation of NRF2 mRNA expression was seen in all treated groups except hexanic A. absinthium pretreated group. Phytochemical screening of the extracts showed the presence of terpenoids, phenols, tannins and flavonoids in the extracts which was further screened in GC-MS/MS or tandem MS analysis. GC-MS/MS study of the extracts revealed the presence of important sesqueterpenoids, phenolics and some quinoxaline as bioactive phytoconstituents in the extracts. Three sesqueterpenoids viz Bourbonene 2631, Eudalene and Capparatriene were found for the first time in A.absinthium. Sesqueterpenoids like Tumerone, ar-Turmerone, Curlone and Atlantone and phenols like Guaiacol (2- Hydroxyanisole Phenol, 2-methoxy) and ethyl ferulate/ ethyl 4'-hydroxy-3'- methoxycinnamate were found in the C.longa. Quinoxaline oxid, an important antimicrobial agent has been detected in any plant for the first time in the current study. These results suggest that A. absinthium and C. longa protect BuMECs from LPS induced inflammation and oxidant insult which might be due to presence of bioactive compounds like terpenoids, phenolics and quinoxaline compounds. However, isolation of these phytoconstituents and the analysis using HPLC and NMR needs to be validated in future studies which would help to deduce the potent anti-inflammatory, antibacterial and antioxidative effects exhibited by them on LPS induced BuMECs.ThesisItem Open Access Studies on Haemato-biochemical profile of yak (Bos grunniens) in Kargil Region of Ladakh(SKUAST Kashmir, 2022) Bilquees Fatima; Sheikh, Bilal AhmadThe establishment of reliable hematologic and biochemical reference values for healthy, free ranging animal population is a challenging task, but very important for providing the baseline information for assessing the populations health and disease. The importance of haemato-biochemical indices cannot be overemphasized for evaluating the health status, chemical evaluation for survey, pathological disorders and diagnostic and prognostic evaluation of different types of diseases in animals. These indices also help to differentiate normal state from state of stress that can be maturational, environmental or physical. Hematobiochemical values have a wide application for determining the systematic relationship and physiological adaptation including the evaluation of general health conditions of animal. Current study was designed to establish the reference intervals for various haemato-biochemical parameters in yaks (Bos grunniens) by using central 90th percentile nonparametric analysis for haematological parameters viz WBCs, RBCs, Hb, HCT, PCV, RDW-CV, RDW-SD, PLT, PDW-CV, PDWSD and 95th parametric analysis for various biochemical parameters: total protein, albumin, glucose, total bilirubin, direct bilirubin, creatinine, uric acid, lipid profile, sodium, potassium, copper, zinc, iron and various enzymes. A total of 720 blood samples comprising 360 male and female clinically health yaks were included in the study. The samples were collected in both summer and winter seasons and in the age group of <1yrs and > 1yrs to obtain base line data. Due to large sample size and uniform sample and handling protocols in the current study, these reference intervals should be applicable to other yak populations.ThesisItem Open Access Investigation on polymorphism in Bovine TLR 1, 2 and Cathelicidin genes in mastitic dairy cows(SKUAST Kashmir, 2020) Sana, Bashir; Ishraq, HussianMastitis is an inflammatory condition of the mammary gland caused by bacterial, chemical, thermal or mechanical injury. Mastitis causes great loss to dairy sector owing to its negative effect on production. Multi-factorial etiology makes use of prophylactic methods for mastitis control difficult. Identification of genetic resistance mechanisms and selection for them is a potential strategy for mastitis control. The present study was done on Holstein Frisian crossbred lactating dairy animals. California mastitis test, pH and electric conductivity of milk were measured to screen animals for mastitis. The milk pH of mastitis affected group was significantly higher than normal animals (p0.05, OR=1.714) was seen between homozygyous/heterozygous genotype and mastitis. Also no significant association (p>0.05, OR=1.238) was seen between alleles “A” or “G” and mastitis. Digestion of TLR2 amplification products with Taq1 revealed two genotypes “TT” and “TG”. A significant association (p<0.05, OR=3.045) was seen between TLR2 polymorphism and mastitis. Homozygous genotype “TT” was significantly higher in normal animals whereas heterozygous “TG” genotype was significantly higher in mastitis. “T” was predominant allele in normal whereas “G” was predominant allele in mastitis affected animals (p<0.05, OR =2.137). Digestion of Cathelicidin 5 amplification product with BccI revealed three genotypes “CC”, “CG” and “GG”. A significant association (p<0.05, OR=0.0875) was seen between Cathelicidin 5 polymorphism and mastitis. Homozygous genotype “CC” was significantly higher in normal animals whereas heterozygous genotype “CG” was significantly higher in mastitis. “C” allele was predominant in normal whereas “G” allele was predominant in mastitis affected animals (p<0.05, OR=3.6667). In conclusion, polymorphism in TLR2 and Cathelicidin genes is associated with mastitis whereas polymorphism in TLR1 gene is not associated with mastitis.ThesisItem Open Access To investigate the Preventive and Therapeutic Role of Zingerone in Adjuvant-induced Arthritis in Experimental Rats(SKUAST Kashmir, 2020) Nazirah, Bashir; Sheikh, Bilal AhmadRheumatoid arthritis (RA) is a chronic systemic inflammatory disorder that preferentially affects the synovial membranes of joints and eventually leads to bone and cartilage destruction. Identification of common dietary substances capable of affording protection or modulating the onset and severity of arthritis may have important health implications. Zingerone (ZGR), a phenolic alkanone isolated from ginger, has been reported to possess various pharmacological activities. Current study was designed to investigate the protective effect of zingerone in complete Freund’s adjuvant (CFA) inoculated rats. Wistar albino rats (150–200g) of either sex were used for the experiment. Animals in group I served as a control group, group II as arthritic group, Group III as arthritic treated with zingerone for 21 days at the dose rate of 25 mg/kg b.wt., from the day of arthritis induction, Group IV as arthritic treated with zingerone for 21 days at the dose rate of 25 mg/kg b.wt., from the day of arthritis onset . Zingerone induced anti-arthritic activity was carried out by using prophylactic model and therapeutic model. Arthritis was induced by single intra-dermal injection of 0.1ml of Complete Freund’s Adjuvant at the base of tail. The protective effects of zingerone on CFA -induced oxidative stress and inflammation were investigated by assaying oxidative stress biomarkers, lipid peroxidation, serum inflammatory marker(CRP), levels of nuclear factor kappa B (NFκB), tumor necrosis factor alpha (TNF-α), transforming growth factor (TGF-β), interleukin-1β ,interleukin-6 and interleukin-10. CFA inoculation significantly increased ankle diameter and paw volume. Rats administered adjuvant alone showed significant increase in the tissue lipid peroxidation markers and a significant decrease in the activities of tissue enzymic antioxidants such as superoxide dismutase, catalase and glutathione peroxidase. Moreover, the increase in the level of inflammatory markers such as NF-κB, TGF-β, TNF-α, IL-1β, IL-6 and CRP and decrease in the level of IL-10 in the serum and joint was observed in CFA inoculated rats. These CFA-induced arthritic changes, cytokine profile, and oxidative stress markers were significantly reversed by zingerone (25mg/kg body weight) supplementation in both the models when compared to arthritic control group. Thus, zingerone, through its antioxidant and anti-inflammatory effects, may be useful in the prevention of onset and severity of arthritis.ThesisItem Open Access Mineral profiling of Dairy Cows in District Srinagar of Jammu and Kashmir(SKUAST Kashmir, 2019) Wani, Irshad Ahmad; Showkeen, MuzamilMinerals are one of the important nutritional components having significant role in health, production, reproduction and immune defence of the animals. Requirement of specific mineral depends on age, body weight, physiological status, breed and bioavailability. Deficiency of minerals affects productive and reproductive health of animal. The present study was carried out in seven Veterinary blocks of District Srinagar of Jammu and Kashmir across different stages of lactation. Before taking blood samples (10 ml) from jugular vein in heparinised tubes, a comprehensive history was taken according to the questionnaire. Plasma samples collected were digested for mineral estimation through Atomic Absorption Spectrophotometer (AAS) and for estimation of biochemical parameters. Present study revealed that in crossbred Holstein Friesian and crossbred Jersey cows, the overall mean plasma calcium levels in mid and late lactation were significantly (P<0.05) elevated as compared to early lactation stage with no significant (P<0.05) difference in mid and late stages of lactation. Similarly, in crossbred Holstein Friesian and Jersey cows, the mean plasma phosphorus and magnesium levels in early lactation were significantly (P<0.05) decreased as compared to mid and late lactation stages while no significant difference was found in plasma phosphorous levels across mid and late lactation stages. Except in block Srinagar, the overall mean zinc levels of both crossbred Holstein Friesian and Jersey cows were non significantly higher in mid lactation followed by late lactation and least in early lactation stage. Except for block Srinagar and Nowhatta, present study revealed that mean plasma copper levels of crossbred Holstein Friesian and Jersey cows were significantly (P<0.05) decreased in early lactation compared to late and mid lactation phases. The overall mean plasma iron levels in both crossbred Holstein Friesian and Jersey cows were non significantly (P<0.05) decreased in mid lactation compared to early and late lactation. The overall mean plasma glucose levels of crossbred H.F and crossbred Jersey cows were significantly elevated in mid and late lactation compared to early lactation phase. The overall mean plasma cholesterol levels in early lactation stage were significantly (P<0.05) elevated compared to mid and late lactation stage with no significant difference in plasma cholesterol levels between mid and late lactation stages among both crossbred H.F and Jersey cows. In both crossbred H.F and Jersey cows significantly (P<0.05) elevated levels of overall mean plasma triglycerides was observed in late and mid lactation stages compared to early lactation stage, while no significant difference was observed in levels of plasma triglycerides in mid and late lactation phases. Among crossbred Jersey cows, significantly elevated levels of blood urea were observed in late lactation stage compared to early and mid in block Palpora, Nowgam and block Nowhatta. Similarly albumin levels were found to decrease with progression of stages of lactation. In crossbred Holstien Friesian and crossbred Jersey cows across all the stages of lactation, a significantly negative correlation was found between milk yield and level of Ca, Mg and P in blood plasma. From the present study it can be concluded that stages of lactation has a significant variation on plasma levels of minerals and serum biochemical parameters. Although we could not observe any significant difference in different blocks of Srinagar district which can be attributed to similar topography and similar meteorological conditions in district. Milk production was found to have direct correlation with calcium, magnesium and phosphorus levels in blood plasma. Further it can be concluded from present study that there is need of formulation of mineral mixture for different stages of lactation for economizing milk production.ThesisItem Open Access Association of Acute Phase Proteins and Polymorphism of Toll-Like Receptor 4 (TLR4) gene with the mastitis in dairy cattle of Kashmir(SKUAST Kashmir, 2015) Bhat, Rahil Razak; Mir, Manzoor ur RahmanIn present study bovine acute phase proteins (APPs) and genetic polymorphism of TLR4 gene were evaluated in normal/control, subclinical and clinical mastitis animals in order to find out their possible association with bovine mastitis so as to find reliable biomarker(s) for early diagnosis of bovine mastitis in local crossbred cattle i.e. Holstein Frisian (HF) crossbred and Jersey crossbred cattle. The animals were selected from two districts of valley i.e. Srinagar and Ganderbal and were diagnosed as mastitis and normal on the basis of clinical examination and diagnostic tests i.e. California mastitis test (CMT), somatic cell count (SCC), milk pH and electric conductivity (EC) of milk. In both selected cross bred cattle, the concentration of mammary associated serum amyloid A3 (M-SAA3) which is a positive APP in milk and serum, was higher in subclinical and clinical cases in milk samples and differ significantly (p 0.05). The mean ± SE of albumin levels in serum for Jersey cross bred cattle in normal, subclinical and clinical cases were 2.426 ± 0.058, 2.303 ± 0.027 and 2.399 ± 0.055 respectively. In case of HF cross bred the values were 2.3592 ± .0877, 2.378 ± 0.0461 and 2.267 ± 0.064 respectively. The study of genetic polymorphism of bovine TLR4 (exon1 and exon2) did not revealed any association of polymorphism with mastitis in Jersey but in HF crossbred cattle only exon1 showed significant polymorphism (p < 0.05) when HaeIII was used as restriction enzyme. For exon2, HinfI and TruI restriction enzymes were used and did not yield any significant difference in the genotypes of both breeds.ThesisItem Open Access Study of Hepato and Nephro Protective Potential of Zingerone in Lead-induced Toxicity in Wistar Rats(SKUAST Kashmir, 2016) Insha, Amin; Ishraq, HussainLead poisoning continues to be a major challenge to global public health posing a great concern. Both animals and humans are affected by lead toxicity to a great extent. Lead causes oxidative stress and leads to derangement of antioxidant status of body system by increasing the levels of ROS and ALA. Liver and kidney are the two important organs where toxic symptoms of lead toxicity can be seen as these two organs are the main routes of lead excretion from the body. Finding the ways to combat lead toxicity is of utmost importance. Zingerone, a phenolic alkanone, is an active constituent present in ginger and that exhibits multiple pharmacological properties like antioxidant, antiinflammatory, anticancer, antimicrobial and antidiabetic. The current study was designed to evaluate the protective effect of zingerone in lead induced toxicity in wistar rats. Animals were randomly divided into five groups (I-V) of five animals in each group. Group I served as normal control and received normal feed and water. Group II was given lead acetate at the dose of 25 mg/kg b.wt. through intraperitoneal route on day 7, 14 and 21 of the experiment. Groups III, IV and V were given lead acetate at the dose of 25 mg/kg b.wt. through intraperitoneal route on day 7, 14 and 21 of the experiment + zingerone orally at the dose rate of 50, 100 and 150 mg/kg b.wt. respectively. Lead intoxication was analysed by estimation of ALAD activity in liver and kidney homogenates. Liver and kidney function parameters were analysed in serum along with lipid profile. Oxidative stress was analysed by estimating lipid peroxidation and antioxidant enzymes, viz., CAT, SOD, GPx and GR in liver and kidney homogenate. These findings revealed that zingerone treatment increased ALAD activity, normalized the increased liver and kidney function parameters, reduced oxidative stress (Lipid peroxidation) and increased activity of antioxidant enzymes (CAT, SOD, GR and GPx). These results suggest that zingerone may be a promising drug candidate for prevention of lead intoxication in animals.ThesisItem Open Access Investigation of variation in cytokines and protein markers in relation to Bovine mastitis(SKUAST Kashmir, 2017) Tabasum, Shaheen; Sheikh, Bilal AhmadMastitis an inflammation of mammary gland is common in lactating cows and a major cause of economic losses in dairy industry worldwide caused by bacteria and its toxins. In this study, we investigated the levels of proinflammatory cytokines IL-1β, IL-2, IL-6and TNF-α, anti-inflammtory cytokines IL-10 and TGF-β, proteins lactoferrin and albumin and milk composition in normal, subclinical and clinical mastitic dairy cows. Clinical mastitic animals showed significant variation in all the proinflammatory, anti-inflammatory levels and milk parameters when compared to normal lactating cows. They also exhibited significant increase in lactoferrin and albumin levels when compared to normal cows.Subclinical lactating cows showed significant alteration in TNF-α, IL-1β when compared to normal lactating cows. There was no significant difference between IL-2 and IL-6 in normal and subclinical lactating cows. The subclincal cows also did not exhibit significant difference in TGF-β, albumin and milk fat, protein and pH when compared with normal cows, however clinical mastitic animals showed significant difference in both TGF-β and IL-10 anti-inflammatory markers when compared with normal lactating cows. Findings in the present study indicate that cytokines can be used as a marker for detection of mastitis.
