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Assam Agricultural University, Jorhat
Assam Agricultural University is the first institution of its kind in the whole of North-Eastern Region of India. The main goal of this institution is to produce globally competitive human resources in farm sectorand to carry out research in both conventional and frontier areas for production optimization as well as to disseminate the generated technologies as public good for benefitting the food growers/produces and traders involved in the sector while emphasizing on sustainability, equity and overall food security at household level.
Genesis of AAU -
The embryo of the agricultural research in the state of Assam was formed as early as 1897 with the establishment of the Upper Shillong Experimental Farm (now in Meghalaya) just after about a decade of creation of the agricultural department in 1882. However, the seeds of agricultural research in today’s Assam were sown in the dawn of the twentieth century with the establishment of two Rice Experimental Stations, one at Karimganj in Barak valley in 1913 and the other at Titabor in Brahmaputra valley in 1923. Subsequent to these research stations, a number of research stations were established to conduct research on important crops, more specifically, jute, pulses, oilseeds etc. The Assam Agricultural University was established on April 1, 1969 under The Assam Agricultural University Act, 1968’ with the mandate of imparting farm education, conduct research in agriculture and allied sciences and to effectively disseminate technologies so generated. Before establishment of the University, there were altogether 17 research schemes/projects in the state under the Department of Agriculture. By July 1973, all the research projects and 10 experimental farms were transferred by the Government of Assam to the AAU which already inherited the College of Agriculture and its farm at Barbheta, Jorhat and College of Veterinary Sciences at Khanapara, Guwahati.
Subsequently, College of Community Science at Jorhat (1969), College of Fisheries at Raha (1988), Biswanath College of Agriculture at Biswanath Chariali (1988) and Lakhimpur College of Veterinary Science at Joyhing, North Lakhimpur (1988) were established. Presently, the University has three more colleges under its jurisdiction, viz., Sarat Chandra Singha College of Agriculture, Chapar, College of Horticulture, Nalbari & College of Sericulture, Titabar. Similarly, few more regional research stations at Shillongani, Diphu, Gossaigaon, Lakhimpur; and commodity research stations at Kahikuchi, Buralikson, Tinsukia, Kharua, Burnihat and Mandira were added to generate location and crop specific agricultural production packages.
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ThesisItem Open Access Development of mucosal vaccine against riemerella anatipestifer based on membrane antigen conjugated with nanoparticle(College of Veterinary Science, Assam Agricultural University, Khanapara Campus, 2023-02) Deka, Naba Jyoti; Kalita, DhrubajyotiDuck plays a significant role to mitigate the poverty by upgrading the socio-economic condition of farmers. Riemerella anatipestifer infection is a contagious bacterial disease of ducks and causes a significant economic loss to duck rearers. The present study was carried out to develop a mucosal vaccine against Riemerella anatipestifer based on membrane antigen which was conjugated calcium phosphate nanoparticle. The outer membrane vesicle (OMVs) was extracted and conjugated with calcium phosphate nanoparticle. The average OMVs yield in terms of protein concentration was found to be 122.33 ± 3.48 mg per litre of BHI broth. In SDS-PAGE analysis, isolated OMVs exhibited presence of 16 distinct protein bands with molecular weight ranging from 142.1 to 12.1 kDa in SDS-PAGE. Among them, seven protein bands of 74.1, 69.3, 55.5, 50.6, 45.6, 25.1 and 13.1 kDa were found relatively more distinct. The major bands detected in our findings were 42 kDa, 37 kDa and 16 kDa that corresponds to OmpA, OmpH, P6 respectively. The mean size (± SD) of nanoparticle was found to be 246.20 ± 0.53 nm and the mean zeta potential (± SD) was found to be -25.60 ± 5.97. The mean size of the nanoparticles was found to be 129.80 ± 11.10 nm in size and spherical morphologies in transmission electron microscopy analysis. The optimum conditions for conjugation of OMV and calcium phosphate nanoparticles were found to be pH=6.00, amount of OMV=1.5 mg, conjugation temperature=10°C and period of conjugation=1 hour. The PD50 or the median protective dose of CAP-OMV nanoparticle was found to be 1881.10 μg of protein. For the bacterin vaccine, the concentration of the vaccine dose was taken as 2 × 109 cfu/ml. The immunization trial was carried out in ducks. Group I birds received 3762 μg of protein (entrapped protein in CAP-OMV nanoparticle) preparation via intra nasal route and it showed the highest serum IgG and secretory IgA level than the other immunized group. Group V which was administered 0.5 ml of inactivated bacterin vaccine by subcutaneous (s/c) route elicited strong immune response after Group I. All the experimental birds were challenged with 10× LD50 (~2 .1×1010 CFU per bird) on 35 days of post primary immunization. Group I and group V showed 100 % survivability, while other vaccine groups showed 0 % survivability. From the present study it can be concluded that CAP-OMV nanoparticle can act as suitable mucosal vaccine delivery system for Riemerella anatipestifer.ThesisItem Open Access Development of a chitosan based packaging film incorporated with zinc oxide nanoparticles and green tea extract : its effect on shelf life of meat and meat product (chicken)(College of Veterinary Science, Assam Agricultural University, Khanapara Campus, 2022-09) Upadhyay, Santosh; Nath, RitaThe present work was aimed at developing an active biopolymeric packaging film based on chitosan with antioxidant and antimicrobial activity and to assess its effect on shelf life of meat and meat product. Zinc oxide nanoparticle (ZnO NP) at a concentration of 2%w/w of chitosan and green tea extract (GTE) at 0.2% w/w of chitosan film was used after determining the antimicrobial activity of ZnO nanoparticles using agar well diffusion assay and antioxidant activity of GTE using DPPH inhibition assay. Four different combinations of films were prepared- F1 (Chitosan alone), F2 (Chitosan+ZnONPs), F3 (Chitosan+GTE) and F4 (Chitosan+ZnONPs+GTE) and compared with commonly used LDPE (control) packaging film for their physicochemical, mechanical, antioxidant and antimicrobial activities. The SEM of the films at resolution of 5000x revealed uniformity of the film in all four different types of combinations confirming the proper dissolution of the added zinc oxide nanoparticles and green tea extract in the film. All the four combinations of films were transparent and of desired thickness. The films F2 and F4 were found to have tensile strength but low elongation at break % as compared to LDPE. The films F2 and F4 showed potent antimicrobial property as compared to other films while F3 and F4 showed potent antioxidant activity. The film F4 possesses both strong antioxidant and antimicrobial activity with good tensile strength and was found to be the best among all the four film combinations. The developed films were then used for wrapping of fresh meat and meat loaf (chicken) and to assess their effect on shelf life at both ambient and refrigeration temperature. Samples without film were used as negative control. The fresh meat wrapped with F4 could maintaining the quality attributes in terms of physicochemical properties like TBARS, pH, colour and microbiological qualities like standard plate count and coliform count below the threshold level up to 12 hours at ambient temperature and up to 9 days of storage at refrigeration temperature. The meat loaf wrapped with F4 could be kept up to 15 hours of storage at ambient temperature and up to 15 days of storage at refrigeration temperature maintaining the quality attributes in terms of pH, TBARS, SPC, colour and textural properties. Sensory panel evaluation also suggested that the meat loaf wrapped with F4 remained acceptable up to 15 days at refrigeration temperature with good sensory scores. A significant difference (p<.01) in crude protein and ether extract contents were observed in the fresh meat and meat loaf without film and those wrapped with F1, F2 and F3. However, minimum decline was observed in samples wrapped with F4. Migration studies on zinc oxide nanoparticles revealed no significant migration of zinc oxide nanoparticles from the film to the food matrix.ThesisItem Open Access TOXICOLOGICAL ANALYSIS OF NANOPARTICLES AND MICROPARTICLES USED AS ORAL VACCINE DELIVERY SYSTEMS FOR POULTRY(College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2021-08) HAZARIKA, DIPANKAR; Tamuly, ShantanuNanoparticles and microparticles offer great applications in the field of biological sciences in terms of oral drug and vaccine delivery systems. The present study was carried out to evaluate in-vitro and in-vivo toxicity associated with chitosan nanoparticles, Gantrez® nanoparticles and poly-lactide co-glycolide (PLG) microparticle in Vero cell line and poultry bird model. The Gantrez® nanoparticles, PLG microparticles and chitosan nanoparticles were administered orally to the week old poultry birds at the limit dose of 2 g per kg body weight for assessment of oral acute toxicity and were found to be safe as the birds did not show any mortality in 24 hours post administration and the birds did not show any clinical signs till 14 days post administration. For assessment of subacute toxicity, the chitosan nanoparticles, PLG microparticles and Gantrez® nanoparticles were administered at the dose rate used in vaccine delivery that is 3 mg/kg, 15 mg/kg and 1.5 mg/kg respectively and in the dose rate 10 times of the former. The second dose was administered after 14 days. No significant elevation of serum AST, ALT, ALP, BUN and creatinine were observed in the treated groups. In addition, the significant influence of the chitosan nanoparticles, PLG microparticles, and Gantrez® nanoparticles on elevation of blood SOD, GPx and catalase were not observed. The significant upregulation of HSP70 gene expression was observed in the spleen of the group treated with PLG microparticle at the dose rate of 150 mg/kg and in the liver of the group treated with Gantrez® nanoparticle at the dose rate of 15 mg/kg. On histopathological investigation, mild changes of congestion and haemorrhage was observed in kidney of the group treated with PLG microparticles at the dose rate of 150 mg/kg body wt. while in case of liver focal aggregation of mononuclear cell was observed in Gantrez® nanoparticles at the dose rate of 15 mg/kg body weight treated group. The congested capillaries in spleen were observed in the group treated PLG microparticles at the dose rate of 150 mg/kg body weight. The groups treated with vaccine dose of chitosan nanoparticles, Gantrez® nanoparticles and PLG microparticles have shown normal cellular architecture. In in vitro study in Vero cell line, the concentration of chitosan nanoparticles and Gantrez® nanoparticles up to 1000 μg/ml did not have any influence in cellular metabolic activities and viability. However, a reduction in the cellular viability and metabolic activities were observed when PLG microparticles were used at 1000 μg/ml. At lower concentrations, all the nanoparticles/microparticles were found to be safe in terms of cytotoxicity.ThesisItem Open Access GREEN SYNTHESIS & CHARACTERISATION OF SILVER NANOPARTICLES AND ASSESSMENT OF ITS ANTIOXIDANT & ANTIMICROBIAL ACTIVITY(College of Veterinary Science, Assam Agricultural University Khanapara, Guwahati-781022, 2017-07) Deka, Naba Jyoti; Nath, RitaThe present study was undertaken to green synthesis of silver nanoparticles using leaves of Neem (Azadirachta indica) and Tulsi (Ocimum tenuiflorum) and to evaluate their antimicrobial & in-vitro antioxidant activities. The plants Neem (Azadirachta indica) and Black Tulsi (Ocimum tenuiflorum) were procured from different nearby places of Guwahati, Kamrup (metro) district, Assam. The dried leaves were pulverized and methanolic and aqueous extracts were prepared. The preliminary qualitative hytochemical analysis of Neem (Azadirachta indica) and Tulsi (Ocimum tenuiflorum) leaves showed the presence of steroid, phenol, tannin, saponin, flavonoid, alkaloid and triterpenes. The mean ± SE of total phenol of Neem and Tulsi were 0.019 ± 0.001 and 0.022 ±0.002 (mg/ml) respectively with no significant difference (P>0.05) between the two plants. The tannin content of Neem and Tulsi were 0.61 ± 0.010 and 0.580 ± 0.000 (mg/ml), flavonoid content were 0.34 ± 0.010 and 0.240 ± 0.010 (%) alkaloid content were 2.67 ± 0.010 and 2.030 ± 0.020 (%) and saponin content were 0.52 ± 0.01 and 0.410 ± 0.01 (%) respectively. Neem showed a significantly (P<0.05) higher concentration of flavonoid, alkaloid and saponin. Though tannin content was higher in Neem than Tulsi but it was found non-significant (P>0.05). The formation of silver nanoparticles were confirmed by the colour change followed by UV-Vis spectrophotometer analysis. The zeta potential measurement of the nanoparticles using Neem and Tulsi leaf extract were found to be -48.9 mV and -18.4 mV respectively. The synthesized nanoparticle were found to be almost spherical in shape in SEM images in both cases. The synthesized nanoparticles showed sensitive response against both gram positive (Staphylococcus aureus) and gram negative bacteria (Escherichia coli) which indicates that it has antimicrobial activity. The antioxidant activity of the methanolic extract, aqueous extract and silver nanoparticles from both Neem and Tulsi were evaluated by using four different standard methods. In DPPH antioxidant assay, the aggregate percent inhibition of aqueous, methanolic and silver nanoparticle of Neem leaf extract was 80.57±0.74, 63.55±0.34 and 85.16±1.44 while that of Tulsi was 68.28±0.59, 67.42±0.26 and 77.12±0.58 respectively. In nitric oxide radical assay ,The percent inhibition of nitric oxide of AgNPs of Neem extract at 50, 80 and 110 ug/ml were 73.29±0.22, 74.32±0.13 and 75.46±0.22 and of Tulsi were 12.61±0.15, 14.67±0.07 and 16.37±0.05 respectively . The aggregate value of super oxide radical scavenging activity of methanolic extract of Neem and Tulsi at different concentration were from 58.04±1.74 and 54.60±1.15 respectively and that of aqueous extract were 72.08±2.04 and 68.98±1.29 respectively .Again, in reductive ability evaluation, both aqueous and methanolic extracts of Neem and Tulsi showed higher reducing power than the standard Butylated hydroxytoluenme .Though silver nanoparticles of both the plants showed reducing power but it was found to be weaker than the standard . It may be concluded that AgNPs derived from Azadirachta indica (Neem) is richer in phytochemicals, antioxidant property and antibacterial activity than AgNPs derived from Ocimum tenuiflorum and can be further exploited for their medicinal and industrial properties.ThesisItem Open Access PHYTOCHEMICAL ANALYSIS OF MEDICINAL PLANTS AND THEIR EFFECT ON OXIDATIVE STATUS, RUMEN ENZYMES AND BIOCHEMICAL PROFILE OF GOAT(Assam Agricultural University, Khanapara, Guwahati, 2017-07) THAKURIA, PURABI; Nath, RitaIn this study, three medicinal plants viz. Artocarpus heterophyllus (Jackfruit), Carica papaya (Papaya) and Terminalia bellerica (Bhumura) were investigated for the presence of phytochemicals and their effect on the rumen enzymes, oxidative status and blood biochemical profile was evaluated. The preliminary phytochemical screening of Artocarpus heterophyllus, Carica papaya and Terminalia bellerica revealed the presence of phytochemicals such as tannin, saponin, flavonoid, terpenoid, alkaloid, glycosides and protein in methanolic extract. Quantitative analysis revealed that the highest concentration of tannin (%) was found in C. papaya (0.638 ± 0.05) followed by T. bellerica (0.562 ± 0.09) and A. heterophyllus (0.253 ± 0.06). However, the flavonoid concentration (mg QE/g) was highest in T. bellerica (0.825 ± 0.001). But there was no significant (P<0.01) difference in saponin concentration among all the three plant leaves and the highest concentration of saponin (mg/100g) was found in T. bellerica (2.67 ± 0.07). The antioxidant activity in terms of DPPH (2, 2- diphenyl-2- picryl hydrazyl hydrates) radical scavenging activity was highest in A. heterophyllus (70.23 ± 0.0.50 %) followed by C. papaya (65.57 ± 0.27 %), T. bellerica (62.70 ± 0.06 %) and Vitamin C (27.04 ± 0.01 %) respectively at concentration of 110 µg/ml. Similarly, in superoxide radical scavenging activity A. heterophyllus showed highest radical scavenging activity (83.35 ± 0.09 %) followed by C. papaya (81.08 ± 0.04 %), T. bellerica (75.36 ± 0.14 %) and vitamin C (14.50 ± 0.05 %) at concentration of 110 µg/ ml. Reductive ability was highest in A. heterophyllus (0.137 ± 0.001) compared to standard Vitamin C (0.134 ± 0.001) respectively at concentration of 110 µg/ml. On the otherhand, the highest nitric oxide free radical scavenging activity (%) was recorded in vitamin C (56.16 ± 0.03) followed by T. bellerica (40.43 ± 0.12), A. heterophyllus (36.05 ± 0.57) and C. papaya (26.01 ± 0.23). The highest concentration of hydroxyl radical scavenging activity (%) was found in C. papaya (76.64 ± 1.11) followed by T. bellerica (83 ± 0.08) and A. heterophyllus (70.36 ± 0.17) when compared with vitamin C (13.12 ± 0.01) at 110 µg/ml concentration. The vitamin C content was highest in C. papaya (39.71 ± 0.24 mg/100g). In the present study, eighteen (18) Assam Hill (local) goats of 3-4 months of age were procured and reared in the Experimental Animal Shed, Department of Veterinary Physiology, C.V.Sc., AAU, Khanapara, Guwahati-22. The animals were divided into three groups. One as control, receiving substrate ration, T1 with plant leaves @ 5% level and T2 @ 10% level of total mixed ration (TMR). The duration of the experiment was 90 days. Rumen liquor analysis revealed that out of the three rumen fibrolytic enzymes, Carboxymethylcellulase showed better enzyme activity in T2 group. The blood glucose level (mg/dl) was within the normal physiological range but apparently decreased in T1 and T2 groups indicating hypoglycemic effect of the plant leaves. The serum total protein level (g/dl) was also within the normal range and the level was significantly (P<0.01) higher in T2 group. The level of blood creatinine (mg/dl) increased significantly (p<0.01) in 0 to 90 days from 1.02 ± 0.08 to 1.80 ± 0.08 in all the experimental groups of animals but the observed values were within the normal range. The values recorded for blood urea nitrogen concentration (mg/dl) in all the groups of animal during different days of experiment were found to be apparently higher but analysis of variance revealed no significant differences during the respective days of experiment. The ALT and AST values (U/L) were found to be apparently higher but the values differed non-significantly amongst all groups during the respective days of experiment and the values were within the normal range indicating no harmful effect of the plant leaves on liver. The values recorded for GGT (U/L) in the present experiment were apparently decreased from 37.10 ± 1.35 to 35.91 ± 1.07 in 0 to 90 days but analysis of variance revealed no significant differences during the respective days of experiment. The overall mean value of GGT in T2 group was insignificantly lower than the values recorded in T1 group which indicated normal hepatic condition of the animal. The serum calcium concentration (mg/dl) recorded during 0 to 90 days of experiment in all the groups of goat increased significantly (p<0.01) from 9.17 ± 0.16 to 11.73 ± 0.15. An increase level of phosphorus (mg/dl) was also recorded in T2 group. The serum sodium concentration (mEq/L) was also within the normal physiological range and no significant differences were observed between the respective days of experiment in both T1 and T2 group. The observed value of serum potassium (mEq/L) increased significantly (p<0.01) from 0 to 90 days of experiment in control, T1 and T2 groups. The iron value ((μg/dl)) recorded in the present experiment increased apparently from 0 to 90 days and insignificantly higher values were observed in T2 group (173.69 ± 1.95 μg/dl) as compared to T1 and control groups. The average body weight (kg per animal) recorded in the present experiment showed significant trend (P<0.01) from 0 to 90 days of experiment. The blood malondialdehyde values (μmol/L) in control group increased significantly (P<0.01) from 0.60 ± 0.06 to 0.80 ± 0.02 in 0 to 90 days of experiment but in T1 and T2 groups, blood MDA value decreased significantly (P<0.01) from 0 to 90 days of experiment. Analysis of variance revealed no significant differences during different days of experiment and also amongst the experimental groups of the animal in respect of blood super oxide dismutase (SOD) and glutathione peroxidase (GPX) concentration. The levels of vitamin C (μg/ml) were apparently higher from 0 to 90 days of experiment but analysis of variance revealed no significant differences. From this study, it can be concluded that three medicinal plants viz., A. heterophyllus, C. papaya, T. bellerica are rich source of phytochemicals with good antioxidant properties and do not have any harmful effect on normal functioning of the rumen, liver and kidney. Thus, the plant leaves can be incorporated in the diet up to 10% of the total mixed ration.ThesisItem Open Access THERAPEUTIC POTENTIAL OF BONE-MARROW DERIVED MESENCHYMAL STEM CELLS IN WOUND HEALING(Assam Agricultural University, Khanapara, Guwahati, 2017-07) HAZORIKA, MOUSUMI; Kalita, Dhruba JyotiThe present experiment was carried out to explore the therapeutic potential of rabbit bone-marrow derived mesenchymal stem cells (rBM-MSCs) in wound healing in New Zealand White rabbits. The experiment was conducted in the Department of Veterinary Biochemistry, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati-781022. Eighteen numbers of clinically healthy male New Zealand white rabbit of 3-4 months with mean body weight of 1.52 kg were randomly divided into three different groups of six animals each viz. Gr-I (Control group), Gr-II (Test group-1) and Gr-III (Test group-2). A total of thirty-six surgical wounds were created on the skin of eighteen rabbits with two in each. The Gr-I rabbits received no treatment and was considered as Control group. Gr-II rabbits were treated with a standard drug i.e. Povidine iodine ointment (Betadine®, USP 10 % w/w) while Gr–III rabbits were treated with cultured BM-MSCs topically. They were raised under standard laboratory conditions and veterinary supervision throughout the experimental period and were given standard pellet feed. The experiment was conducted in a 2x2 factorial design for a period of 30 days. Bone marrow was collected from rabbits to culture and characterize the BM-MSCs. Blood was collected from all the animals of each group on days 0, 3, 7, 15 and 30 to estimate haemato-biochemical constituents viz. haemoglobin, PCV, ESR, TEC, TLC and DLC; serum glucose, total protein, triglycerides, SGOT, SGPT, alkaline phosphatase, uric acid, Ca2+, P, Mg2+, Na+, K+, Cl-, iron, copper, zinc, cytokine proteins: TNF-α, IL-1β, IL-4, IL-6, IL-10 and to study cytokine gene expression pattern during wound healing. Wound area was measured on days 1, 3, 7, 15, 21 and 30 of each animal, of different experimental groups. Tissue was collected on day 10 and day 20 of surgery from each animal of different experimental groups to estimate certain granulation tissue markers viz. LPO, NO; SOD, CAT, GSH, vitamin A, vitamin C, vitamin E; MPO; protein and DNA; and connective tissue markers viz. HYP, HXA, HUA, collagen and elastin. The tissue was also collected on days 1, 10 and 20 of surgery to observe the histopathological changes during wound healing. The rBM-MSCs was isolated, cultured and characterized as per the standards set by ISCT. The rBM-MSC treated group showed complete healing of wound by day 21 whereas complete healing of wound was observed on day 30 in both Gr-I and Gr-II. All the haemato-biochemical constituents were found within the normal range throughout the experimental period. However, the levels of certain parameters significantly increase or decrease amongst the groups with the advancement of time. The rBM-MSCs significantly increased the antioxidants’ levels and reduced the generation of free radicals’ and acute inflammatory marker on day 10 and day 20 of surgery. Significant increase observed in the content of granulation tissue biochemical markers (protein and DNA) and connective tissue markers (HYP, HXA, HUA, collagen and elastin). Histopathologically, the BM-MSCs treated group showed early tissue changes compared to other groups and approximately resembles the adjacent normal skin on day 20. On the basis of macroscopic, biochemical and histopathological evaluation of wounds, it can be concluded that the rBM-MSCs can be used successfully for the treatment of surgical wounds without any significant adverse effects on the animal.ThesisItem Open Access EXPLORATION OF CYTOKINE PROFILES FOR PREDICTION OF SUCCESSFUL PREGNANCY AND LACTATION AND MANIPULATION OF CYTOKINE LEVEL USING CERTAIN STIMULANT IN CROSS BRED COWS OF ASSAM(Assam Agricultural University, Khanapara, Guwahati, 2017-07) BARKAKATI, JURI; Kalita, Dhruba JyotiThe present study was conducted to explore the cytokine profile of crossbred cattle of Assam for the prediction of successful pregnancy and lactation and to manipulate the level of cytokine by using certain cytokine stimulants/ regulators/plant leaves. Selected animals was divided into five groups keeping six in each in accordance with the feeding of different cytokine stimulator/regulator. All cows included for the experiment was in similar managemental practices and were offered same type of ration throughout the study. Group I was control without any treatment. Group II was treated with neem leaves (Azadirachta indica) at the rate of 3gm per kg body weight, Group III was fed with bogori leaves (Zizyphus mauritiana) at the rate of 300 mg per kg body weight orally. Group IV and group V were fed Restobal, a herbal immunomodulator at the dose rate of 50 ml orally and bamboo leaves (Bambusa bambos) at the rate of 3gm per kg body weight daily from the last three months of pregnancy to three months after parturition. Inflammatory indices, liver specific biomarkers, energy metabolism indices, protein metabolism index, oxidative stress response, different serum minerals etc. and different hematological parameters were estimated. Estimation of different cytokine profiles was carried out namely TNF-α, IL-1β. IL-6, IL-10 in serum. And record of daily milk yield and growth performance of calves with different cytokine stimulator/regulator/plant leaves. A non significant rise in the level of RBC count was seen in group IV (Restobal treated) in the late gestation than that of the control group. All the groups showed a similar trend of increase in RBC count till late gestation and then a decline on the day of calving and thereafter. Overall mean showed that there was a higher level of haemoglobin in group II followed by group IV, group III and II. All the groups showed an apparent rise in the level of haemoglobin than that of control group. Overall mean of WBC count in Group II and III were found to be higher than that of the control group but the increase was not significant. No significant difference was observed between the control and treated groups. A higher level of protein was found in group II and group V as compared to the control group which was non significant. A significantly higher (P < 0.0001) level of protein was recorded 30 days after parturition than that of the day of calving. Group II (Neem fed) and group IV (Restobal treated) had a significantly higher (P <0.001) level of globulin as compared to that of the control group. A non significant reduction in the level of creatinine in group IV after parturition . Almost all the groups showed a higher level of albumin during the late gestation which showed a slight decline around calving and on the day of calving and then there was gradual increase in the level of albumin after around one month post parturition. Lower level of total bilirubin was recorded in the experimental groups as compared to the control group. The pre partum values of the paraoxonase enzyme was found to be higher than the level at calving and at postpartum .No significant difference in the overall mean between the experimental groups and that of the control group was recorded. The experimental groups showed a higher level of GGT enzyme activity upto the day of calving and then thereafter showed a reduced level at postpartum . There was a non significant decrease in the level of ALP in the experimental groups as compared to that of the control group. Significant difference (P<0.01) was found in between group I and III and in between group I and V. (P<0.05).The groups showed a higher level of SGOT at parturition which declined after calving. There was non significant decrease in the level of SGOT in the neem treated group.Lower level of NEFA was found in group III and IV as compared to the control group. The groups showed a higher level of NEFA during early lactation. Group I, II and III showed a lower level of beta hydroxyl butyric acid as compared to the control group. However, an increase in the level of BHBA was recorded in group V than that of the control group. Group II (neem leaves fed) and III (Bogori leaves fed) showed a higher level of SOD as compared to the control group. Antioxidant enzyme activity was found to be higher at prepartum, then showed a reduction in the level at calving and thereafter there was gradual increase in the activity of enzyme at early lactation. Enhanced antioxidant activity was observed in the treated groups as compared to the control group. Dietary Neem leaf meal did not significantly (P>0.05) affect the calcium, sodium and potassium levels but significantly (P<0.05) decreased the phosphate level while significantly (P<0.05) and steadily, increasing the chloride level as its level increased. Group II that was fed with neem leaves showed a high preparum level of TNF -α and the peak level (1248.33±226.53 pg/ml) on the day of parturition. The increased level of tumor necrosis factor alpha (TNF-alpha) might be due to the cytokine and immune system stimulating property of neem plants. Significantly lower level of IL-6 was recorded in group III (Bogori leaves fed) and group V (bamboo leaves fed) than that of control. IL-6 level increased non significantly in the Restobal treated and Neem treated group. The level of IL-10 showed an increased level up to the day calving and then a decline was recorded approximately 15 days after calving. Significantly higher level of IL- 1β was recorded in group II (neem treated) (P<0.001) and in group V (P<0.01) than that of the control group. The level of (IL-1β) showed a high level during the advanced stage of pregnancy.Overall mean level of milk yield (litres) was found to be 9.25± 0.115 in group 1, 9.7 ± 0.137 in group II, 9.3±0.139 in group III, 9.6± 0.133 and 8.63±0.085 in group IV and V respectively.All the experimental animals differed significantly from that of the control group, except group IV .The highest average milk yield was recorded in group II (neem fed ) followed by group IV (Restobal treated) and then group III (Bogori leaves ). A lower level of milk yield was found in group V (bamboo leaves fed). The highest body weight was recorded in group IV ( Restobal treated ) followed by group II (neem fed) and then group III and V. supplementation of neem leaves , Restobal, Bogori leaves has a positive effect on the performance of the animals. The cytokine regulators/plant leaves used in the study can be given to the animals during pregnancy which might play an important role in enhancing immunity, maintenance of pregnancy and a successful lactation period.ThesisItem Open Access EXPLORING SEASON RELATED BIOCHEMICAL INDICES IN CROSS-BRED AND LOCAL CATTLE(Assam Agricultural University, Khanapara, Guwahati, 2016-07) Chetia, Mayuri; Sarma, SatyaThe present experiment was conducted to study the significant changes of various physiological, biochemical, hormonal and HSP gene expression profiles in Cross-bred (HF X Jersey) cattle under the agro-climatic condition of Assam and Local cattle of Assam were included in the experiment as reference experimental animals. The experiment was aimed at determining the status of some of the important season related thermal stress biomarkers with an idea to get some clues about the heat tolerance and the adaptability status of the Cross-bred cattles compared to the Local breeds of Assam. The experiment comprised of a total twelve numbers of healthy cattle of ages between 2-3 years, which were divided into two groups each containing six numbers of Cross-bred and six numbers of Assam Local cattle being reared under the semi-intensive system in Instructional Livestock Farm (Cattle) of College of Veterinary Science Khanapara, Assam Agricultural University and Experimental Animal Shed, Department of Veterinary Physiology, College of Veterinary Science Khanapara, Assam Agricultural University. The animals were kept under standard feeding and management. The monthly ambient temperature (°C), relative humidity (%) were recorded from the automatic weather station and temperature humidity index (THI) were calculated during the summer and winter season using standard formula. The physiological parameters such as the rectal temperature, respiration rate and pulse rate were recorded daily twice for the two seasons. The different biochemical parameters such as serum total protein, serum glucose, serum electrolytes - Na+, K+, Cl-, Ca2+, P and Mg2+, erythrocytic oxidative enzymes - superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), serum Lactate Dehydrogenase (LDH), serum hormonal parameters namely, Cortisol, T3 and T4 and mRNA expression profile of HSP 40, HSP 60, HSP 70, HSP 90 genes were studied in both the breeds of cattle. The Temperature Humidity Index (THI) during the study period was found to be indicative of thermal stress to the experimental animals during the summer season compared to winter season. The different physiological parameters viz. rectal temperature, respiration rate and pulse rate were found to increase in both the breeds during summer compared to winter season. All the physiological parameters were increased in the evening hours in both the cattle types compared to the morning hours. The serum total protein and serum glucose concentrations were significantly lower (P<0.001) during the summer season as compared to the winter season in both cross-bred and Assam local cattle. Similarly, the serum electrolytes, such as Na+, K+, Cl-, Ca2+, P and Mg2+ concentrations were significantly (P<0.001) lower in the summer as compared to winter season in both cross-bred and local cattle types of Assam. A significant difference was observed in serum total protein, serum glucose, serum Na+, K+, Cl-, Ca2+ and Mg2+ concentration between the breeds.The activity of erythrocytes SOD, GSH-Px, CAT and serum LDH was found to be significantly (P<0.001) higher in the summer season in both the breeds compared to the winter season. In addition, a significant (P<0.001) difference on the concentration of different erythrocytic and serum enzymes was observed between the two breeds. The concentrations of Cortisol, T3 and T4 were found to be significantly (P<0.001) different between the seasons in both cross-bred and local cattle of Assam. The cortisol level significantly increased during the summer season in both the breeds compared to the winter season while T3 and T4 level significantly decreased in the summer season in both the cattle types. A significant (P<0.001) difference in the hormonal concentration of Cortisol, T3 and T4 was found between the breeds. The present experiment shows a wide variation in the expression of different HSP genes during summer and winter season. The mRNA expression of HSP 40, 70 and 90 genes were significantly higher (P<0.01) during summer season as compared to winter season. The mRNA expression of HSP40, HSP 60, HSP70 and HSP90 was significantly differed between the two breeds of cattle types. HSP 70 has been found to be the potential biomarkers of heat stress in both the breeds of cattle and based on the relative expression, the ranking of genes from higher to lower abundance in cross-bred cattle were in the following order: HSP70>HSP90>HSP40>HSP60 and in local cattle, it is HSP70>HSP60>HSP90>HSP40. All the physiological and biochemical alterations were observed in local and cross-bred animals in the different seasons. However, these alterations were quite more prominent in case of cross-bred cattle in comparison to the local cattle. Hence, the local cattle have better heat tolerance than those of the cross-bred cattle under the agro-climatic condition of Assam.ThesisItem Open Access EVALUATION OF EFFICACY OF WHOLE OUTER MEMBRANE PROTEIN OF Salmonella Typhimurium ADJUVANTED WITH CALCIUM PHOSPHATE NANOPARTICLES AS VACCINE CANDIDATE AGAINST SALMONELLOSIS IN CHICKEN(Assam Agricultural University, Khanapara, Guwahati, 2015-06) DEKA, SURAKSHA SUBEDI; Sarma, SatyaSalmonella sp can infect a broad array of animals, causing diseases ranging from gastroenteritis to life threatening systemic infections. Substantial economic loss is manifested through mortality and poor growth of infected birds. Salmonella enterica serovar Typhimurium is the most frequently isolated serovar causing global food-borne outbreaks and chicken alone accounts for 10.4% of the total isolates. The control of salmonellosis can be accomplished either by vaccination or medication. Antibiotic resistance and issue of antibiotic residue is a major hurdle in medication. Thus control through immunization is the most efficient and economic method. The present study was conducted to study the efficacy of whole outer membrane protein adjuvanted with calcium phosphate nanoparticles (CAP-OMP) as vaccine candidate against salmonellosis in chicken. The study was carried out in Kamrupa birds which are dual purpose breed. The OMP was extracted from Salmonella Typhimurium (MTCC - 98) strain and confirmed by SDS-PAGE. CAP-OMP vaccine was then synthesized by method that is based on co-precipitation of calcium phosphate and OMP. The amount of protein entrapped in the complex was determined and the formulation was used to immunise the chicks on 14th day of life followed by booster dose after two weeks. The humoral immune response of the target vaccine was compared with aluminium hydroxide adjuvanted OMP and unadjuvanted OMP by indirect ELISA. Blood was collected from all the birds at 0 before vaccination and days 7, 14, 21, 28, 35 45 and 60 after booster vaccination. Some of the birds from each group were challenged on 60th day with either S. Typhimurium or with S. Gallinarum. The humoral response of the unchallenged birds was studied from 60th to 90th day. The CAP-OMP vaccine was able to elicit significantly higher antibody titres compared to other two groups up to 60th day post booster vaccination. However antibody titre level fell thereafter in all the groups. Mortality was not observed in any of the challenged birds but some clinical symptoms were exhibited by the control group. The challenged birds were slaughtered after one week and total CFU of either S. Typhimurium or S. Gallinarum per gram of liver was determined. It was observed that none of the vaccines gave total protection against challenge organisms but the Salmonella isolates recovered from the birds immunized with target vaccine was significantly lower than that of control group. The other two formulations viz; aluminium hydroxide –OMP vaccine and unadjuvanted OMP vaccine also decreased the bacterial count in liver tissue. Some important serum biochemical parameters were also studied from the vaccinated birds and it was found that the total protein and globulin content was significantly higher in CAP-OMP vaccinated group than the control groups. Creatine kinase level was significantly higher during first 21 days post booster vaccination birds given Aluminium hydroxide -OMP indicating some tissue damage at the site of injection caused by Aluminium hydroxide. Other parameters like alkaline phosphatase and creatinine showed non-significant fluctuations throughout the experiment. From this study, it can be concluded that CAP-OMP (S.Typhimurium) vaccine can be an effective vaccine candidate against salmonellosis.
