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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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20212
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Subject: Animal Reproduction, Gynaecology and Obstetrics × End date: 2021 × Start date: 2021 × Type: Thesis ×
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    ThesisItemOpen Access
    Effect of cryopreservation on semen biochemical parameters including lipid profile in beetal and Assam hill goat
    (2021-01) Das, Prasanta Kumar; Sinha, Sudip
    A total of 72 ejaculates comprising six ejaculates from each of 12 bucks (six Beetal and six Assam Hill Goat) were used to study the effect of cryopreservation on the physical and biochemical characteristics of semen including sperm ultrastructure. The physical characteristics of fresh semen viz., ejaculate volume, mass activity, initial sperm motility, live sperm, sperm concentration, cold shock resistance index, acrosomal integrity, HOSTreacted sperm and sperm abnormalities were studied by conventional methods. The biochemical characteristics viz., sodium, potassium, calcium, total cholesterol, total lipid, AST, ALT and lipid profile were studied in seminal plasma, and total cholesterol, total lipid and lipid profile were also studied in spermatozoa of fresh and frozen semen. Semen was extended in Optixcell extender and frozen in 0.25 ml straws using liquid nitrogen vapour and stored in liquid nitrogen. Each sample was evaluated on the following day for sperm motility, live sperm, acrosomal integrity and HOST-reacted sperm. The biochemical characteristics studied after freezing in extracellular medium were the same as in case of seminal plasma. Sperm ultrastructure was studied both in fresh and frozenthawed spermatozoa. Physical and biochemical characteristics of semen of Beetal and Assam Hill Goats were within normal range and differed significantly between breed for ejaculate volume (P<0.001), sperm concentration (P<0.001), cold shock resistance (P<0.05), Host-reacted sperm (P<0.01), sodium (P<0.001), potassium (P<0.001) and total lipid (P<0.001). Postthaw sperm parameters viz. sperm motility, live sperm, acrosomal integrity and HOST– reacted sperm, and content of biochemical constituents viz. sodium, potassium, calcium, total cholesterol and total lipid decreased after freezing in semen of both the breeds, while AST and ALT increased. Thirteen and fourteen fatty acids were identified in the seminal plasma of Beetal and Assam Hill Goat bucks, respectively. Pentanedioic acid was identified only in seminal plasma of Assam Hill Goat. In the extracellular medium of frozen Beetal semen, all the fatty acids of seminal plasma of fresh semen were present, however, in Assam Hill Goat bucks both Dodecadienoic acid and Pentanedioic acid which were present in seminal plasma of fresh semen were found to be absent in frozen semen. Breed variation was observed in respect of Pentanedioic acid of fresh semen. Freezing had significant effect in alteration of membrane stabilizing fatty acids viz. Erucic acid, carboceric acid, linoleic acid, tricontanoic acid, arachidic acid, eicosadienoic acid, margaric acid, mentanic acid, tetradecadienoate, tridecanoic acid, heneicosylic acid, cerotic acid and tricosanoic acid. Major ultrastructural changes in spermatozoa after freezing were separating and ruptured plasma membrane, fusion of plasma membrane with outer acrosomal membrane, swelling of acrosome and loss of acrosomal content. Based on the physical and biochemical parameters studied it could be concluded that cryopreservation of goat semen has deleterious effects on fatty acid profile of seminal plasma and sperm plasma membrane, and also on sperm ultrastructure in both Beetal and Assam Hill Goat. The proportion of loss of plasma membrane fatty acids after freezing was lower in Assam Hill Goat as compared to that of Beetal goat. Post-thaw semen quality in Assam Hill Goat was superior to that of Beetal goat.
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    ThesisItemOpen Access
    CHARACTERIZATION OF MEMBRANE PROTEINS IN FRESH AND FROZEN SPERMATOZOA IN ASSAM HILL GOAT
    (College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2021-04) Dutta, Mitali; Sinha, Sudip
    Thirty six pooled ejaculates from nine Assam Hill Goat bucks aged 2 to 2.5 years collected by artificial vagina method were used to study the fresh and frozen semen characteristics, characterization of seminal plasma proteins, characterization of sperm membrane proteins in fresh and frozen semen and to study the effect of supplementation of three membrane stabilizers, each at two different concentrations viz. 50 and 80 mM sucrose, 50 and 100 mM trehalose, and 100 and 150ng/ml IGF-1 to tris-citric acid fructose egg yolk glycerol extender (TCFEYG) on post-thaw semen characteristics and on sperm membrane proteins of frozen semen. Characterization of two fertility related membrane proteins viz. ADAM1 and ADAM2 were also done in fresh and frozen spermatozoa of Assam Hill goat by western blotting and immunolocalization using anti ADAM1 and anti ADAM2 antibodies raised in rabbit respectively. The mean per cent progressive sperm motility, HOST-reacted sperm and intact acrosome was significantly (p<0.01) higher in fresh semen than in semen frozen in different extenders. The mean per cent post-thaw progressive motility, HOST-reacted sperm and intact acrosome differed significantly (p<0.01) between the different extenders. However, no significant difference was observed in the mean per cent HOST-reacted sperm between TCFEYG supplemented with 100mM trehalose and TCFEYG supplemented with 100ng/ml IGF-1 and no significant difference was observed in the mean per cent intact acrosome between TCFEYG supplemented with 100mM trehalose and TCFEYG supplemented with 100ng/ml IGF-1 and between TCFEYG supplemented with 50mM sucrose and TCFEYG supplemented with 50mM trehalose. The mean per cent post-thaw progressive sperm motility, HOST-reacted sperm and intact acrosome in frozen semen was found to be the highest in TCFEYG supplemented with 150ng/ml IGF-1. SDS- PAGE of seminal plasma and sperm membrane extract of fresh semen revealed the presence of 20 and 24 protein bands respectively with molecular weights ranging from10 kDa to 240 kDa. The SDS-PAGE electrophoretogram of sperm membrane proteins of semen frozen using TCFEYG and TCFEYG supplemented with 50mM sucrose (TCFEYG + 50mM S) and, 80mM sucrose (TCFEYG + 80mM S) revealed 21 protein bands with molecular weights ranging from 10 kDa to 240 kDa. The 21 protein bands were same with that observed in the sperm membrane of fresh spermatozoa, except three protein bands. These three proteins of molecular weights 23 kDa (~Phosphatidyl-ethanolamine-binding protein), 29 kDa (~Proacrosin binding protein) and 42 kDa (~tyrosine- phosphorylated SPACA1) were absent in TCFEYG and TCFEYG supplemented with 50mM, and 80mM sucrose. The SDS-PAGE electrophoretogram of sperm membrane proteins of semen frozen using TCFEYG supplemented with 50mM trehalose (TCFEYG + 50mM T), and 100mM trehalose (TCFEYG + 100mM T) revealed 22 protein bands with molecular weights ranging from 10 kDa to 240 kDa. The 22 protein bands were same with that observed in the sperm membrane of fresh spermatozoa, except two protein bands. These two proteins of molecular weights 29 kDa (~Proacrosin binding protein) and 42 kDa (~tyrosine- phosphorylated SPACA1) were absent in TCFEYG supplemented with 50mM, and 100mM trehalose. The supplementation of trehalose to the basic TCFEYG extender at 50mM and 100mM concentrations, however, had a protective effect on the sperm membrane protein of 23kDa when compared to the basic TCFEYG extender and TCFEYG supplemented with 50 and 80mM sucrose. The SDS-PAGE electrophoretogram of sperm membrane proteins of semen frozen using TCFEYG supplemented with 100ng/ml IGF-1 (TCFEYG + 100ng/ml IGF-1), and 150ng/ml IGF-1 (TCFEYG + 150ng/ml IGF-1) revealed 21 protein bands with molecular weights ranging from 10 kDa to 240 kDa. The 21 protein bands were same with that observed in the sperm membrane of fresh spermatozoa, except three protein bands. These three proteins of molecular weights 29 kDa (~Proacrosin binding protein), 130 kDa (~ lipoprotein binding protein) and 240 kDa (~golgi-associated retrograde protein) were absent in TCFEYG supplemented with 100ng/ml, and 150ng/ml IGF-1. Supplementation of the basic tris extender with IGF1 at concentrations of 100ng/ml and 150ng/ml resulted in protection of sperm membrane proteins of molecular weights 23kDa (~Phosphatidyl-ethanolamine-binding protein) and 42 kDa (~tyrosine- phosphorylated SPACA1) during the freeze-thaw process when compared to the basic tris extender and tris extender supplemented with sucrose. The 23 kDa protein was however, also found to be protected in the tris extender supplemented with trehalose. ADAM1 was detected as three bands of ~ 25kDa, 66kDa and~ 90kDa in the sperm membrane extract of fresh sperm. However, in the sperm membrane extract of semen frozen using TCFEYG, TCFEYG + 80mM S, TCFEYG + 50mM S, TCFEYG + 50mM T, TCFEYG + 100mM T, TCFEYG + 100ng/ml IGF-1 and TCFEYG + 150ng/ml IGF-1 extenders it was detected as ~25kDa, 66kDa and ~90kDa; ~25kDa, 66kDa and ~90kDa; ~25kDa, ~49kDa,~66kDa, ~90kDa and ~110kDa; ~25kDa,66kDa and ~90kDa; ~25kDa, 66kDa and ~90kDa; ~25kDa, ~49kDa,~66kDa, ~90kDa and ~110kDa ; and ~25kDa, ~49kDa,~66kDa, ~90kDa and ~110kDa respectively. In the present study, freeze-thaw process and supplementation of tris extender used for freezing of semen with membrane stabilizers such as sucrose, trehalose and IGF-1 has been found to result in certain variations in the molecular weight of ADAM1. However, reactive protein bands of 25kDa, 66kDa and 90kDa were found to be consistently present in the fresh sperm as well as sperm frozen in different extenders. ADAM2 was detected as two bands of ~ 80kDa and~ 130kDa in the sperm membrane extract of fresh sperm. However, in the sperm membrane extract of sperm frozen using TCFEYG, TCFEYG + 80mM S, TCFEYG + 50mM S, TCFEYG + 50mM T, TCFEYG + 100mM T, TCFEYG + 100ng/ml IGF-1 and TCFEYG + 150ng/ml IGF-1 extenders it was detected as ~70kDa, ~80kDa, ~100kDa and ~130kDa; ~70kDa, 80 kDa and ~130kDa; ~70kDa, ~80kDa,~90kDa, ~100kDa and ~130kDa; ~70kDa, ~80kDa, and ~100kDa; ~70kDa, ~80kDa, and ~100kDa; ~70kDa, ~80kDa, and ~100kDa; and ~70kDa, ~80kDa, and ~100kDa respectively. In the present study, freeze-thaw process and supplementation of tris extender used for freezing of semen with membrane stabilizers such as sucrose, trehalose and IGF1 has been found to result in certain variations in the molecular weight of ADAM2. However, a protein band of 80 kDa was found to be consistently present in the fresh sperm as well as sperm frozen in different extenders. Immunolocalization of the ADAM1 and ADAM2 proteins revealed the presence of the proteins in the acrosomal region of sperm cells in both fresh and frozen semen. Present study revealed no change in the localization of ADAM1 and ADAM2 post freezing thereby indicating that there is no effect of freezing on the distribution of these two proteins. It was concluded that cryopreservation of Assam Hill Goat semen resulted in alterations in sperm membrane proteins, however, supplementation of membrane stabilizers exerted protective effects. Based on post-thaw semen characteristics and study on membrane proteins it was found that IGF-1 @ 150ng/ml was superior to other membrane stabilizers in maintaining post-thaw semen quality.