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ThesisItem Open Access Biotic agents for the management of American serpentine leaf miner, Liriomyza trifolii(Burgess) (Diptera:Agromyzidae)(Department of agricultural entomology, College of horticulture, Vellanikkara, 2014) Jyothi Sara, Jacob; KAU; Maicykutty P, MathewA study on “Biotic agents for the management of American serpentine leaf miner, Liriomyza trifolii (Burgess) (Diptera: Agromyzidae)” was carried out at the Department of Agricultural Entomology, College of Horticulture, K.A.U., Vellanikkara during 2011-2013 with the objectives of collection and identification of indigenous natural enemies and to assess the pathogenicity of the entomopathogens to explore the feasibility of utilizing them for its management. Surveys were conducted in the vegetable fields for the collection and identification of natural enemies associated with L. trifolii in three districts, namely, Thrissur, Ernakulam and Kottayam from January to March, 2011. The surveys revealed the occurrence of nine species of hymenopteran parasitoids. The per cent parasitism varied from 10.96 to 58.99 per cent among the crops surveyed. Three species of eulophids, namely, Cirrospilus acadius Narendran, C. brevicorpus Shafee & Rizvi and Aprostocetus sp. as well as the braconid, Toxares sp. are new reports for India. Among the parasitoids, Closterocerus spp. were the dominant group followed by Chrysonotomyia sp. All parasitoids were solitary, larval endoparasitoids except Toxares sp. which was larval-pupal in nature. One species each of small ants (Formicidae) and a dipteran fly (Dolichopodidae) were observed as predators on L. trifolii. In the study, no entomopathogens were observed from L. trifolii. Considering the level of pesticide consumption in vegetable crops that undermine the potential of insect parasitoids and also that no entomopathogens could be observed during the survey, it was decided to evaluate entomopathogenic nematodes (EPNs) as biocontrol agents against L. trifolii. Isolation of EPNs from 72 soil samples from Thrissur, Ernakulam and Kottayam districts yielded four isolates of Steinernema carpocapsae. Bioefficacy studies carried out on these four isolates along with Steinernema bicornutum and Heterorhabditis indica showed that S. carpocapsae Isolate - 1 had the lowest LC 50 , LC 90 and LT values indicating their higher effectiveness against the maggots of the pest. 50 Pot culture study conducted to compare the potential of S. carpocapsae Isolate - 1 with other treatments showed that azadirachtin 1 EC at 0.005% was the most effective causing 84.51 per cent mortality to the maggots of L. trifolii. This was followed by the foliar application of H. indica at 32 infective juveniles (IJs)/ maggot which caused 18.98 per cent mortality. Application of Beauveria bassiana at 1×10 7 spores/ ml was not effective. In the field evaluation, fipronil 5 SC at 0.002% was found to be the most effective treatment for controlling L. trifolii followed by azadirachtin 1 EC at 0.005%. Compatibility of the IJs of the S. carpocapsae Isolate - 1, S. bicornutum and H. indica was studied with ten commonly used insecticides in the laboratory by direct exposure method. Chlorantraniliprole 18.5 SC at 0.005% was found to be the most compatible insecticide with S. carpocapsae isolate - 1 causing only 0.17 per cent mortality to IJs at 72 hours after treatment (HAT). Quinalphos 25 EC at 0.05% and chlorpyriphos20 EC at 0.05% were highly incompatible, causing 96.17 and 92.87 per cent mortality of the nematodes. Dimethoate 30 EC at 0.04% was the most compatible insecticide with S. bicornutum and caused only 0.60 per cent mortality at 72 HAT and was followed by azadirachtin 1 EC at 0.005% with 0.78 per cent mortality to the IJs. Quinalphos 25 EC at 0.05% caused 99.93 per cent mortality at 72 HAT. Heterorhabditis indica was compatible with all insecticides except quinalphos 25 EC at 0.05% which was moderately toxic resulting in 39.6 per cent mortality. The virulence, pathogenicity and multiplication of the survived IJs were not affected by the insecticide treatments. Parasitoids and EPNs were observed as potential candidates for the management of L. trifolii. Hence future studies on the bio-ecology and mass production of dominant parasitoids and standardization of methods to improve the efficacy of EPNs are suggested for the successful control of L. trifolii in polyhouses as well as in the field.ThesisItem Open Access Cryopreservation of chethikoduveli (Plumbago rosea L.) and assessment of genetic fidelity of regenerated plantlets using molecular markers(Department of Plant Biotechnology, College of Agriculture, Vellayani, 2014) Anand, Vishnu Prakash; KAUInvestigations on “Cryopreservation of Chethikoduveli (Plumbago rosea L.) and assessment of genetic fidelity of regenerated plantlets using molecular markers” were carried out at the Department of Plant Biotechnology, College of Agriculture, Vellayani during 2011-2013. Plumbago rosea var. Agni plants were collected from AMPRS, Odakkali, Ernakulam and maintained at the Department of Plant Biotechnology, College of Agriculture, Vellayani as source of explant during the course of the study. The objectives of the present study was to standardise cryopreservation protocol by encapsulation dehydration technique for long term conservation of P. rosea and genetic fidelity assessment of plantlets recovered and regenerated from cryostorage using molecular markers. The project was carried out in two phases viz., in vitro regeneration and in vitro conservation by cryopreservation of P. rosea. In vitro regeneration protocol was optimised for P. rosea var. Agni. Various steps of in vitro regeneration viz., surface sterilization, axillary shoot proliferation, in vitro rooting and acclimatization and planting out has been standardised. For surface sterilizing, single nodal explants (3-4 cm long) were subjected to fungicide treatment with 0.1 per cent carbendazim 50 per cent W. P. (for 30 min) followed by aseptic sterilisation dip with absolute alcohol. Further, the explants were surface sterilised with 0.2 per cent mercuric chloride (for 5 min) which gave 100 per cent survival without any contamination. Enhanced release of axillary buds from single nodal explants, with maximum shoot proliferation (5.28 shoots/culture) was obtained in the medium, MS + BA 1.5 mg l-1 + IAA 1.0 mg l-1. The best response (10.67 roots/culture) of in vitro rooting of plantlets was obtained in the medium, MS + NAA 1.0 mg l-1. In vitro rooted plants gave a maximum survival rate of 76 per cent and 72 per cent, when planted out in potting media consisting of red soil and coir pith (3:1) and red soil and coir pith (2:1) supplemented with VAM respectively at 25 per cent shade. In cryopreservation studies, preconditioning treatment (sucrose 0.5 M for 7 days) recorded maximum shoot proliferation (2.67 shoots/culture) when nodal segments with single axillary bud were cultured on MS + BA 1.5 mg l-1 + IAA 1.0 mg l-1 medium. Among different encapsulation treatments, maximum shoot proliferation of (2.31 shoots/culture) was obtained in beads formed with sodium alginate 2.5 per cent and calcium chloride 100 mM, when cultured on the medium, MS + BA 1.5 mg l-1 + IAA 1.0 mg l-1. Pre-culture medium supplemented with sucrose 0.5 M for 3days gave maximum shoot proliferation (3.44 shoots/culture) when cultured on the medium, MS + BA 1.5 mg l-1 + IAA 1.0 mg l-1. A desiccation duration of 5 h at 18.13 per cent moisture level was found to be most effective giving 66.67 per cent survival and 62.50 per cent regeneration on thawing and culturing on the recovery medium MS + BA 1.5 mg l-1 + IAA 1.0 mg l-1. The beads when stored in liquid nitrogen for different duration and cultured on recovery medium did not show any significant variation with respect to survival per cent. RAPD markers were tried to study the genetic fidelity of the regenerated plantlets from encapsulated and cryopreserved axillary buds. Six primers were screened and RAPD banding patterns of the cryoregenerated plantlets and control plants were compared. Polymorphism was not found with any of the primers tested. RAPD profiles of cryoregenerated plantlets were identical to those of the control. The in vitro regeneration protocol optimized included surface sterilization of single node cuttings with 0.2 per cent HgCl2 for 5 min, axillary shoot proliferation in MS medium supplemented with BA 1.5 mg l-1 and IAA 1.0 mg l-1, in vitro rooting in MS medium supplemented with NAA 1.0 mg l-1 and planting out in potting medium, red soil and coir pith (3:1). The protocol for encapsulation dehydration technique of cryopreservation was standardised for the axillary buds of P. rosea with preconditioning in semi solid MS medium supplemented with sucrose 0.5 M for 7 days, encapsulation using sodium alginate 2.5 per cent and calcium chloride 100 mM followed by pre-culture in liquid MS supplemented with sucrose 0.5 M for 3 days and 5 h dehydration (MC 18.13 %), rapid freezing in LN for at least 2 h and recovery in the medium MS + BA 1.5 mg l-1 + IAA 1.0 mg l-1. The cryopreservation protocol using encapsulation-dehydration technique standardised could be utilised for long-term conservation of P. rosea.ThesisItem Open Access Bioactivity of carotenoids from shrimp shell waste(Department of Processing Technology,College of Fisheries,Panangad, 2010) Sindhu, S; KAU; Sherief, P MShrimp processing waste is the single largest industrial waste in the country causing diverse environmental problems. A study was carried out to assess the extractability of astaxanthin from shrimp waste in different organic solvents and vegetable oils. Extraction was tried using wet and dried waste, with and without deproteinisation. Waste was subjected to deproteinisation using alkali and enzyme (pancreatin). The different solvent systems tried were ether:acetone:water (15:75:10 v/v/v), acetone, hexane:isopropanol (3:2 v/v) and 90% acetone v/v. Astaxanthin in the extract was quantified by measuring the OD at 470 nm in hexane. Extraction was also done using vegetable oils viz. coconut oil, soybean oil and sunflower oil. Quantification of astaxanthin in pigmented oil was done by measuring the absorbance at 485 nm using 2155 as extinction coefficient. Astaxanthin yields from deproteinised samples were significantly lower than those from non deproteinised samples. The highest astaxanthin yield of 87.14 ± 4.55μg/g was obtained with non deproteinised wet waste extracted using acetone. The astaxanthin yield was significantly lower when oil was used as the extraction medium. Of the three oils coconut oil gave the highest yield. The results showed that acetone is the best solvent for extracting astaxanthin from shrimp shell waste in wet condition. The astaxanthin content in Aristeus alcocki shell waste is double that of Pandalus borealis shell waste, which is currently used as the commercial source of astaxanthin. The deep sea species Aristeus alcocki can thus be considered as a better source of astaxanthin for commercial exploitation than Pandalus borealis. TLC analysis of the shell waste extract showed that it contains free astaxanthin, astaxanthin monoester and astaxanthin diester in the ratio 1:1:2. GLC identification of the fatty acids esterified with astaxanthin revealed that saturated fatty acids, MUFA and PUFA are in the ratio 5:3:2 in monoester, whereas in diester they are in the ratio 4:3:3. The main fatty acids in monoester and diesters are palmitic acid, oleic acid, stearic acid and PUFAs: DHA and EPA. The in vitro antioxidant activity of the astaxanthin extract showed significant hydroxyl radical scavenging activity, superoxide anion scavenging activity and inhibition of lipid peroxidation. The IC50 values obtained were 56.43 ± 1.06 ng/ml, 27.91 ± 0.54 ng/ml and 26.54 ± 0.42 ng/ml, respectively. The antioxidant activity of astaxanthin from Aristeus alcocki was obtained at nanogram levels. This powerful antioxidant function may be due to the unique molecular structure of astaxanthin and synergistic effect of astaxanthin and PUFAs present in the astaxanthin monoester and diester fractions. The astaxanthin extract from shrimp shell waste significantly reduced carageenan induced paw edema in mice, percentage inhibition being 47.83 and 67.11 percent at astaxanthin concentrations of 0.5 mg/kg body weight and 1.0 mg/kg body weight, respectively. The inhibition of inflammation at 1.0mg/kg body weight was greater than that produced by the standard reference drug diclofenac. Cardioprotective effect of astaxanthin was examined in isoproterenol induced myocardial infarction in rats. Levels of diagnostic marker enzymes, LDH, CPK, GOT, GPT, CK, CK-MB in plasma, lipid peroxides, ascorbic acid, reduced glutathione and the activities of glutathione-dependent antioxidant enzymes GPx, GR, GST and antiperoxidate enzymes CAT, SOD and the membrane bound enzyme Na+ - K+ ATPase in the heart tissues of experimental groups of rats were determined. The prior administration of astaxanthin @ 10mg/kg feed for 45 days significantly prevented the isoproterenol-induced elevation in the levels of diagnostic marker enzymes in plasma, induction of lipid peroxidation and alterations in the level of reduced glutathione and in the activities of glutathione dependent antioxidant enzymes and antiperoxidative enzymes of experimental rats. Feeding astaxanthin caused a decrease in the inhibition of Na+ - K+ ATPase activity against isoproterenol induced myocardial infarction. The powerful cardioprotective effect of astaxanthin can be attributed to the multiple independent mechanisms viz. antioxidant effects, singlet oxygen quenching ability and inhibition of lipid peroxidation of membranes, increased functional gap junctional intercellular communication, anti-inflammatory effects etc. Immunostimulatory action of astaxanthin extract was evaluated in experimental mice. Astaxanthin administration was found to enhance the proliferation of spleen cells and bone marrow cells. Esterase activity was found to be enhanced in bone marrow cells indicating increased maturation of cells of lymophoid linkage. Astaxanthin also enhanced number of antibody forming cells and circulating antibody titre. Thus astaxanthin exhibits strong immunomodulating properties. A significant reduction in the viability of ascites tumour cells DLA in vitro was noted in the current study. The % viability was reduced to 4.34 % at a concentration of 15μg astaxanthin/ml. The cytotoxic action of astaxanthin against DLA may be through induction of apoptosis or through a different pathway. Antitumour activity of astaxanthin was studied by ascite and solid tumour models in mice. An increase in life span of about 67 % was noted in DLA bearing mice administered with astaxanthin at 5 mg/kg body weight. The tumour volume and tumour weight were significantly lower in mice injected with 5 mg/kg body weight astaxanthin. In vitro studies revealed that astaxanthin from shrimp shell waste of Aristeus alcocki inhibited the proliferation of cervical cancer cells HeLa in a dose dependent manner.ThesisItem Open Access Formulation of low fat beef burger with fat replacers(Department of Livestock Products Technology, College of Veterinary and Animal Sciences,Mannuthy, 2010) Govande Premanand, Laxmanrao; KAU; George Oommen, THealth conscious meat consumers prefer low fat meat products due to increasing incidents of high fat induced diseases. Manufacturing meat products with fat replacers (FR) enable to reduce fat and to alleviate the problems with the reduction of fat in products. Therefore, the present study was undertaken with the objectives of developing a palatable and economic formulary for low fat beef burger (LFBB) with carrageenan (CG), tapioca starch (TS), pregelatinised pork skin collagen (PSC) and their blends as FR and to assess its pH, cooking characteristics, proximate composition, nutritional value, textural and organoleptic qualities and shelf life under aerobic (AP) and vacuum packaging (VP) at 0-4oC and -20oC and its cost of production. Beef burgers (BB) are formulated at two different fat levels, viz., full fat (FF) 20 per cent and low fat (LF) 5 per cent as controls. Seven formulations of LFBB with 5 per cent fat are prepared with 0.5 per cent CG, 1.5 per cent TS, 2 per cent PSC and their blends, viz., CG-TS - 0.5% CG & 1.5% TS; CG-PSC - 0.5% CG & 2.0% PSC; TS-PSC - 1.5% TS & 2.0% PSC; CG-TS-PSC - 0.5% CG, 1.5% TS & 2.0% PSC as FR. BB are prepared as per the formularies with minced lean beef trimmings, tallow, salt, spices and condiments, rusk, ice flakes and FR. They are packaged aerobically in HDPE and in vacuum in polyethylene-polyamide (PEPA) pouches. pH, cook yield (CY), cook loss (CL), fat retention percentage (FRP), moisture retention percentage (MRP), dimensional shrinkage (DS), water holding capacity (WHC), Warner-Bratzler Shear Force (WBSF), Hunter L*, a*, b* colour values, proximate and mineral composition and nutritional value, purge loss (PL), Thiobarbituric Acid Reactive Substances (TBARS) value and sensory qualities are assessed on d 0, 10, 20 and 30 of storage at 0-4oC and -20oC or till spoilage, whichever is earlier. Six trials of the experiment were conducted. Cooking reduced the acidity of all the burgers. By the addition of FR a significantly (P< 0.05) very low acid cooked LFBB could be prepared. CY of burgers with CG-TS-PSC was significantly (P< 0.05) the highest with 85.84 per cent. LFBB with blends of FR significantly (P< 0.05) increased CY and correspondingly reduced CL. The DS in LFBB with CG-TS-PSC was significantly (P< 0.05) the lowest with 13.21 per cent. Addition of blends of FR holds water and fat in LFBB and reduces DS during cooking. FRP and MRP in CG-TS-PSC formulation was significantly (P< 0.05) the highest with 97.66 and 74.36 per cent, respectively due to blends of CG, TS and PSC. The WHC of LFBB with CG-TS-PSC was 95.36 per cent and WBSF value 5.30 N comparable to FF and the burgers were significantly (P< 0.05) most succulent, juicy and tender with the addition of blends of FR compared to tougher BB without FR. According to Hunter L*, a*, b* values, LFBB with blends of FR, especially CG-TS-PSC was lighter, less reddish (more bluish) and less yellowish (more greenish) and comparable to FF burger. Fat content in the beef trimmings and PSC were < 1.76 per cent. Cooking significantly (P< 0.05) reduced moisture content with a corresponding increase in the protein, fat, carbohydrate and ash. The percentage total calorific value of LFBB ranged from 6.36 to 7.18 of the Recommended Dietary Allowance (RDA). The contribution of fat to RDA of calorific value was from 2.22 to 2.42 per cent only, which was below the recommended 30 per cent. More than one third of the daily requirement of protein is obtained from 100g of LFBB. LFBB with FR are good sources of Na, K and P but not of Ca. Blends of FR in LFBB, especially CG-TS-PSC, were more efficient in significantly (P< 0.05) reducing PL and TBARS value on storage at 0-4oC for 10 days and at -20oC for 30 days in AP and VP. TBARS values were lower than the acceptable range of 1mg malonaldehyde/kg for oxidative rancidity. The low fat content and the presence of onion containing antioxidants in the formulary would have synergistically acted with CG in reducing the TBARS. On sensory evaluation on zero day, the LFBB with CG-TS-PSC scored significantly higher (P< 0.05) values of 7.00 and above for very good appearance and colour, very intense flavour, very desirable texture, juiciness, practically nil mouth coating and very acceptable overall acceptability similar to FF burger. But saltiness was very desirable than in FF. The LFBB with CG-TS-PSC in AP and VP retained all the sensory attributes and proximate composition even on storage. The very acceptable nature of CG-TS-PSC formulation might be due to the synergistic effect of fat replacers. The LFBB with 5 per cent fat and CG (0.5%), TS (1.5%), PSC (2%) and their blends as FR are developed economically with very acceptable overall acceptability, CY, nutritional quality, reduced PL and oxidative rancidity and shelf life up to 10 days at 0-4oC and 30 days at -20oC under AP and VP. The best LFBB with overall acceptability was CG-TS-PSC followed by CG-TS, CG-PSC, TS-PSC, PSC, CG and TS. Blends of FR are better than single FR, particularly CG-TS-PSC, as they increased CY, FRP, MRP, WHC, sensory attributes and decreased pH, CL, DS, WBSF, PL and TBARS. Further investigations with production of large quantities are required for calculation of cost of production at commercial scale.ThesisItem Open Access Genetic characterization, controlled breeding and development of transgenic varieties of puntius denisonii (day, 1865).(Department of Aquaculture, College of Fisheries, Panangad, 2010) Manoj, C K; KAU; Mohanakumaran Nair, CPuntius denisonii, a beautiful ornamental fish indigenous to the Western Ghats, which has been indiscriminately exploited from the different rivers of Kerala has been recently declared to be vulnerable by the IUCN. The population structure and genetic diversity of P. denisonii has not yet been studied and documented. Many previous attempts to breed this fish in captivity have yielded negative results. The increasing demand for this fish to decorate aquariums worldwide could be satisfied only by developing controlled breeding techniques and larval rearing of its fry. In the present study, the present population structure of P. denisonii has been studied combining both phenotypic and genotypic techniques. Fishes were collected from Irrity, Chaliyar and Periyar rivers of Kerala. Truss network analysis was conducted and the size adjusted morphometric variables were subjected to Principal Component Analysis and Canonical Variance Analysis. Scatter diagram and Dendrogram was plotted using PCA and CVA loadings. The Irrity and the Chaliyar populations were grouped on the positive sector of the PC and CV component showing morphological similarities between the two populations while the Periyar population was placed in the negative sector of the component separated far from the other two. The PC scores were used to find out the variables showing maximum variation between fishes collected from different rivers. RAPD PCR was conducted after isolating DNA from the fins of different populations of P. denisonii. Universal random primers were screened and the primers that produced reproducible bands were selected. Popgene analysis of the binary data yielded the genetic structure of different populations of P. denisonii. Number and percentage of polymorphic loci, Nei's (1973) gene diversity, Shannon's Information index Lewontin (1972), Nei's Unbiased Measures of Genetic Identity and Genetic distance and Dendrogram Based Nei's (1978) Genetic distance using UPGMA --Modified from NEIGHBOR procedure of PHYLIP Version 3.5 were studied. The results obtained supports the truss analysis in that the Irrity and Chaliyar populations in Northern Kerala are genetically more similar while that of Periyar population in Central Kerala are distinct. P. denisonii was successfully induced bred under controlled conditions with synthetic hormone preparations Ovaprim and WOVA-FH. Stress during transport and handling was minimized and live feed was supplemented to enhance maturation of the broodstock. The whole developmental sequence starting from fertilized eggs to hatching was photographed and documented. It took 29-30 hours for the eggs to hatch at 280C. Rearing of fry was successfully accomplished under laboratory conditions. In an attempt to develop transgenic varieties of P. denisonii, pCMV-GFP was electroporated into newly fertilized eggs, maintained in hypoosmolar electroporation buffer. The electroporation parameters that yielded best results were 20V, 3 bursts at 1 second interval. Fin clips were taken from the transgenic individuals reared for a period of 6 weeks. Dot blot test was positive showing integration of the GFP gene in P. denisonii, eventhough expression was not detected under blue or UV light. The genetic and phenotypic data of P. denisonii populations in the present study will aid as a base line for formulating conservation procedures to protect the genetic diversity of wild ones. Stock identification studies are recommended for more concise information on each population. Moreover, the larval rearing and controlled breeding techniques along with the genetic diversity studies will help to design captive breeding programs and enhance the production of hatchery bred ones to meet increasing demand. Further research is recommended for generating transgenic lines with uniform GFP expression.ThesisItem Open Access Heterosis breeding in sesame (Sesamum indicum L.).(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2011) Gayathri, G; KAU; Dijee, BastianThe study entitled ‘Heterosis breeding in sesame (Sesamum indicum L.)’ was undertaken at the Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara. The objectives of the study were to collect and evaluate different genotypes of sesame for morphological traits and yield attributes, to identify useful parents producing heterotic crosses and developing hybrids in sesame. The study also intended to develop male sterile lines in sesame through interspecific hybridization with Sesamum malabaricum. Sesamum indicum and Sesamum malabaricum accessions were collected from Kerala and Tamil Nadu and evaluated for their morphological traits. Wide range of variation was noticed for characters like plant height, number of days to flowering and seed yield per plant which contributed maximum to genetic divergence. The genotypes studied were grouped into six clusters. High genotypic coefficient of variation (GCV) was recorded for number of capsules per plant, plant height, seed yield per plant and number of branches per plant. High heritability with high genetic advance as per cent of mean was recorded for number of days to flowering, plant height, number of branches per plant, number of capsules per plant and seed yield per plant. This indicates that the characters are governed by additive gene effects and selection for these traits will be effective. Association analysis revealed that seed yield per plant was correlated to plant height, number of capsules per plant and number of days to flowering. Path coefficient analysis indicated maximum positive direct effect by number of capsules per plant, capsule length, plant height and 1000 seed weight on seed yield per plant. In order to develop hybrids, fourteen parents were selected based on the per se performance of the genotypes. They were crossed in line X tester mating design. Forty eight hybrid combinations obtained were raised in the field along with the parents and evaluated for their heterosis and combining ability effects. Parental genotypes AVTS-06-5, AVTS-06-10, IVTS-06-12, KYM-1, Tilak and TMV-6 were identified as high combiners based on general combining ability (gca) effects. Two combinations viz. AVTS-06-5 X KYM-1 and IVTS-06-12 X TMV-3 had significant values of per se performance, specific combining ability (sca) effects and standard heterosis for seed yield per plant. They can be evaluated for their hybrid vigour over locations and seasons. The crosses AVTS-06-5 X TMV-3, AVTS-06-5 X TMV-6 and TCR 3279A X KYM-1 have been identified as potential cross combinations for isolation of promising segregants as the parents involved in these crosses had high significant gca effects for seed yield per plant but the hybrids recorded non significant sca effects. Interspecific hybridization between S.malabaricum and S.indicum was attempted to develop male sterile lines. Seed set was noticed in three interspecific hybrids which failed to germinate due to embryo abortion. Hence these embryos were rescued and raised in vitro to obtain the hybrids.ThesisItem Open Access Invitro propagation in ashoka : saraca asoca (Roxb.) de wilde.(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2011) Brindha Devi, I; KAU; Sreenivasan, EAsoka (Saraca asoca) is an important medicinal and ornamental tropical tree currently facing the threat of extinction due to overexploitation of trees occurring in forests and other natural habitats. Unscientific and destructive extraction of bark from trees has lead to acute shortage of raw bark by ayurvedic industries. Hence, the International Union for Conservation of Nature and Natural Resources (IUCN) has listed this species under ‘globally vulnerable’ category. It is also enlisted among the 36 threatened and endangered medicinal plants of India. It is considered as the sacred tree of buddhists and Hindus. Literally the term ‘asoca’ means ‘sorrow-less’ and the tree is believed to remove the grief and unhappiness. The tree has immense medicinal properties. Its bark is considered as the primary medicinal part. Due to its acute short supply compared to its demand, various development and research activities are being prioritized to conserve, utilize and improve this species. It is mainly propagated by seeds. Due to heterozygous and cross pollinated nature of the species, it never gives a true to type progeny. Therefore the present study was undertaken to standardize the technique of in vitro propagation of Saraca asoca. Standardization of suitable explants, surface sterilization procedures and culture establishment protocol, Induction of multiple shoots and Elongation of root, hardening and planting out are the major objectives of the study. Nodal segment, Internodal segment and shoot tip were the three explants tried. Various surface sterilization procedures were tried using Chloramphenicol, ethyl alcohol, 0.1% mercuric chloride and combination of ethyl alcohol and mercuric chloride in various concentration and duration, using nodal segments as explant. Surface sterilization using 70% ethyl alcohol for 3 minutes followed by 0.1% mercuric chloride for minutes proved to be the best, which gave the maximum survival percentage of 80. The next part of the study was standardisation of suitable explants for culture establishment. Among the three explants, Nodal segments gave maximum response of 60 per cent in ½ MS medium with BAP 0.5mg/l. This was followed by shoot tips in the same medium, which gave 10 per cent response. Internodal segments did not respond in any of the media used. Standardisation of basal media for culture establishment was done using nodal segments as the explants. Three media supplemented with BA 0.5 mg.l-1 were tried viz. MS, Half strenght MS, Woody plant media. Among the three, ½ MS media was identified as the best basal medium followed by MS medium. No response was seen in WPM medium. Culture establishment as well as Shoot bud initiation was attempted in ½ MS and MS media with various growth regulator combinations. Maximum response of 60 per cent was obtained in ½ MS medium containing BAP 0.5 mg/l followed by 30 percent in the same medium containing BAP 1.5 mg/l. There were no response with 2,4-D. The response obtained was callusing in all cases. Induction of multiple shooting was tried in ½ MS medium supplemented with BAP, and Kn alone as well as combinations of BAP, IAA at various concentration. Here highest response of 30 per cent of single shoots was recorded in ½ MS media containing BAP 0.5 mg/l. Response was in the form of single shoot. The single shoots with a mean length of about 1.5mm after one week of growth was obtained. With BAP 2.0 mg/l, single shoots were produced in about 5% of cultures within 54 days. Effect of Kn in various concentration ranging from 0.5 to 2.0 mg/l was found to be low in shoot induction. The maximum length of shoot of about 1.6 cm was recorded in combination of BAP 0.5 mg/l and IAA0.5 mg/l. Various combinations of IAA and IBA at different concentrations were tried for rooting of in vitro shoots. However there was no response in any of the combinations tried.ThesisItem Open Access Induction of parturition and evaluation of postpartum fertility in crossbred cows(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2010) Sheeja, S; KAU; Aravinda Ghosh, K NA preliminary study was conducted by collecting data regarding gestation length and details of calving among crossbred cattle of the University Livestock Farm and local breeds belonging to “ICAR Scheme on Conservation of Germplasm of Vechur Cattle”. The mean gestation length of crossbred cattle of ULF was 274 ± 0.48 days and that of Vechur scheme was 282 ± 0.98 days. The average birth weight of new born calf at ULF was 26.52 ± 0.39 kg and that of Vechur was 10.43 ± 0.12 kg. The sex ratio of male and female was 1: 0.9 and 1: 1.2 for ULF and Vechur scheme respectively The main experiment was undertaken to develop a suitable protocol for induction of parturition in crossbred cattle with prolonged gestation and to assess the postpartum fertility of these animals. The study was performed in 24 pregnant animals of the University Livestock Farm and private farms near by Mannuthy during the period from December 2008 to February 2010. In all animals in group I, II and III, the drug was administered for inducing parturition on 286th day of gestation. In group 1, 24 mg of dexamethsone, in group II 500µg of prostaglandin analogue (cloprostenol) and in group III, a combination of 12 mg of dexamehasone and 250 µg cloprostenol was administered intramuscularly and group IV acted as control. The mean time taken in hours for induction of parturition in group I to III was 39.50 ± 1.26, 30.50 ± 2.17 and 26.90 ± 1.80 respectively and the least time was recorded in combination group. The duration for first stage of labour in groups I to IV was 4.00 ± 0.16, 3.12 ± 0.15, 3.24 ± 0.02, 4.49 ± 0.12 hours respectively and for second stage was 1.27 ± 0.02, 1.12 ± 0.14, 1.21 ± 0.12, 1.53 ± 0.10 hours respectively. The mean time for the expulsion of placenta was 6.67 ± 0.33, 6.35 ± 1.87, 3.02 ± 0.13 2.74 ± 0.14 hours respectively. The mean weight of the placenta for the groups was 2.87 ± 0.43, 3.50 ± 0.54, 3.00 ± 0.28, 3.60 ± 0.25 kg and the mean number of cotyledons were 89.50 ± 0.76, 91.20 ± 0.60, 90.5 ± 0.84, and 91.50 ± 0.76 respectively. The incidence of dystocia in groups I to IV was 50, 0, 33.33 and 50 per cent respectively. The incidence of retention of foetal membranes in groups I to IV was 50, 33.33, 16.66 and 16.66 per cent respectively. In group I, the incidence of postpartum prolapse of genital organs, downer cow and mastitis were recorded as 16.66 per cent each. The sex ratio for the groups I to IV was 1:1, 0.57: 1, 1:1 and 1:1. The mean birth weight in kg for the male calves was 29.33 ± 1.2, 26.65 ±6.5, 31.5 ± 3.40, 34.66 ± 2.03 respectively. Similarly the birth weight of female calves were 30.00 ± 1.15, 27.25 ± 1.97, 23.33 ± 2.40, 32.33 ± 1.20 kg respectively. There was steady increase in body weight of calves as age advanced in experimental and control groups, however there was no significant difference between groups in mean body weight gain. The mean peak yield in the present lactation for the experimental and control animals was 9.57 ± 0.58, 11.33 ±1.17, 11.67 ±1.54 and 13.17± 0.75 liters respectively. The corresponding values in the previous lactation for the experimental and control groups were 11.48 ± 0.48, 11.60 ± 0.75, 12.70 ± 0.47 and 13.50 ± 0.65 liters respectively. The day of peak yield in the present lactation was 25.00 ± 0.63, 21.66 ± 0.61, 22.33 ± 1.05 and 19.16 ± 0.79 days and the corresponding values in previous lactation were 19.80 ± 0.95, 18.70 ± 0.67, 20.30 ± 1.28 and 19.30 ± 0.63 days respectively. The disappearance of lochial discharge for the experimental and control groups was 20.16 ± 1.04, 17.31 ± 1.13, 17.17 ± 0.87, 21.00 ± 1.26 days respectively. The first postpartum oestrus was observed at 33.20 ± 1.25, 30.70 ± 0.88, 29.50 ± 0.76 days for experimental animals and for control animals it was 31.60 ± 0.76 days. Similarly, the second postpartum oestrus was on 59.00 ± 1.22, 51.83 ± 0.83, 48.33 ± 1.99, 53.83 ± 0.94 days respectively. Conception rate for the first AI in group I to IV was 0, 50, 50, and 33.33 respectively where as the overall conception rate for these animals was 16.66, 66.66, 83.33, 66.66 per cent respectively. The highest conception rate was obtained in group III. The mean calving to conception interval for the experimental and control animals was 91.50 ± 15, 77.66 ± 9.38, 74.00 ± 7.00 and 82.00 ± 13.97 days respectively Premature induction of parturition was carried out in four downer cows presented at Veterinary College Hospital, Mannuthy which failed with routine medical treatment and having the gestation length of 253, 285, 270 and 275 days. In the first two animals parturition was induced with prostaglandin and the time taken for induction was 48.30 and 31.20 hours respectively. In dexamethasone treated animal calving occurred at 51 hours after administration of drug. Similarly the time taken for induction in animal which treated with combination of dexamethasone and prostaglandin was 29.45 hours. All the three animals in which prostaglandin and its combination with dexamethasone, recovered from recumbent stage after delivery and had regained normal feeding habits. The animal treated with dexamethasone had not regained the normal condition and was advised disposal. But all the four calves survived in this experiment. The present study revealed that induction of parturition with prostaglandin alone in normal dose and its combination with dexamethasone at a lower dose were equally useful for successful induction of parturition in animals with prolonged gestation with least reproductive complications. When parturition was induced with dexamethasone milk yield was found to be reduced during early stages of lactation while when prostaglandin and its combination with dexamethasone were used reduction in milk yield was negligible. In animals in which parturition was induced with prostaglandin and its combination had normal disappearance of lochial discharge, early involution of uterus and had early normal postpartum oestrus and had fairly good overall conception rate. Further it is recommended that premature induction of parturition in downer cows when all other medical treatments have failed, prostaglandin or its combination with dexamethasone could ideally be used to induce premature induction of parturition to save the life of mother and new born.ThesisItem Open Access Pesticide use pattern and monitoring of residues in cardamom in Idukki district(Department of Agricultural Entomology, College of Agriculture, Vellayani, 2013) Seena, S M; KAU; Naseema Beevi, SThe field survey conducted among the farmers of Idukki district revealed that major pest infesting cardamom were shoot and capsule borer and cardamom thrips. For the timely management of these pests, farmers are following strict plant protection measures at an interval of 15 to 40 days with conventional insecticides. Farmers are widely applying heavy doses of chemicals especially the organophosphorus insecticides like phorate, chlorpyriphos, quinalphos, profenophos, methyl parathion and synthetic pyrethroids like cypermethrin and lambda cyhalothrin. Majority of the farmers resort to prophylactic spraying of plant protection chemicals rather than remedial measures. Adoption of IPM strategies are also negligible. Most of the farmers used their own spraying schedules for pest management. The pesticide use pattern in cardamom growing tracts of Idukki district shows that the farmers are applying plant protection chemicals aggressively and the liberal and continual use of pesticides has disturbing consequences on the ecosystem. In multiresidue mehod validation cardamom samples were spiked at five different levels viz. 0.01 µg g-1, 0.05 µg g-1 , 0.10 µg g-1 0.50 µg g-1 and 1 µg g-1 and extraction was carried out using various solvent/ solvent system and the modified QuEChERS method which gave 69.7–110% per cent recovery with RSD < 20 was selected and the same method was adopted for the estimation of pesticide residues from cardamom samples. In order to assess the residue level and to study the extend of contamination due to pesticides in cardamom, samples were collected from the cardamom growing plantations of Idukki district. Three major cardamom growing zones were selected namely Vandanmedu, Udumbanchola and Poopara in Idukki district and ten samples were collected from each location for a period of six months. Data on monitoring of pesticide residues in cardamom samples collected from the study regions for a period of six months revealed varying level of residues of several pesticides. Out of the total 180 samples analyzed, residues were detected in 173 samples and only seven samples were free of residues. Out of the 173 samples detected with pesticide residues, 160 contained multiple residues of pesticides whereas only 13 contained residues of single pesticide. Cardamom capsules contained residues of 16 different pesticide molecules belonging to organochlorines, organophosphates and synthetic pyrethroids. The most common contaminant was quinalphos which was detected in 121 out of 180 samples analysed. Other major contaminants include lambda cyhalothrin (104), cypermethrin (100), chlorpyriphos (87) and profenophos (64). Pesticides detected in cardamom which have no label claim in cardamom include Beta cyfluthrin (5), bifenthrin (3), fenpropathrin (4), fenvalerate (5), lambda cyhalothrin (104), methyl parathion (64) and triazophos (4) . A field experiment was carried out in order to study the curing process on removal of residues of quinalphos, chlorpyriphos, triazophos, cypermethrin, lambda cyhalothrin and imidacloprid. Curing process removed the residues of pesticides at varying levels. Processing factor was worked out for each chemical. Extent of removal of residues as a result of curing were: quinalphos (61.78-67.78%), chlorpyriphos (70.23-76.66%), triazophos (49.62-55.02%), cypermethrin (65.71-67.63%), lambda cyhalothrin (13.15-40.00%) and imidacloprid (75.56-77.32%).
