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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, Mathew
A 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.
Open Access
Bioactivity of carotenoids from shrimp shell waste
(Department of Processing Technology,College of Fisheries,Panangad, 2010) Sindhu, S; KAU; Sherief, P M
Shrimp 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.
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, C
Puntius 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.
Open Access
Heterosis breeding in sesame (Sesamum indicum L.).
(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2011) Gayathri, G; KAU; Dijee, Bastian
The 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.
Open Access
Development of F1 hybrids of indeterminate tomato (Solanum lycopersicum L.) for protected cultivation
(Department of Olericulture, College of Agriculture, Vellayani, 2015) Lekshmi, S L; KAU; Celine, V A
The present investigation entitled “Development of F1 hybrids of indeterminate tomato (Solanum lycopersicum L.) for protected cultivation” was conducted at the Department of Olericulture, College of Agriculture, Vellayani, from 2013 to 2015 with the objectives of identifying superior varieties and developing F1 hybrids of indeterminate tomato suited for protected cultivation. The study consisted of two experiments conducted in the naturally ventilated polyhouse of size 50 m x 20 m located at the Instructional Farm, Vellayani. In the first part of the first experiment, 40 tomato genotypes were evaluated for two consecutive years in an RBD with three replications. As the second part, 12 commercial hybrids were evaluated. The second experiment consisted of a 9 x 9 half diallel analysis laid out in an RBD with three replications. Analysis of variance showed significant differences between the genotypes for all the characters for two crops. Pooled analysis revealed that, LE 1 recorded the highest yield (2443.43 g) and fruit weight (108.13 g) followed by LE 7. LE 53 had maximum number of fruits per plant (65.00). In the present study, genotypes had wide variation for quality parameters. Fruits of LE 14 recorded highest TSS with a mean of 5.74 ºBrix. LE 7 had maximum ascorbic acid (30.13 mg/100g) and lycopene content (13.09 mg/100g). Beta carotene value was maximum in LE 16 (184.15 mg/100g). There was minimum incidence of pests, diseases and physiological disorders under protected conditions. Among the 12 hybrids evaluated, INDAM 9802 was the highest yielder (1444.40 g) followed by F1 T 30 (1412.22 g). F1 T 30 recorded maximum fruits per plant (35.66) which was on par with F1 Queen (35.55). Genetic parameters like phenotypic and genotypic coefficients of variation, heritability and genetic advance were studied to assess the genetic variability among the genotypes. High heritability coupled with high genetic advance were observed for characters like truss per plant, fruits per truss, fruit weight, fruits per plant, yield per plant and yield per plot. Path analysis revealed highest positive direct effect for fruit weight (0.3956), truss per plant (0.3558) and fruits per plant (0.3381). Based on D2 analysis the 40 genotypes were grouped into eight clusters. Cluster I was the largest with twenty four genotypes followed by cluster II with ten genotypes. Diallel analysis was carried out using nine parents selected based on genetic divergence and per se performance. The parents were crossed in a diallel fashion excluding reciprocals to obtain 36 F1 hybrids. The study revealed that P5 x P9 had the highest yield (3114.03 g) which was on par with P6 x P8 (3074.37 g) and P1 x P5 (3077.58 g). P1 x P5 had the maximum fruits per plant (103.93). The magnitude of relative heterosis, heterobeltiosis and standard heterosis varied considerably. For yield, relative heterosis ranged from -32.40 to 92.72, heterobeltiosis from -47.14 to 89.54 and standard heterosis from 2.91 to 160.95. The σ2gca and σ2sca ratio indicated that non-additive gene action was predominant for all traits. Among the nine parents, P9 (LE 1), P5 (LE 20) P1 (LE 2), and P2 (LE 7) were superior for yield and yield attributes. The estimates of sca effects indicated that P5 x P9 (LE 20 x LE 1), P6 x P8 (LE 39 x LE 38) and P1 x P5 (LE 2 x LE 20) were the most promising hybrids for protected cultivation. The present study revealed that the genotypes LE 1 and LE 7 and the hybrids INDAM 9802 and F1 T 30 were superior for yield and yield attributes under protection. Based on the mean performance, standard heterosis and sca effects the three potential crosses viz., P5 x P9, P6 x P8 and P1 x P5 could be adjudged as suitable indeterminate tomato hybrids for protected cultivation.
Open Access
Detergent potential of enzymes of dairy microflora and their effect on the shelf life of milk products
(Department of Dairy Science, College of Veterinary and Animal Sciences, Mannuthy, 2010) Beena, A K; KAU; Geevarghese, P I
A study was conducted to assess the detergent potential of a spoilage protease enzyme obtained from the microflora of dairy plant environment. An attempt was also made to study the impact of selected enzyme producers on the shelf life of curd (dahi) and sterilised skim milk. A total of 71 bacterial isolates obtained from dairy environment were screened for their ability to produce spoilage enzymes like proteases lipases and lecithinases. Based on the spoilage potential, Pseudomonas aeruginosa (P12) isolated from pasteurised milk and Bacillus cereus (S4) isolated from sterilized skim milk were selected for further work. The influence of spoilage enzymes on selected physico-chemical characteristics of curd (dahi) and sterilized skim milk was evaluated by preparing the products from milk precultured with isolate P12 and S4. In general, proteolysis of milk was found to have an adverse effect on the quality of products. The stimulatory effect of proteolytic products of P12 and S4 on curd starters was evident from the higher values of acidity, firmness and syneresis in treated curd. The spoilage enzymes adversely affected the overall quality and shelf life of curd. In treated sterilised milk, tyrosine and NPN values were highly elevated. A linear correlation was found to exist between off-flavour and proteolysis. Curd and sterilised skim milk prepared from milk precultured with proteolytic organism were significantly different from that of control. The possibility of exploiting an alkaline protease from spoilage organism in dairy plant sanitation was also looked into. Environmental conditions for the production of alkaline protease by a psychrotrophic strain of Bacillus cereus (S4) was optimised in whey based medium. The protease used in this trial preferred an alkaline medium to remain stable. The enzyme was found to be stable over a wide temperature range of -10°C to 80°C and a pH range of 7.0 to 12.0. The metal ions Ca++, Mg++, Zn++ and Hg++ enhanced the enzyme activity. Lack of inhibition by Hg++ suggested lack of disulphide bonds in the active site of enzyme. Significant inhibition of activity by serine inhibitors indicated an essential serine residue in the active site of enzyme. The deleterious effect of EDTA on enzyme activity showed the supportive role of divalent cations. Marked residual activity on treatment with β-mercaptoethanol indicated the absence of cysteine residue for the enzyme. Enhancement of protease activity in the presence of surfactants and stability in the presence of H2O2 signified its potential to be used as detergent additive. Qualitative assessment of cleaning efficiency of inbuilt formulation substantiated the superiority of enzyme based formulations. Ammonium sulphate fractionation, dialysis and gel filtation using seralose 4B and Seralose 6B were effective in purifying the protease preparation by 141.31 fold. The purified protease was found to be a homogenous preparation of molecular weight of 50.5 kDa as determined by SDS PAGE.
Open Access
Development and evaluation of different vaccines against duck pasteurellosis
(Department of Dairy Science, College of Veterinary and Animal Sciences, Mannuthy, 2010) Jesto, George; KAU; Krishnan Nair
This study was undertaken to develop biofilm vaccines against duck pasteurellosis using oil, saponin and aluminium hydroxide as adjuvants and to experimentally evaluate their immunogenicity in ducks. Identity of P. multocida serotype A: 1 (DP1) used for study was confirmed by biochemical tests and by PM-PCR and pathogenicity was established in Swiss albino mice before vaccine production. The LD50 (11 w) of the DP1 isolate determined was 10 CFU / bird and MDT was 23.75 h in 11 week old ducklings when a high dose of 3 X 10 6 CFU of P. multocida per bird was given. At 11 Weeks age MDT gradually increased as the dose of inoculum decreased. In 21 week old ducks, the LD50 (21w) of the isolate DP1 was estimated to be 3 ×108 CFU of P. multocida and it showed that the Kuttanad duck had decreased susceptibility to pasteurellosis with age. On light microscopic studies planktonic cells appeared to be Gram negative coccobacillary, while biofilm cells were Gram negative and pleomorphic. Electron microscopic studies revealed that P. multocida could form well differentiated classic biofilm and 0.32 per cent TSB media supplemented 0.5 per cent chitin seemed to be excellent medium for biofilm formation by P. multocida. Four different vaccines viz. OV, OBV, SV and SAV were prepared and all of them were found to be sterile and safe. The oil adjuvanted vaccines (OV and OBV) offered better protection compared to saponin and aluminium hydroxide adjuvanted vaccine groups (SV and SAV) following primary vaccination, up to seven weeks. The modified oil adjuvanted vaccine prepared was not only found to be homogenous, but also more efficient in stimulating a humoral immune response and hence may be recommended. The oil adjuvanation gave better protection than saponin and aluminium hydroxide adjuvanation. The SAV gave better protection than SV which might be due to the presence of aluminium hydroxide which potentiated the immunostimulating ability of saponin. The combined vaccine (SAV) although was found to be better than single vaccine (SV) they cannot be used as a substitute to oil adjuvanted vaccines. The booster vaccination was found to have added advantageous effect on protection and is a must, to prevent losses. Pasteurella biofilms although found to be weak in inducing a primary immune response had the potency to evoke a more powerful secondary response compared to planktonic cells. Vaccination done at six weeks age followed by booster vaccination at 16 weeks age seemed to be a better modification of existing schedule and may be recommended. In histopathological studies, lymphoid hyperplasia was observed in spleen in survived control birds and in SV and SAV vaccine groups that did not survive challenge test, which indicated the persistence of Pasteurella organisms through mild infection in them following experimental challenge. Lymphoid depletion was observed in caecal tonsil in experimental pasteurellosis as in spleen. As the survived vaccinated birds following challenge test showed normal intact caecal tonsil, the course of disease and lesions might be less prominent in vaccinated birds during infection process. The well developed bursa observed in OV and OBV birds that survived challenge test indicated that the humoral immune response was well induced in them compared to other groups. The designed primers E1 and E2 amplified the gene E and hence, this pair of primers could be used for the production of amplified Gene E sequences for further studies on recombinant ghost system. In conclusion, 0.32 per cent TSB media supplemented 0.5 per cent chitin seemed to be an excellent medium that support classical biofilm formation by P. multocida. Booster vaccination definitely had added advantageous effect on protection. Immunization at 6 weeks of age with OV followed by booster vaccination at 16 weeks age with OBV seemed to be a better modification of existing schedule and may be recommended. In histopathological studies, the lesions were less prominent in vaccinated birds than control birds which indicated that the vaccines were effective.
Open Access
Sulphur and boron nutrition and their foliar diagnosis in sesame
(Department of Soil Science and Agricultural Chemistry,College of Agriculture, Vellayani, 2010) Jeena, Mathew; KAU; Sumam, George
A laboratory cum field experiment was conducted to study the effect of S and B on the growth, yield and quality of sesame var. Thilarani and to standardize the foliar diagnosis of these elements in Onattukara sandy loam soil. The study included an incubation study and two field experiments. The treatments comprising the different levels of S and B laid out in 42factorial RBD. The treatments were T1(S0B0), T2(S0B1), T3(S0B2), T4(S0B3), T5(S1B0), T6(S1B1), T7(S1B2), T8(S1B3), T9(S2B0), T10(S2B1), T11(S2B2), T12(S2B3), T13(S3B0), T14(S3B1), T15(S3B2), T16(S3B3). The different levels of S were S0 (0 kg ha-1), S1 (7.5 kg ha-1), S2, (15 kg ha-1) and S3 (30 kg ha-1) and B0 (0 kg ha-1), B1(2.5 kg ha- 1), B2 (5 kg ha-1) and B3 (7.5 kg ha-1). The incubation study was conducted at College of Agriculture, Vellayani to understand the dissolution and release pattern of S and B from their sources gypsum and borax respectively in Onattukara sandy soil. The results revealed that the release of S and B was maximum at the 30th DOI. Increasing levels of S and B has a positive influence on the S content of the soil. T16 (S3B3) recorded the highest value at all the sampling stages for S whereas in the case of B, the treatment combinations which received B at the highest levels in combination with S3 or S2 showed the highest value. The field experiments were laid out at ORARS, Kayamkulam in 42 factorial RBD having two replications using Thilarani as the test crop. It was observed that application of S and B favourably influenced the yield and yield attributes of sesame. T16 was found to be the treatment which gave the highest grain yield and oil yield in both the years and was found to be on par with T14 (S3B1). S3 was the superior S level. As for the different levels of B, B1 can be inferred as the best level. The content of saturated fatty acids such as palmatic and stearic acid showed a decreasing trend with increasing levels of S and B whereas the content of the unsaturated fatty acids showed an increasing trend. The quality attributes of oil such as acid value, iodine value and saponification value was also studied and it was found that there is a decreasing trend with regard to acid and saponification value and an increasing trend for iodine number. The grain protein content also showed an increasing trend with the increase in rate of application of S and B. Regarding the content and uptake of N, P, K, S, B, Fe, Mn, Cu and Zn, a favourable influence for the different levels of S and B was recorded. Results regarding the S and B use efficiency and their apparent recovery showed that with increase in levels of S, an increasing trend was observed for S. In the case of B, increase was noticed up to B1 (2.5 kg ha-1) and there after showed a decreasing trend. This positive influence was also reflected on the available nutrient status of the soil such as organic carbon content, available N, P, K, S, B and DTPA extractable micronutrients. Correlation studies conducted to standardize the part and stage of sampling for the foliar diagnosis of sesame showed petiole at 30 DAS and 20 DAS in the case of S and B respectively. The same stages were found for the soil sampling also for both the nutrients. The critical nutrient level in the part standardised for these two nutrients were standardized using the graphical method proposed by Cate and Nelson (1965). In the case of S, it had been standardized as 0.088 per cent and for B, it had been found to be 28 mg kg-1. The critical nutrient level in soil was also estimated using the scatter diagram technique and was found to be 23 kg ha-1 at 30 DAS for S and 1.4 ppm at 20 DAS for B. Hence the application of S @ 30 kg ha-1 and B @ 2.5 kg ha-1 could faourably enhance growth of sesame with regard to the growth characters, yield and yield attributes and the quality aspects. Moreover, analysis of the plant and soil samples at the critical stages fixed for the respective nutrients will provide the necessary data for the sustainable management of the crop in Onattukara sandy loam soil.
Open Access
Variability in asoka (Saraca asoca(Roxb.) de wilde)
(Department of Plant Breeding and Genetics, College of Horticulture, Vellanikkara, 2010) Vidhu Francis, Palathingal; KAU; Radhakrishnan, V V
Asoka (Saraca asoca) is a sacred tree among the Buddhists and Hindus. It is called sorrow-less tree as it removes the grief. The tree has immense medicinal properties. Its bark is mainly used for correcting uterine problems. The well-known Ayurvedic preparations of asoka bark are Asokarishtam and Asokaghrutham. Due to over exploitation of this tree for its bark, this has now become almost extinct. 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. 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. Therefore the present study was undertaken to assess morphological variations in existing germplasm and to study the reproductive biology of S. asoca. Collection of seeds and evaluation of seed and seedling traits, evaluation of therapeutical components and molecular characterisation of asoka were the other objectives of the study. Variability studies for morphological traits of asoka indicated that height of plant and stem girth have high correlation with bark yield as well as higher direct effect. These traits can be used for identifying better genotypes for higher bark yield. In discriminant function analysis, the selection index involving height of plant along with bark yield constituted for selection criteria among asoka genotypes. Using this selection index, accessions IC566463, IC566489, IC566488, IC566482 were selected as better accessions for higher bark yield. The 43 accessions maintained in germplasm of asoka at AICRP on M&AP were grouped into two major clusters based on morphological traits. These clusters further formed 6 clusters. Accession IC566488 grouped alone in one cluster indicated that this accession was different from all other accessions. In all the other 5 clusters, accessions from Thrissur was included. This indicated that accessions did not follow geographical distributions. Some of the accessions from Thrissur may have relationship with accessions from other districts. Reproductive biology of asoka was studied. It indicated that the reproductive traits like floral biology, anthesis, pollen morphology, its viability have variability among eight trees studied in KAU campus. Among the eight trees studied, ‘KAU8’ have better reproductive traits compared to the rest. Preliminary studies were carried out in pollination system and agents for pollination in asoka. The studies indicated that ants may be one of the pollinating agents. Seed and seedling behaviour of asoka were studied on seeds collected from 80 trees located in different districts of Kerala. Seed breadth, seed volume, height of plant and stem girth were identified as selection traits for better seedlings. Hence at seed stage, bigger sized seeds with higher seed volume will result into better seedlings. Vigorous seedlings is produced from tall seedlings with higher stem girth and in mature plants for higher bark yield, height of plant and stem girth can be selection traits. Thus the selection parameters were worked out in three stages in asoka. Based on seed and seedling selection parameters, OKL4, OKL2 from Odakkali, KKL2 from Kottakkal and TVM2 from Thiruvananthapuram were selected as better mother plants for higher bark yielding accessions. The biochemical constituents (phenol and tannin content) imparting medicinal properties were estimated among the different age groups of asoka. Both phenol and tannin contents were higher in bark compared to flower and leaves. About 50 per cent of therapeutical constituents are available in flowers and hence at non destructive level, flowers of asoka can be substituted for bark. The trees KAU8, KAU7, KAU6 and among the accessions IC566474, IC566467 and among the seedlings VKA6, VKA7, VKA8, KMK3 indicated higher phenol and tannin content compared to rest. Molecular characterisation among the selected asoka accessions representing the different districts of Kerala were studied. RAPD analysis was attempted among ten accessions selected. Dendrogram was constituted based on pooled RAPD data. The ten selected accessions were grouped into two major clusters as done in morphological grouping. In grouping at molecular level, the accession IC566488 kept apart as a single group indicating the same trend at its morphological level.The clustering pattern based on molecular characterisation did not follow geographical distribution of accessions.

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