Development and characterization of cell lines derived from rohu, Labeo rohita (Hamilton), and catla, Catla catla (Hamilton)

Two new cell lines, designated RE and CB, were derived from the eye of rohu, Labeo rohita , and the brain of catla, Catla catla , respectively. The cell lines were maintained in Leibovitz's L-15 supplemented with 20% foetal bovine serum. The RE cell line was sub-cultured for more than 70 passages and the CB cell line for more than 35 passages. The RE cells are rounded and consist predominantly of epithelial cells. The CB cell line consists of predominantly fibroblastic-like cells. Both cell lines are able to grow at temperatures between 25 and 32 °C with an optimum of 28 °C. The growth rate of the cells increased as the foetal bovine serum concentration increased from 2% to 20% at 28 °C, with optimum growth at concentrations of 15% or 20% foetal bovine serum. The cells were successfully cryopreserved and revived at different passage levels. The cell lines were not susceptible to four marine fish viruses. Extracellular products from Aeromonas sp . were toxic to the cell lines. When the cells were transfected with plasmid eukaryotic green fluorescent protein (pEGFP [Clontech, Carlsbad, CA, USA]) vector DNA, a significant fluorescent signal was observed suggesting that these cell lines could be a useful tool for transgenic and genetic manipulation studies. Polymerase chain reaction amplification of mitochondrial 12S rRNA from rohu and catla confirmed that the cell lines originated from these fish species. The cell lines were further characterized by immunocytochemistry using confocal laser scanning microscopy.     

Productivity of radiata pine (Pinus radiata D. Don.) clones in monoclonal and clonal mixture plots at age 12 years

Productivities of monoclonal plots and clonal mixtures of 10 radiata pine (Pinus radiata D. Don.) clones were compared in a trial established in 1993 at Dalethorpe, Canterbury, New Zealand. Ten monoclonal and one mixture of the 10 clones were planted in a complete randomised block design with three replications using 40-tree plots (un-thinned, pruned to 2.5 m, stocking of 1250 stems per hectare). The study was conducted to determine if mode of deployment (monoclonal versus clonal mixture) affected overall productivity and how or if each clone was affected by mode of deployment.     

Effects of dietary arachidonic acid on final maturation,spawning and composition of gonad of black sea urchin Diadema setosum (Leske, 1778)

This study investigated the effect of dietary arachidonic acid (ARA, 20:4n‐6) on final maturation, spawning performance and composition of the gonad of black sea urchin (Diadema setosum). Four different dietary ARA levels of 0.3 g/kg (Control diet), 1.4 g/kg (ARA 1.4), 2.5 g/kg (ARA 2.5) and 3.7 g/kg (ARA 3.7) were prepared and tested in this feeding trial for 90 days. The result showed that growth in terms of weight was not relatively affected by dietary ARA. Gonadosomatic Index (GSI) of animals were gradually increased in all the diets throughout the trial. The highest GSI level (25.7 ± 3.16%) was observed in animals fed with the diet ARA 1.4. ARA content in the gonad proportionally increased with dietary ARA levels. The highest larval survival rate (85.5 ± 5.67%) was found in diet ARA 2.5. The results of this study indicate that ARA plays an important role in the reproduction functions of black sea urchin and supplementing an appropriate level of ARA in the broodstock diet could induce the final maturation and improve reproduction of black sea urchin, but a higher level of ARA (3.7 g/kg) seems to have a negative effect on both growth rate and GSI.     

An artificial neural network model for estimating Mentha crop biomass yield using Landsat 8 OLI

Yield forecasting is essential for management of the food and agriculture economic growth of a country. Artificial Neural Network (ANN) based models have been used widely to make precise and realistic forecasts, especially for the nonlinear and complicated problems like crop yield prediction, biomass change detection and crop evapo-transpiration examination. In the present study, various parameters viz. spectral bands of Landsat 8 OLI (Operational Land Imager) satellite data and derived spectral indices along with field inventory data were evaluated for Mentha crop biomass estimation using ANN technique of Multilayer Perceptron. The estimated biomass showed a good relationship (R²?=?0.762 and root mean square error (RMSE)?=?2.74 t/ha) with field-measured biomass.

     

Discovery and detection of single nucleotide polymorphism (SNP) in coding and genomic sequences in chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

Chickpea genetic mapping has been hampered by insufficient amplicon length polymorphism for sequence based markers. To develop an alternative source of polymorphic markers, we determined naturally abundant single nucleotide polymorphism (SNP) in coding and genomic regions between FLIP 84-92C (C. arietinum) and PI 599072 (C. reticulatum) and identified an inexpensive method to detect SNP for mapping. In coding sequences, 110 single base changes or substitutions (47% transitions and 53% transversions) and 18 indels were found; while 50 single base changes (68% transitions and 33% transversions) and eight indels were observed in genomic sequences. SNP frequency in coding and genomic regions was 1 in 66 bp and 1 in 71 bp, respectively. In order to effectively use this high frequency of polymorphism, we used Cleaved Amplified Polymorphic Site (CAPS) and derived CAPS (dCAPS) marker systems to identify a restriction site at SNP loci. In this study, we developed six CAPS and dCAPS markers and fine mapped QTL1, a region previously identified as important for ascochyta blight resistance. One of the CAPS markers from a BAC end was identified to account for 56% of the variation for ascochyta blight resistance in chickpea. Conversion of naturally abundant SNPs to CAPS and dCAPS for chickpea mapping, where absence of amplicon length polymorphism is a constraint, has potential to generate high-density maps necessary for map-based cloning and integration of physical and genetic maps.     

Transgene integration - an analysis in autotransgenic Labeo rohita Hamilton (Pisces: Cyprinidae)

Transgenic Labeo rohita founder population was analyzed for the presence of autotransgene having histone 3 promoter and growth hormone (GH) cDNA (LRH3-GHcDNA) or total GH gene (LRH3-GH2.8) by PCR with transgene specific primers. Transgene specific amplification was seen with LRH3-GHcDNA in five out of seven individuals and all three fishes with LRH3-GH2.8, indicating their transgenic nature. Transgene integration was also studied by Southern hybridization of DNA isolated from blood of the transgenic fishes with two different probes (histone 3 promoter and cDNA of L. rohita). Autotransgene integration was confirmed in all PCR positive transgenic individuals. The site of integration of the transgene in the genome of the four transgenic fish could be determined by inverse PCR. Two individuals showed integration at the same site whereas in the remaining two individuals the integration sites were different.     

Evaluation of Bacillus subtilis as a probiotic to Indian major carp Labeo rohita (Ham.)

Bacillus subtilis, a Gram‐positive, aerobic, endospore‐forming bacterium, was evaluated for its probiotic potential in Indian major carp, Labeo rohita. Labeo rohita (15±2 g) were fed a feed containing B. subtilis in three concentrations for 2 weeks, e.g., 0.5 (T₂), 1.0 (T₃) and 1.5 (T₄) × 10⁷ CFU g^?1 feed. The control group (T₁) was fed feed without B. subtilis for the same period. Haematological and serum parameters were monitored at weekly intervals. The response variables were total erythrocyte count, total leucocyte count (TLC), haemoglobin, total protein, albumin, globulin, albumin–globulin ratio, alkaline phosphatase activity, alanine aminotransferase activity and aspartate aminotransferase activity. Fish were challenged intraperitoneally with a virulent strain of Aeromonas hydrophila after 2 weeks of feeding to the treatment groups and positive control group, while the negative control group was challenged with phosphate‐buffered saline only. Clinical signs and symptoms, and mortality/survival percentage were noted in each group. The haematological and serum parameters were monitored each week and during post challenge on the third and tenth day. The B. subtilis‐treated fish (T₄, 1.5 × 10⁷ CFU g^?1 feed) showed maximum per cent survival (87.50%), weight gain (35.5%), TLCs (3.23 × 10⁴ cells mm^?3), haemoglobin content (7.4 g%), total protein (2.37 gdL^?1) and globulin content (1.28 g dL^?1) during the pre‐challenge. Enzymes showed higher activities during post challenge (P<0.05). The result suggests that B. subtilis can be used effectively as a commercial product for use in aquaculture.     

Phytochemical and termiticidal study of Lantana camara var. aculeata leaves

Extracts of Lantana camara var. aculeata leaves were studied for their phytochemical constituents and termiticidal effects against adult termite workers. The 5% chloroform extract was found to be significantly effective against termite workers.     

Economic impact of <Emphasis Type="Italic">Przhevalskiana silenus</Emphasis> infestation in native goats of Northern India

Economic losses incurred by Przhevalskiana silenus in goats of Jammu province of Jammu and Kashmir (North India) were assessed from July 2005 to June 2006. Abattoir studies revealed holes in hides for 112 days (mid-December 2005 to first week of April 2006) and trimming of carcass for 243 days (July 2005 to February 2006) during a lifecycle, leading to annual losses of Rs. 7,391,432 and Rs. 10,510,837, respectively. An effect on production traits (weight gain and milk) was assessed on naturally P. silenus-infested Beetal goats. The mean body weight loss of warbled goats was 2.40 kg (equal to 1.096 kg of meat, assuming 45.7% dressing percentage of goat) leading to an annual loss of Rs. 35,473,265. The mean milk yield loss per warbled goat per day was 101.7 g resulting in losses to the tune of Rs. 19,879,012. It can be concluded that P. silenus infestation resulted in an overall loss of Rs. 73,254,547, i.e. 73.26 million annually (equivalent to US 1.62 million; US1.62 million; US 1 = Rs. 45.24), where reduced growth alone contributed 49% of the total losses. A study recommends that a control scheme against P. silenus infestation in goats using an effective insecticide should be followed to prevent huge economic losses in hide, milk and meat production. Economic losses associated with P. silenus infestation in goats were estimated for the first time from India.