Development of chitosan conjugated DNA vaccine against nodavirus in Macrobrachium rosenbergii (De Man, 1879)

The protective efficacy of a DNA construct containing extra small virus antisense (XSVAS) gene of nodavirus encapsulated with chitosan nanoparticles (NPs) was investigated in giant freshwater prawn Macrobrachium rosenbergii (De Man, 1879). The delivery was carried out using oral and immersion methods. A plasmid concentration of 100 ng μL^?1 when conjugated with chitosan NPs was found to be more effective in increasing the survivability of the infected prawn. The particle mean size, zeta potential and loading efficiency percentage were 297 nm, 27 mV and 85%, respectively. The ability of the chitosan to form a complex with the plasmid was studied by agarose gel electrophoresis. The NPs were characterized by atomic force microscopy (AFM). Persistence study showed the presence of the DNA construct up to 30th day post‐treatment. The oral treatment was found to be better than the immersion treatment for delivery of the chitosan‐conjugated DNA construct. This is probably the first report on the delivery of nanoconjugated DNA construct in M. rosenbergii, against nodavirus.     

DDT and HCH Residues in Basmati Rice (Oryza sativa) Cultivated in Dehradun (India)

Organochlorine pesticides were used earlier for agricultureproduction. Their residues may still be present in soil and mayaccumulate in food crops, posing potential health problems to consumers. DDT, HCH, their isomers and metabolites were analyzedin samples of soil and rice plants collected from ten differentvillages of a well-known Basmati rice growing area in Dehradun.Residues of both pesticides were found in all samples ofsoil and different parts of rice plants except for a few grainsamples. Maximum residue was observed in husk and minimum ingrains. The average concentration of DDT in soil ranged from0.013 to 0.238 ppm. p,p′-DDE was the major metabolite (>63%). Theaverage concentration of DDT in rice grain varied from 0.002 to 0.040 ppm. o,p′-DDT was the main isomer (>93%). Theaverage concentration of HCH in soil ranged from 0.122 to 0.638 ppm. β-HCH was the predominant (43%) isomerfollowed by α-HCH (21%). The average HCH concentrationin rice grain ranged between 0.013 and 0.113 ppm. All four isomers were present in grains. The levels of DDT and CHCin grains were similar in magnitude as those from differentIndian states, but well below the maximum residue limit of 0.1 ppm for DDT and 0.05 ppm for HCH prescribed by the Government ofIndia and WHO/FAO. As such, the pesticide residue levels in thisexport commodity are not of hazardous nature.     

Antimicrobial activity of saponins from Acacia auriculiformis

Acaciaside A and B, two acylated bisglycoside saponins originally isolated from the funicles of Acacia auriculiformis, are known to have antihelminthic activity. Their antifungal and antibacterial activities were investigated. Complete inhibition of conidial germination of Aspergillus ochraceous and Curvularia lunata was recorded at 300 microg/ml or less whereas to inhibit the growth of Bacillus megaterium, Salmonella typhimurium and Pseudomonas aeruginosa 700 microg/ml or higher concentrations of the mixture was required. Two catabolic enzymes, phosphofructokinase and isocitrate dehydrogenase, responded differentially in fungi and bacteria against sublethal concentrations of the compound when assayed from their cell free extracts. An increased specific activity of the enzymes in bacteria and a decrease activity in fungi indicate a possible different mechanism of inhibition of saponins on the organisms tested.     

罗氏沼虾谷氨酰胺合成酶基因的克隆与表达

谷氨酰胺合成酶(glutamine synthetase, GS)是一种广泛分布在动植物体内的酶,并参与细胞多种代谢调控。在甲壳动物中,GS在能量代谢和渗透压调节过程中起着重要作用。实验克隆了罗氏沼虾GS基因。GS基因cDNA全长1 965 bp,开放读码框(ORF)为1 086 bp,编码361个氨基酸(aa),分子量大小为40.75 ku,等电点为5.81。进化树分析发现,罗氏沼虾GS基因与凡纳滨对虾、斑节对虾和中国明对虾GS聚为一支,氨基酸相似性达到95%。氨基酸多序列比对分析结果显示,罗氏沼虾GS属于无脊椎动物分支的GSⅡ群,有5个保守区域。实验对罗氏沼虾GS蛋白进行表达并制备了多克隆抗体。采用荧光定量PCR技术(qRT-PCR)和免疫印迹(Western blot)对罗氏沼虾蜕壳前后的不同组织GS表达进行检测。qRT-PCR结果显示,GS基因在所检测的8个组织中均有表达;蜕壳前,其相对表达量顺序为:肝胰脏肌肉胃肠鳃心脏脑血淋巴。蜕壳后和蜕壳前GS基因在组织中的差异表达比较结果显示,除了在肝胰脏表达下调外,其他7个组织都表达升高,其相对表达量的差异顺序为:脑鳃胃肠肌肉心脏血淋巴。Westernblot结果显示,蜕壳后GS蛋白在鳃和肌肉组织表达量上调,与其mRNA表达一致。此外,罗氏沼虾蜕壳后肝胰脏GS酶的活性和谷氨酰胺的含量下调,而其在鳃、肌肉和血淋巴中上调,结果与其基因表达一致。研究表明,GS基因在不同组织中表达的差异可能和罗氏沼虾的能量代谢、渗透压调节有关。     

Investigating external effects of shrimp farming on rice farming in southern Thailand: a technical efficiency approach

Rice and shrimp farming are two important economic activities in rural southern Thailand that compete each other in land allocation leading to land use conflicts. Moreover, it can be observed that the conversion of rice fields to shrimp farms is already extended from coastal areas to the inlands. These conversions are making more area be saline which is essential condition for shrimp cultivation but deleterious for rice farming. We are looking at the unidirectional externalities originating from the conversion which brings the productivity of adjacent rice fields further down. This study attempts to quantify the external effects on rice yield and efficiency of rice farming by constructing three models. Model I, we assess the rice yield function by using the multiple linear regression while in Model II, we attempt to delineate the technical efficient frontier of rice farms by using an input oriented non-parametric approach, so called data envelopment analysis. Model III, we probe into the factors influencing technical efficiency scores using the Tobit regression. Database of the study is constituted by primary survey data from 120 rice farms in Songkhla province for the crop year 2004/2005. The 30 out of 120 sample farms have been affected by externality of shrimp farming. The empirical results of all three models show that shrimp farming externalities caused a production drop of 467–515 kg paddy rice per ha and significantly affect the technical efficiency of rice farms. The measures of internalizing this externality are discussed.     

Histopathological changes in giant freshwater prawn Macrobrachium rosenbergii (de Man 1879) fed with probiotic Bacillus licheniformis upon challenge with Vibrio alginolyticus

The effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the histopathological changes in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g) challenged with known pathogenic strain of Vibrio alginolyticus are reported. Two isocaloric basal diets supplemented with probiotic bacteria B. licheniformis (1.0 × 10⁹ cfu/g feed) and other without probiotic supplementation were fed to the M. rosenbergii juveniles for 45 days. The histological observations revealed no significant changes in the hepatopancreas and gut tissues of both the experimental and the control groups which indicate that the present bacterium is a safe candidate probiont for the host. Prawns were challenged with V. alginolyticus after 45 days of feeding with probiotic diet. The histopathological studies of the hepatopancreas revealed that M. rosenbergii fed with probiotic‐supplemented diet showed less changes as compared to the prawns fed with control diet on second and fourth day of post‐experimental challenge with V. alginolyticus. Histopathological observations revealed that the gills of the prawns fed with control diet were severely affected in comparison to the prawns fed with probiotic‐supplemented diet after challenging with V. alginolyticus. Results from this study revealed the improved protection by dietary incorporation of B. licheniformis in reducing the histopathological manifestations due to V. alginolyticus infection in freshwater prawn.     

Comparative performance of four multi-purpose trees associated with four grass species in the humid regions of Southern India

The results of a field trial conducted at the Livestock Research Station, Thiruvazhamkunnu, Kerala to study the compatibility of different components in a silvo-pastoral system revealed that growth and yield of fodder species were significantly influenced by the tree components only after tree canopy formation. The fodder species such as Pennisetum purpureum Schum., Panicum maximum Jacq., Brachiaria ruziziensis Griseb. and Euchlaena mexicana Schrad. grown in association with Casuarina equisetifolia J. R. & G. Forst. and Ailanthus malabarica DC recorded comparatively higher forage yield even after canopy formation. However, forage crops grown in combination with Acacia auriculiformis A. Cunn. ex Benth. and Leucaena leucocephala (Lam.) de Wit. registered relatively lower values for growth and yield. Increased light infiltration into the understorey due to the cladophyllous canopy can be attributed as reason for the higher fodder productivity under Casuarina. Due to the combination of crown size and shape, tree height and spacing the amount of light intercepted by Ailanthus also was very low. Among the four multi-purpose trees used, Acacia recorded the maximum growth rate followed by Casuarina, Ailanthus and Leucaena. Forage productivity of the four species was in the order: Pennisetum purpureum > Panicum maximum > Brachiaria ruziziensis > Euchlaena mexicana. Casuarina with Pennisetum/ Panicm were found to be optimal tree — forage combinations for silvo-pastoral systems.     

Doubled haploids in tropical maize: <Emphasis Type="SmallCaps">II</Emphasis>. Quantitative genetic parameters for testcross performance

Single crosses (SC) of elite inbreds and open-pollinated populations (OP) are suitable source germplasm for doubled haploid (DH) line development in hybrid maize breeding, given that they combine a high population mean ([`(x)] \overline{x} ) for testcross performance with adequate response to selection ( \Updelta G \Updelta G ). This is the first study reporting testcross grain yield (TCGY) and dry matter content (TCDMC) evaluations of 131 DH lines developed from ten tropical source germplasm comprising five OP (OP1–OP5) and five SC (SC1–SC5). Gene diversity (d) and the average number of alleles (a <sub>r</sub> ) per locus was estimated for DH lines based on 24 simple sequence repeat markers. Analysis across three environments revealed no significant differences between [`(x)] \overline{x} of OP- and SC-derived DH lines for TCGY and TCDMC. Significant genetic variance for both traits was only detected among OP-derived DH lines which may be explained by a larger number of segregating quantitative trait loci (QTL) as suggested by higher d and a _r values than in SC-derived DH lines. The usefulness criterion ( U = [`(x)] + \Updelta G U = \overline{x} + \Updelta G ) was higher for OP-derived DH lines for TCDMC, but higher for SC-derived DH lines for TCGY. DH lines from OP1, OP2, and OP3 showed high TCGY, suggesting that they may be useful in tropical hybrid breeding. We conclude that tropical OP represent a valuable source of untapped genetic variation that can efficiently be exploited with DH technology for hybrid maize breeding.     

Effect of fortifying concentrate supplement with fibrolytic enzymes on nutrient utilization, milk yield and composition in lactating goats

Thirty lactating Beetle‐sannen crossbred goats were randomly divided into 3 groups based on first fortnight milk yield, and then adjusted to body weights to study the effect of fibrolytic enzyme supplementation on digestibility and milk production. Goats in the control group (C) were fed 500 g of concentrate supplement (CS) without exogenous fibrolytic enzymes (control; CG) or CS fortified with cellulase and xylanases @ of 4000 and 12500 (EG₁) or 8000 and 18 750 IU/kg (EG₂), respectively. CS was offered in two equal proportions just after milking at 7 and 17 h. After feeding CS, goats were fed 1200 g of wheat straw as roughage in two equal parts at 08.00 and 18.00 hours. Milk yield was studied for 3½ months. Milking was done by hand. Changes in body weight were also recorded. After a preliminary period of 60 days of feeding, a 7‐day digestibility trial was conducted in two intervals with 5 animals from each group at a time. The increase in either wheat straw or NDF intake due to the enzyme mixture was only 3%. There was significant improvement (P < 0.05) in the diet digestibility of DM, OM, CP, NDF, ADF and total carbohydrates (TC) between the control and EG₁ and EG₂ goats. Improvements were greater (P < 0.05) in the latter group. Microbial protein (MBP) was estimated based on purine derivatives and creatinine excreted in spot urine samples. MBP increased about 10 and 15% in EG₁ and EG₂, but these values were statistically invalid. Phenomenally milk yield, fat and SNF were significantly (P < 0.001) changed with progression of lactation. On a temporal basis, fortnightly changes in FCM yield of this study could be grouped as 1 month each of first and last and 1½ of middle phase of lactation. Repeated measure analysis showed 180 to 260 mL/day of higher FCM and 16 to 20 g/day of higher SNF yield (P < 0.001) in EG₂ than control or even EG₁ during 5 to 7^th fortnight. However, no significant impact (P > 0.05) was noticed on TF. Feed intake, g/kg FCM yield was also 7% less in EG₂ than either CG or EG₁. The study concluded that apart from increased digestibility, fortification of CS with cellulase and xylanases enzymes improved FCM yield in the last quarter of lactation in goats, and improved body weights by 5% (P > 0.05). While the digestibility of the diet was higher in EG₁ and EG₂, the impact on FCM and SNF yield was noticed only on EG₂. Our study indicated that CS of lactating goats may be fortified with 8000 and 18 500 IU/kg CS in the last quarter of lactation.