Detection and molecular characterization of infectious spleen and kidney necrosis virus from major ornamental fish breeding states in Peninsular Malaysia

‘Gold standard’ OIE reference PCR assay was utilized to detect the presence of infectious spleen and kidney necrosis virus (ISKNV) in freshwater ornamental fish from Malaysia. From total of 210 ornamental fish samples representing 14 species, ISKNV was detected in 36 samples representing 5 fish species. All positive cases did not show any clinical signs of ISKNV. Three restriction enzymes analyses showed that the fish were infected by identical strains of the same virus species within Megalocytivirus genus. Major capsid protein (MCP) genes of 10 ISKNV strains were sequenced and compared with 9 other reference nucleotide sequences acquired from GenBank. Sequence analysis of MCP gene showed that all strains detected in this study were closely related to the reference ISKNV with nucleotide sequence identity that was ranging from 99.8% to 100%. In addition, phylogenetic analysis of MCP gene revealed that viruses from genus Megalocytivirus can be divided into three genotypes: genotype 1 include reference ISKNV and all other strains that were detected in this study, genotype 2 include viruses closely related to red sea bream iridovirus (RSIV), and genotype 3 include viruses closely related turbot reddish body iridovirus (TRBIV).     

Use of spectral radiance for correcting nitrogen deficiencies and estimating soil test variability in an established bermudagrass pasture

The use of variable rate technology has become increasingly popular for applying plant nutrient elements. The most widely used method for determining variable fertilizer rates is presently based on soil testing and yield mapping. Three field studies (Bumeyville 1995, Burneyville 1996, and Ardmore 1996) were initiated in established Midland bermudagrass [Cynodon dacrylon (L) Pers.] pastures to determine the relationship between spectral radiance at specific wavelengths with forage nitrogen (N) removal and biomass, and to determine field variability of soil test parameters. Variable N (applied to 1.5 × 2.4 m subplots within 2.4 × 45.7 m main plots), fixed N and check treatments were evaluated at each location. Spectral radiance readings were taken in the red (671±6 nm), green (570±6 nm), and near infrared (NIR) (780±6 nm) wavelengths. The normalized difference vegetation index (NDVI) was calculated as NIR‐red/NIR+red. Variable N rates were applied based on NDVI. The highest fixed variable N rate was set at 224, 336, and 672 kg N ha^‐1 for Burneyville, 1995, 1996, and Ardmore, 1996, respectively. At Bumeyville, soil samples were collected in all variable rate plots (1.5 × 2.4 m) and analyzed for various soil test characteristics. NDVI, red, green, and NIR spectral radiance readings were correlated with bermudagrass forage N removal and yield. Correlation of forage yield and N removal with red, NIR, and NDVI were best with maximum forage production, however, when forage production levels were low correlation decreased dramatically for the red wavelength compared with NIR and NDVI. Forage yield and forage N removal in variable rate treatments increased when compared to the check while being equal to the half‐fixed and fixed rates where higher N rates were applied. Also, variability about the mean in variable rate plots was significantly lower than half‐fixed and fixed rates which supports adjusting N rates based on indirect NDVI measurements. Variable N rate plots reduced fertilizer inputs by 60% and produced the same yield as fixed rate plots, while fixed and half‐fixed rates did not increase N content in the forage over that of the variable rate treatment. Soil sample data collected from small consecutive plots (<4 m²) was extremely variable indicating that intense sampling would be needed if variable fertilizer application were to be based on soil test results.     

Performance and Kinetic Study on Bioremediation of Diazo Dye (Reactive Black 5) in Wastewater Using Spent GAC–Biofilm Sequencing Batch Reactor

Combinations of sequential anaerobic and aerobic process enhance the treatment of textile wastewater. The aim of this study was to investigate the treatment of diazo dye Reactive Black 5 (RB5)-containing wastewater using granular activated carbon (GAC)–biofilm sequencing batch reactor (SBR) as an integration of aerobic and anaerobic process in a single reactor. The GAC–biofilm SBR system demonstrated higher removal of COD, RB5 and aromatic amines. It was observed that the RB5 removal efficiency improved as the concentration of co-substrate in the influent increased. The alternative aeration introduced into the bioreactor enhanced mineralization of aromatic amines. Degradation of RB5 and co-substrate followed second-order kinetic and the constant (k ₂) values for COD and RB5 decreased from 0.002 to 0.001 and 0.004 to 0.001 l/mg h, respectively, as the RB5 concentration increased from 100 to 200 mg/l in the GAC–biofilm SBR system.     

Optimization of Agrobacterium-mediated transformation parameters for Melastomataceae spp. using green fluorescent protein (GFP) as a reporter

Agrobacterium-mediated transformation for both Melastoma malabathricum and Tibouchina semidecandra were optimized using green fluorescent protein (GFP) as a reporter. The binary vector pCAMBIA1304 harboring the modified green fluorescent protein (mgfp) gene driven by the CaMV 35S promoter was used. Parameters optimized were bacterial strain, bacterial concentration, pre-culture period, co-cultivation period, immersion time, acetosyringone concentration and wounding type. Results obtained were based on the percentage of GFP expression which was observed 3 days post-transformation. Agrobacterium tumefaciens strain LBA4404 and EHA105 at concentration 1 × 10⁷ cfu ml⁻¹ (OD_600nm 0.8) showed the highest virulence on M. malabathricum and T. semidecandra, respectively. Four days of pre-culture and 2 days of co-cultivation were optimum for M. malabathricum transformation, while 3 days of pre-culture and co-cultivation for T. semidecandra. Result also showed that 60 min of immersion and addition of 200 μM acetosyringone gave the highest percentage of positive transformants for both M. malabathricum and T. semidecandra. Mild wounding also significantly increased the efficiency of M. malabathricum transformation.     

Tree and crop productivity in gliricidia/maize/pigeonpea cropping systems in southern Malawi

This study examined the hypothesis that incorporation of Gliricidia sepium (Jacq.) Walp.) (gliricidia), a fast-growing, nitrogen-fixing tree, into agroforestry systems in southern Malawi may be used to increase the input of organic fertilizer and reduce the need for expensive inorganic fertilizers. The productivity of maize (Zea mays L.), pigeonpea (Cajanus cajan L.) and gliricidia grown as sole stands or in mixed cropping systems was examined at Makoka Research Station (latitude 15° 30′ S, longitude 35° 15′ E) and a nearby farm site at Nazombe between 1996 and 2000. Treatments included gliricidia intercropped with maize, with or without pigeonpea, and sole stands of gliricidia, maize and pigeonpea. Trees in the agroforestry systems were pruned before and during the cropping season to provide green leaf manure. Maize yields and biomass production by each component were determined and fractional light interception was measured during the reproductive stage of maize. Substantial quantities of green leaf manure (2.4 to 9.0 Mg ha⁻¹ year⁻¹) were produced from the second or third year after tree establishment. Green leaf manure and fuelwood production were greatest when gliricidia was grown as unpruned sole woodlots (c. 8.0 and 22 Mg ha⁻¹ year⁻¹ respectively). Improvements in maize yield in the tree-based systems also became significant in the third year, when c. 3.0 Mg ha⁻¹ of grain was obtained. Tree-based cropping systems were most productive and exhibited greater fractional light interception (c. 0.6 to 0.7) than cropping systems without trees (0.1 to 0.4). No beneficial influence of pigeonpea on maize performance was apparent either in the presence or absence of gliricidia at either site in most seasons. However, as unpruned gliricidia provided the greatest interception of incident solar radiation (>0.9), coppicing may be required to reduce shading when gliricidia is grown together with maize. As pigeonpea production was unaffected by the presence of gliricidia, agroforestry systems containing gliricidia might be used to replace traditional maize + pigeonpea systems in southern Malawi. This revised version was published online in June 2006 with corrections to the Cover Date.