The Value of Tissue Imprint Hybridization for Rapid Detection of Infectious Bursal Disease Virus from Field Outbreaks

The use of a peroxidase labelled PCR generated probe followed by enhanced chemiluminescence hybridization assay detected infectious bursal disease virus directly from bursal imprints on a nylon membrane. Tissue imprint hybridization proved to be a simple, rapid and safe means of detecting IBD virus for screening large numbers of field samples. The PCR generated probe was highly specific for IBD virus and did not hybridize with cellular nucleic acids in control imprints. Tissue imprint hybridization was found to be a more sensitive method than conventional antigen detection assays.     

Quantifying the dietary biotin requirement of the catfish,Clarias batrachus

Asian catfish, Clarias batrachus, were fed semi-purified basaldiets containing 0, 0.1, 0.5, 1, 3 and 5 mg biotin kg^–1diet for 60 days. Fish fed the control diet (no biotin) showed(P < 0.05) higher mortality, lower weight gain, specificgrowth rate (SGR), feed efficiency ratio (FER) and protein efficiencyratio (PER) than in fish fed diets supplemented with biotin. The highestweight gain, SGR, FER and PER were noticed in fish fed 1 mg biotinkg^–1, followed by 0.5, 5, 3 and 0.1 mg biotinkg^–1, except for PER (followed by 0.5, 5, 0.1 and 3 mgbiotin kg^–1). Quadratic analysis showed that the optimumdietary biotin requirements for maximal weight gain, PER and PER were2.49, 2.54 and 2.52 mg kg^–1, respectively. Liver biotinconcentrations were influenced by levels of biotin in the diet.Concentration of liver biotin increased as level of dietarysupplementation increased and no biotin was detected in the liver of thecontrol fish. Liver pyruvate carboxylase and acetyl CoA carboxylaseactivities were higher in fish fed biotin-supplemented diets than incontrols. Biotin concentrations, pyruvate carboxylase and acetyl CoAcarboxylase activities in liver associated with normal growth rangedfrom 10.59 to 10.66 g g^–1, 147.97 to 148.18 units mgprotein^–1 and 12.76 to 12.78 units mg protein^–1, respectively. Biotin deficiency symptoms such as anorexia, darkskin colour and convulsions were observed in fish fed the control diet.The optimum dietary biotin requirement for maximal growth of C.batrachus is about 2.49 mg kg^–1 diet.     

Influence of a vesicular-arbuscular mycorrhizal fungus on the competitive ability of Bradyrhizobium spp. for nodulation of cowpea Vigna unguiculata (L.) Walp in non-sterilized soil

Summary We examined the influence of a vesicular-arbuscular (VAM) fungus (Glomus pallidum Hall) on the competitive ability of introduced and native Bradyrhizobium strains to nodulate cowpeas [Vigna unguiculata (L) Walp]. Our experiments in non-sterilized soil revealed that in the presence of VAM fungus, introduced Bradyrhizobium spp. strains become more competitive than native rhizobia. For example, strain JRC29 occupied 59.2% of the total nodules when inoculated alone, but this figure increased to 71.2% when JRC29 was used in dual inoculations with VAM fungus. A similar pattern of enhanced competitiveness for nodule formation was observed with the two other strains in the presence of the VAM fungus. Our results suggest that the competitiveness of rhizobia can be enhanced by co-inoculating with a selected strain of a VAM fungus.     

Resistance of pearlspot larvae,Etroplus suratensis,to redspotted grouper nervous necrosis virus by immersion challenge

Viral nervous necrosis (VNN) affects more than 120 species mostly belonging to the order Perciformes. However, none of the brackishwater species belonging to the family Cichlidae under the order Perciformes are reported to be susceptible. Hence, the present experiment was undertaken to study the susceptibility of the brackishwater cichlid, pearlspot, Etroplus suratensis to NNV. Thirty‐day‐old pearlspot larvae were infected with NNV by immersion. Mortality was recorded till 14 days post‐infection, and the infected larvae were subjected to nested RT‐PCR and histology. The virus was isolated from infected larvae using SSN‐1 cells. To study the replication of the virus in vitro, primary cultured brain cells of E. suratensis and IEK cells were infected with NNV. No mortality was observed in any of the control or experimentally infected larvae. However, the experimentally infected larvae were positive for NNV by nested RT‐PCR and the virus was isolated using SSN‐1 cells. Further, the infected pearlspot brain cells and IEK cells showed cytopathic effect at second and third passage of the virus and they were positive for NNV by nested RT‐PCR. Pearlspot is relatively resistant to VNN although the virus could replicate in the larvae and in cell culture.     

Calcium phosphate coupled Newcastle disease vaccine elicits humoral and cell mediated immune responses in chickens

Calcium phosphate (CaP) particles were coupled with inactivated Newcastle disease virus (NDV) vaccine. The surface morphology of CaP particles coupled to NDV was found to be spherical, smooth and with a tendency to agglomerate. The mean (± SE) size of CaP particles was found 557.44 ± 18.62 nm. The mean percent encapsulation efficiency of CaP particles coupled to NDV assessed based on total protein content and haemagglutination (HA) activity in eluate was found to be 10.72 ± 0.89 and 12.50 ± 2.09, respectively. The humoral and cell mediated immune responses induced by CaP coupled NDV vaccine were assessed in comparison to a commercial live vaccine (RDV ‘F’). CaP coupled NDV vaccine elicited prolonged haemagglutination inhibition (HI) and enzyme linked immunosorbent assay (ELISA) titres in the serum even at fourth and fifth week post-vaccination (PV), unlike RDV ‘F’ inoculated chickens whose titres declined to insignificant levels by this time. CaP coupled NDV vaccine could stimulate HI antibodies in tracheal washings and tears from second and first week PV, respectively. IgA ELISA antibodies were also seen in tracheal washings of these birds from third week PV and in tears from second week PV. CaP coupled NDV vaccine elicited cell mediated immune responses (CMI) from two to four weeks PV. The stimulation indices obtained after stimulation with specific antigen was not significantly different between CaP coupled antigen and live NDV virus except on first week PV. However, CaP coupled antigen did not cause suppression of lympo proliferation as indicated by statistically similar responses to mitogen, concanavalin A between the two groups. Overall, CaP coupled NDV vaccine elicited stronger and prolonged immune responses in comparison to the commercial live vaccine. No increase in the serum calcium and phosphorous levels were seen in CaP coupled NDV vaccine inoculated chickens.     

Discharge of Escherichia Coli from Agricultural Surface and Subsurface Drainage Water: Tillage Effects

Drainage water from agricultural fields with applied manure can degrade the bacterial quality of surface and groundwater. The impact of conventional tillage (CT) and zero tillage (ZT) practices on Escherichia coli (E.coli) discharge through artificially drained soils is not well understood. Consequently, two field trials were conducted during 2002–2004. The first trial involved fall applications of beef manure while the second involved spring applications of dairy manure. Both surface and subsurface drainage water were monitored in the first trial while only subsurface drainage water was monitored in the second. Under fall applied beef manure (trial 1), no differences (p?>?0.05) were observed in E.coli concentrations (cfu/100 ml) in combined drainage water under both tillage systems. However, during 2003–2004, subsurface drainage water under ZT had higher E.coli concentrations and loads than drainage water under CT. When the combined (surface + subsurface) annual E.coli loads were considered, CT loads were greater than ZT during 2002–2003 with an opposite situation during 2003–2004. Overall, annual E.coli loads were similar under ZT (4.7?×?10¹⁰ cfu/ha) and CT (4.8?×?10¹⁰ cfu/ha). Spring dairy manure application (trial 2) produced significant (p?>?0.03) tillage effect on E.coli loads in subsurface drainage water only during the second year. During the study period, ZT plots (1.55?×?10¹⁰ cfu/ha) discharged 5× more E.coli than CT (0.23?×?10¹⁰ cfu/ha). A longer duration of ZT practices resulted in higher subsurface flow volumes and subsequently greater loads of E.coli discharge in both trials.     

Phosphatase activity and cytokinin content in cowpeas (Vigna unguiculata) inoculated with a vesicular-arbuscular mycorrhizal fungus

We examined the effect of a vesicular-arbuscular mycorrhizal (VAM) fungus Glomus pallidum Hall on the phosphatase activity and cytokinin concentration in cowpea [Vigna unguiculata (L.) Walp] roots at successive stages of plant growth. Both acid and alkaline phosphatase activity were significantly (P=0.05) higher in mycorrhizal than in non-mycorrhizal roots 30 days after inoculation. Similarly, the cytokinin content was significantly increased in mycorrhizal roots compared to non-mycorrhizal roots. Our study suggests that these biochemical changes may improve the growth of mycorrhizal cowpea plants.     

Co-selection of compatible rhizobia and vesicular-arbuscular mycorrhizal fungi for cowpea in sterilized and non-sterilized soils

Summary We selected two isolates of Rhizobium for cowpea (Vigna unguiculata) with sterilized soil tests and two different isolates by non-sterilized soil testing. The four rhizobia were then paired individually with either Glomus pallidum, Glomus aggregatum, or Sclerocystis microcarpa in separate, sterilized, or non-sterilized soil experiments. The purpose of the experiments was to determine the effect of soil sterilization on the selection of effective cowpea rhizobia, and to see whether these rhizobia differed in their effects on cowpea growth when paired with various vesicular-arbuscular mycorrhizal (VAM) fungi. Our experiments showed that the rhizobia selected in sterilized soil tests produced few growth responses in the cowpea compared to the other introduced rhizobia, irrespective of pairing with VAM fungi in sterilized or non-sterilized soil. In contrast, the two rhizobia initially selected by non-sterilized soil testing significantly improved cowpea growth in non-sterilized soil, especially when paired with G. pallidum. Our results suggest that it is important to select for effective rhizobia in non-sterilized soil, and that pairing these rhizobia with specific, coselected VAM fungi can significantly improve the legume growth response.     

Assessment of the new generation aeration systems efficiency and water current flow rate,its relation to the cost economics at varying salinities for Penaeus vannamei culture

The selection of aerators and correct numbers can play an essential role in reducing the cost of production in aquaculture. The new generation aerators, namely spiral leaf, air‐jet, submersible and impeller, used in aquaculture, were assessed for its aeration efficiency and energy cost compared with the commonly used paddle wheel aerator. Of the aerators tested, the impeller had the highest aeration efficiency of 2.098 kg O₂/kW hr, followed by paddle wheel with 1.436 kg O₂/kW hr at 20‰ salinity. The spiral and air‐jet aerators had maximum aeration efficiency of 1.326 and 1.419 kg O₂/kW hr, respectively, at 35‰ salinity. The submersible aerator was not efficient as its maximum efficiency was 0.380 kg O₂/kW hr. The water flow by paddle wheel was 3 ft/sec and also provides better coverage than other types of aerators. The efficiency of aerators was high in optimum salinities (20‰ and 35‰) than the low or high saline condition. The average energy cost of shrimp pond aeration per hectare was lowest for impeller, followed by paddle wheel aerator. The study provided economic comparisons of vannamei culture using different aeration systems by keeping a uniform set of economic assumptions. Shrimp farms with impellors can give 14%–25% high returns across salinities, whereas spiral leaf can provide 5% high returns at 35‰ salinity. The combination of the type of aerators and the calculated use based on the salinity of the culture systems can result in energy‐saving and also a reduction in the production cost.