Studies on "Sappe" disease of the silkworm, Bombyx mori : II. Effect of age of larvae on the manifestation of the disease

The effect of age of the larvae on the manifestation of the "Sappe" disease of the silkworm by oral inoculation of different pathogens, viz., Aerobacter cloacae, Pseudomonas boreopolis, Escherichia freundii, Achromobacter delmarvae, A. Superficialis, Pseudomonas ovalis, and Staphylococcus albus was tested. It was found that the reaction of the larva to the pathogen was influenced by its age. Some, e.g., Escherichia freundii, were more lethal when introduced at early stages whereas certain others, e.g., Aerobacter cloacae and Staphylococcus albus, caused maximum damage when invading older larvae. Irrespective of the age of infection, death of the worms mainly occurred during molting and before spinning. The studies also indicated that growth and mortality of the larvae were affected differentially by the pathogens.

Queensland Spanner Crab Fishery: Commercial quota setting for June 2016 – May 2018

The Queensland (QLD) fishery for spanner crabs primarily lands live crab for export overseas, with gross landings valued around A$5 million per year. Quota setting rules are used to assess and adjust total allowable harvest (quota) around an agreed target harvest of 1631 t and capped at a maximum of 2000 t. The quota varies based on catch rate indicators from the commercial fishery and a fishery independent survey. Quota management applies only to ‘Managed Area A’ which includes waters between Rockhampton and the New South Wales (NSW) border. This report has been prepared to inform Fisheries Queensland (Department of Agriculture and Fisheries) and stakeholders of catch trends and the estimated quota of spanner crabs in Managed Area A for the forthcoming annual quota periods (1 June 2016–31 May 2018). The quota calculations followed the methodology developed by the crab fishery Scientific Advisory Group (SAG) between November 2007 and March 2008. The QLD total reported spanner crab harvest was 1170 t for the 2015 calendar year. In 2015, a total of 55 vessels were active in the QLD fishery, down from 262 vessels at the fishery’s peak activity in 1994. Recent spanner crab harvests from NSW waters average about 125 t per year, but fell to 80 t in 2014–2015. The spanner crab Managed Area A commercial standardised catch rate averaged 0.818 kg per net-lift in 2015, 22.5% below the target level of 1.043. Compared to 2014, mean catch rates in 2015 were marginally improved south of Fraser Island. The NSW–QLD survey catch rate in 2015 was 20.541 crabs per ground-line, 33% above the target level of 13.972. This represented an increase in survey catch rates of about four crabs per groundline, compared to the 2014 survey. The QLD spanner crab total allowable harvest (quota) was set at 1923 t in the 2012-13 and 2013-14 fishing years, 1777 t in 2014-15 and 1631 t in 2015-16. The results from the current analysis rules indicate that the quota for the next two fishing years be retained at the base quota of 1631 t.

Queensland Spanner Crab Fishery : Commercial quota setting for June 2015 – May 2016

The Australian fishery for spanner crabs is the largest in the world, with the larger Queensland (QLD) sector’s landings primarily exported live overseas and GVP valued ~A$5 million per year. Spanner crabs are unique in that they may live up to 15 years, significantly more than blue swimmer crabs (Portunus armatus) and mud crabs (Scylla serrata), the two other important crab species caught in Queensland. Spanner crabs are caught using a flat net called a dilly, on which the crabs becoming entangled via the swimming legs. Quota setting rules are used to assess and adjust total allowable harvest (quota) around an agreed target harvest of 1631 t and capped at a maximum of 2000 t. The quota varies based on catch rate indicators from the commercial fishery and a fishery-independent survey from the previous two years, compared to target reference points. Quota management applies only to ‘Managed Area A’ which includes waters between Rockhampton and the New South Wales (NSW) border. This report has been prepared to inform Fisheries Queensland (Department of Agriculture and Fisheries) and stakeholders of catch trends and the estimated quota of spanner crabs in Managed Area A for the forthcoming quota period (1 June 2015–31 May 2016). The quota calculations followed the methodology developed by the crab fishery Scientific Advisory Group (SAG) between November 2007 and March 2008. The total reported spanner crab harvest was 917 t for the 2014 calendar year, almost all of which was taken from Managed Area A. In 2014, a total of 59 vessels were active in the QLD fishery, the lowest number since the peak in 1994 of 262 vessels. Recent spanner crab harvests from NSW waters have been about 125 t per year. The spanner crab Managed Area A commercial standardised catch rate averaged 0.739 kg per net-lift in 2014, 24% below the target level of 1.043. Mean catch rates declined in the commercial fishery in 2014, although the magnitude of the decreases was highest in the area north of Fraser Island. The NSW–QLD survey catch rate in 2014 was 16.849 crabs per ground-line, 22% above the target level of 13.972. This represented a decrease in survey catch rates of 0.366 crabs per ground-line, compared to the 2013 survey. The Queensland spanner crab total allowable harvest (quota) was set at 1923 t in 2012 and 2013. In 2014, the quota was calculated at the base level of 1631 t. However, given that the 2012 fisheryindependent survey was not undertaken for financial reasons, stakeholders proposed that the total allowable commercial catch (TACC) be decreased to 1777 t; a level that was halfway between the 2012/13 quota of 1923 t and the recommended base quota of 1631 t. The results from the current analysis indicate that the quota for the 2015-2016 financial year be decreased from 1777 t to the base quota of 1631 t.

Expression of Human Growth Hormone in Silkworm Larvae through RecombinantBombyx moriNuclear Polyhedrosis Virus

We have generated a recombinantBombyx morinuclear polyhedrosis virus, vBmhGH, harboring the full-length human growth hormone gene (2.4-kb genomic DNA, with four introns and the signal peptide sequences) under the control of the polyhedrin promoter. BmN cells in culture infected with the recombinant virus showed the presence of RNA corresponding to the authentic growth hormone mRNA as well as its incompletly processed precusor. Electrophoretic analysis and immunoprecipitation of proteins of recombinant virus-infected BmN cells revealed the presence of the growth hormone protein. Infection of silkworm larvae with vBmhGH led to the synthesis and efficient secretion of the protein into hemolymph. The recombinant human growth hormone was biologically active in a radioreceptor competition binding assay. The secreted protein was isolated and purified to homogeneity by a single step immunoaffinity chromatography, to a specific activity of 2.4 × 104U/mg. The recombinant hGH retained the immunological and biolological properties of the native peptide. We conclude that BmNPV vectors can be used successfully for expressing chromosomal genes harboring multiple introns.

Baculovirus mediated high-level expression of luciferase in silkworm cells and larvae

Bombyx mori nuclear polyhedrosis virus (BmNPV)-based baculovirus expression system exploits silkworm larvae as an economical alternative to large-scale cell cultures for production of biomolecules. To generate recombinant BmNPV at high efficiency, we have achieved high efficiency transfection of B. mori cells, BmN, through lipofection. Optimal conditions for lipofection were standardized by quantification of the transient expression level of firefly luciferase (luc) reporter gene under control of an immediate early gene promoter of BmNPV Lipofection was 50-fold and 100-fold more efficient than the calcium phosphate method for transfecting BmN and Sf9 cells, respectively. Lipofection enabled us to generate a recombinant BmNPV (vBmluc), harboring luc under control of the strong polyhedrin promoter On infection with vBmluc, luciferase was expressed at very high levels, 170 mu g/10(6) BmN cells or 13 mg/larva. Expression of luciferase in vBmluc-infected larvae was visualized by luminescence emission instantaneously following luciferin injection generating ''glowing silkworms''.

Effects of temperature on complexes I and II mediated respiration, ROS generation and oxidative stress status in isolated gill mitochondria of the mud crab Scylla serrata

Effects of fluctuations in habitat temperature (18-30 degrees) on mitochondrial respiratory behavior and oxidative metabolic responses in the euryhaline ectotherm Scylla serrate are not fully understood. In the present study, effects of different temperatures ranging from 12 to 40 degrees C on glutamate and succinate mediated mitochondrial respiration, respiratory control ratio (RCR), ATP generation rate, ratio for the utilization of phosphate molecules per atomic oxygen consumption (P/O), levels of lipid peroxidation and H2O2 in isolated gill mitochondria of S. serrata are reported. The pattern of variation in the studied parameters was similar for the two substrates at different temperatures. The values recorded for RCR ( >= 3) and P/O ratio (1.4-2.7) at the temperature range of 15-25 degrees C were within the normal range reported for other animals (3-10 for RCR and 1.5-3 for P/O). Values for P/O ratio, ATP generation rate and RCR were highest at 18 degrees C when compared to the other assay temperatures. However, at low and high extreme temperatures, i.e. at 12 and 40 degrees C, states III and IV respiration rates were not clearly distinguishable from each other indicating that mitochondria were completely uncoupled. Positive correlations were noticed between temperature and the levels of both lipid peroxidation and H2O2. It is inferred that fluctuations on either side of ambient habitat temperature may adversely influence mitochondria respiration and oxidative metabolism in S. serrata. The results provide baseline data to understand the impacts of acute changes in temperature on ectotherms inhabiting estuarine or marine environments. (C) 2014 Elsevier Ltd. All rights reserved.

A new genus and species of arboreal toad with phytotelmonous larvae, from the Andaman Islands, India (Lissamphibia, Anura, Bufonidae)

A new bufonid amphibian, belonging to a new monotypic genus, is described from the Andaman Islands, in the Bay of Bengal, Republic of India, based on unique external morphological and skeletal characters which are compared with those of known Oriental and other relevant bufonid genera. Blythophryne gen. n. is distinguished from other bufonid genera by its small adult size (mean SVL 24.02 mm), the presence of six presacral vertebrae, an absence of coccygeal expansions, presence of an elongated pair of parotoid glands, expanded discs at digit tips and phytotelmonous tadpoles that lack oral denticles. The taxonomic and phylogenetic position of the new taxon (that we named as Blythophryne beryet gen. et sp. n.) was ascertained by comparing its 12S and 16S partial genes with those of Oriental and other relevant bufonid lineages. Resulting molecular phylogeny supports the erection of a novel monotypic genus for this lineage from the Andaman Islands of India.

The mechanical effects of water flow on fish eggs and larvae

The impact of mechanical stresses upon ichthyoplankton entrained in power plant cooling systems has long been considered negligible. Arguments and evidence exist, however, to show that such a supposition is not universally true, especially in nuclear power plants. The mechanisms of mechanical damage can be detailed in terms of pressure change, acceleration, and shear stress with in the fluid flow field. Laboratory efforts to quantify the effects of mechanical stress have been very sparse. A well-planned bioassay is urgently needed. (PDF has 11 pages.)

Effects of shear on eggs and larvae of striped bass, morone saxatilis, and white perch, M. americana

Shear stress, generated by water movement, can kill fish eggs and larvae by causing rotation or deformation. Through the use of an experimental apparatus, a series of shear (as dynes/cm2)-mortality equations for fixed time exposures were generated for striped bass and white perch eggs and larvae. Exposure of striped bass eggs to a shear level of 350 dynes/cm2 kills 36% of the eggs in 1 min; 69% in 2 min, and 88% in 4 min; exposure of larvae to 350 dynes/cm2 kills 9.3% in 1 min, 30.0% in 2 min, and 68.1% in 4 min. A shear level of 350 dynes/cm2 kills 38% of the white perch eggs in 1 min, 41% in 2 min, 89% in 5 min, 96% in 10 min, and 98% in 20 min. A shear level of 350 dynes/cm2 applied to white perch larvae destroys 38% of the larvae in 1 min, 52% in 2 min, and 75% in 4 min. Results are experimentally used in conjunction with the determination of shear levels in the Chesapeake and Delaware Canal and ship movement for the estimation of fish egg and larval mortalities in the field.