Is acetyl/butyrylcholine specificity a marker for insecticide‐resistance mutations in insect acetylcholinesterase?

Substrate specificity has been widely studied in vertebrate cholinesterases and it has been shown that two phenylalanines in the acyl pocket of acetylcholinesterase govern the acceptance of the acetyl/butyryl moiety of the choline esters. As an insecticide‐resistance mutation has been evidenced in the acyl pocket of Drosophila melanogaster and Musca domestica acetylcholinesterase we investigated the possibility of linking changes in acetyl/butyrylthiocholine specificity with mutations in insect acetylcholinesterase. We thus analyzed the effect of 28 mutations in Drosophila enzyme on acetyl/butyrylthiocholine, N‐methyl/N‐propyl‐carbamates and ethyl/methyl‐paraoxon preference. It appeared that the highest changes on acetyl/butyrylthiocholine and N‐propyl/N‐methyl‐carbamates preference were due to mutations in the acyl pocket. Nevertheless, other insecticide‐resistance mutations, not located in the acyl pocket, also modified these substrate preferences. Moreover, the effect of mutations in the acyl pocket was hidden when some other insecticide‐resistance mutations were combined in the enzyme. Consequently, acetyl/butyrylthiocholine preference alteration cannot be used as a marker to localize a mutation in the insect AChE. © 2000 Society of Chemical Industry     

Evaluation of Degree of Elasticity and Other Mechanical Properties of Wheat Kernels

Three samples were selected representing bread, soft, and durum wheat. Uniaxial compression and stress relaxation tests were performed on wheat kernels. Force‐deformation curves from intact wheat grain typically exhibited at least two points of inflection (PI) at ≈0.1 and 0.2 mm displacement. The first PI is related to the mechanical properties of all the bran layers. The second PI (0.2 mm) seems to be the endosperm boundary near the aleurone layer. These structures had higher degree of elasticity (DE) compared to the inner endosperm (0.5–0.6 mm). Besides wheat class and specific structures of the caryopsis, moisture content is a prominent factor affecting the mechanical strength of kernels. Stress relaxation tests show that bread wheat kernels with 69.2% DE at 13% moisture decreased to 31.6% DE with additional 6% moisture content. Soft wheat kernels DE of 61.0% at 13% moisture decreased to 22.7% at 19.7% moisture. Stress relaxation revealed pronounced time‐dependence. However, the differences of stress values at 120–180 sec were not significant in all wheat classes and moisture contents evaluated. The stress values after 120 sec might be attributed to the elastic deformation of the kernels.     

Potential of indigenous Bacillus spp. as probiotic feed supplements in an extruded low‐fish‐meal diet for juvenile olive flounder,Paralichthys olivaceus

A 12‐week feeding trial was designed to assess the probiotic potential of indigenous Bacillus amyloliquefaciens and/or Bacillus subtilis singly or in combination with Bacillus licheniformis in an extruded feed for olive flounder (Paralichthys olivaceus) juveniles. A high fish meal (FM) diet (control) and a low‐FM diet containing an alternative protein blend (30% FM replacement, FM30) were formulated. Three other experimental diets were prepared by inclusion of B. amyloliquefaciens (BA), B. subtilis (BS), or a mixture of B. amyloliquefaciens, B. subtilis, and B. licheniformis (BASL) into FM30 diet, with a final concentration of 10⁶ CFU/g diet. Results indicated that the FM30 diet was well tolerated by flounder, and the overall performance was not affected by dietary treatments. Lysozyme activity and total immunoglobulin level were significantly reduced in flounders when fed with the FM30 diet compared with the BASL and BA diets, respectively. The Bacillus additives neither enriched the relative abundance of the corresponding Bacillus spp. in the relevant gut microbiota of olive flounder nor modulated the presumptive gene functions of the gut microbiome. Despite the absence of growth‐promoting effect, the tested probiotics could still be economically viable for use as immunostimulants in commercial flounder diets with partial FM replacement.     

Intimal thickening of coronary arteries in the rabbitfish,Chimaera monstrosa L. (Chondrichthyes: Holocephali)

This study is the first to report the occurrence of intimal thickening of coronary arteries in a holocephalan, namely the rabbitfish, Chimaera monstrosa. The sample studied consisted of five hearts from rabbitfish with body weights ranging between 12 g and 1116 g. The specimens were fixed in formalin, in methanol:acetone:water (MAW), or in paraformaldehyde and were examined by brightfield and polarization light microscopy. Coronary arteries from three larger animals displayed focal intimal thickenings, which were located in the left main coronary artery trunk, at the level of the cono‐ventricular junction, and in several intramyocardial ventricular arteries. The vascular changes were characterized by myointimal proliferation, breakage or absence of the inner elastic lamina, and, in one case, by increased collagen within the myointimal proliferation. Taking into account the severity of the coronary arterial changes, their location, and the body weight of the affected animals, we conclude that (1) these changes are age‐related in the rabbitfish and (2) local intense mechanical tension seems to be an important factor in their formation in this species.     

Mud crab pen culture: replacement of fish feed requirement and impacts on mangrove community structure

Brackishwater pond culture has been a major factor in mangrove loss in Southeast Asia, hence, the need to develop environment‐friendly technologies such as mud crab Scylla (Portunidae) culture in mangrove pens exists. This study evaluated the effects of mud crab netpen systems in central Philippines on mangrove macroflora, and the replacement of dietary fish with low‐cost pellets. Wild or hatchery‐sourced Scylla olivacea and Scylla serrata were stocked at 0.5–0.8 m⁻² in 167–200 m² nylon netpens (2.3 cm stretched mesh) in Avicennia‐dominated mangrove habitats. The feeding treatments were: (A) Zarraga: (1) no feeding (natural productivity), (2) no feeding for 1 month+supplementary feeding, (3) fish biomass and (4) low‐cost pellets, and (B) Batan: (1) fish biomass and (2) pellets+fish biomass. Feeds were given ad libitum twice daily. Growth and survival rates of S. olivacea in Zarraga pens were not significantly different among treatments, although crabs fed fish biomass had the highest survival, body weight and production. Similarly, growth and survival of S. serrata were not significantly different between the Batan treatments. Economic analysis of the latter gave a 38.5% return on investment (ROI) and 2.6 years payback period (PP) for pellets+fish biomass treatment compared with 27.5% ROI and 3.6 years PP for fish alone. Sensitivity analysis showed an improved economic performance of the pellets+fish biomass treatment by increasing the survival rate. Evaluation of mangrove community structure showed that crab culture reduced species diversity, numbers and biomass of seedlings and saplings, but not of mangrove trees. Therefore, mud crab pen culture is recommended for mangrove sites with mature trees, but not seedlings and saplings, and low‐cost pellets can reduce dependence on fish biomass.     

Long‐term survival and colony growth of Acropora palmata fragments transplanted by volunteers for restoration

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The potential of pet‐grade poultry by‐product meal to replace fish meal in the diet of the juvenile spotted rose snapper Lutjanus guttatus (Steindachner, 1869)

A 12‐week feeding trial was conducted to examine the replacement of fish meal with pet‐grade poultry by‐product meal (PBM‐PG) in the spotted rose snapper Lutjanus guttatus diet. Five experimental diets were formulated to contain graded levels of PBM‐PG at proportion of 250, 500, 75 or 900 g kg^?1. The control diet contained sardine fish meal as the main protein source. Four groups of 15 randomly assigned L. guttatus juveniles were fed to satiation 3 times day^?1. Except for the fish fed the PBM‐PG90 diet, the growth performance, survival and feed utilization efficiency of the experimental fish were not significantly lower than those of the control fish. The dietary level of PBM‐PG did significantly affect the haematological characteristics (P < 0.05). The dietary dry matter and protein apparent digestibility coefficients (ADCs) decreased with increasing dietary PBM‐PG. High values for lipid ADCs were observed in all diets, with significant differences among the dietary treatments. The fish whole‐body protein, moisture, lipid and ash contents were not affected by the inclusion of dietary PBM. These results indicate that high‐quality terrestrial PBM can successfully replace more than half of the marine fish meal protein in the L. guttatus diet.     

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).