Developmental changes in structure and polypeptide profile of the androgenic gland of the freshwater prawn Macrobrachium rosenbergii

Although the mechanism of sex-differentiation in crustaceans has yet to be defined, the androgenic gland (AG) is thought to be the exclusive organ that produces the androgenic hormone (AH) which induces male sexual development. This paper presents results of light and transmission electron microscopy and total protein analysis of androgenic glands from three male morphotypes (orange-claw, orange-blue-claw and blue-claw) of the freshwater prawn, Macrobrachium rosenbergii. Highest protein content (76 µg//AG) was found in the blue-claw morphotype as compared to the orange-blue-claw (45 µg/AG) and the orange-claw morphotype (19 µg/AG). Sodiumdodecyl sulfate polyacrylamide gel electrophoresis (SDS-page) analysis of the cell free extract of the AG from the three morphotypes revealed four polypeptides (16, 18, 23 and 26 Kd) which quantitatively increase from the sexually immature orange-claw to the sexually mature blue-claw morphotype. The 16 and 18 Kd polypeptides could be the AHs.     

Comparative evaluation of molecular diagnostic tests for Nucleospora salmonis and prevalence in migrating juvenile salmonids from the Snake River, USA

Nucleospora salmonis is an intranuclear microsporidian that primarily infects lymphoblast cells and contributes to chronic lymphoblastosis and a leukemia-like condition in a range of salmonid species. The primary goal of this study was to evaluate the prevalence of N. salmonis in out-migrating juvenile hatchery and wild Chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss from the Snake River in the U.S. Pacific Northwest. To achieve this goal, we first addressed the following concerns about current molecular diagnostic tests for N. salmonis: (1) nonspecific amplification patterns by the published nested polymerase chain reaction (nPCR) test, (2) incomplete validation of the published quantitative PCR (qPCR) test, and (3) whether N. salmonis can be detected reliably from nonlethal samples. Here, we present an optimized nPCR protocol that eliminates nonspecific amplification. During validation of the published qPCR test, our laboratory developed a second qPCR test that targeted a different gene sequence and used different probe chemistry for comparison purposes. We simultaneously evaluated the two different qPCR tests for N. salmonis and foundthat both assays were highly specific, sensitive, and repeatable. The nPCR and qPCR tests had good overall concordance when DNA samples derived from both apparently healthy and clinically diseased hatchery rainbow trout were tested. Finally, we demonstrated that gill snips were a suitable tissue for nonlethal detection of N. salmonis DNA in juvenile salmonids. Monitoring of juvenile salmonid fish in the Snake River over a 3-year period revealed low prevalence of N. salmonis in hatchery and wild Chinook salmon and wild steelhead but significantly higher prevalence in hatchery-derived steelhead. Routine monitoring of N. salmonis is not performed for all hatchery steelhead populations. At present, the possible contribution of this pathogen to delayed mortality of steelhead has not been determined.     

Genetic analysis of tropical quality protein maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm

Maize (Zea mays L.) is an important source of carbohydrates and protein in the diet in sub-Saharan Africa. The objectives of this study were to (i) estimate general (GCA) and specific combining abilities (SCA) of 13 new quality protein maize (QPM) lines in a diallel under stress and non-stress conditions, (ii) compare observed and predicted performance of QPM hybrids, (iii) characterize genetic diversity among the 13 QPM lines using single nucleotide polymorphism (SNP) markers and assess the relationship between genetic distance and hybrid performance, and (iv) assess diversity and population structure in 116 new QPM inbred lines as compared to eight older tropical QPM lines and 15 non-QPM lines. The GCA and SCA effects were significant for most traits under optimal conditions, indicating that both additive and non-additive genetic effects were important for inheritance of the traits. Additive genetic effects appeared to govern inheritance of most traits under optimal conditions and across environments. Non-additive genetic effects were more important for inheritance of grain yield but additive effects controlled most agronomic traits under drought stress conditions. Inbred lines CKL08056, CKL07292, and CKL07001 had desirable GCA effects for grain yield across drought stress and non-stress conditions. Prediction efficiency for grain yield was highest under optimal conditions. The classification of 139 inbred lines with 95 SNPs generated six clusters, four of which contained 10 or fewer lines, and 16 lines of mixed co-ancestry. There was good agreement between Neighbor Joining dendrogram and Structure classification. The QPM lines used in the diallel were nearly uniformly spread throughout the dendrogram. There was no relationship between genetic distance and grain yield in either the optimal or stressed environments in this study. The genetic diversity in mid-altitude maize germplasm is ample, and the addition of the QPM germplasm did not increase it measurably.     

Determination of residues of isofenphos and its phosphoramidate analogue

A method is described for the gas chromatographic determination of residues of isofenphos (BAY 92114, O-ethyl O-2-isopropoxycarbonylphenyl isopropylphosphor-amidothioate) and its phosphoramidate which is a potential metabolite. When potato, cabbage, rapeseed and a sandy loam soil were fortified with 0.01, 0.1 and 1.0 mg/kg and analysed by the method described, the recoveries ranged from 76 to 98%. The limits of detection of the method were 0.5 and 1.0 ng for isofenphos and isofenphos oxygen analogue respectively. Degradation under field conditions of isofenphos to its oxygen analogue was demonstrated in a sample of field-treated potatoes.