The extent of genetic diversity among Vanilla species: Comparative results for RAPD and ISSR

Vanilla is a large genus of about 110 species in the orchid family (Orchidaceae), including the species Vanilla planifolia from which commercial vanilla flavoring is derived. Since most species of vanilla are considered rare and endangered there is an urgent need to conserve them through genetic analysis and propagation/conservation studies on this crop.The present study investigated the genetic diversity among nine leafy- and leaf-less Vanilla species employing 30 decamer RAPD primers and 10 ISSR primers. The species under study were diverse and displayed a range of variability (0–66% and 0–81% for RAPD and ISSR, respectively). A total of 154 RAPD polymorphic markers (83.24%, h = 0.378) and 93 ISSR polymorphic markers (86.11%, h = 0.363) were used to generate a genetic similarity matrix followed by the cluster analysis. Specific groupings were revealed by each cluster analysis with slight variation between two different markers. Among the nine species studied, V. planifolia, Vanilla aphylla and Vanilla tahitensis revealed very low level of variation within their collections, thus indicating a narrow genetic base. The large genetic distance of Vanilla andamanica from other species suggests its different origin. A close genetic affinity was observed between the pairs V. planifolia, V. tahitensis and Vanilla albida, V. aphylla. These are the first comparative results for RAPD and ISSR reporting inter-relationship among nine cultivated, wild and hybrid Vanilla species.     

Development and evaluation of an indirect enzyme-linked immunosorbent assay for detection of foot-and-mouth disease virus nonstructural protein antibody using a chemically synthesized 2B peptide as antigen.

Forty peptides were synthesized corresponding to hydrophilic clusters of amino acids within the sequences of foot-and-mouth disease virus (FMDV) nonstructural proteins (NSP). Six peptides were studied in more detail and the most promising, a 2B peptide, was evaluated in enzyme-linked immunosorbent assay (ELISA) using sera from naive, vaccinated, and vaccinated-and-challenged cattle as well as bovine sera from field outbreaks. The performance of the new NSP peptide ELISA was compared to that of 4 commercial NSP ELISA kits. Antibody to 2B was detectable from the end of the first week to the second week after infection in most of the nonvaccinated animals and by the second to third week in vaccinated-and-challenged animals. The sensitivity of the 2B peptide ELISA was comparable to the 3ABC Ceditest (Ceditest FMDV-NS, Cedi Diagnostics B.V.; Chung et al., 2002). With some modification and further validation, this 2B test could be useful as a screening or conformational NSP test in postvaccination surveillance for FMD.     

Fruit infestation of Acacia tortilis (Forsk) hyne by Bruchidius andrewesi Pic. (Coleoptera: Bruchidae) in the Thar desert

Bruchidius andrewesi Pic. has been recorded as a serious pest of pods and seeds of Acacia tortilis in the Thar desert of India. Pest infestation on developing pods and its relationship with morphological traits is reported. Pod infestation varied from 5 to 19% with 5–29% infestation of seeds. Infestation of pods is directly related to infestation of seed (r = 0.72^**), and both pod and seed infestation are also directly correlated with loss in seed biomass (r = 0.79^** and R = 0.88^**). The infestation of pods starts in November and increases steady until harvest. Seeds kept in the laboratory for further studies were found to be 100% infested with B. andrewesi, as the insect multiplied faster under these conditions. The heavy infestation is damaging not only to A. tortilis but also to other leguminous trees of the desert. Bruchidius andrewesi has also been found on pods and seeds of Prosopis cineraria, an important indigenous tree of the region.     

Genetic divergence of kingfish from Japan, Australia and New Zealand inferred by microsatellite DNA and mitochondrial DNA control region markers

ABSTRACT: Genetic polymorphism in kingfish, collected from coastal waters of Japan, Australia and New Zealand, were examined using microsatellite (MS) DNA and mitochondrial DNA (mtDNA) control region markers. Sixteen to 25.7 alleles per locus were observed in three MS markers, while the average observed (and expected) heterozygosities were 0.782 (0.918), 0.750 (0.809) and 0.650 (0.888) for Australian, Japanese and New Zealand kingfish, respectively. Twelve mtDNA haplotypes were detected by the digestion of control region sequences with five endonucleases: Hae III, Hinf I Mbo I, Rsa I and Taq I. Significant genetic divergence was observed between the kingfish population from Japan and those from Australia–New Zealand. There was no significant differentiation among the Australian and New Zealand population samples.     

Diofenolan: a novel insect growth regulator in common citrus butterfly, <Emphasis Type="Italic">Papilio demoleus</Emphasis>

Penultimate and last instar larvae of the common citrus swallowtail butterfly, Papilio demoleus L. (Papilionidae: Lepidoptera), were treated with different doses of a novel juvenoid, diofenolan. Several deformities were observed as a result of topical administration of diofenolan which include delay in larval–larval and larval–pupal ecdysis, ecdysial failure, mortality, severe reduction in pupation, deformed pupae and complete inhibition of adult emergence. The juvenoid diofenolan severely hampers the normal growth, development and metamorphosis of P. demoleus and can be incorporated into integrated pest management programs for successful control of this important pest of citrus all over the world.     

Ozone treatment of spent medium from Auricularia polytricha cultivation for enzymatic saccharification and subsequent ethanol production

We examined the enzymatic saccharification and ethanol fermentation of the spent media (SMs) from Auricularia polytricha cultivation using wood meals of Falcataria moluccana, Shorea sp., and Tectona grandis. Although the hydrolysis weight decrease and reducing sugar yield were higher in SM of F. moluccana, the ethanol yield was higher in SM of Shorea sp. Ozone treatment of SM further increased the hydrolysis weight decrease, reducing sugar, and ethanol yields in Shorea sp. These results indicate that SM of A. polytricha is a suitable biomass material to produce fermentable sugars for ethanol production, and that ozone treatment is a suitable method for increasing the ethanol yield.     

An investigation of the possible causes for the loss of productivity in genetically improved farmed tilapia strain in Fiji: inbreeding versus wild stock introgression

Four microsatellite markers and a mitochondrial DNA (mtDNA) fragment were used to investigate two possible explanations for a reported decline in productivity of genetically improved farmed tilapia (GIFT) in Fiji: (i) a decline in genetic diversity (GD) and (ii) genetic introgression from feral tilapia populations. Genetic diversity was estimated using θ and allelic richness, while Bayesian clustering was used to assign individuals to genetic groups (K=2 or 3) to test for introgression. Differentiation among groups was estimated using F_ST analysis. Results indicate that genetic diversity had declined compared with a GIFT reference stock from WorldFish Centre, while there was little evidence for introgression from feral tilapia populations. Loss of genetic diversity most probably resulted from practices that have not actively managed genetic resources in the hatchery. While GIFT is considered to be an improved line of Nile tilapia (Oreochromis niloticus), mtDNA analysis here revealed haplotypes assigned previously to three discrete Oreochromis species (O. niloticus, Oreochromis mossambicus and Oreochromis aureus) in both the Fijian strain and the WorldFish Centre strain. Possible sources for the three divergent lineages are discussed. Results have implications for the management and future expansion of the tilapia culture industry in Fiji as well as in other Pacific island nations.     

Effects of dietary protein on growth performance,nutrient utilization,digestive enzymes and physiological status of grey mullet,Mugil cephalus L. fingerlings reared in inland saline water

A 60 days feeding trial was conducted to illustrate the effect of graded levels of protein on the growth and metabolic enzymes of grey mullet (Mugil cephalus L.) fingerlings reared in inland saline water (ISW). Six isoenergetic (16 MJ/kg) and isolipidic (60 g/kg) diets containing 240, 260, 280, 300, 320 and 340 g crude protein (CP)/kg diet were formulated and fed to triplicate. Weight gain %, specific growth rate, protein utilizing efficiency, feed efficiency and RNA:DNA ratio were significantly higher (p < .05) in 320 and 300 g CP/kg diets. Fish fed with 240 g CP/kg diet showed significantly higher (p < .05) feed intake, whole‐body lipid content, hepatosomatic index value and liver glycogen content. Transaminase enzymes and malate dehydrogenase activities were elevated in fish fed 340 g CP/kg diet. Protease activity increased with increasing dietary CP level, but amylase activities showed an inverse relationship. No significant (p > .05) variations were observed for lactate dehydrogenase, oxidative stress enzymes, blood parameters and serum osmolality among all the treatment groups, but red blood cell count increases with increasing dietary CP levels. Based on the results, feeding dietary protein level of 300 g CP/kg is economically viable for rearing of grey mullet in ISW.     

Reconstructing historical trends of small pelagic fish in the Java Sea using standardized commercial trip based catch per unit of effort

Pelagic fish stocks in the Java Sea have been exploited intensively since the beginning of the 1970s. However, due to the effect of increased fishing power of the fleets and the changing nature of spatial and temporal distribution of the fishing effort, assessment of stock trends based on commercial data of catch and effort requires the standardization of both effort and catch data. Here we present the first attempt to reconstruct a standardized time series of commercial catch per unit of effort (CPUE) for the main pelagic species exploited by the purse seine fleet in the Java Sea. The results showed that all the stocks analysed have largely declined since the beginning of the 1990s. For bigeye scad, Indian mackerel and sardine, current estimates are only between 3 and 19% of the maximum observed value while round scads and spotted sardine biomass estimates lie between 18 and 34%. However, our assumptions about the effect of lamp power and level of creeping and the fact that the influence of fish density on catchability and the effect of targeting were not taken into account thus observed decline is a rather conservative estimate of the real decline of the stocks. If effective management actions are not put in place as a matter of urgency in the Java Sea for small pelagic fisheries, one runs the risk of adding those species to the long list of overexploited stocks of the world oceans.