Two independent LAMP assays for rapid identification of the serpentine leafminer, Liriomyza huidobrensis (Blanchard, 1926) (Diptera: Agromyzidae) in Australia.

Liriomyza huidobrensis is a leafminer fly and significant horticultural pest. It is a quarantine listed species in many countries and is now present as an established pest in Australia. Liriomyza huidobrensis uses a broad range of host plants and has potential for spread into various horticultural systems and regions of Australia. Rapid in-field identification of the pest is critically needed to assist efforts to manage this pest. Morphological identification of the pest is effectively limited to specialist examinations of adult males. Generally, molecular methods such as qPCR and DNA barcoding for identification of Liriomyza species require costly laboratory-based hardware. Herein, we developed two independent and rapid LAMP assays targeted to independently inherited mitochondrial and nuclear genes. Both assays are highly sensitive and specific to L. huidobrensis. Positive signals can be detected within 10 min on laboratory and portable real-time amplification fluorometers. Further, we adapted these assays for use with colorimetric master mixes, to allow fluorometer free in-field diagnostics of L. huidobrensis. Our LAMP assays can be used for stand-alone testing of query specimens and are likely to be essential tools used for rapid identification and monitoring of L. huidobrensis.

Molecular identification of hymenopteran parasitoids and their endosymbionts from agromyzids.

Three polyphagous pest Liriomyza spp. (Diptera: Agromyzidae) have recently invaded Australia and are damaging horticultural crops. Parasitic wasps are recognized as effective natural enemies of leafmining species globally and are expected to become important biocontrol agents in Australia. However, the hymenopteran parasitoid complex of agromyzids in Australia is poorly known and its use hindered due to taxonomic challenges when based on morphological characters. Here, we identified 14 parasitoid species of leafminers based on molecular and morphological data. We linked DNA barcodes (5' end cytochrome c oxidase subunit I (COI) sequences) to five adventive eulophid wasp species (Chrysocharis pubicornis (Zetterstedt), Diglyphus isaea (Walker), Hemiptarsenus varicornis (Girault), Neochrysocharis formosa (Westwood), and Neochrysocharis okazakii Kamijo) and two braconid species (Dacnusa areolaris (Nees) and Opius cinerariae Fischer). We also provide the first DNA barcodes (5' end COI sequences) with linked morphological characters for seven wasp species, with three identified to species level (Closterocerus mirabilis Edwards & La Salle, Trigonogastrella parasitica (Girault), and Zagrammosoma latilineatum Ubaidillah) and four identified to genus (Aprostocetus sp., Asecodes sp., Opius sp. 1, and Opius sp. 2). Phylogenetic analyses suggest C. pubicornis, D. isaea, H. varicornis, and O. cinerariae are likely cryptic species complexes. Neochrysocharis formosa and Aprostocetus sp. specimens were infected with Rickettsia. Five other species (Cl. mirabilis, D. isaea, H. varicornis, Opius sp. 1, and Opius sp. 2) were infected with Wolbachia, while two endosymbionts (Rickettsia and Wolbachia) co-infected N. okazakii. These findings provide background information about the parasitoid fauna expected to help control the leafminers.

Meschia brevirostris sp. nov., from New Caledonia (Hemiptera: Heteroptera: Lygaeoidea: Meschiidae).

A new species, Meschia brevirostris sp. nov. (Hemiptera: Heteroptera: Lygaeoidea: Meschiidae), is described from New Caledonia. Photographs and SEM micrographs of the male and female habitus, genital structures and selected morphological structures are presented.

Revision of a unique Australian leafhopper genus Stenopsoides Evans (Hemiptera: Cicadellidae: Idiocerinae: Macropsini).

The Australian genus Stenopsoides Evans (Idiocerinae: Macropsini) is revised. The type species, S. turneri Evans, is redescribed and three new species are added: S. newi Semeraro sp. nov., S. punctatus Semeraro sp. nov. and S. truncatus Semeraro sp. nov.. Species photographic plates, a distribution map and an illustrated key to the four species are provided. The possibility of parthenogenetic reproduction occuring in this genus is discussed.

Body weight, egg production, and egg quality traits of gray, brown, and white varieties of Japanese quail (Coturnix coturnix japonica) in coastal climatic condition of Odisha.

AIM: The present study was conducted to evaluate the performance of gray, brown, and white varieties of Japanese quail (Coturnix coturnix japonica) with respect to body weight, egg production, and egg quality traits in the coastal climatic condition of Odisha. MATERIALS AND METHODS: A total of 500-day-old straight run Japanese quail chicks of three varieties, viz., gray, brown, and white were randomly selected and reared in deep litter system at Central Poultry Development Organization, Eastern Region, Bhubaneswar. The weekly body weight of the birds was recorded till their egg production stage (up to 6 weeks of age). The average egg production was recorded every biweekly from 6(th) to 20(th) week. Exterior and interior quality of eggs from each variety was determined at 6 weeks of age. RESULTS: The initial average weekly body weight of three varieties did not differ (p>0.05) among the varieties. However, from 1(st) to 6(th) week significantly higher body weight was observed in gray than white and brown. Brown varieties had reached 50% egg production 1 week earlier than gray and white. Brown had higher peak hen day (HD) production or hen-housed egg production followed by white and gray. External quality such as: Egg weight, egg length, egg width, volume, shape index, shell weight, shell thickness depicted no significant difference among the varieties except circumference length and circumference width, which were significantly higher (p≤0.05) in gray varieties than brown varieties. Internal egg characteristics such as: Albumen length, albumen width, albumen height, albumen index, yolk length, yolk width, yolk height, yolk index, albumen weight, yolk weight, Haugh unit revealed no significance difference among the varieties. CONCLUSION: It may be summarized from the findings that gray excelled in body weight followed by white and brown. Egg production potential in terms of hen house egg production or HD egg production was higher for brown followed by white and gray in the coastal climatic condition of Odisha.

Barcoding Queensland Fruit Flies (Bactrocera tryoni): impediments and improvements.

Identification of adult fruit flies primarily involves microscopic examination of diagnostic morphological characters, while immature stages, such as larvae, can be more problematic. One of the Australia's most serious horticultural pests, the Queensland Fruit Fly (Bactrocera tryoni: Tephritidae), is of particular biosecurity/quarantine concern as the immature life stages occur within food produce and can be difficult to identify using morphological characteristics. DNA barcoding of the mitochondrial Cytochrome Oxidase I (COI) gene could be employed to increase the accuracy of fruit fly species identifications. In our study, we tested the utility of standard DNA barcoding techniques and found them to be problematic for Queensland Fruit Flies, which (i) possess a nuclear copy (a numt pseudogene) of the barcoding region of COI that can be co-amplified; and (ii) as in previous COI phylogenetic analyses closely related B. tryoni complex species appear polyphyletic. We found that the presence of a large deletion in the numt copy of COI allowed an alternative primer to be designed to only amplify the mitochondrial COI locus in tephritid fruit flies. Comparisons of alternative commonly utilized mitochondrial genes, Cytochrome Oxidase II and Cytochrome b, revealed a similar level of variation to COI; however, COI is the most informative for DNA barcoding, given the large number of sequences from other tephritid fruit fly species available for comparison. Adopting DNA barcoding for the identification of problematic fly specimens provides a powerful tool to distinguish serious quarantine fruit fly pests (Tephritidae) from endemic fly species of lesser concern.

Improvement of the anti-C3 activity of compstatin using rational and combinatorial approaches.

Compstatin is a 13-residue cyclic peptide that has the ability to inhibit the cleavage of C3 to C3a and C3b. The effects of targeting C3 cleavage are threefold, and result in hindrance of: (i) the generation of the pro-inflammatory peptide C3a, (ii) the generation of opsonin C3b (or its fragment C3d), and (iii) further complement activation of the common pathway (beyond C3) with the end result of the generation of the membrane attack complex. We will report on our progress on: (i) rational design of more active compstatin analogues based on the three-dimensional structure of compstatin, (ii) experimental combinatorial design based on the generation of a phage-displayed peptide library partially randomized with the implementation of structure-induced restraints, and (iii) theoretical combinatorial design based on a novel computational optimization method, structure-induced restraints and flexible structural templates. All three approaches have resulted in analogues with improved activities. Currently, the lead analogue has the sequence acetyl-I[CVYQDWGAHRC]T-NH(2) (where the brackets denote cyclization), and is 16-fold more active than the parent peptide. We will also report on our progress towards understanding the dynamic character of compstatin using molecular dynamics simulations. The identification of an ensemble of interconverting conformers of compstatin with variable populations is a first step towards the incorporation of dynamic elements in the design of new analogues using dynamics-activity relationships in addition to structure-activity relationships.

Discrete charge carrier transfer sites on proteins? A D.C. conductivity study on ultrathin protein films.

D.C. electrical conduction in thin protein films (200 mg/cm2; approximately 2 mu in thickness) of bovine serum albumin and its dinitrophenylated derivatives with different stoichiometric composition was investigated at 17, 20 and 23% relative humidities and at room temperature. Statistically significant decrease in conductivity due to derivatization was observed even at protein-2,4-dinitrophenol stoichiometry as low as 1:2. A charge injection mechanism based upon discrete charge carrier transfer sites could account for the observations. Analogous events may operate in cellular signaling and coding.

Charge-transfer complexes of some linear conjugated polyenes.

On adsorption of some electron-acceptor molecules on the solid films of all-trans-beta-carotene, beta-apo-8'-carotenal, astacene and methylbixin a new absorption band appears on the longer-wavelength side of the spectrum in addition to the original bands. The position of this new band is dependent on the electron affinity (EA) of the acceptor molecules, and the intensity of this band increases with the amount of adsorbed acceptor molecules. A linear relationship between the vmax. of the new band and EA was observed. The value of the ionization potential of the polyenes estimated from such linear relationship agrees satisfactorily with the value obtained by other methods. It has been concluded that the polyenes behave as electron donor and first form molecular charge-transfer complexes (of type [polyene . I2] with iodine) with electron acceptors, these finally dissociating to yield ionic complexes (of type [polyene . I+] with iodine).