The fishery and reproductive biology of Barking crayfish, Linuparus trigonus (Von Siebold, 1824) along Queenslands east coast

This paper describes the fishery and reproductive biology for Linuparus trigonus obtained from trawl fishermen operating off Queensland’s east coast, Australia. The smallest mature female lobster measured 59.8 mm CL, however, 50% maturity was reached between 80 and 85 mm CL. Brood fecundity (BF) was size dependent and ranged between 19,287 and 100,671 eggs in 32 females from 59.8 to 104.3 mm CL. The relationship was best described by the power equation BF = 0.1107*CL to the power of 2.9241 (r to the power of 2 = 0:74). Egg size ranged from 0.96 to 1.12 mm in diameter (mean = 1:02 (+or-) 0:01 mm). Egg weight and size were independent of lobster size. Length frequencies displayed multi-modal distributions.The percentage of female to male lobsters was relatively stable for small size classes (30 to 70 mm CL; 50.0 to 63.6% females), but female proportions rose markedly between 75 and 90 mm (72.2 to 85.4%) suggesting that at the onset of sexual maturity female growth rates are reduced. In size classes greater than 95 mm, males were numerically dominant. A description of the L. trigonus fishery in Queensland is also detailed.

Cytochrome P450 aromatase in grey mullet: cDNA and promoter isolation; brain, pituitary and ovarian expression during puberty.

In a study towards elucidating the role of aromatases during puberty in female grey mullet, the cDNAs of the brain (muCyp19b) and ovarian (muCyp19a) aromatase were isolated by RT-PCR and their relative expression levels were determined by quantitative real-time RT-PCR. The muCyp19a ORF of 1515 bp encoded 505 predicted amino acid residues, while that of muCyp19b was 1485 bp and encoded 495 predicted amino acid residues. The expression level of muCyp19b significantly increased in the brain as puberty advanced; however, its expression level in the pituitary increased only slightly with pubertal development. In the ovary, the muCyp19a expression level markedly increased as puberty progressed. The promoter regions of the two genes were also isolated and their functionality evaluated in vitro using luciferase as the reporter gene. The muCyp19a promoter sequence (650 bp) contained a consensus TATA box and putative transcription factor binding sites, including two half EREs, an SF-1, an AhR/Arnt, a PR and two GATA-3s. The muCyp19b promoter sequence (2500 bp) showed consensus TATA and CCAAT boxes and putative transcription binding sites, namely: a PR, an ERE, a half ERE, a SP-1, two GATA-binding factor, one half GATA-1, two C/EBPs, a GRE, a NFkappaB, three STATs, a PPAR/RXR, an Ahr/Arnt and a CRE. Basal activity of serially deleted promoter constructs transiently transfected into COS-7, [alpha]T3 and TE671 cells demonstrated the enhancing and silencing roles of the putative transcription factor binding sites. Quinpirole, a dopamine agonist, significantly reduced the promoter activity of muCyp19b in TE671. The results suggest tissue-specific regulation of the muCyp19 genes and a putative alternative promoter for muCyp19b.

Concordant molecular and phenotypic data delineate new taxonomy and conservation priorities for the endangered mountain yellow-legged frog

The mountain yellow-legged frog Rana muscosa sensu lato, once abundant in the Sierra Nevada of California and Nevada, and the disjunct Transverse Ranges of southern California, has declined precipitously throughout its range, even though most of its habitat is protected. The species is now extinct in Nevada and reduced to tiny remnants in southern California, where as a distinct population segment, it is classified as Endangered. Introduced predators (trout), air pollution and an infectious disease (chytridiomycosis) threaten remaining populations. A Bayesian analysis of 1901 base pairs of mitochondrial DNA confirms the presence of two deeply divergent clades that come into near contact in the Sierra Nevada. Morphological studies of museum specimens and analysis of acoustic data show that the two major mtDNA clades are readily differentiated phenotypically. Accordingly, we recognize two species, Rana sierrae, in the northern and central Sierra Nevada, and R. muscosa, in the southern Sierra Nevada and southern California. Existing data indicate no range overlap. These results have important implications for the conservation of these two species as they illuminate a profound mismatch between the current delineation of the distinct population segments (southern California vs. Sierra Nevada) and actual species boundaries. For example, our study finds that remnant populations of R. muscosa exist in both the southern Sierra Nevada and the mountains of southern California, which may broaden options for management. In addition, despite the fact that only the southern California populations are listed as Endangered, surveys conducted since 1995 at 225 historic (1899-1994) localities from museum collections show that 93.3% (n=146) of R. sierrae populations and 95.2% (n=79) of R. muscosa populations are extinct. Evidence presented here underscores the need for revision of protected population status to include both species throughout their ranges.

Molecular and serological detection of A. centrale- and A. marginale-infected cattle grazing within an endemic area

A reverse line blot hybridization (RLB) one-stage nested PCR (nPCR) for Anaplasma centrale and a nested PCR for Anaplasma marginale were used to detect infected cattle grazing within an endemic region in Israel. A novel set of PCR primers and oligonucleotide probes based on a 16S ribosomal RNA gene was designed for RLB detection of both Anaplasma species, and the performance of the molecular assays compared. The immunofluorescent antibody test (IFA) was used to detect antibodies to both Anaplasma species, whereas, a highly sensitive and specific competitive enzyme-linked immunosorbent assay (cELISA) was used to detect antibodies in A. centrale-vaccinated cattle. The RLB and the nested PCR procedures showed bacteremia with sensitivity of 50 infected erythrocytes per milliliter. Up to 93% of the A. centrale vaccinates carried specific antibodies that were detected by cELISA, and up to 71% of the vaccinated cattle were found to be naturally infected with A. marginale according to the PCR and the RLB assays. Nevertheless, no severe outbreaks of A. marginale infection occurred among vaccinated herds in this endemic region. It appears that both, molecular tools and serology are useful for evaluation of the vaccine efficacy. In the light of wide natural field infection with A. marginale, strong recommendations to continue the A. centrale vaccination program regime will continue until a new generation of non-blood-based vaccine will be developed.

Innovative management of herbicide resistant weeds - seed banks and soil biology

The project will evaluate seed bank depletion of key northern herbicide resistant weeds under different environments, cropping systems, crop agronomies and non-chemical control tactics. The project will also evaluate soil biology and seed bank relationships to explain differences in seed bank persistence.

Effects of light conditions and plant density on growth and reproductive biology of Cascabela thevetia (L.) Lippold

Cascabela thevetia (L.) Lippold (Apocynaceae) is an invasive woody weed that has formed large infestations at several locations in northern Australia. Understanding the reproductive biology of C. thevetia is vital to its management. This paper reports results of a shade house experiment that determined the effects of light conditions (100% or 30% of natural light) and plant densities (one, two, four or eight plants per plot) on the growth, time to flowering and seed formation, and monthly pod production of two C. thevetia biotypes (peach and yellow). Shaded plants were significantly larger when they reached reproductive maturity than plants grown under natural light. However, plants grown under natural light flowered earlier (268 days compared with 369 days) and produced 488 more pods per pot (a 5-fold increase) over 3 years. The yellow biotype was slightly taller at reproductive maturity but significantly taller and with significantly greater aboveground biomass at the end of the study. Both biotypes flowered at a similar time under natural light and low plant densities but the yellow biotype was quicker to seed (478 versus 498 days), produced significantly more pods (364 versus 203 pods) and more shoot growth (577 g versus 550 g) than the peach biotype over 3 years. Higher densities of C. thevetia tended to significantly reduce the shoot and root growth by 981 g and 714 g per plant across all light conditions and biotypes over 3 years and increase the time taken to flower by 140 days and produce seeds by 184 days. For land managers trying to prevent establishment of C. thevetia or to control seedling regrowth once initial infestations have been treated, this study indicates that young plants have the potential to flower and produce seeds within 268 and 353 days, respectively. However, with plant growth and reproduction most likely to be slower under field conditions, annual surveillance and control activities should be sufficient to find and treat plants before they produce seeds and replenish soil seed banks. The most at-risk part of the landscape may be open areas that receive maximum sunlight, particularly within riparian habitats where plants would consistently have more favourable soil moisture conditions.

Synthesis of Silica Vesicles with Controlled Entrance Size for High Loading, Sustained Release, and Cellular Delivery of Therapeutical Proteins

A rationally designed two-step synthesis of silica vesicles is developed with the formation of vesicular structure in the first step and fine control over the entrance size by tuning the temperature in the second step. The silica vesicles have a uniform size of ≈50 nm with excellent cellular uptake performance. When the entrance size is equal to the wall thickness, silica vesicles after hydrophobic modification show the highest loading amount (563 mg/g) towards Ribonuclease A with a sustained release behavior. Consequently, the silica vesicles are excellent nano-carriers for cellular delivery applications of therapeutical biomolecules.

Gene expression evidence for off-target effects caused by RNA interference-mediated gene silencing of Ubiquitin-63E in the cattle tick Rhipicephalus microplus.

Knowledge of cattle tick (Rhipicephalus (Boophilus) microplus; Acari: Ixodidae) molecular and cellular pathways has been hampered by the lack of an annotated genome. In addition, most of the tick expressed sequence tags (ESTs) available to date consist of similar to 50% unassigned sequences without predicted functions. The most common approach to address this has been the application of RNA interference (RNAi) methods to investigate genes and their pathways. This approach has been widely adopted in tick research despite minimal knowledge of the tick RNAi pathway and double-stranded RNA (dsRNA) uptake mechanisms. A strong knockdown phenotype of adult female ticks had previously been observed using a 594 bp dsRNA targeting the cattle tick homologue for the Drosophila Ubiquitin-63E gene leading to nil or deformed eggs. A NimbleGen cattle tick custom microarray based on the BmiGI.V2 database of R. microplus ESTs was used to evaluate the expression of mRNAs harvested from ticks treated with the tick Ubiquitin-63E 594 bp dsRNA compared with controls. A total of 144 ESTs including TC6372 (Ubiquitin-63E) were down-regulated with 136 ESTs up-regulated following treatment. The results obtained substantiated the knockdown phenotype with ESTs identified as being associated with ubiquitin proteolysis as well as oogenesis, embryogenesis, fatty acid synthesis and stress responses. A bioinformatics analysis was undertaken to predict off-target effects (OTE) resulting from the in silico dicing of the 594 bp Ubiquitin-63E dsRNA which identified 10 down-regulated ESTs (including TC6372) within the list of differentially expressed probes on the microarrays. Subsequent knockdown experiments utilising 196 and 109 bp dsRNAs, and a cocktail of short hairpin RNAs (shRNA) targeting Ubiquitin-63E, demonstrated similar phenotypes for the dsRNAs but nil effect following shRNA treatment. Quantitative reverse transcriptase PCR analysis confirmed differential expression of TC6372 and selected ESTs. Our study demonstrated the minimisation of predicted OTEs in the shorter dsRNA treatments (similar to 100-200 bp) and the usefulness of microarrays to study knockdown phenotypes.