Gene Expression Profiling of the Cephalothorax and Eyestalk in Penaeus Monodon during Ovarian Maturation.

In crustaceans, a range of physiological processes involved in ovarian maturation occurs in organs of the cephalothorax including the hepatopancrease, mandibular and Y-organ. Additionally, reproduction is regulated by neuropeptide hormones and other proteins released from secretory sites within the eyestalk. Reproductive dysfunction in captive-reared prawns, Penaeus monodon, is believed to be due to deficiencies in these factors. In this study, we investigated the expression of gene transcripts in the cephalothorax and eyestalk from wild-caught and captive-reared animals throughout ovarian maturation using custom oligonucleotide microarray screening. We have isolated numerous transcripts that appear to be differentially expressed throughout ovarian maturation and between wild-caught and captive-reared animals. In the cephalothorax, differentially expressed genes included the 1,3-beta-D-glucan-binding high-density lipoprotein, 2/3-oxoacyl-CoA thiolase and vitellogenin. In the eyestalk, these include gene transcripts that encode a protein that modulates G-protein coupled receptor activity and another that encodes an architectural transcription factor. Each may regulate the expression of reproductive neuropeptides, such as the crustacean hyperglycaemic hormone and molt-inhibiting hormone. We could not identify differentially expressed transcripts encoding known reproductive neuropeptides in the eyestalk of either wild-caught or captive-reared prawns at any ovarian maturation stage, however, this result may be attributed to low relative expression levels of these transcripts. In summary, this study provides a foundation for the study of target genes involved in regulating penaeid reproduction.

Follicle stimulating hormone secretion and dominant follicle growth during treatment of Bos indicus heifers with intra-vaginal progesterone releasing devices, oestradiol benzoate, equine chorionic gonadotrophin and prostaglandin F-2 alpha

The aim of this study was to investigate the effects on follicle stimulating hormone (FSH) secretion and dominant follicle (OF) growth, of treatment of Bos indicus heifers with different combinations of intra-vaginal progesterone releasing devices (IPRD), oestradiol benzoate (ODB), PGF(2 alpha), and eCG. Two-year-old Brahman (BN; n=30) and Brahman-cross (BNX; n=34) heifers were randomly allocated to three IPRD-treatments: (i) standard-dose IPRD [CM 1.56 g; 1.56 g progesterone (P-4); n = 17]; (ii) half-dose IPRD (CM 0.78 g; 0.78 g p(4); n=15); (iii) half-dose IPRD + 300 IU eCG at IPRD removal (CM 0.78 g+G; n=14); and, (iv) non-IPRD control (2 x PGF(2 alpha); n=18) 500 mu g cloprostenol on Days -16 and -2. IPRD-treated heifers received 250 mu g PGF(2 alpha) at IPRD insertion (Day 10) and IPRD removal (Day -2) and 1 mg ODB on Day -10 and Day -1. Follicular dynamics were monitored daily by trans-rectal ultrasonography from Day -10 to Day 1. Blood samples for determination of P-4 were collected daily and samples for FSH determination were collected at 12 h intervals from Day -9 to Day -2. A significant surge in concentrations of FSH was observed in the 2 x PGF(2 alpha), treatment 12 h prior and 48 h after follicular wave emergence, but not in the IPRD-treated heifers. Estimated mean concentrations of total plasma P-4 during the 8 days of IPRD insertion was greater (P<0.001) in the CM 1.56 g P-4 treated heifers compared to the CM 0.78 g P-4 treated heifers (18.38 ng/ml compared with 11.09 ng/ml, respectively). A treatment by genotype interaction (P=0.036) was observed in the mean plasma P4 concentration in heifers with no CL during IPRD insertion, whereby BN heifers in the CM 1.56 g treatment had greater plasma P-4 than the BNX heifers on Days-9, -7, -6, -5, and -4. However, there was no genotype effect in the CM 0.78 g +/- G or the 2 x PGF(2 alpha) treatment. Treatment had no effect on the DF growth from either day of wave emergence (P=0.378) or day of IPRD removal (P=0.780) to ovulation. This study demonstrates that FSH secretion in B. indicus heifers treated with a combination of IPRD's and ODB to synchronise ovulation was suppressed during the period of IPRD insertion but no significant effect on growth of the DF was observed. (C) 2013 Elsevier B.V. All rights reserved.

Maturation studies of pecan nuts grown in Queensland

Changes in chemical composition, physical and sensory characteristics were followed in two pecan cultivars Wichita and Western Schley harvested from a commercial orchard at Gatton in Queensland seven times during 1996.Testa colour of both pecan cultivars darkened and opalescence decreased as the nuts matured. Bitterness of Western Schley pecans decreased with maturity. Colour of shuck, shell and kernel of both cultivars developed as the nuts matured. Wichita pecans were larger than Western Schley at all harvest times. Both nut-in-shell and kernel moisture decreased with maturity, whereas oil and sucrose contents increased. Both pecan cultivars had reached advanced maturation by the first harvest on March 18.

Regulation of Gonadotropin-Releasing Hormone (GnRH)-Receptor Gene Expression in Tilapia: Effect of GnRH and Dopamine

The present work was designed to study certain aspects of the endocrine regulation of gonadotropin-releasing hormone receptor (GnRH-R) in the pituitary of the teleost fish tilapia. A GnRH-R was cloned from the pituitary of hybrid tilapia (taGnRH-R) and was identified as a typical seven-transmembrane receptor. Northern blot analysis revealed a single GnRH-R transcript in the pituitary of approximately 2.3 kilobases. The taGnRH-R mRNA levels were significantly higher in females than in males. Injection of the salmon GnRH analog (sGnRHa; 5–50 μg/kg) increased the steady-state levels of taGnRH-R mRNA, with the highest response recorded at 25 μg/kg and at 36 h. At the higher dose of sGnRHa (50 μg/kg), taGnRH-R transcript appeared to be down-regulated. Exposure of tilapia pituitary cells in culture to graded doses (0.1–100 nM) of seabream (sbGnRH = GnRH I), chicken II (cGnRH II), or salmon GnRH (sGnRH = GnRH III) resulted in a significant increase in taGnRH-R mRNA levels. The highest levels of both LH release and taGnRH-R mRNA levels were recorded after exposure to cGnRH II and the lowest after exposure to sbGnRH. The dopamine-agonist quinpirole suppressed LH release and mRNA levels of taGnRH-R, indicating an inhibitory effect on GnRH-R synthesis. Collectively, these data provide evidence that GnRH in tilapia can up- regulate, whereas dopamine down-regulates, taGnRH-R mRNA levels.

Spatio-temporal patterns in maturation of the chokka squid (Loligo vulgaris reynaudii) off the coast of South Africa

Knowledge of the temporal and spatial characteristics of chokka squid (Loligo vulgaris reynaudii) biology in South African waters is limited, so the possibility of there being a geographically fragmented stock was examined by investigating the distribution of maturity patterns for the species, covering all known spawning areas and using both historical and recent data. Gonadosomatic indices (GSI) varied between year-round consistency and apparent seasonal peaks in both summer and winter; there was no clear spatial pattern. Monthly percentage maturity provided further evidence for two peak reproductive periods each year, although mature squid were present throughout. Sex ratios demonstrated great variability between different areas and life history stages. Male-biased sex ratios were only apparent on the inshore spawning grounds and ranged between 1.118:1 and 4.267:1. Size at sexual maturity was also seasonal, squid maturing smaller in winter/spring than in summer/autumn. Also, squid in the east matured smaller than squid in the west. Although the results from the present study do not provide conclusive evidence of distinct geographic populations, squid likely spawn over a significantly larger area of the Agulhas Bank than previously estimated, and squid on the west coast of South Africa may return to spawn on the western portion of the Agulhas Bank. It remains likely, however, that the east and west coast populations are a single stock and that migration of juveniles to the west coast and their subsequent return as sub-adults is an integral but non-essential and variable part of the life history.

Impact of resistance on the efficacy of binary combinations of spinosad, chlorpyrifos-methyl and s-methoprene against five stored-grain beetles

Laboratory experiments were conducted to determine the efficacy of spinosad (a biopesticide), chlorpyrifos-methyl (an organophosphorus compound (OP)) and s-methoprene (a juvenile hormone analogue) applied alone and in binary combinations against five stored-grain beetles in wheat. There were three strains of Rhyzopertha dominica, and one strain each of Sitophilus oryzae, Tribolium castaneum, Oryzaephilus surinamensis and Cryptolestes ferrugineus. These strains were chosen to represent a range of possible resistant genotypes, exhibiting resistance to organophosphates, pyrethroids or methoprene. Treatments were applied at rates that are registered or likely to be registered in Australia. Adults were exposed to freshly treated wheat for 2 weeks, and the effects of treatments on mortality and reproduction were determined. No single protectant or protectant combination controlled all insect strains, based on the criterion of >99% reduction in the number of live F1 adults relative to the control. The most effective combinations were spinosad at 1 mg kg-1+chlorpyrifos-methyl at 10 mg kg-1 which controlled all strains except for OP-resistant O. surinamensis, and chlorpyrifos-methyl at 10 mg kg-1+s-methoprene at 0.6 mg kg-1 which controlled all strains except for methoprene-resistant R. dominica. The results of this study demonstrate the difficulty in Australia, and potentially other countries which use protectants, of finding protectant treatments to control a broad range of pest species in the face of resistance development.

Seed production and maturation of Limnocharis flava (L.) Buchenau in the field and glasshouse.

The aquatic herb Limnocharis flava, native to tropical America, is the target of an eradication program in Queensland but little is known about its reproductive biology. Their field and glasshouse studies showed that seedlings exhibited relatively high survival (64%) and that fruits containing over 1000 seeds could be produced on young plants within 46 days, at any time of the year. Mature fruits, follicles and seeds were also buoyant. The authors findings were incorporated into the eradication program and influenced the frequency of infestation monitoring.

Uneven application can influence the efficacy of s-methoprene against Rhyzopertha dominica (F.) in wheat.

The Juvenile Hormone analogue s-methoprene is used to protect stored grain from pests such as the lesser grain borer, Rhyzopertha dominica (F.). The possibility that uneven application influences s-methoprene efficacy against this species was investigated in the laboratory. Adults of methoprene-susceptible strains were exposed for 14 days to wheat treated at doses of up to 0.6 mg kg-1, or to mixtures of treated and untreated wheat giving equivalent average doses. Adult mortality after exposure to treated wheat was negligible in all cases (3.3%) and there was no significant effect of either average dose or evenness of application. In contrast, the number of adult progeny depended on both the average dose and evenness of application. Average doses of 0.3 and 0.6 mg kg-1 reduced the number of live F1 adults by 99-100% relative to the untreated wheat and no effect of evenness of application was detected. At lower doses, however, efficacy tended to decrease with increasing unevenness of application. When adults from the parental generation were transferred to untreated wheat for another 14 days neither the average dose nor evenness of application in the wheat from which they came had any significant effect on reproduction of these adults. This study demonstrates that uneven application can reduce the efficacy of s-methoprene against R. dominica, but that this is unlikely to influence the performance of s-methoprene against susceptible populations at target doses likely to be used in practice (e.g. 0.6 mg kg-1 in Australia). However, the possibility that uneven application leads to underdosing and selects for resistance should be investigated.

Quantifying differences between treated and untreated coir substrate

The aim of this project was to quantify differences between treated and untreated coir (coconut industrial residues) products and to identify differences in growth, yield and quality of cut flowers grown in different coir products. This has been brought about largely by the concern that some coir products, washed in low quality (saline) water may have detrimental effects on plant productivity and quality. There is concern in the flower production industry and among media suppliers, that lower quality products are favoured due to price alone, which as this project shows is a false economy. Specifically the project examined: • Differences in physical and chemical properties of treated and untreated coir along with another commonly used growing media in the flower industy; • Potential improvements in yield and quality of Gerbera (Gerbera jamesonii); • Potential differences in vase life of Gerbera as a result of the different growing media; and • Cost-benefit implications of treated (more expensive) coir substrate products versus untreated (less expensive) coir including any subsequent differences in yield and quality. By first examining the physical and some chemical properties of different coir substrates and other industry standard media, the researchers have been able to validate the concerns raised about the potential quality issues in coir based growing media. There was a great deal of variation in both the electrical conductivity and sodium contents. Physical properties were also variable as expected since manufacturers are able to target the specific physical preferences of plants through manipulation of the particle size distribution. A field trial was conducted under protected cropping practices in which three growing media were compared in terms of total productivity and also flower quality parameters such as stem length, flower diameter and vase life. The trial was a completely randomised design with the three growing media comprising treated coir discs, untreated coir discs and a pine bark coir mix. Four cultivars of Gerbera were assessed: Balance®; Carambole®; Dune® and Picobello®, all new products from Florist de Kwakel B.V., Denmark. Initial expansion from tissue culture was conducted at the Highsun Express Facility, Ormiston, Queensland. The trial included 12 replications of each cultivar in each media (a total of 144 plants) to ensure all data collected, and the derived conclusions were statistically rigorous. The coir supplied with no pre-treatment or buffering produced significantly less flowers than those grown in a pine bark coir mix or the pre-treated coir. Interestingly, the pine bark coir mix produced a greater number of flowers. However, the flowers produced in the pine bark coir mix were generally a shorter length stem. Productivity data, combined with flower quality data and component costs were all analysed through a cost/benefit economic model which showed that the greater revenue from better stem length outweighed the stem numbers, giving a cost benefit ratio of 2.58 for treated coir, 2.49 for untreated coir and 2.52 for pine bark coir mix. While this does not seem a large difference, when considering the number of plants a producer maintains can be upwards of 50,000 the difference in revenue would be, at a minimum $60,000 in this example. In conclusion, this project has found that there are significant effects on plant health, growth, yield and quality between those grown in treated and untreated coir. The outcome being growers can confidently invest in more expensive treated products with the assurance that benefits will outweigh initial cost. It is false economy to favour untreated coir products based on price alone. Producers should ensure they fully understand the production processes when purchasing growing media. Rather than targeting lower priced materials, it is recommended that quality be the highest priority in making this management decision. In making recommendations for future research and development it was important to consider conclusions from other researchers as well as those of the current project. It has been suggested that the media has greater longevity, which although not captured in this study could also lead to further cost efficiencies. Assessment of the products over a longer time period, and using a wider range of plant species are the major recommendations for further research to ensure greater understanding as to the importance in choosing the right growing media to meet specific needs.

Termite resistance of DMDHEU-treated wood.

Four field trials were conducted with wood modified with dimethyloldihydroxy-ethyleneurea (DMDHEU) in contact with subterranean termites. Trials 1 to 3 were conducted with Coptotermes acinaciformis (Froggatt); 1 and 2 in south-east Queensland, and 3 in northern Queensland, Australia. Trial 4 was conducted in northern Queensland with Mastotermes darwiniensis (Froggatt). Four timber species (Scots pine, beech, Slash pine and Spotted gum) and two levels (1.3 M and 2.3 M) of DMDHEU were used. The tests were validated. DMDHEU successfully prevented damage by C. acinaciformis in south-east Queensland, but not in northern Queensland. It also did not protect the wood against M. darwiniensis. Except for beech in trial 4, DMDHEU led to reduced mass losses caused by termite attack compared to the unmodified feeder stakes. Slash pine (in trials 1 and 3) and Spotted gum (in trial 1) presented low mass losses. Modification of Scots pine was more effective against termite damage than the modification of beech.

Potential of the neonicotinoid imidacloprid and the oxadiazine indoxacarb for controlling five coleopteran pests of stored grain.

The potential for using imidacloprid (a neonicotinoid) and indoxacarb (an oxadiazine) as grain protectants was investigated in bioassays against resistant strains of five stored grain beetles. The species investigated were Rhyzopertha dominica (F.) (the lesser grain borer), Sitophilus oryzae (L.) (the rice weevil), Tribolium castaneum (Herbst) (the rust-red flour beetle), Oryzaephilus surinamensis (L.) (the saw tooth flour beetle), and Cryptolestes ferrugineus (Stephens) (the flat grain beetle). Each of these species has developed resistance to one or more protectants, including organophosphorus insecticides, synthetic pyrethroids and the juvenile hormone analogue methoprene. Mortality and reproduction after a 2-week exposure of adults to treated wheat depended on species, dose and insecticide. Imidacloprid had no effect on S. oryzae at any dose, but none of the other species produced any live progeny at 10 mg/kg. Indoxacarb had no effect on T. castaneum at any dose, but none of the other species produced any live progeny at 5 mg/kg. The results show that although both imidacloprid and indoxacarb can control at least four of the five key pests tested at doses comparable to those used for organophosphorus protectants, more potent neonicotinoid or oxadiazine insecticides would be needed than either of these to provide broad spectrum protection of stored grain.

Animal-Level Factors Affecting Ovarian Function in Bos indicus Heifers Treated to Synchronize Ovulation with Intravaginal Progesterone-Releasing Devices and Oestradiol Benzoate

The primary objective of this study was to investigate the impact of animal-level factors including energy balance and environmental/management stress, on the ovarian function of Bos indicus heifers treated to synchronize ovulation. Two-year-old Brahman (BN) (n = 30) and BN-cross (n = 34) heifers were randomly allocated to three intravaginal progesterone-releasing device (IPRD) treatment groups: (i) standard-dose IPRD [Cue-Mate (R) (CM) 1.56 g; n = 17]; (ii) half-dose IPRD [0.78 g progesterone (P4); CM 0.78 g; n = 15]; (iii) half-dose IPRD + 300 IU equine chorionic gonadotrophin at IPRD removal (CM 0.78 g + G; n = 14); (iv) and a control group, 2x PGF2a [500 mu g prostaglandin F2a (PGF2a)] on Day -16 and -2 (n = 18). Intravaginal progesterone-releasing device-treated heifers received 250 mu g PGF2a at IPRD insertion (Day -10) and IPRD removal (Day -2) and 1 mg oestradiol benzoate on Day -10 and -1. Heifers were managed in a small feedlot and fed a defined ration. Ovarian function was evaluated by ultrasonography and plasma P4 throughout the synchronized and return cycles. Energy balance was evaluated using plasma insulin-like growth factor 1 (IGF-I) and glucose concentrations. The impact of environmental stressors was evaluated using plasma cortisol concentration. Heifers that had normal ovarian function had significantly higher IGF-I concentrations at commencement of the experiment (p = 0.008) and significantly higher plasma glucose concentrations at Day -2 (p = 0.040) and Day 4 (p = 0.043), than heifers with abnormal ovarian function. There was no difference between the mean pre-ovulatory cortisol concentrations of heifers that ovulated or did not ovulate. However, heifers that ovulated had higher cortisol concentrations at Day 4 (p = 0.056) and 6 (p = 0.026) after ovulation than heifers that did not ovulate.

Alginate encapsulation of shoot tips and nodal segments for short-term storage and distribution of the eucalypt Corymbia torelliana × C. citriodora

A protocol was developed for short-term preservation and distribution of the plantation eucalypt, Corymbia torelliana × C. citriodora, using alginate-encapsulated shoot tips and nodes as synthetic seeds. Effects of sowing medium, auxin concentration, storage temperature and planting substrate on shoot regrowth or conversion into plantlets were assessed for four different clones. High frequencies of shoot regrowth (76–100%) from encapsulated explants were consistently obtained in hormone-free half- and full-strength Murashige and Skoog (MS) sowing media. Conversion into plantlets from synthetic seeds was achieved on half-strength MS medium by treating shoot tips or nodes with 4.9–78.4 μM IBA prior to encapsulation. Pre-treatment with 19.6 μM IBA provided 62–100% conversion, and 95–100% of plantlets survived after acclimatisation under nursery conditions. Synthetic seeds containing explants pre-treated with IBA were stored for 8 weeks much more effectively at 25°C than at 4°C, with regrowth frequencies of 50–84% at 25°C compared with 0–4% at 4°C. To eliminate the in vitro culture step after encapsulation, synthetic seeds were allowed to pre-convert before sowing directly onto a range of ex vitro non-sterile planting substrates. Highest frequencies (46–90%) of plantlet formation from pre-converted synthetic seeds were obtained by transferring shoot tip-derived synthetic seeds onto an organic compost substrate. These plantlets exhibited almost 100% survival in the nursery without mist irrigation. Pre-conversion of non-embryonic synthetic seeds is a novel technique that provides a convenient alternative to somatic embryo-derived artificial seeds.

Cadmium stress alters the redox reaction and hormone balance in oilseed rape (Brassica napus L.) leaves

In order to understand the physiological response of oilseed rape (Brassica napus L.) leaves to cadmium (Cd) stress and exploit the physiological mechanisms involved in Cd tolerance, macro-mineral and chlorophyll concentrations, reactive oxygen species (ROS) accumulation, activities of enzymatic antioxidants, nonenzymatic compounds metabolism, endogenous hormonal changes, and balance in leaves of oilseed rape exposed to 0, 100, or 200 μM CdSO4 were investigated. The results showed that under Cd exposure, Cd concentrations in the leaves continually increased while macro-minerals and chlorophyll concentrations decreased significantly. Meanwhile, with increased Cd stress, superoxide anion (O 2 • − ) production rate and hydrogen peroxide (H2O2) concentrations in the leaves increased significantly, which caused malondialdehyde (MDA) accumulation and oxidative stress. For scavenging excess accumulated ROS and alleviating oxidative injury in the leaves, the activity of enzymatic antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), was increased significantly at certain stress levels. However, with increased Cd stress, the antioxidant enzyme activities all showed a trend towards reduction. The nonenzymatic antioxidative compounds, such as proline and total soluble sugars, accumulated continuously with increased Cd stress to play a long-term role in scavenging ROS. In addition, ABA levels also increased continuously with Cd stress while ZR decreased and the ABA/ZR ratio increased, which might also be providing a protective role against Cd toxicity.

Polybridge Technical Report

This study examined the physical and chemical properties of a novel, fully-recirculated prawn and polychaete production system that incorporated polychaete-assisted sand filters (PASF). The aims were to assess and demonstrate the potential of this system for industrialisation, and to provide optimisations for wastewater treatment by PASF. Two successive seasons were studied at commercially-relevant scales in a prototype system constructed at the Bribie Island Research Centre in Southeast Queensland. The project produced over 5.4 tonnes of high quality black tiger prawns at rates up to 9.9 tonnes per hectare, with feed conversion of up to 1.1. Additionally, the project produced about 930 kg of high value polychaete biomass at rates up to 1.5 kg per square metre of PASF, with the worms feeding predominantly on waste nutrients. Importantly, this closed production system demonstrated rapid growth of healthy prawns at commercially relevant production levels, using methods that appear feasible for application at large scale. Deeper (23 cm) PASF beds provided similar but more reliable wastewater treatment efficacies compared with shallower (13 cm) beds, but did not demonstrate significantly greater polychaete productivity than (easier to harvest) shallow beds. The nutrient dynamics associated with seasonal and tidal operations of the system were studied in detail, providing technical and practical insights into how PASF could be optimised for the mitigation of nutrient discharge. The study also highlighted some of the other important advantages of this integrated system, including low sludge production, no water discharge during the culture phase, high ecosystem health, good prospects for biosecurity controls, and the sustainable production of a fishery-limited resource (polychaetes) that may be essential for the expansion of prawn farming industries throughout the world. Regarding nutrient discharge from this prototype mariculture system, when PASF was operating correctly it proved feasible to have no water (or nutrient) discharge during the entire prawn growing season. However, the final drain harvest and emptying of ponds that is necessary at the end of the prawn farming season released 58.4 kg ha-1 of nitrogen and 6 kg ha-1 of phosphorus (in Season 2). Whilst this is well below (i.e., one-third to one-half of) the current load-based licencing conditions for many prawn farms in Australia, the levels of nitrogen and chlorophyll a in the ponds remained higher than the more-stringent maximum limits at the Bribie Island study site. Zero-net-nutrient discharge was not achieved, but waste nutrients were low where 5.91 kg of nitrogen and 0.61 kg of phosphorus was discharged per tonne of prawns produced. This was from a system that deployed PASF at 14.4% of total ponded farm area which treated an average of 5.8% of pond water daily and did not use settlement ponds or other natural or artificial water remediation systems. Four supplemental appendices complement this research by studying several additional aspects that are central to the industrialisation of PASF. The first details an economic model and decision tool which allows potential users to interactively assess construction and operational variables of PASF at different scales. The second provides the qualitative results of a prawn maturation trial conducted collaboratively with the Commonwealth Scientific and Industrial Research Organisation (CSIRO) to assess dietary inclusions of PASF-produced worms. The third provides the reproductive results from industry-based assessments of prawn broodstock produced using PASF. And the fourth appendix provides detailed elemental and nutritional analyses of bacterial biofilm produced by PASF and assesses its potential to improve the growth of prawns in recirculated culture systems.