Sex change strategy and the aromatase genes

5′ flanking regions of CYP19A1/A2 genes are reported for three sex changing fish.

Identification of the Sex Pheromone of Holotrichia Reynaudi

The male attractant pheromone of the scarab beetle Holotrichia reynaudi, an agricultural pest native to southern India, was extracted from abdominal glands of females with hexane and analyzed by gas chromatography– mass spectrometry. Field testing of the candidate chemicals, indole, phenol, and anisole, both alone and as binary mixtures, led us to conclude that anisole was the major component of the sex pheromone. Neither male nor female beetles were attracted to indole or phenol on their own. Similarly, when indole and anisole were combined, the attractiveness of the solution did not increase over that obtained with anisole alone. However, combination of phenol and anisole did alter the attractiveness of anisole, with fewer male beetles attracted to the binary mixture than to anisole on its own. The behavior of female beetles was not altered by any of the chemicals tested. Anisole is also the sex pheromone of H. consanguinea, making this the first known example of two melolonthine scarabs sharing the same pheromone.

(Z)-11-Hexadecenal and (3Z,6Z,9Z)-Tricosatriene: Sex Pheromone Components of the Red Banded Mango Caterpillar Deanolis sublimbalis

The sex pheromone of the red banded mango caterpillar, Deanolis sublimbalis (Lepidoptera: Crambidae), a serious pest of the mango Mangifera indica (Anacardiaceae) in India and Southeast Asia and a recent invader into northern Australia, has been identified. Three candidate compounds were identified from pheromone gland extracts of female moths, using gas chromatography (GC), GC-electroantennographic detection and GC-mass spectrometric analyses, in conjunction with dimethyldisulfide derivatization. Field bioassays established that both (Z)-11-hexadecenal (Z11-16:Ald) and (3Z,6Z,9Z)-tricosatriene (3Z,6Z,9Z-23:Hy) were required for attraction of male D. sublimbalis moths, and 1,000 μg of a 1:1 mix of Z11-16:Ald and 3Z,6Z,9Z-23:Hy was more attractive to male moths than caged virgin females. However, the binary blend was only attractive when the isomeric purity of the monounsaturated aldehyde was >99%, suggesting that the (E)-isomer was inhibitory. Although (Z)-11-hexadecen-1-ol (Z11-16:OH) was tentatively identified in gland extracts, the addition of this compound to the binary blend did not increase the numbers of moths captured. The pheromone can now be used in integrated pest management strategies.

Sexing Sirenians: Validation of Visual and Molecular Sex Determination in Both Wild Dugongs (Dugong dugon) and Florida Manatees (Trichechus manatus latirostris)

Sexing wild marine mammals that show little to no sexual dimorphism is challenging. For sirenians that are difficult to catch or approach closely, molecular sexing from tissue biopsies offers an alternative method to visual discrimination. This paper reports the results of a field study to validate the use of two sexing methods: (1) visual discrimination of sex vs (2) molecular sexing based on a multiplex PCR assay which amplifies the male-specific SRY gene and differentiates ZFX and ZFY gametologues. Skin samples from 628 dugongs (Dugong dugon) and 100 Florida manatees (Trichechus manatus latirostris) were analysed and assigned as male or female based on molecular sex. These individuals were also assigned a sex based on either direct observation of the genitalia and/or the association of the individual with a calf. Individuals of both species showed 93 to 96% congruence between visual and molecular sexing. For the remaining 4 to 7%, the discrepancies could be explained by human error. To mitigate this error rate, we recommend using both of these robust techniques, with routine inclusion of sex primers into microsatellite panels employed for identity, along with trained field observers and stringent sample handling.

Best management practices for sustainable and safe playing surface of Australian Football League sports fields

In 2002, AFL Queensland and the Brisbane Lions Football Club approached the Department of Primary Industries and Fisheries (Queensland) for advice on improving their Premier League sports fields. They were concerned about player safety and dissatisfaction with playing surfaces, particularly uneven turf cover and variable under-foot conditions. They wanted to get the best from new investments in ground maintenance equipment and irrigation infrastructure. Their sports fields were representative of community-standard, multi-use venues throughout Australia; generally ‘natural’ soil fields, with low maintenance budgets, managed by volunteers. Improvements such as reconstruction, drainage, or regular re-turfing are generally not affordable. Our project aimed to: (a) Review current world practice and performance benchmarks; (b) Demonstrate best-practice management for community-standard fields; (c) Adapt relevant methods for surface performance testing; (d) Assess current soils, and investigate useful amendments; (e) Improve irrigation system performance; and (e) Build industry capacity and encourage patterns for ongoing learning. Most global sports field research focuses on elite, sand-based fields. We adjusted elite standards for surface performance (hardness, traction, soil moisture, evenness, sward cover/height) and maintenance programs, to suit community-standard fields with lesser input resources. In regularly auditing ground conditions across 12 AFLQ fields in SE QLD, we discovered surface hardness (measured by Clegg Hammer) was the No. 1 factor affecting player safety and surface performance. Other important indices were turf coverage and surface compaction (measured by penetrometer). AFLQ now runs regularly audits affiliated fields, and closes grounds with hardness readings greater than 190 Gmax. Aerating every two months was the primary mechanical practice improving surface condition and reducing hardness levels to < 110 Gmax on the renovated project fields. With irrigation installation, these fields now record surface conditions comparable to elite fields. These improvements encouraged many other sporting organisations to seek advice / assistance from the project team. AFLQ have since substantially invested in an expanded ground improvement program, to cater for this substantially increased demand. In auditing irrigation systems across project fields, we identified low maintenance (with < 65% of sprinklers operating optimally) as a major problem. Retrofitting better nozzles and adjusting sprinklers improved irrigation distribution uniformity to 75-80%. Research showed that reducing irrigation frequency to weekly, and preparedness to withhold irrigation longer after rain, reduced irrigation requirement by 30-50%, compared to industry benchmarks of 5-6 ML/ha/annum. Project team consultation with regulatory authorities enhanced irrigation efficiency under imposed regional water restrictions. Laboratory studies showed incorporated biosolids / composts, or topdressed crumb rubber, improved compaction resistance of soils. Field evaluations confirmed compost incorporation significantly reduced surface hardness of high wear areas in dry conditions, whilst crumb rubber assisted turf persistence into early winter. Neither amendment was a panacea for poor agronomic practices. Under the auspices of the project Trade Mark Sureplay®, we published > 80 articles, and held > 100 extension activities involving > 2,000 participants. Sureplay® has developed a multi-level curator training structure and resource materials, subject to commercial implementation. The partnerships with industry bodies (particularly AFLQ), frequent extension activities, and engagement with government/regulatory sectors have been very successful, and are encouraged for any future work. Specific aspects of sports field management for further research include: (a) Understanding of factors affecting turf wear resistance and recovery, to improve turf persistence under wear; (b) Simple tests for pinpointing areas of fields with high hardness risk; and (c) Evaluation of new irrigation infrastructure, ‘water-saving’ devices, and irrigation protocols, in improving water use and turf cover outcomes.

Objective `Safe' Grazing Capacities for South-West Queensland Australia: Model Application and Evaluation

This paper describes the employment of two experienced graziers as consultants to apply and evaluate a model for calculating 'safe' long-term grazing capacities of individual properties. The model was based on ecological principles and entailed estimates of average annual forage grown (kglha) on the different land systems on each property and the calculation of the number of livestock (dry sheep equivalents, DSE) required to 'safely' utilise this forage. The grazier consultants applied and evaluated the 'safe' grazing capacity model on 20 properties of their choosing. For evaluation, model results were compared with; (a) the Department of Lands rated carrying capacities for those properties and (b) the grazing capacity assessed independently by the owners of those properties. For the 20 properties, the average 'safe' grazing capacity calculated by the model (21.0 DSE/kmZ) was 8% lighter than the average of the owner assessed capacities (22.7 DSE/kmZ), which in tum was 37% lighter than the average of the pre-1989 Department of Lands rated carrying capacity (31.0 DSE/kmZ). The grazing land management and administrative implications of these results and the role graziers played as consultants are discussed.

Objective `Safe' Grazing Capacities for South-West Queensland Australia: Development of a Model for Individual Properties

Few tools are available to assist graziers, land administrators and financiers in making objective grazing capacity decisions on Australian rangelands, despite existing knowledge regarding stocking rate theory and the impact of stocking rates on land condition. To address this issue a model for objectively estimating 'safe' grazing capacities on individual grazing properties in south-west Queensland was developed. The method is based on 'safe' levels of utilisation (15%-20%) by domestic livestock of average annual forage grown for each land system on a property. Average annual forage grown (kglha) was calculated as the product of the rainfall use efficiency (kglhdmm) and average annual rainfall (mm) for a land system. This estimate included the impact of tree and shrub cover on forage production. The 'safe' levels of forage utilisation for south- west Queensland pastures were derived from the combined experience of (1) re-analysis of the results of grazing trials, (2) reaching a consensus on local knowledge and (3) examination of existing grazing practice on 'benchmark' grazing properties. We recognise the problems in defining, determining and using grazing capacity values, but consider that the model offers decision makers a tool that can be used to assess the grazing capacity of individual properties.

Cuelure but not zingerone make the sex pheromone of male Bactrocera tryoni (Tephritidae: Diptera) more attractive to females

In tephritid fruit flies of the genus Bactrocera Macquart, a group of plant derived compounds (sensu amplo ‘male lures’) enhance the mating success of males that have consumed them. For flies responding to the male lure methyl eugenol, this is due to the accumulation of chemicals derived from the male lure in the male rectal gland (site of pheromone synthesis) and the subsequent release of an attractive pheromone. Cuelure, raspberry ketone and zingerone are a second, related group of male lures to which many Bactrocera species respond. Raspberry ketone and cuelure are both known to accumulate in the rectal gland of males as raspberry ketone, but it is not known if the emitted male pheromone is subsequently altered in complexity or is more attractive to females. Using Bactrocera tryoni as our test insect, and cuelure and zingerone as our test chemicals, we assess: (i) lure accumulation in the rectal gland; (ii) if the lures are released exclusively in association with the male pheromone; and (iii) if the pheromone of lure-fed males is more attractive to females than the pheromone of lure-unfed males. As previously documented, we found cuelure was stored in its hydroxyl form of raspberry ketone, while zingerone was stored largely in an unaltered state. Small but consistent amounts of raspberry ketone and β-(4-hydroxy-3-methoxyphenyl)-propionic acid were also detected in zingerone-fed flies. Males released the ingested lures or their analogues, along with endogenous pheromone chemicals, only during the dusk courtship period. More females responded to squashed rectal glands extracted from flies fed on cuelure than to glands from control flies, while more females responded to the pheromone of calling cuelure-fed males than to control males. The response to zingerone treatments in both cases was not different from the control. The results show that male B. tryoni release ingested lures as part of their pheromone blend and, at least for cuelure, this attracts more females.

Identification of the sex pheromone of the tree infesting Cossid Moth Coryphodema tristis (Lepidoptera: Cossidae)

The cossid moth (Coryphodema tristis) has a broad range of native tree hosts in South Africa. The moth recently moved into non-native Eucalyptus plantations in South Africa, on which it now causes significant damage. Here we investigate the chemicals involved in pheromone communication between the sexes of this moth in order to better understand its ecology, and with a view to potentially develop management tools for it. In particular, we characterize female gland extracts and headspace samples through coupled gas chromatography electro-antennographic detection (GC-EAD) and two dimensional gas chromatography mass spectrometry (GCxGC-MS). Tentative identities of the potential pheromone compounds were confirmed by comparing both retention time and mass spectra with authentic standards. Two electrophysiologically active pheromone compounds, tetradecyl acetate (14:OAc) and Z9-tetradecenyl acetate (Z9-14:OAc) were identified from pheromone gland extracts, and an additional compound (Z9-14:OH) from headspace samples. We further determined dose response curves for the identified compounds and six other structurally similar compounds that are common to the order Cossidae. Male antennae showed superior sensitivity toward Z9-14:OAc, Z7-tetradecenyl acetate (Z7-14:OAc), E9-tetradecenyl acetate (E9-14:OAc), Z9-tetradecenol (Z9-14:OH) and Z9-tetradecenal (Z9-14:Ald) when compared to female antennae. While we could show electrophysiological responses to single pheromone compounds, behavioral attraction of males was dependent on the synergistic effect of at least two of these compounds. Signal specificity is shown to be gained through pheromone blends. A field trial showed that a significant number of males were caught only in traps baited with a combination of Z9-14:OAc (circa 95 of the ratio) and Z9-14:OH. Addition of 14:OAc to this mixture also improved the number of males caught, although not significantly. This study represents a major step towards developing a useful attractant to be used in management tools for C. tristis and contributes to the understanding of chemical communication and biology of this group of insects.