A mitochondrial species identification assay for Australian blacktip sharks (Carcharhinus tilstoni, C. limbatus and C. amblyrhynchoides) using real-time PCR and high-resolution melt analysis

Tropical Australian shark fisheries target two morphologically indistinguishable blacktip sharks, the Australian blacktip (Carcharhinus tilstoni) and the common blacktip (C. limbatus). Their relative contributions to northern and eastern Australian coastal fisheries are unclear because of species identification difficulties. The two species differ in their number of precaudal vertebrae, which is difficult and time consuming to obtain in the field. But, the two species can be distinguished genetically with diagnostic mutations in their mitochondrial DNA ND4 gene. A third closely related sister species, the graceful shark C. amblyrhynchoides, can also be distinguished by species-specific mutations in this gene. DNA sequencing is an effective diagnostic tool, but is relatively expensive and time consuming. In contrast, real-time high-resolution melt (HRM) PCR assays are rapid and relatively inexpensive. These assays amplify regions of DNA with species-specific genetic mutations that result in PCR products with unique melt profiles. A real-time HRM PCR species-diagnostic assay (RT-HRM-PCR) has been developed based on the mtDNA ND4 gene for rapid typing of C. tilstoni, C. limbatus and C. amblyrhynchoides. The assay was developed using ND4 sequences from 66 C. tilstoni, 33. C. limbatus and five C. amblyrhynchoides collected from Indonesia and Australian states and territories; Western Australia, the Northern Territory, Queensland and New South Wales. The assay was shown to be 100% accurate on 160 unknown blacktip shark tissue samples by full mtDNA ND4 sequencing.

Genetic Options for Drought Management in Groundnut

Groundnut is one of the principal oilseeds in the world. It is cultivated on 24.8 million ha with a total production of 32.8 million t and an average productivity of 1.32 t ha-'. Developing countries account for 96.9% of the world groundnut area and 93.8% of total production. Production is concentrated in Asia (56.8% area and 66.5% production of the world) and Africa (38.0% area and 24.7% production). The groundnut productivity in Africa is only 0.86 t ha-' compared with 1.55 t hx1 of Asia. The world groundnut economy-facts, trends and outlook are desaibed in detail by Freeman et al., 1999. Briefly, in medium-term (i.e. up to 2010), 'groundnut production and consumption is likely to shift increasingly to developing countries; production will grow in all regions but most rapidly in Asia, slowly in sub-Saharan Africa and decline in Latin America; and utilizationwill continue to shift away from groundnut oil toward groundnut meal, specially confectionery products'.

Bats: Important reservoir hosts of emerging viruses

Bats (order Chiroptera, suborders Megachiroptera and Microchiroptera) are abundant, diverse, and geographically widespread. These mammals provide us with resources, but their importance is minimized and many of their populations and species are at risk, even threatened or endangered. Some of their characteristics (food choices, colonial or solitary nature, population structure, ability to fly, seasonal migration and daily movement patterns, torpor and hibernation, life span, roosting behaviors, ability to echolocate, virus susceptibility) make them exquisitely suitable hosts of viruses and other disease agents. Bats of certain species are well recognized as being capable of transmitting rabies virus, but recent observations of outbreaks and epidemics of newly recognized human and livestock diseases caused by viruses transmitted by various megachiropteran and microchiropteran bats have drawn attention anew to these remarkable mammals. This paper summarizes information regarding chiropteran characteristics and information regarding 66 viruses that have been isolated from bats. From these summaries, it is clear that we do not know enough about bat biology, that we are doing too little in terms of bat conservation, and that there remain a multitude of questions regarding the role of bats in disease emergence.

New microsatellite loci for Carcharhinid sharks (Carcharhinus tilstoni and C. sorrah) and their cross-amplification in other shark species

Twelve microsatellite DNA markers were isolated in the spot-tail shark (Carcharhinus sorrah) and nine were isolated in Australian black-tip shark (Carcharhinus tilstoni). These loci plus 18 others developed for sharks from the genera Negaprion, Ginglymostoma, Carcharodon and Isurus were tested for amplification success on four species of Carcharhinus (including C. sorrah and C. tilstoni) and four other species representing three diverse families. Cross-amplification was most common within families. Five loci were subsequently tested for polymorphism on 50 C. sorrah and 60 C. tilstoni. The number of alleles per locus was two to 24 and the average heterozygosity was 0.54 (range 0.16-0.87) for C. sorrah and 0.64 (range 0.44-0.78) for C. tilstoni. These loci may be useful tools for genetic analyses of the Carcharhinidae.

The introduction and release of Chiasmia inconspicua and C. assimilis (Lepidoptera: Geometridae) for the biological control of Acacia nilotica in Australia

Two geometrid moths Chiasmia inconspicua and Chiasmia assimilis, identified as potential biological control agents for prickly acacia Acacia nilotica subsp. indica, were collected in Kenya and imported into quarantine facilities in Australia where laboratory cultures were established. Aspects of the biologies of both insects were studied and CLIMEX® models indicating the climatically favourable areas of Australia were developed. Host range tests were conducted using an approved test list of 74 plant species and no-choice tests of neonate larvae placed on both cut foliage and potted plants. C. inconspicua developed through to adult on prickly acacia and, in small numbers, Acacia pulchella. C. assimilis developed through to adult on prickly acacia and also in very small numbers on A. pulchella, A. deanei, A. decurrens, and A. mearnsii. In all experiments, the response on prickly acacia could be clearly differentiated from the responses on the non-target species. Both insects were approved for release in Australia. Over a three-year period releases were made at multiple sites in north Queensland, almost all in inland areas. There was no evidence of either insect's establishment and both colonies were terminated. A new colony of C. assimilis was subsequently established from insects collected in South Africa and releases of C. assimilis from this new colony were made into coastal and inland infestations of prickly acacia. Establishment was rapid at one coastal site and the insect quickly spread to other infestations. Establishment at one inland area was also confirmed in early 2006. The establishment in coastal areas supported a CLIMEX model that indicated that the climate of coastal areas was more suitable than inland areas.

Spatial requirements of animals: Allometry and beyond

Space allowance is a major factor influencing animal welfare. For livestock, at least, it plays a critical role in profitability, yet there is little information on the amount of space that animals require. The amount of space an animal occupies as a consequence of its shape and size can be estimated using allometry; linear dimensions (L) can be expressed as L = kW1/3 and surface area (S) as S = kW2/3, where k = a constant and W = the weight of the animal. Such equations have been used to determine the amount of space needed by standing (area [m2] = 0.019W0.66) and lying (area [m2] = 0.027W0.67) animals. Limited studies on the lying down and standing up behaviors of pigs and cattle suggest that the amount of space required can be estimated by area (m2) = 0.047W0.66. Linear space required per animal for behaviors such as feeding or drinking from a trough can be estimated from 0.064W0.33, but in groups this requirement will be affected by social interactions among group members and the amount of competition for the resource. Determining the amount of space for groups of animals is complex, as the amount of useable space can vary with group size and by how group members share space in time. Some studies have been conducted on the way in which groups of domestic fowl use space, but overall, we know very little about the ways in which livestock time-share space, synchronicity in the performance of behaviors, and the effects of spatial restrictions on behavior and welfare.

Spatial patterns of sub-tidal seagrasses and their tissue nutrients in the Torres Strait, northern Australia: Implications for management

The distribution and nutritional profiles of sub-tidal seagrasses from the Torres Strait were surveyed and mapped across an area of 31,000 km2. Benthic sediment composition, water depth, seagrass species type and nutrients were sampled at 168 points selected in a stratified representative pattern. Eleven species of seagrass were present at 56 (33.3%) of the sample points. Halophila spinulosa, Halophila ovalis, Cymodocea serrulata and Syringodium isoetifolium were the most common species and these were nutrient profiled. Sub-tidal seagrass distribution (and associated seagrass nutrient concentrations) was generally confined to northern-central and south-western regions of the survey area (mean water depth was relatively shallow (approximately 13 m below MSL) and where sediments were comprised primarily muddy sand to gravelly sand. Seagrass nitrogen and starch content, the most important nutrients for marine herbivores, were significantly correlated with species and with the plant component (above or below ground). For all seagrass species, the above-ground component (shoots and leaves) possessed greater nitrogen concentrations than the below-ground component (roots and rhizomes), which possessed greater starch concentrations. S. isoetifolium had the highest total nitrogen concentrations (1.40±0.05% DW). However, it also had higher fibre concentrations (38.2±0.68% DW) relative to the other four species. H. ovalis possessed the highest starch concentrations (2.76±0.12% DW) and highest digestibility (83.24±0.66% DW) as well as the lowest fibre (27.2±0.66% DW). The high relative abundance (found at 55% of the sites that had seagrass) and nutrient quality characteristics of H. ovalis make it an important source of energy to marine herbivores that forage sub-tidally in the Torres Strait. There were two regions in Torres Strait (north-central and south-western) where sub-tidal seagrass meadows were prevalent and of relatively higher nutritional value. This spatial and nutritional information can be used by local agencies to manage and to protect the ecological, economic and cultural values of the sub-tidal seagrass ecosystems and associated fisheries of the Torres Strait.

Space allowances for confined livestock and their determination from allometric principles

The amount of space provided to animals governs important elements of their behaviour and, hence, is critical for their health and welfare. We review the use of allometric principles and equations to estimate the static space requirements of animals when standing and lying, and the space required for animals to feed, drink, stand-up and lie-down. We use the research literature relating to transportation and intensive housing of sheep and cattle to assess the validity of allometric equations for estimating space allowances. We investigated these areas because transportation and intensive housing provide points along a continuum in terms of the duration of confinement, (from hours to months) and spatial requirements are likely to increase with increasing duration of confinement, as animals will need to perform a greater behavioural repertoire for long-term survival, health and welfare. We find that, although there are theoretical reasons why allometric relationships to space allowances may vary slightly for different classes of stock, space allowances that have been demonstrated to have adverse effects on animal welfare during transportation correlated well with an inability to accommodate standing animals, as estimated from allometry. For intensive housing, we were able to detect a space allowance below which there were adverse effects on welfare. For short duration transportation during which animals remain standing, a space allowance per animal described by the allometric equation: area (m^2) = 0.020W^0.66, where W = liveweight (kg), would appear to be appropriate. Where it is desirable for all animals to lie simultaneously, then a minimum space allowance per animal described by the allometric equation: area (m^2) = 0.027W^0.66 appears to permit this, given that animals in a group time-share space. However, there are insufficient data to determine whether this allowance onboard a vehicle/vessel would enable animals to move and access food and water with ease. In intensive housing systems, a minimum space allowance per animal described by the allometric equation: area (m^2) = 0.033W^0.66 appears to be the threshold below which there are adverse effects on welfare. These suggested space allowances require verification with a range of species under different thermal conditions and, for transportation, under different conditions of vehicular/vessel stability. The minimum length of trough per animal (L in m) required for feeding and drinking can be determined from L = 0.064W^0.33, with the number of animals required to feed/drink simultaneously taken into account, together with any requirement to minimise competition. This also requires verification with a range of species. We conclude that allometric relationships are an appropriate basis for the formulation of space allowances for livestock.

In vitro propagation of Corymbia torelliana x C. citriodora (Myrtaceae) via cytokinin-free node culture

Hybrids between Corymbia torelliana (F.Muell.) K.D.Hill & L.A.S.Johnson and C. citriodora subsp. variegata (F.Muell.) A.R.Bean & M.W.McDonald are used extensively to establish forestry plantations in subtropical Australia. Methods were developed for in vitro seed germination, shoot multiplication and plantlet formation that could be used to establish in vitro and ex vitro clone banks of juvenile Corymbia hybrids. Effects of sodium hypochlorite concentration and exposure time on seed contamination and germination, and effects of cytokinin and auxin concentrations on shoot multiplication and subsequent rooting, were assessed. A two-step surface sterilisation procedure, involving 70% ethanol followed by 1% sodium hypochlorite, provided almost no contamination and at least 88% germination. A novel method of cytokinin-free node culture proved most effective for in vitro propagation. Lateral bud break of primary shoots was difficult to induce by using cytokinin, but primary shoots rooted prolifically, elongated rapidly and produced multiple nodes in the absence of exogenous cytokinin. Further multiplication was obtained either by elongating lateral shoots of nodal explants in cytokinin-free medium or by inducing organogenic callus and axillary shoot proliferation with 2.2 µm benzyladenine. Plantlets were produced using an in vitro soil-less method that provided extensive rooting in sterile propagation mixture. These methods provide a means for simultaneous laboratory storage and field-testing of clones before selection and multiplication of desired genotypes.

Mortality and blood loss by blue swimmer crabs (Portunus pelagicus) after simulated capture and discarding from gillnets

Two laboratory experiments were carried out to quantify the mortality and physiological responses of juvenile blue swimmer crabs (Portunus pelagicus) after simulated gillnet entanglement, air exposure, disentanglement, and discarding. In both experiments, all but control blue swimmer crabs were entangled in 1-m(2) gillnet panels for 1 h, exposed to air for 2 min, subjected to various treatments of disentanglement ranging between the forceful removal of none, one, two, and four appendages, then "discarded" into individual experimental tanks and monitored for 10 d. In Experiment 1, mortalities were associated with the number of appendages removed and the occurrence of unsealed wounds. In Experiment 2, live blue swimmer crabs were sampled for blood at 2 min and 6, 24, and 72 h post-discarding to test for the effects of disentanglement and appendage removal on total haemocyte counts, clotting times, protein levels (by refractive index), and blood ion concentrations. Compared with blue swimmer crabs that had sealed or no wounds, those with unsealed wounds had lower total haemocyte counts, protein, and calcium concentrations and increased clotting ties and magnesium and sodium levels. Induced autotomy, as opposed to the arbitrary, forceful removal of a appendages has the potential to minimize the mortality and stress of discarded, juvenile blue swimmer crabs.

Quality of handling and holding yard environment, and beef cattle temperament: 1. Relationships with flight speed and fear of humans.

Numerous tests have been used to measure beef cattle temperament, but limited research has addressed the relationship between such tests and whether temperament can be modified. One-hundred-and-forty-four steers were given one of three human handling and yarding experiences on six occasions during a 12-month grazing period post-weaning (backgrounding): Good handling/yarding, Poor handling/yarding and Minimal handling/yarding. At the end of this phase the cattle were lot-fed for 78 days, with no handling/yarding treatments imposed, before being transported for commercial slaughter. Temperament was assessed at the start of the experiment, during backgrounding and lot-feeding by flight speed (FS) and a fear of humans test, which measured the proximity to a stimulus person (zone average; ZA), the closest approach to the person (CA) and the amount the cattle moved around the test arena (total transitions; TT). During backgrounding, FS decreased for all treatments and at the end of backgrounding there was no difference between them. The rate of decline, however, was greatest in the Good group, smallest in the Minimal group with the Poor intermediate. In contrast, ZA was affected by treatment, with a greater reduction for the Good group than the others (P = 0.012). During lot-feeding, treatment did not affect FS, but all groups showed a decrease in ZA, with the greatest change in the Poor group, the least in the Good and the Minimal intermediate (P = 0.052). CA was positively correlated with ZA (r = 0.18 to 0.66) and negatively with TT (r = -0.180 to -0.659). FS was consistently correlated with TT only (r = 0.17 to 0.49). These findings suggest that FS and TT measure a similar characteristic, as do ZA and CA, but that these characteristics are different from one another, indicating that temperament is not a unitary trait, but has different facets. FS and TT measure one facet that we suggest is general agitation, whilst ZA and CA measure fear of people. Thus, the cattle became less agitated during backgrounding, but the effect was not permanently influenced by the quantity and quality of handling/yarding. However, Good handling/yarding reduced fearfulness of people. Fear of people was also reduced during lot-feeding, probably as a consequence of frequent exposure to humans in a situation that was neutral or positive for the cattle.

Salinity tolerance of twelve hybrid Bermudagrass (Cynodon Dactylon (L.) Pers. x C. Transvaalensis Burtt Davy) genotypes

Salinity is an increasingly important issue in both rural and urban areas throughout much of Australia. The use of recycled/reclaimed water and other sources of poorer quality water to irrigate turf is also increasing. Hybrid Bermudagrass (Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt Davey), together with the parent species C. dactylon, are amongst the most widely used warm-season turf grass groups. Twelve hybrid Bermudagrass genotypes and one accession each of Bermudagrass (C. dactylon), African Bermudagrass (C. transvaalensis) and seashore paspalum (Paspalum vaginatum Sw.) were grown in a glasshouse experiment with six different salinity treatments applied hydroponically through the irrigation water (ECW = <0.1, 6, 12, 18, 24 or 30 dSm-1) in a flood-and-drain system. Each pot was clipped progressively at 2-weekly intervals over the 12-week experimental period to determine dry matter production; leaf firing was rated visually on 3 occasions during the last 6 weeks of salinity treatment. At the end of the experiment, dry weights of roots and crowns below clipping height were also determined. Clipping yields declined sharply after about the first 6 weeks of salinity treatment, but then remained stable at substantially lower levels of dry matter production from weeks 8 to 12. Growth data over this final 4-week experimental period is therefore a more accurate guide to the relative salinity tolerance of the 15 entries than data from the preceding 8 weeks. Based on these data, the 12 hybrid Bermudagrass genotypes showed moderate salinity tolerance, with FloraDwarfM, 'Champion Dwarf', NovotekM and 'TifEagle' ranking as the most salt tolerant and 'Patriot', 'Santa Ana', 'Tifgreen' and TifSport M the least tolerant within the hybrid group. Nevertheless, Santa Ana, for example, maintained relatively strong root growth as salinity increased, and so may show better salt tolerance in practice than predicted from the growth data alone. The 12 hybrid Bermudagrasses and the single African Bermudagrass genotype were all ranked above FloraTeXM Bermudagrass in terms of salt tolerance. However, seashore paspalum, which is widely acknowledged as a halophytic species showing high salt tolerance, ranked well above all 14 Cynodon genotypes in terms of salinity tolerance.

Factors contributing to wear tolerance of Bermudagrass (Cynodon dactylon (L.) Pers., C. dactylon x C. transvaalensis Burtt-Davey) on a sand-based profile under simulated sports field conditions

Wear resistance and recovery of 8 Bermudagrass (Cynodon dactylon (L.) Pers.) and hybrid Bermudagrass (C. Dactylon x C. transvaalensis Burtt-Davey) cultivars grown on a sandbased soil profile near Brisbane, Australia, were assessed in 4 wear trials conducted over a two year period. Wear was applied on a 7-day or a 14-day schedule by a modified Brinkman Traffic Simulator for 6-14 weeks at a time, either during winter-early spring or during summer-early autumn. The more frequent wear under the 7-day treatment was more damaging to the turf than the 14-day wear treatment, particularly during winter when its capacity for recovery from wear was severely restricted. There were substantial differences in wear tolerance among the 8 cultivars investigated, and the wear tolerance rankings of some cultivars changed between years. Wear tolerance was associated with high shoot density, a dense stolon mat strongly rooted to the ground surface, high cell wall strength as indicated by high total cell wall content, and high levels of lignin and neutral detergent fiber. Wear tolerance was also affected by turf age, planting sod quality, and wet weather. Resistance to wear and recovery from wear are both important components of wear tolerance, but the relative importance of their contributions to overall wear tolerance varies seasonally with turf growth rate.

Morphological and developmental comparisons of seven greens quality hybrid bermudagrass (Cynodon dactylon (L.)Pers.x C transvaalensis Burtt-Davy

Fine-textured hybrid bermudagrass [Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt-Davy] cultivars have been widely used for golf putting greens and lawn bowls greens in warm-climate areas for more than 40 years. During the past decade, the choice of cultivar for professional turfgrass managers has been expanded by a range of secondgeneration hybrid bermudagrasses, which differ from the first-generation cultivars ‘Tifgreen’ and ‘Tifdwarf ’ in their management requirements. In this paper, we present comparative morphological and developmental data for seven cultivars (Champion Dwarf, FloraDwarf, MS-Supreme, Novotek, Tifdwarf, TifEagle, Tifgreen) grown in spaced plant and sward experiments at Cleveland, Australia (27º32’S lat, 153º15’E long, 25 masl). The four ‘ultradwarf ’ cultivars (Champion Dwarf, MS-Supreme, FloraDwarf, TifEagle) showed slower vertical extension and produced fewer inflorescences than Tifdwarf, Tifgreen, and Novotek. However, in terms of the length of stolon internodes and their overall rate of lateral spread, Champion Dwarf, FloraDwarf, and TifEagle were comparable to Tifdwarf; MS-Supreme (with longer internodes) spread faster laterally, though slower than Tifgreen (which had the longest stolon internodes). In unmown swards, the four ultradwarfs produced shorter leaves than Tifgreen, Tifdwarf, and Novotek, but only Champion Dwarf produced significantly narrower leaves than Tifgreen, Tifdwarf, and Novotek, with TifEagle leaves also significantly narrower than those of Tifgreen and Novotek. Minimum threshold temperatures for growth were approximately 9° to 10°C (air temperature) and 15° to 16°C at 10 cm soil depth.

Selectivity of chlorantraniliprole to parasitoid wasps.

BACKGROUND: Chlorantraniliprole is a novel anthranilic diamide insecticide registered for use in vegetables, fruits, grains and turf against a variety of insect pests. The objective of this article is to summarize results of acute toxicity testing of chlorantraniliprole on seven species of parasitic wasps with wide geographic distribution and relevance to different crops and integrated pest management (IPM) programmes. RESULTS: Tier-1, worst-case laboratory studies evaluated wasp survival and reproduction following different exposure concentrations and scenarios to chlorantraniliprole (i.e. fresh-dried spray deposits on glass plates, direct contact, ingestion, egg card, dipped leaf residue bioassays, sprayed mummies). No statistically significant effects on adult survival, percentage parasitism or emergence were observed following exposures to chlorantraniliprole compared with controls. CONCLUSION: Chlorantraniliprole was harmless to the parasitoid wasp species tested according to IOBC classification criteria (<30% effects) and may be a useful tool in IPM programmes.