Application of a species-specific PCR-RFLP to identify Ancylostoma eggs directly from canine faeces

Apart from their veterinary importance, the hookworms Ancylostoma caninum, Ancylostoma braziliense and Ancylostoma caninum are also capable of causing zoonotic disease in humans. A highly sensitive and species-specific PCR-RFLP technique was utilised to detect and differentiate the various canine Ancylostoma spp directly from eggs in faeces. This technique was utilised to screen 101 canine faecal samples from parasite endemic tea growing communities in Assam, India, as part as an ongoing epidemiological investigation into canine parasitic zoonoses. The prevalence of hookworms in dogs was found to be 98% using a combination of PCR and conventional microscopy. Overall, 36% of dogs were found positive for single hookworm infections with A. caninum, 24% positive for single infections with A. braziliense and 38% had mixed infections with both A. caninum and A. braziliense. No dogs were found positive for A. ceylanicum in the community under study. The high prevalence of A. caninum and A. braziliense in dogs in this community may account for the high incidence of cutaneous larva migrans (CLM) observed among the human population residing at the tea estates. The PCR-RFLP technique described herein allows epidemiological screening of canine hookworms to be conducted rapidly, with ease and accuracy, and has the potential to be applied to a number of different clinical, pharmacological and epidemiological situations. (C) 2004 Elsevier B.V. All rights reserved.

Improving wheat simulation capabilities in Australia from a cropping systems perspective: water and nitrogen effects on spring wheat in a semi-arid environment

Systems approaches can help to evaluate and improve the agronomic and economic viability of nitrogen application in the frequently water-limited environments. This requires a sound understanding of crop physiological processes and well tested simulation models. Thus, this experiment on spring wheat aimed to better quantify water x nitrogen effects on wheat by deriving some key crop physiological parameters that have proven useful in simulating crop growth. For spring wheat grown in Northern Australia under four levels of nitrogen (0 to 360 kg N ha(-1)) and either entirely on stored soil moisture or under full irrigation, kernel yields ranged from 343 to 719 g m(-2). Yield increases were strongly associated with increases in kernel number (9150-19950 kernels m(-2)), indicating the sensitivity of this parameter to water and N availability. Total water extraction under a rain shelter was 240 mm with a maximum extraction depth of 1.5 m. A substantial amount of mineral nitrogen available deep in the profile (below 0.9 m) was taken up by the crop. This was the source of nitrogen uptake observed after anthesis. Under dry conditions this late uptake accounted for approximately 50% of total nitrogen uptake and resulted in high (>2%) kernel nitrogen percentages even when no nitrogen was applied,Anthesis LAI values under sub-optimal water supply were reduced by 63% and under sub-optimal nitrogen supply by 50%. Radiation use efficiency (RUE) based on total incident short-wave radiation was 1.34 g MJ(-1) and did not differ among treatments. The conservative nature of RUE was the result of the crop reducing leaf area rather than leaf nitrogen content (which would have affected photosynthetic activity) under these moderate levels of nitrogen limitation. The transpiration efficiency coefficient was also conservative and averaged 4.7 Pa in the dry treatments. Kernel nitrogen percentage varied from 2.08 to 2.42%. The study provides a data set and a basis to consider ways to improve simulation capabilities of water and nitrogen effects on spring wheat. (C) 1997 Elsevier Science B.V.

Engineering closure of an open pit gold operation in a semi-arid climate

Along with material characteristics and geometry, the climate in which a mine is located can have a dramatic effect on the appropriate options for rehabilitation. The paper outlines the setting, mining, milling and waste disposal at Kidston Gold Mine's open pit operations in the semi-arid climate of North Queensland, Australia, before focusing on the engineering aspects of the rehabilitation of Kidston. The mine took a holistic and proactive approach to rehabilitation, and was prepared to demonstrate a number of innovative approaches, which are described in the paper. Engineering issues that had to be addressed included the geotechnical stability and deformation of waste rock dumps, including a 240 m high in-pit dump: the construction and performance monitoring of a “store and release” cover over potentially acid forming mineralised waste rock; erosion from the side slopes of the waste rock dumps; the in-pit co-disposal of waste rock and thickened tailings; the geotechnical stability of the tailings dam wall; the potential for erosion of bare tailings; the water balance of the tailings dam; direct revegetation of the tailings; and the pit hydrology. The rehabilitation of the mine represents an important benchmark in mine site rehabilitation best practice, from which lessons applicable worldwide can be shared.

An evolutionary radiation of beeflies in semi-arid Australia: systematics of the Exoprosopini (Diptera : Bombyliidae)

Almost half of the 4822 described beeflies in the world belong to the subfamily Anthracinae, with most of the diversity found in three cosmopolitan tribes: Villini, Anthracini, and Exoprosopini. The Australian Exoprosopini previously contained three genera, Ligyra Newman, Pseudopenthes Roberts and Exoprosopa Macquart. Pseudopenthes is an Australian endemic, with two species including Ps. hesperis, sp. nov. from Western Australia. Two new species of the exoprosopine Atrichochira Hesse, Atr. commoni, sp. nov. and Atr. paramonovi, sp. nov., are also described from Australia, extending the generic distribution from Africa. Cladistic analysis clarified the phylogenetic relationships between the recognised groups of the Exoprosopini and determined generic limits on a world scale. Inclusion of 18 Australian exoprosopines placed the Australian species in the context of the world fauna. The Exoprosopini contains six large groups. The basal group I contains species previously included in Exoprosopa to which the name Defilippia Lioy is applied. Group II contains Heteralonia Rondani, Atrichochira, Micomitra Bowden, Pseudopenthes, and Diatropomma Bowden. Colossoptera Hull is newly synonymised with Heteralonia. Group III is a paraphyletic assemblage of Pterobates Bezzi and Exoprosopa including the Australian Ex. sylvana ( Fabricius). Ligyra is paraphyletic, forming two well-separated clades. The African clade is described as Euligyra Lambkin, gen. nov., which, together with Litorhina Bezzi and Hyperalonia Rondani, form group IV. The Australian group V is true Ligyra. The remaining monophyletic lineage of exoprosopines, group VI, the Balaana-group of genera, shows evidence of an evolutionary radiation of beeflies in semi-arid Australia. Phylogenetic analysis of all 42 species of the Balaana-group of genera formed a basis for delimiting genera. Seven new genera are described by Lambkin & Yeates: Balaana, Kapua, Larrpana, Munjua, Muwarna, Palirika and Wurda. Four non-Australian species belong to Balaana. Thirty two new Australian species are described: Bal. abscondita, Bal. bicuspis, Bal. centrosa, Bal. gigantea, Bal. kingcascadensis, K. corusca, K. irwini, K. westralica, Lar. collessi, Lar. zwicki, Mun. erugata, Mun. lepidokingi, Mun. paralutea, Mun. trigona, Muw. vitreilinearis, Pa. anaxios, Pa. basilikos, Pa. blackdownensis, Pa. bouchardi, Pa. cyanea, Pa. danielsi, Pa. decora, Pa. viridula, Pa. whyalla, W. emu, W. impatientis, W. montebelloensis, W. norrisi, W. patrellia, W. skevingtoni, W. windorah, and W. wyperfeldensis. The following new combinations are proposed: from Colossoptera: Heteralonia latipennis (Brunetti); from Exoprosopa: Bal. grandis (Pallas), Bal. efflatounbeyi (Paramonov), Bal. latelimbata ( Bigot), Bal. obliquebifasciata ( Macquart), Bal. tamerlan (Portschinsky), Bal. onusta ( Walker), Def. busiris (Jaennicke), Def. efflatouni ( Bezzi), Def. eritreae (Greathead), Def. gentilis ( Bezzi), Def. luteicosta ( Bezzi), Def. minos (Meigen), Def. nigrifimbriata ( Hesse), Def. rubescens ( Bezzi), K. adelaidica ( Macquart), Lar. dimidiatipennis ( Bowden), Muw. stellifera ( Walker), and Pa. marginicollis ( Gray); from Ligyra: Eu. enderleini ( Paramonov), Eu. mars ( Bezzi), Eu. monacha (Klug), Eu. paris ( Bezzi), Eu. sisyphus ( Fabricius), and Eu. venus (Karsch).

A simulation model of cereal-legume intercropping systems for semi-arid regions I. Model development

Cereal-legume intercropping plays an important role in subsistence food production in developing countries, especially in situations of limited water resources. Crop simulation can be used to assess risk for intercrop productivity over time and space. In this study, a simple model for intercropping was developed for cereal and legume growth and yield, under semi-arid conditions. The model is based on radiation interception and use, and incorporates a water stress factor. Total dry matter and yield are functions of photosynthetically active radiation (PAR), the fraction of radiation intercepted and radiation use efficiency (RUE). One of two PAR sub-models was used to estimate PAR from solar radiation; either PAR is 50% of solar radiation or the ratio of PAR to solar radiation (PAR/SR) is a function of the clearness index (K-T). The fraction of radiation intercepted was calculated either based on Beer's Law with crop extinction coefficients (K) from field experiments or from previous reports. RUE was calculated as a function of available soil water to a depth of 900 mm (ASW). Either the soil water balance method or the decay curve approach was used to determine ASW. Thus, two alternatives for each of three factors, i.e., PAR/SR, K and ASW, were considered, giving eight possible models (2 methods x 3 factors). The model calibration and validation were carried out with maize-bean intercropping systems using data collected in a semi-arid region (Bloemfontein, Free State, South Africa) during seven growing seasons (1996/1997-2002/2003). The combination of PAR estimated from the clearness index, a crop extinction coefficient from the field experiment and the decay curve model gave the most reasonable and acceptable result. The intercrop model developed in this study is simple, so this modelling approach can be employed to develop other cereal-legume intercrop models for semi-arid regions. (c) 2004 Elsevier B.V. All rights reserved.

A simulation model of cereal-legume intercropping systems for semi-arid regions II. Model application

Smallholder farmers in Africa practice traditional cropping techniques such as intercropping. Intercropping is thought to offer higher productivity and resource milisation than sole cropping. In this study, risk associated with maize-bean intercropping was evaluated by quantifying long-term yield in both intercropping and sole cropping in a semi-arid region of South Africa (Bloemfontein, Free State) with reference to rainfall variability. The crop simulation model was run with different cultural practices (planting date and plant density) for 52 summer crop growing seasons (1950/1951-2001/2002). Eighty-one scenarios, consisted of three levels of initial soil water, planting date, maize population, and bean population, were simulated. From the simulation outputs, the total land equivalent ratio (LER) was greater than one. The intercrop (equivalent to sole maize) had greater energy value (EV) than sole beans, and the intercrop (equivalent to sole beans) had greater monetary value (MV) than sole maize. From these results, it can be concluded that maize-bean intercropping is advantageous for this semi-arid region. Soil water at planting was the most important factor of all scenario factors, followed by planting date. Irrigation application at planting, November/December planting and high plant density of maize for EV and beans for MV can be one of the most effective cultural practices in the study region. With regard to rainfall variability, seasonal (October-April) rainfall positively affected EV and MV, but not LER. There was more intercrop production in La Nina years than in El Nino years. Thus, better cultural practices may be selected to maximize maize-bean intercrop yields for specific seasons in the semi-arid region based on the global seasonal outlook. (c) 2004 Elsevier B.V. All rights reserved.

Molecular characterization of Drosophila C virus isolates

Reverse transcription coupled with polymerase chain reaction and restriction enzyme analysis was used to characterize 12 Drosophila C virus isolates from geographically different regions. A 1.2-kb fragment was amplified from cDNA and profiles from digestion with 20 restriction enzymes were generated. Analysis of the restriction fragment data gave estimates of nucleotide divergence of 0-10% between isolates. The isolates were grouped on the basis of genetic distance estimates derived from the restriction data. For the isolates from which a single genotype could be purified, a geographical pattern in the distribution of viral genotypes was identified. The 4 Moroccan isolates were very closely related to each other, differing in only 1 restriction profile. The 2 Australian isolates were each other's closest relatives, as were the 2 isolates first recovered in France. The PCR-RFLP technique used in this study has provided us with a simple procedure which can be used to characterize DCV isolates. A single enzyme, Tag I, generated 5 distinct and diagnostic restriction fragment patterns, which allowed easy assignment of isolates to one of the five viral genotypes identified in this study. (C) 1999 Academic Press.

Speciation and distribution of the members of the Anopheles punctulatus (Diptera : Culicidae) group in Papua New Guinea

Mosquito collections were made throughout the mainland of Papua New Guinea to identify the members of the Anopheles punctulatus group present and to determine their distribution. Identification was made using morphology, DNA hybridization, and polymerase chain reaction (PCR)-RFLP analysis. Nine members of the group were identified: An. farauti s.s. Laveran, An.farauti 2, An. koliensis Owen, and An. punctulatus Donitz, were common and widespread; An. farauti 4 was restricted to the north of the central ranges where it was common; An. farauti 6 was found only in the highlands above 1,000 m; and An. farauti 3, An. sp. near punctulatus and An. clowi Rozeboom & Knight were uncommon and had restricted distributions. Identification of An. koliensis and An. punctulatus using proboscis morphology was found to be unreliable wherever An. farauti 4 occurred. The distribution and dispersal of the members of the An. punctulatus group is discussed in regard to climate, larval habitats, distance from the coast, elevation, and proximity to human habitation.

Development and evaluation of a species diagnostic polymerase chain reaction-restriction fragment-length polymorphism procedure for cryptic members of the Culex sitiens (Diptera : Culicidae) subgroup in Australia and the southwest Pacific

Members of the Culex sitiens subgroup are important vectors of arboviruses, including Japanese encephalitis virus, Murray Valley encephalitis virus and Ross River virus. Of the eight described species, Cx. annulirostris Skuse, Cx. sitiens Wiedemann, and Cx. palpalis Taylor appear to be the most abundant and widespread throughout northern Australia and Papua New Guinea (PNG). Recent investigations using allozymes have shown this subgroup to contain cryptic species that possess overlapping adult morphology. We report the development of a polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) procedure that reliably separates these three species. This procedure utilizes the sequence variation in the ribosomal DNA ITS1 and demonstrates species-specific PCR-RFLP profiles from both colony and field collected material. Assessment of the consistency of this procedure was undertaken on mosquitoes sampled from a wide geographic area including Australia, PNG, and the Solomon Islands. Overlapping adult morphology was observed for Cx. annulirostris and Cx. palpalis in both northern Queensland and PNG and for all three species at one site in northwest Queensland.