Real-time PCR as a surveillance tool for the detection of Trichinella infection in muscle samples from wildlife

Trichinella nematodes are the causative agent of trichinellosis, a meat-borne zoonosis acquired by consuming undercooked, infected meat. Although most human infections are sourced from the domestic environment, the majority of Trichinella parasites circulate in the natural environment in carnivorous and scavenging wildlife. Surveillance using reliable and accurate diagnostic tools to detect Trichinella parasites in wildlife hosts is necessary to evaluate the prevalence and risk of transmission from wildlife to humans. Real-time PCR assays have previously been developed for the detection of European Trichinella species in commercial pork and wild fox muscle samples. We have expanded on the use of real-time PCR in Trichinella detection by developing an improved extraction method and SYBR green assay that detects all known Trichinella species in muscle samples from a greater variety of wildlife. We simulated low-level Trichinella infections in wild pig, fox, saltwater crocodile, wild cat and a native Australian marsupial using Trichinella pseudospiralis or Trichinella papuae ethanol-fixed larvae. Trichinella-specific primers targeted a conserved region of the small subunit of the ribosomal RNA and were tested for specificity against host and other parasite genomic DNAs. The analytical sensitivity of the assay was at least 100 fg using pure genomic T. pseudospiralis DNA serially diluted in water. The diagnostic sensitivity of the assay was evaluated by spiking log of each host muscle with T. pseudospiralis or T. papuae larvae at representative infections of 1.0, 0.5 and 0.1 larvae per gram, and shown to detect larvae at the lowest infection rate. A field sample evaluation on naturally infected muscle samples of wild pigs and Tasmanian devils showed complete agreement with the EU reference artificial digestion method (k-value = 1.00). Positive amplification of mouse tissue experimentally infected with T. spiralis indicated the assay could also be used on encapsulated species in situ. This real-time PCR assay offers an alternative highly specific and sensitive diagnostic method for use in Trichinella wildlife surveillance and could be adapted to wildlife hosts of any region. (C) 2012 Elsevier B.V. All rights reserved.

The microbiological status of prawns from retail and wholesale outlets in the Brisbane region

Cooked prawns from retail and wholesale outlets in the Brisbane region included a large proportion with high bacterial counts (10(6)-10(8)/g) indicating a short remaining shelflife. Microbially inferior product entering the wholesale and retail trade indicated poor handling and storage practices from point of capture all along the distribution chain to the retail outlets. Current handling practices need to be upgraded and HACCP principles adopted.

Distribution of barnacle epizoites of the crab Portunus pelagicus in the Moreton Bay region, eastern Australia

The parasites of some decapod crustaceans are known to cause sterilisation of their hosts, and can thus have an important impact on the population dynamics of infested species. Blue swimmer crabs (Portunus pelagicus) collected in three areas around Moreton Bay, Australia were examined for the presence of epizoic barnacles in their branchial chambers and on their carapace. Of the 952 crabs inspected 92% were infested with Octolasmis spp. The mean number of barnacles (predominantly Octolasmis warwickii) per carapace and gill chamber (mainly O. angulata) were 2.35 and 71.1, respectively. Barnacle infestation of gills was found to differ significantly by area, season and sex with the deeper offshore areas exhibiting the highest number of barnacles. The distribution within the hosts showed barnacles were more likely to be distributed in areas closer to the inhalant aperture. Highest abundances were found on the proximal surface of the hypobranchial side of gills 3, 4 and 5. Host moult stage and parasitism by Sacculina granifera were also found to affect the abundance of epizoic barnacles in some areas.

Microbiological status of piggery effluent from 13 piggeries in the south east Queensland region of Australia

Aims: To assist in the development of safe piggery effluent re-use guidelines by determining the level of selected pathogens and indicator organisms in the effluent ponds of 13 south-east Queensland piggeries. Methods and Results: The numbers of thermotolerant coliforms, Campylobacter jejuni/coli, Erysipelothrix rhusiopathiae, Escherichia coli, Salmonella and rotavirus were determined in 29 samples derived from the 13 piggeries. The study demonstrated that the 13 final effluent ponds contained an average of 1Æ2 · 105 colony-forming units (CFU) 100 ml)1 of thermotolerant coliforms and 1Æ03 · 105 CFU 100 ml)1 of E. coli. The Campylobacter level varied from none detectable (two of 13 piggeries) to a maximum of 930 most probable number (MPN) 100 ml)1 (two of 13 piggeries). Salmonella was detected in the final ponds of only four of the 13 piggeries and then only at a low level (highest level being 51 MPN 100 ml)1). No rotavirus and no Erysip. rhusiopathiae were detected. The average log10 reductions across the ponding systems to the final irrigation pond were 1Æ77 for thermotolerant coliforms, 1Æ71 for E. coli and 1Æ04 for Campylobacter. Conclusions: This study has provided a baseline knowledge on the levels of indicator organisms and selected pathogens in piggery effluent. Significance and Impact of the Study: The knowledge gained in this study will assist in the development of guidelines to ensure the safe and sustainable re-use of piggery effluent.

The reliability of supply of feed grains in the northern region

Reliability of supply of feed grain has become a high priority issue for industry in the northern region. Expansion by major intensive livestock and industrial users of grain, combined with high inter-annual variability in seasonal conditions, has generated concern in the industry about reliability of supply. This paper reports on a modelling study undertaken to analyse the reliability of supply of feed grain in the northern region. Feed grain demand was calculated for major industries (cattle feedlots, pigs, poultry, dairy) based on their current size and rate of grain usage. Current demand was estimated to be 2.8Mt. With the development of new industrial users (ethanol) and by projecting the current growth rate of the various intensive livestock industries, it was estimated that demand would grow to 3.6Mt in three years time. Feed grain supply was estimated using shire scale yield prediction models for wheat and sorghum that had been calibrated against recent ABS production data. Other crops that contribute to a lesser extent to the total feed grain pool (barley, maize) were included by considering their production relative to the major winter and summer grains, with estimates based on available production records. This modelling approach allowed simulation of a 101-year time series of yield that showed the extent of the impact of inter-annual climate variability on yield levels. Production estimates were developed from this yield time series by including planted crop area. Area planted data were obtained from ABS and ABARE records. Total production amounts were adjusted to allow for any export and end uses that were not feed grain (flour, malt etc). The median feed grain supply for an average area planted was about 3.1Mt, but this varied greatly from year to year depending on seasonal conditions and area planted. These estimates indicated that supply would not meet current demand in about 30% of years if a median area crop were planted. Two thirds of the years with a supply shortfall were El Nino years. This proportion of years was halved (i.e. 15%) if the area planted increased to that associated with the best 10% of years. Should demand grow as projected in this study, there would be few years where it could be met if a median crop area was planted. With area planted similar to the best 10% of years, there would still be a shortfall in nearly 50% of all years (and 80% of El Nino years). The implications of these results on supply/demand and risk management and investment in research and development are briefly discussed.

Antibiotic resistance in Campylobacter jejuni and Campylobacter coli isolated from poultry in the South-East Queensland region

Objectives: The aim of this study was to determine the antimicrobial resistance patterns of 125 Campylobacter jejuni and 27 Campylobacter coli isolates from 39 Queensland broiler farms. Methods: Two methods, a disc diffusion assay and an agar-based MIC assay, were used. The disc diffusion was performed and interpreted as previously described (Huysmans MB, Turnidge JD. Disc susceptibility testing for thermophilic campylobacters. Pathology 1997; 29: 209–16), whereas the MIC assay was performed according to CLSI (formerly NCCLS) methods and interpreted using DANMAP criteria. Results: In both assays, no C. jejuni or C. coli isolates were resistant to ciprofloxacin or chloramphenicol, no C. coli were resistant to nalidixic acid, and no C. jejuni were resistant to erythromycin. In the MIC assay, no C. jejuni isolate was resistant to nalidixic acid, whereas three isolates (2.4%) were resistant in the disc assay. The highest levels of resistance of the C. jejuni isolates were recorded for tetracycline (19.2% by MIC and 18.4% by disc) and ampicillin (19.2% by MIC and 17.6% by disc). The C. coli isolates gave very similar results (tetracycline resistance 14.8% by both MIC and disc; ampicillin resistance 7.4% by MIC and 14.8% by disc). Conclusions: This work has shown that the majority of C. jejuni and C. coli isolates were susceptible to the six antibiotics tested by both disc diffusion and MIC methods. Disc diffusion represents a suitable alternative methodology to agar-based MIC methods for poultry Campylobacter isolates.

Subsoil constraints to grain production in the cropping soils of the north-eastern region of Australia: an overview

In dryland agricultural systems of the subtropical, semi-arid region of north-eastern Australia, water is the most limiting resource. Crop productivity depends on the efficient use of rainfall and available water stored in the soil during fallow. Agronomic management practices including a period of fallow, stubble retention, and reduced tillage enhance reserves of soil water. However, access to stored water in these soils may be restricted by the presence of growth-limiting conditions in the rooting zone of the crop. These have been termed as subsoil constraints. Subsoil constraints may include compacted or gravel layers (physical), sodicity, salinity, acidity, nutrient deficiencies, presence of toxic elements (chemical) and low microbial activity (biological). Several of these constraints may occur together in some soils. Farmers have often not been able to obtain the potential yield determined by their prevailing climatic conditions in the marginal rainfall areas of the northern grains region. In the past, the adoption of soil management practices had been largely restricted to the top 100 mm soil layer. Exploitation of the subsoil as a source of water and nutrients has largely been overlooked. The key towards realising potential yields would be to gain better understanding of subsoils and their limitations, then develop options to manage them practically and economically. Due to the complex nature of the causal factors of these constraints, efforts are required for a combination of management approaches rather than individual options, with the aim to combat these constraints for sustainable crop production, managing natural resources and avoiding environmental damage.

Grower group case study on new farming practices in the Herbert region, Australia

Growers working together have proven to be a successful method for improving the utilization of farm resources and accelerating the adoption of the Sugar Yield Decline Joint Venture principles (SYDJV). The Pinnacle Precision Farming Group was formed in 2004 with the aim to bring together the ideas, knowledge and resources of growers in the Herbert region. Along with their common interest in controlled traffic, minimal tillage and crop rotations, the grower group utilize a farm machinery contractor to provide some of their major farming operations. This paper provides an insight into the changes made by the Pinnacle Precision Farming Group and their journey to adopt the new farming system practices. This paper also details the changes made by the group machinery contractor and a comparison of the old and new farming systems used by a group member. A focus point of the document is the impact of the new farming system on the economic, social and environmental components of the farming business. Analysis of the new farming system with a legume crop rotation revealed an increase in the farm gross margin by AU$22 024 and, in addition, a reduction in tractor operation time by 38% across the whole farm. This represents a return on marginal capital of 14.68 times the original capital outlay required by the group member. Using the new farming system without a legume crop will still improve the group members whole of farm gross margin by AU$6 839 and reduce tractor operation time by 43% across the whole farm. The Pinnacle Precision Farming group recognize the need to continually improve their farming businesses and believe that the new farming system principles are critical for the long term viability of the industry. [U$1 = AU$1.19].

Assessment and rationalization of genetic diversity of Papua New Guinea taro (Colocasia esculenta) using SSR DNA fingerprinting

Taro (Colocasia esculenta) accessions were collected from 15 provinces of Papua New Guinea (PNG). The collection, totalling 859 accessions was collated for characterization and a core collection of 81 accessions (10%) was established on the basis of characterization data generated on 30 agro-morphological descriptors, and DNA fingerprinting using seven SSR primers. The selection of accessions was based on cluster analysis of the morphological data enabling initial selection of 20% accessions. The 20% sample was then reduced and rationalized to 10% based on molecular data generated by SSR primers. This represents the first national core collection of any species established in PNG based on molecular markers. The core has been integrated with core from other Pacific Island countries, contributing to a Pacific regional core collection, which is conserved in vitro in the South Pacific Regional Germplasm Centre at Fiji. The core collection is a valuable resource for food security of the South Pacific region and is currently being utilized by the breeding programmes of small Pacific Island countries to broaden the genetic base of the crop.

High resolution genetic and physical mapping of the I-3 region of tomato chromosome 7 reveals almost continuous microsynteny with grape chromosome 12 but interspersed microsynteny with duplications on Arabidopsis chromosomes 1, 2 and 3

The tomato I-3 gene introgressed from the Lycopersicon pennellii accession LA716 confers resistance to race 3 of the fusarium wilt pathogen Fusarium oxysporum f. sp. lycopersici. We have improved the high-resolution map of the I-3 region of tomato chromosome 7 with the development and mapping of 31 new PCR-based markers. Recombinants recovered from L. esculentum cv. M82 × IL7-2 F2 and (IL7-2 × IL7-4) × M82 TC1F2 mapping populations, together with recombinants recovered from a previous M82 × IL7-3 F2 mapping population, were used to position these markers. A significantly higher recombination frequency was observed in the (IL7-2 × IL7-4) × M82 TC1F2 mapping population based on a reconstituted L. pennellii chromosome 7 compared to the other two mapping populations based on smaller segments of L. pennellii chromosome 7. A BAC contig consisting of L. esculentum cv. Heinz 1706 BACs covering the I-3 region has also been established. The new high-resolution map places the I-3 gene within a 0.38 cM interval between the molecular markers RGA332 and bP23/gPT with an estimated physical size of 50-60 kb. The I-3 region was found to display almost continuous microsynteny with grape chromosome 12 but interspersed microsynteny with Arabidopsis thaliana chromosomes 1, 2 and 3. An S-receptor-like kinase gene family present in the I-3 region of tomato chromosome 7 was found to be present in the microsyntenous region of grape chromosome 12 but was absent altogether from the A. thaliana genome.

Harmonising GAP programs for fresh produce in ASEAN region

Many forces are driving the global demand for assurance that fruit and vegetables are safe to eat and of the right quality, and are produced and handled in a manner that does not cause harm to the environment and the health, safety and welfare of workers. The impact of these driving forces is that retailer requirements for suppliers to comply with Good Agricultural Practice (GAP) is increasing and governments are strengthening legal requirements for food safety, environmental protection, and worker health, safety and welfare. The implementation of GAP programs currently within the ASEAN (Association of South East Asian Nations) region varies, with some countries having government certified systems and others beginning the journey with awareness programs for farmers. Under a project funded by the ASEAN Australia Development Cooperation Program, a standard for ASEAN GAP has been developed to harmonise GAP Programs in the region. The goal is to facilitate trade between ASEAN countries and to global markets, improve viability for farmers, and help sustain a safe food supply and the environment. ASEAN GAP is an umbrella standard that individual member countries will benchmark their national programs against to gain equivalence. The scope of ASEAN GAP covers the production, harvesting and postharvest handling of fresh fruit and vegetables on farm and postharvest handling in locations where produce is packed for sale. ASEAN GAP consists of four modules covering food safety, environmental management, worker health, safety and welfare, and produce quality. Each module can be used alone or in combination with other modules. This enables progressive implementation of ASEAN GAP, module by module based on individual country priorities.

A contemporary assessment of land condition in the Northern Gulf region of Queensland

A framework using assessments of soil condition, pasture composition and woodland density was applied to describe 14 grazing land types as being in A (100% of original carrying capacity), B (75%), C (45%) or D (20%) condition. We assessed the condition of 260 sites, principally along public and some station roads, to provide a benchmark for current land condition. Land types were also assigned relative grazing values between 10 (best) and 0, reflecting soil fertility and potential biomass production. The method identifies particular, 'at-risk' land types for priority investment of resources, while the rationale behind assessments might point to management interventions to improve the condition of those land types. Across all land types, 47% of sites were in A condition, 34% in B condition, 17% in C condition and only 2% in D condition. Seventy-five percent of land types with grazing values >5 were in A or B condition, compared with 88% for those with grazing values ?5. For Georgetown granites, only 27% of sites were in A or B condition, with values for other land types being: alluvials 59%, black soils 64% and red duplex soils 57%, suggesting that improving management of these land types is a priority issue. On land types with high grazing value, the major discounting factor was pasture composition (72% of sites discounted), while increasing woodland density was the main discount (73% of sites discounted) on low grazing value land types.

Capsicum chlorosis virus infecting Capsicum annuum in the East Kimberley region of Western Australia

Capsicum chlorosis virus (CaCV) was detected in field grown Capsicum annuum from Kununurra in northeast Western Australia. Identification of the Kununurra isolate (WA-99) was confirmed using sap transmission to indicator hosts, positive reactions with tospovirus serogroup IV-specific antibodies and CaCV-specific primers, and amino acid sequence comparisons that showed >97% identity with published CaCV nucleocapsid gene sequences. The reactions of indicator hosts to infection with WA-99 often differed from those of the type isolate from Queensland. The virus multiplied best when test plants were grown at warm temperatures. CaCV was not detected in samples collected in a survey of C. annuum crops planted in the Perth Metropolitan area.

Weed species richness, density and relative abundance on farms in the subtropical grain region of Australia

Weed management is one of the most important economic and agronomic issues facing farmers in Australia's grain regions. Weed species occurrence and abundance was monitored between 1997 and 2000 on 46 paddocks (sites) across 18 commercial farms located in the Northern Grain Region. The sites generally fell within 4 disjunct regions, from south to north: Liverpool Plains, Moree, Goondiwindi and Kingaroy. While high species richness was found (139 species or species groups), only 8 species occurred in all 4 regions and many (56 species) only occurred at 1 site or region. No species were observed at every site but 7 species (Sonchus spp., Avena spp., Conyza spp., Echinochloa spp., Convolvulus erubescens, Phalaris spp. and Lactuca serriola) were recorded on more than 70% of sites. The average number of species observed within crops after treatment and before harvest was less than 13. Species richness tended to be higher in winter pulse crops, cotton and in fallows, but overall was similar at the different sampling seasons (summer v. winter). Separate species assemblages associated with the Goondiwindi and Kingaroy regions were identified by correspondence analysis but these appeared to form no logical functional group. The species richness and density was generally low, demonstrating that farmers are managing weed populations effectively in both summer and winter cropping phases. Despite the apparent adoption of conservation tillage, an increase in opportunity cropping and the diversity of crops grown (13) there was no obvious effect of management practices on weed species richness or relative abundance. Avena spp. and Sonchus spp. were 2 of the most dominant weeds, particularly in central and southern latitudes of the region; Amaranthus spp. and Raphanus raphanistrum were the most abundant species in the northern part of the region. The ubiquity of these and other species shows that continued vigilance is required to suppress weeds as a management issue.

Survey of weed flora and management relative to cropping practices in the north-eastern grain region of Australia

The main weeds and weed management practices undertaken in broad acre dryland cropping areas of north-eastern Australia have been identified. The information was collected in a comprehensive postal survey of both growers and agronomists from Dubbo in New South Wales (NSW) through to Clermont in central Queensland, where 237 surveys were returned. A very diverse weed flora of 105 weeds from 91 genera was identified for the three cropping zones within the region (central Queensland, southern Queensland and northern NSW). Twenty-three weeds were common to all cropping zones. The major common weeds were Sonchus oleraceus, Rapistrum rugosum, Echinochloa spp. and Urochloa panicoides. The main weeds were identified for both summer and winter fallows, and sorghum, wheat and chickpea crops for each of the zones, with some commonality as well as floral uniqueness recorded. More genera were recorded in the fallows than in crops, and those in summer fallows exceeded the number in winter. Across the region, weed management relied heavily on herbicides. In fallows, glyphosate and mixes with glyphosate were very common, although the importance of the glyphosate mix partner differed among the cropping zones. Use and importance of pre-emergence herbicides in-crop varied considerably among the zones. In wheat, more graminicides were used in northern NSW than in southern Queensland, and virtually none were used in central Queensland, reflecting the differences in winter grass weed flora across the region. Atrazine was the major herbicide used in sorghum, although metolachlor was also used predominantly in northern NSW. Fallow and inter-row cultivation were used more often in the southern areas of the region. Grazing of fallows was more prominent in northern NSW. High crop seeding rates were not commonly recorded indicating that growers are not using crop competition as a tool for weed management. Although many management practices were recorded overall, few growers were using integrated weed management, and herbicide resistance has been and continues to be an issue for the region.