Field evaluation of anthelmintic drug sensitivity using in vitro egg hatch and larval motility assays with Necator americanus recovered from human clinical isolates

A field-applicable assay for testing anthelmintic sensitivity is required to monitor for anthelmintic resistance. We undertook a study to evaluate the ability of three in vitro assay systems to define drug sensitivity of clinical isolates of the human hookworm parasite Necator americanus recovered from children resident in a village in Madang Province, Papua New Guinea. The assays entailed observation of drug effects on egg hatch (EHA), larval development (LDA), and motility of infective stage larvae (LMA). The egg hatch assay proved the best method for assessing the response to benzimidazole anthelmintics, while the larval motility assay was suitable for assessing the response to ivermectin. The performance of the larval development assay was unsatisfactory on account of interference caused by contaminating bacteria. A simple protocol was developed whereby stool samples were subdivided and used for immediate egg recovery, as well as for faecal culture, in order to provide eggs and infective larvae, respectively, for use in the egg hatch assay and larval motility assay systems. While the assays proved effective in quantifying drug sensitivity in larvae of the drug-susceptible hookworms examined in this study, their ability to indicate drug resistance in larval or adult hookworms remains to be determined. (c) 2005 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Comparison of cultures from rectoanal-junction mucosal swabs and feces for detection of Escherichia coli 0157 in dairy heifers

Fecal culture for Escherichia coli 0157:H7 was compared to rectoanal mucosal swab (RAMS) culture in dairy heifers over a 1-year period. RAMS enrichment culture was as sensitive as fecal culture using immunomagnetic separation (IMS) (P = 0.98, as determined by a chi-square test). RAMS culture is less costly than fecal IMS culture and can yield quantitative data.

Long-term persistence of multi-drug-resistant Salmonella enterica serovar Newport in two dairy herds

Objective - To evaluate the association between maintaining joint hospital and maternity pens;and persistence of multi-drug-resistant (MDR) Salmonella enterica serovar Newport on 2 dairy farms. Design - Observational study. Sample Population - Feces and environmental samples from 2 dairy herds. Procedure - Herds were monitored for fecal shedding of S enterica Newport after outbreaks of clinical disease. Fecal and environmental samples were collected approximately monthly from pens housing sick cows and calving cows and from pens containing lactating cows. Cattle shedding the organism were tested serially on subsequent visits to determine carrier status. One farm was resampled after initiation of interventional procedures, including separation of hospital and maternity pens. Isolates were characterized via serotyping, determination of antimicrobial resistance phenotype, detection of the CMY-2 gene, and DNA fingerprinting. Results - The prevalence (32.4% and 33.3% on farms A and B, respectively) of isolating Salmonella from samples from joint hospital-maternity pens was significantly higher than the prevalence in samples from pens housing preparturient cows (0.8%, both farms) and postparturient cows on Farm B (8.8%). Multi-drug-resistant Salmonella Newport was isolated in high numbers from bedding material, feed refusals, lagoon slurry, and milk filters. One cow excreted the organism for 190 days. Interventional procedures yielded significant reductions in the prevalences of isolating the organism from fecal and environmental samples. Most isolates were of the C2 serogroup and were resistant to third-generation cephalosporins. Conclusions and Clinical Relevance - Management practices may be effective at reducing the persistence of MDR Salmonella spp in dairy herds, thus mitigating animal and public health risk.

Development of an animal repellent - selection, efficacy and presentation

Under some circumstances it may be desirable to repel herbivorous pests (e. g. goats and wallabies) from plants (e. g. horticultural or forestry seedlings) rather than to kill them. These circumstances may include using these animals as a resource at a later time, and there may also be welfare, public perception or legislative issues where repelling animals from particular plants or area is preferable to killing them. The first experiment tested the efficacy of 5 different materials (tiger fecal extract, maggot brew-the liquid exudate from a flyblown goat, capsaicin, dog urine, and burnt goat hair, skin and carcass+fat) to repel groups of goats in a feedlot pen from 1 trough of feed when another trough of feed was available to them to eat. Each repellent was tested on 5 groups of mixed age female goats for periods of 3 days. All 5 groups were exposed to the 5 repellents. Both tiger fecal extract and maggot brew reduced intake of feed from the 'protected' feed trough and significantly (P < 0.05) delayed 'normal' feeding behaviour by more than 5 h from that trough. The repellents became less effective with repeated use even with different groups of goats. The reduction in effectiveness was indicated by higher intakes of feed and earlier feeding from the protected trough. This habituation to the repellents, where the effectiveness is reduced with repeated exposure, is of concern. In the second experiment, we selected the most effective repellent (tiger fecal extract) and tested its efficacy under similar conditions, after the repellent had been mixed with a carrier (bentonite) in an endeavour to increase the duration of its effectiveness. This repellent was significantly more effective in repelling groups of 3 goats in a feedlot pen from 1 trough of feed for 3 days, when another trough of feed was available to them to eat.

Healthy plantations. A field guide to pests and pathogens of Acacia, Eucalyptus and Pinus in Vietnam.

This book is a resource for those involved ‘on-the-ground’ with growing plantation trees in Vietnam, identifying the pests and diseases found on them, and managing the impacts of these organisms. The book, supported by AusAID’s Vietnam CARD (Cooperation for Agricultural and Rural Development) Program, and draws on the collective, long-standing experience of forest health scientists in Vietnam, Australia and South Africa. The book provides illustrations and information on 23 pests and 25 diseases of Acacia, Eucalyptus and Pinus for Vietnam; four of these species are important biosecurity threats not yet present in Vietnam.

Organic inputs, tillage and rotation practices influence soil health and suppressiveness to soilborne pests and pathogens of ginger.

A field experiment was established in which an amendment of poultry manure and sawdust (200 t/ha) was incorporated into some plots but not others and then a permanent pasture or a sequence of biomass-producing crops was grown with and without tillage, with all biomass being returned to the soil. After 4 years, soil C levels were highest in amended plots, particularly those that had been cropped using minimum tillage, and lowest in non-amended and fallowed plots, regardless of how they had been tilled. When ginger was planted, symphylans caused severe damage to all treatments, indicating that cropping, tillage and organic matter management practices commonly used to improve soil health are not necessarily effective for all crops or soils. During the rotational phase of the experiment, the development of suppressiveness to three key pathogens of ginger was monitored using bioassays. Results for root-knot nematode (Meloidogyne javanica) indicated that for the first 2 years, amended soil was more suppressive than non-amended soil from the same cropping and tillage treatment, whereas under pasture, the amendment only enhanced suppressiveness in the first year. Suppressiveness was generally associated with higher C levels and enhanced biological activity (as measured by the rate of fluorescein diacetate (FDA) hydrolysis and numbers of free-living nematodes). Reduced tillage also enhanced suppressiveness, as gall ratings and egg counts in the second and third years were usually significantly lower in cropped soils under minimum rather than conventional tillage. Additionally, soil that was not disturbed during the process of setting up bioassays was more suppressive than soil which had been gently mixed by hand. Results of bioassays with Fusarium oxysporum f. sp. zingiberi were too inconsistent to draw firm conclusions, but the severity of fusarium yellows was generally higher in fumigated fallow soil than in other treatments, with soil management practices having little impact on disease severity. With regard to Pythium myriotylum, biological factors capable of reducing rhizome rot were present, but were not effective enough to suppress the disease under environmental conditions that were ideal for disease development.

Integrating pests and pathogens into the climate change/food security debate

While many studies have demonstrated the sensitivities of plants and of crop yield to a changing climate, a major challenge for the agricultural research community is to relate these findings to the broader societal concern with food security. This paper reviews the direct effects of climate on both crop growth and yield and on plant pests and pathogens and the interactions that may occur between crops, pests, and pathogens under changed climate. Finally, we consider the contribution that better understanding of the roles of pests and pathogens in crop production systems might make to enhanced food security. Evidence for the measured climate change on crops and their associated pests and pathogens is starting to be documented. Globally atmospheric [CO(2)] has increased, and in northern latitudes mean temperature at many locations has increased by about 1.0-1.4 degrees C with accompanying changes in pest and pathogen incidence and to farming practices. Many pests and pathogens exhibit considerable capacity for generating, recombining, and selecting fit combinations of variants in key pathogenicity, fitness, and aggressiveness traits that there is little doubt that any new opportunities resulting from climate change will be exploited by them. However, the interactions between crops and pests and pathogens are complex and poorly understood in the context of climate change. More mechanistic inclusion of pests and pathogen effects in crop models would lead to more realistic predictions of crop production on a regional scale and thereby assist in the development of more robust regional food security policies.

Application of nox-restriction fragment length polymorphism for the differentiation of Brachyspira intestinal spirochetes isolated from pigs and poultry in Australia

Sixty-nine intestinal spirochetes isolated from pigs and poultry in eastern Australia were selected to evaluate the effectiveness of a species-specific PCR-based restriction fragment length polymorphism (RFLP) analysis of the Brachyspira nox gene. For comparative purposes, all isolates were subjected to species-specific PCRs for the pathogenic species Brachyspira hyodysenteriae and Brachyspira pilosicoli, and selected isolates were examined further by sequence analysis of the nox and 16S ribosomal RNA genes. Modifications to the original nox-RFLP method included direct inoculation of bacterial cells into the amplification mixture and purification of the PCR product, which further optimized the nox-RFLP for use in a veterinary diagnostic laboratory, producing sufficient product for both species identification and future comparisons. Although some novel profiles that prevented definitive identification were observed, the nox-RFLP method successfully classified 45 of 51 (88%) porcine and 15 of 18 (83%) avian isolates into 5 of the 6 recognized species of Brachyspira. This protocol represents a significant improvement over conventional methods currently used in veterinary diagnostic laboratories for rapid specific identification of Brachyspira spp. isolated from both pigs and poultry.

Prevalence, severity, and heritability of Stephanofilaria lesions on cattle in central and southern Queensland, Australia

Observations of cattle in central and southern Queensland are collated to de. ne the prevalence and area of Stephanofilaria lesions associated with infestations of the buffalo fly, Haematobia irritans exigua. The observations were made on herds that were being used for other purposes. In a survey of similar to 1500 animals at Belmont in central Queensland in 1982, 98% of cows and 70% of calves had lesions. Most lesions were on the neck and dewlap and 10% were raw and weeping at the time of sampling. The total area of lesions per animal was strongly related to cattle breed and age. Old Bos taurus animals had the greatest area of lesions, whereas young Bos indicus had the least. Heritability estimates were low, averaging 0.01 for calves and 0.18 for cows. A smaller survey of cows and steers at Craighoyle in central Queensland in 1986 showed a higher numbers of lesions and positive correlations between the total lesion area and animal size. The lesion area increased with tick survival, suggesting that tick-resistant animals are also resistant to Stephanofilaria infection. Steers had smaller areas of lesions than cows, as found previously with cattle ticks. Long-term monitoring observations in central and southern Queensland between 1981 and 1986 showed that the total area of lesions was seasonal with a peak in late summer, consistent with the seasonal incidence of buffalo fly. Animals segregated into Low and High lesion herds maintained their differences over time. The lesions penetrated the dermis of the cattle hides and rendered the affected area unusable, but few lesions occurred on valuable parts of the hide so such economic effects are likely to be insignificant. One animal nearly died of a haemorrhage from a lesion on the dewlap and had to be treated. The results can inform policy on buffalo fly control, and biosecurity preparations in relation to the potential establishment of the OldWorld screw-worm fly, Chrysomyia bezziana, in Australia, which will be facilitated by the lesions. The results emphasise the significant animal welfare and biosecurity risks posed by the lesions in northern Australia.

Biological Control of Eucalypt Pests Overseas and Australia

This project aims to address the growing need for a coordinated approach to research into the biological control of Australian eucalypt insect pests by scoping the formation of a Centre in Australia which would (a) coordinate the evaluation and provision of biological control agents (initially to South Africa and Brazil, but in future years more widely), (b) research the role natural enemies play in pest population regulation in Australian eucalypt plantations and how this may be enhanced as a management tool, and (c) form a network focussed on forest biosecurity with an emphasis on eucalypt pests and pathogens.

ANIMAL POPULATION ECOLOGY AND CONTROL FUNDAMENTALS

Expensive, extensive and apparently lethal control measures have been applied against many species of pest vertebrates and invertebrates for decades. In spite of this, few pests have been annihilated, and in many cases the stated goals have become progressively more modest, so that now we speak of saving foliage or a crop, rather than extermination. It is of interest to examine the reasons why animals are so difficult to exterminate, because this matter, of course, has implications for the type of control policy we pursue in the future. Also, it has implications for the problem of evaluating comparatively various resource management strategies. There are many biological mechanisms which could, in principle, enhance the performance of an animal population after control measures have been applied against it. These are of four main types: genetic, physiological, populationa1, and environmental. We are all familiar with the fact that in applying a control measure, we are, from the pest's point of view, applying intense selection pressure in favor of those individuals that may be preadapted to withstand the type of control being used. The well-known book by Brown (1958) documents, for invertebrates, a tremendous number of such cases. Presumably, vertebrates can show the same responses. Not quite so familiar is the evidence that sub-lethal doses of a lethal chemical may have a physiologically stimulating effect on population performance of the few individuals that happen to survive (Kuenen, 1958). With further research, we may find that this phenomenon occurs throughout the animal kingdom. Still less widely recognized is the fact that pest control elicits a populational homeostatic mechanism, as well as genetic and physiological homeostatic mechanisms. Many ecologists, such as Odum and Allee (1950, Slobodkin (1955), Klomp (1962) and the present author (1961, 1963) have pointed out that the curve for generation survival, or the curve for trend index as a function of last generations density is of great importance in population dynamics.