Nematode assemblages from Avicennia marina leaf litter in a temperate mangrove forest in south-eastern Australia

Meiofauna from Avicennia marina leaf litter in a temperate mangrove forest was enumerated, and the nematode assemblages compared on the bases of leaf colour (used as a guide to leaf age) and shore horizon where samples were collected. Twenty-one putative nematode species were collected from 48 leaf litter samples. Univariate analyses indicated that neither the colour of the leaf nor the shore horizon significantly affected abundance of nematodes. However, of the four (222) treatment groups, rarefaction curves revealed highest diversity on brown leaves from under the shade of the tree canopy (H'=0.751-0.126 SE, n=17). Species diversity of leaf litter nematodes was lower in this temperate mangrove system than reported from tropical mangrove studies. ANOSIM tests confirmed a significant effect of shore horizon on nematode assemblages. The dominant feeding group among nematodes was non-selective deposit feeders (7/21 species, but 77% of all nematodes). Epigrowth grazers were represented by 8/21 species of nematodes, but only 19% of the total number. Excised leaves became skeletonised by about 15 weeks. Shorter temporal scales of life cycles of nematodes compared with leaf degradation, and the dynamic nature of epibiontic assemblages, probably explain the similar assemblage structure on yellow and brown leaves.

Description of new species of Setosabatieria and Desmolaimus (Nematoda: Monhysterida) and a checklist of New Zealand free-living marine nematode species

Two new marine nematode species are described from intertidal sediments in southern New Zealand. Setosabatieria australis sp. nov. is characterised by an amphideal fovea with 3.5 turns, 7–9 small pre-cloacal supplements, and 2–5 sub-cephalic setae per file. Setosabatieria australis sp. nov. differs from all known species of the genus in having L-shaped spicules with a hollow median region, in contrast to the typical curved shape. Desmolaimus courti sp. nov. is characterised by the presence of four long (15 µm) cephalic setae, two cuticularised rings in the buccal cavity, and a lightly cuticularised pharyngeal lumen. It differs from other species of the genus by having eight sub-cephalic setae. A total of 116 marine species are included in the checklist of New Zealand free-living nematodes, with the majority of records from the coast of Campbell and Auckland islands from the early part of the 20th century. Most records were from habitats such as seaweed or sponge, which may explain the high proportion (>50%) of cosmopolitan species recorded in New Zealand to date. Little is known about the sediment nematode fauna, particularly in deeper waters (>120 m), from where there are no records to date.

Screening for resistance to potato cyst nematode in Australian potato cultivars and alternative solanaceous hosts

The potato cyst nematodes (PCN), Globodera rostochiensis (Woll.) and G. pallida (Stone), are major pests of ware and seed potato (Solanum tuberosum L.) crops worldwide and severely impact the movement of potatoes around the globe through quarantine restrictions. In Australia, only G. rostochiensis has been discovered, on four separate occasions between 1986 and 2008. The infested areas are the subject of strict regulation and quarantine procedures and while they are considered to be contained, managing nematode populations remains a priority. This study has identified the G. rostochiensis Ro1 resistance-status of potato cultivars currently grown by Australian potato growers, and new cultivars emerging from the Australian Potato Breeding Program. Resistance was assessed by a simple and robust procedure carried out in a purpose-built quarantine facility. Of the 24 potato cultivars grown in the affected Koo Wee Rup district in 2004, 10 were resistant to nematode infestation, including the locally important cultivar Atlantic. Other cultivars important to the Victorian and Australian potato industry, such as Kennebec, Desiree, Sebago and Coliban, were classified as susceptible. Importantly, this study provided evidence that the Koo Wee Rup PCN population was able to complete its lifecycle on the native plant species, S. aviculare (kangaroo apple), potentially acting as an alternate host and spreading PCN among potato crops.

Colonisation by epibionts and meiofauna of real and mimic pneumatophores in a cool temperate mangrove habitat

The size and pace of change in meiofaunal assemblages suggest that meiofauna make excellent subjects for testing theories about how ecological communities change. A field experiment was performed in which the  abundance and composition of epibionts and meiofauna on natural,  transplanted and mimic pneumatophores were monitored over a 47 wk period. Meiofaunal density increased with growth of algal epibionts, both reaching maximum values after 24 wk, at the end of winter. At this time the assemblages from the 3 substrata were similar, although the combined abundances of meiofauna on transplants and mimics were only 28% of the average on natural pneumatophores. Meiofaunal abundance on all substrata decreased rapidly during spring as algal cover declined due to desiccation. Twenty-three species of nematode were recorded from mimics compared with 8 and 7 from transplants and pneumatophores, respectively. A temporal sequence of feeding groups occurred in the order of epigrowth feeders, deposit feeders, and omnivore/predators, with the latter 2 adding to rather than replacing earlier trophic groups. Scavengers were found only on natural pneumatophores. The turnover rates of nematode species between all census times were similar, peaking at 63%, but there was no trend in the turnover rates with time. We conclude that mimics are more suitable than transplanted pneumatophores for colonisation studies because of their greater persistence and more easily standardised surface area. Moreover, the composition of colonising assemblages on them closely resembled assemblages on natural pneumatophores at the time of peak meiofaunal abundance.

A genomic approach to understanding the cause and effect of annual ryegrass toxicity

Annual Ryegrass Toxicity (ARGT) is a potentially lethal disease affecting livestock grazing on pastures or consuming fodder that include annual ryegrass (Lolium rigidum) contaminated with corynetoxins. The corynetoxins (CTs), among the most lethal toxins produced in nature, are produced by the bacterium Rathayibacter toxicus that uses a nematode vector to attach to and infect the seedheads of L.rigidum. There is little known of the factors that control toxin production. Several studies have speculated that a bacteriophage specific to R.toxicus may be implicated in CT production. We have developed a PCR-based assay to test for both bacterium and phage in ryegrass material and results indicate that there is a correlation between phage and bacterial presence in all toxic ryegrass samples tested so far. This PCR-based technique may ultimately allow for a rapid, high-throughput screening assay to identify potentially toxic pastures and feed in the field. Currently, ~80% of the 45 Kb genome has been sequenced an investigation to further elucidate its potential role in toxin production.Furthermore, specific alterations in gene expression as a result of exposure to CTs or the closely related tunicamycins (TMs), which are commercially available and considered biologically indistinguishable from CTs, will be evaluated for use as biomarkers of exposure. The effects of both toxins will be analysed in vitro using a rat hepatocyte cell line and screened on a low-density DNA micro array “CT-Chip” that contains <100 selected rat hepatic genes. The results are expected to further define the bioequivalence of CTs and TMs and to identify levels of exposure that are related to specific toxic effects or have no adverse effect on livestock.

Trapping and imaging of embryonic organisms using dielectrophoresis

Development of dielectrophoretic (DEP) arrays for real-time imaging of embryonic organisms is described. Microelectrode arrays were used for trapping both embryonated eggs and larval stages of Antarctic nematode Panagrolaimus davidi. Ellipsoid single-shell model was also applied to study the interactions between DEP fields and developing multicellular organisms. This work provides proof-of-concept application of chip-based technologies for the analysis of individual embryos trapped under DEP force.

Trapping and imaging of micron-sized embryos using dielectrophoresis

Development of dielectrophoretic (DEP) arrays for real-time imaging of embryonic organisms is described. Microelectrode arrays were used for trapping both embryonated eggs and larval stages of Antarctic nematode Panagrolaimus davidi. Ellipsoid single-shell model was also applied to study the interactions between DEP fields and developing multicellular organisms. This work provides proof-of-concept application of chip-based technologies for the analysis of individual embryos trapped under DEP force.

The influence of stocking rate and mixed grazing of Angora goats and Merino sheep on animal and pasture production in southern Australia. 4. Gastrointestinal parasitism

Gastrointestinal nematodes limit the growth, production and welfare of goats but there are few reliable sources of information for recommending management practices across flocks. The effects of animal species (Angora goat, Merino sheep, mixed-grazed goats and mixed-grazed sheep at the ratio of 1:1) and stocking rate (SR: 7.5, 10, 12.5 animals/ha) on gastrointestinal parasitism were determined in a replicated experiment on improved annual temperate pastures in southern Australia, from 1981 to 1984. Detailed monitoring of gastrointestinal nematodes was undertaken on animals before, during (five times per year) and at the conclusion of studies using faecal strongyle egg counts (WEC) and total worm counts. Sheep had a greater proportion of nematodes as Teladorsagia spp. and goats a greater incidence of Trichostrongylus spp. Both goats and sheep developed resistance to Nematodirus spp. during the experiment. WEC was similar in goats and sheep at the start of the experimental period but, thereafter, was consistently greater in goats than in sheep. While WEC was highly related to total worm count, the regressions for sheep and goats were different. Increasing the SR increased the WEC of goats and mixed-grazed goats but not of sheep. During the experiment, WEC declined at 7 and 10 animals/ha but increased at 12.5/ha. Mixed grazing with goats provided beneficial effects for sheep at all stocking rates, but the effects for goats were dependent on the stocking rate, being beneficial at 7.5 and 10/ha but harmful at 12.5/ha. The WEC of separately grazed goats were generally higher than the WEC of mixed grazed goats. The WEC of mixed sheep were lower than those of separately grazed sheep. During the experiment, the WEC of mixed grazed sheep declined faster than the WEC of separately grazed sheep but the WEC of separately grazed goats at 12.5/ha and of mixed grazed goats at 10 and 12.5/ha increased. Under the environmental and pastoral conditions examined, Angora wether goats should not be grazed at SR above those recommended for wether sheep. In the present study, the impact of gastrointestinal-nematode infections in goats was reduced at lower SR. Further, mixed grazing of Angora wether goats with wether sheep at or below the recommended SR resulted in reduced gastrointestinal parasitism for both sheep and goats, compared with monospecific grazing conditions. Goats did not represent a gastrointestinal-nematode hazard to sheep. © 2014 CSIRO.

Screening of the 'pathogen box' identifies an approved pesticide with major anthelmintic activity against the barber's pole worm

There is a substantial need to develop new medicines against parasitic diseases via public-private partnerships. Based on high throughput phenotypic screens of largely protozoal pathogens and bacteria, the Medicines for Malaria Venture (MMV) has recently assembled an open-access 'Pathogen Box' containing 400 well-curated chemical compounds. In the present study, we tested these compounds for activity against parasitic stages of the nematode Haemonchus contortus (barber's pole worm). In an optimised, whole-organism screening assay, using exsheathed third-stage (xL3) and fourth-stage (L4) larvae, we measured the inhibition of larval motility, growth and development of H. contortus. We also studied the effect of the 'hit' compound on mitochondrial function by measuring oxygen consumption. Among the 400 Pathogen Box compounds, we identified one chemical, called tolfenpyrad (compound identification code: MMV688934) that reproducibly inhibits xL3 motility as well as L4 motility, growth and development, with IC50 values ranging between 0.02 and 3 μM. An assessment of mitochondrial function showed that xL3s treated with tolfenpyrad consumed significantly less oxygen than untreated xL3s, which was consistent with specific inhibition of complex I of the respiratory electron transport chain in arthropods. Given that tolfenpyrad was developed as a pesticide and has already been tested for absorption, distribution, excretion, biotransformation, toxicity and metabolism, it shows considerable promise for hit-to-lead optimisation and/or repurposing for use against H. contortus and other parasitic nematodes. Future work should assess its activity against hookworms and other pathogens that cause neglected tropical diseases.