Non-invasive assessment of parasitic nematode species diversity in wild Soay sheep using molecular markers

Considerable effort has been put into detecting and identifying parasitic nematodes in live ruminants, but to date most studies are limited to a small group of nematodes and/or to experimentally infected sheep. In this study, a PCR-based assay using species-specific primer pairs, located in the second internal transcribed spacer ribosomal DNA, was developed to identify nine different species from six different families of parasitic nematodes in a wild, unmanaged and naturally infected population of sheep. Each primer pair was tested for its specificity and sensitivity and it exclusively amplified the species it was designed for and exhibited a high degree of sensitivity. The method was applied to eggs and cultured larvae to identify the parasitic nematodes present in a pooled faecal sample from several host individuals with unknown parasite burden. To test detection reliability, a faecal sample from an individual with known parasite burden (through post-mortem analysis) was also examined. All species present could be correctly identified by PCR, but detecting very low levels and/or early stages of infection proved to be difficult. The method was also tested for its applicability to high through-put screening of faecal samples.     

Aquatic nematodes from Ethiopia III

Six species of the genus Eumonhystera, one of which is new to science, are described from bottom samples of Lake Tana and two associated rivers, River Abbay and R. Gelda, Ethiopia. E. geraerti n.sp. can be identified by being the only species in the genus that combines very posteriorly positioned vulva [V% = 68.2 ± 0.7 (67.3–69.3)] with very anteriorly positioned amphids (Amph-ABE/LRW = 0.8). The other five species encountered, E. dispar (Bastian, 1865) Andrássy, 1981, E. filiformis (Bastian, 1865) Andrássy, 1981, E. pseudobulbosa (von Daday, 1896) Andrássy, 1981, E. mwerazii (Meyl, 1957) Andrássy, 1981, and E. vulgaris (de Man, 1880), Andrássy, 1981, are described in detail. The identities of some of the already described populations of E. dispar, E. filiformis and E. vulgaris are questioned and clearer and better ways of defining each of these three species are suggested. A peritrophic-like membrane on the outer side of the brush-border of the intestine of E. mwerazii, a species reported here for the first time out of its type locality, is reported. Also, scanning electronmicrographs of sectioned and entire specimens of E. mwerazii are presented. All species are reported from Ethiopia for the first time.     

Maggot therapy: a review of the therapeutic applications of fly larvae in human medicine, especially for treating osteomyelitis

In traditional medical practice, the larvae of some Diptera: Calliphoridae, notably Lucilia illustris (Meigen), L.sericata (Meigen) and Phormia regina (Meigen), have been employed for maggot therapy, i.e. to help clean lesions antiseptically, especially for treatment of chronic osteomyelitis. This mode of treatment remains appropriate for cases where antibiotics are ineffective and surgery impracticable.     

Merino ewes bred for parasite resistance reduce larval contamination onto pasture during the peri-parturient period

The peri-parturient period is crucial for controlling worms as the acquired immunity of ewes is disrupted, resulting in an increase in faecal worm egg counts. Two hypotheses were tested in this experiment - that ewes bred for worm resistance would have lower faecal worm egg counts than unselected control ewes, during late pregnancy and lactation, under similar but separate grazing areas; and also that numbers of infective nematode larvae would be lower on pastures grazed by resistant ewes than pastures grazed by unselected control ewes. Faecal samples were collected from resistant and unselected ewes in late pregnancy and early lactation, during the winter rainfall season, and analysed for numbers of Trichostrongylus colubriformis and Teladorsagia circumcincta. Pasture samples were taken 1 week before and 7 weeks after lambing started and analysed for infective larvae. In all sheep, worm egg counts rose 2 weeks prior to lambing and continued into lactation. Worm egg counts were significantly lower in the resistant ewes from 1 week before lambing to 2 weeks after lambing. There were no differences in egg counts between single- and twin-bearing ewes in the resistant line. However, twin-bearing control ewes had significantly higher egg counts than single-bearing control ewes. Following lactation, plots grazed by resistant ewes had substantially less contamination with T. colubriformis larvae, but there were no differences in numbers of T. circumcincta larvae. Our results demonstrate that sheep bred for worm resistance has lower worm burdens during the peri-parturient phase and that lambs born to resistant ewes face a lower larval challenge during their introduction to grazing. In our environment, selection for low worm egg counts has produced sheep highly resistant to T. colubriformis, but has had less impact on resistance towards T. circumcincta.     

Hoplolaimoidea (Nematoda: Tylenchida) from the Aleutian Islands with Descriptions of Four New Species

Four new species of hoplolaimoid nematodes (Merlinius adakensis, Pratylenchoides megalobatus, Pratylenchus pratensisobrinus, and Pratylenchus ventroprojectus) are described from Adak Island in the Aleutian chain. M. adakensis n. sp. is separated from other species by body length (0.96-1.3 mm), stylet length (32-36 μm), number of tail annules (49-68), and c'' (3.1-4.1). P. megalobatus n. sp. differs from all known Pratylenchoides spp. by having a very long esophageal gland lobe (b'' = 2.4-3.3, overlap 3-6 times the body width). P. pratensisobrinus n. sp. closely resembles P. pratensis (de Man) Filipjev, but has a longer stylet (15-17 μm), a longer tail (c = 12-15; c'' = 2.8-3.7), and more tail annules (23-37). P. ventroprojectus n. sp. is distinguished by body length (392-475 μm), three lip annules, low and flattened cephalic capsule, and presence of terminal subventral projection. Pratylenchoides variabilis Sher, Helicotylenchus amplius Anderson &Eveleigh, and H. spitsbergensis Loof are also reported from Adak and Amchitka Islands.     

Characterisation of a 34 kD protein from potato cyst nematodes, using monoclonal antibodies with potential for species diagnosis

Two monoclonal antibodies, which differentially recognise the two species of potato cyst nematodes (PCN), Globodera pallida and G. rostochiensis, are described. They have been shown to have potential for quantification of these two species, recognising proteins of the same molecular weight (34 kD) in both species. Further investigation showed these proteins to have isoelectric points at pH values of 5.7 in G. pallida and 5.9 in G. rostochiensis, in common with the proteins used by Fleming & Marks (1983) to differentiate the species of PCN. They are likely to be structurally very similar, with the same physiological function (and therefore similar concentrations) in the two species. In cross-reactivity tests with a wide range of soil nematode species, the antibodies reacted strongly only with species of the genus Globodera, and thereby confirmed their potential as the basis of a quantitative immunoassay likely to be useful in management of PCN populations.     

Effects of Caenorhabditis elegans (Nematoda: Rhabditidae) on the spread of the bacterium Pseudomonas tolaasii in mushrooms (Agaricus bisporus)

The effects of mass-produced saprobic rhabditid nematodes, Caenorhabditis elegans on the spread of the bacterial blotch pathogen, Pseudomonas tolaasii , were studied in mushroom growth chambers. C. elegans significantly reduced the intensity of blotch on sporophores. Repeated isolations of the bacterial flora from the gut of C. elegans recovered from mushroom sporophores during cropping, revealed the presence of Pseudomonas fluorescens biovar reactans . All the isolates of P. fluorescens biovar reactans isolated from nematodes were antagonists of P. tolaasii .
C. elegans produced much larger populations in monoxenic cultures with P. fluorescens biovar reactans than with P. tolaasii . It is suggested that as C. elegans selects P. fluorescens biovar reactans rather than P. tolaasii as a food substrate it probably spreads the antagonist in the mushroom crop and may contribute to the control of bacterial blotch.     

In vitro characterization bioassays of the nematophagous fungus Purpureocillium lilacinum: Evaluation on growth,extracellular enzymes,mycotoxins and survival in the surrounding agroecosystem of tomato

The effects of water stress and temperature on in vitro growth and enzymatic activity of Purpureocillium lilacinum (Sordariomycetes, Hypocreales, Ophiocordycipitaceae) isolates with demonstrated capacity to control Nacobbus aberrans (Secernentea, Tylenchida, Pratylenchidae) were evaluated in this study. Also, saprophytic and endophytic colonization in tomato plants were determined. P. lilacinum was able to grow under the evaluated levels of osmotic and matric stress, but the increase in water stress caused reductions in radial growth rates. Moreover, the fungal isolates produced chitinases, proteases, and leucinostatins under inductive conditions. The nematophagous fungi were able to develop saprophytically (10⁴ CFU g^?1 of soil). Meanwhile, only P. lilacinum SR38 demonstrated endophytic capacity. The results suggest that P. lilacinum can be effectively applied as biocontrol agents of phytoparasitic nematodes in tomatoes under variable agroecological conditions.