Halioticida noduliformans infection in eggs of lobster (Homarus gammarus) reveals its generalist parasitic strategy in marine invertebrates.

A parasite exhibiting Oomycete-like morphology and pathogenesis was isolated from discoloured eggs of the European lobster (Homarus gammarus) and later found in gill tissues of adults. Group-specific Oomycete primers were designed to amplify the 18S ribosomal small subunit (SSU), which initially identified the organism as the same as the 'Haliphthoros' sp. NJM 0034 strain (AB178865.1) previously isolated from abalone (imported from South Australia to Japan). However, in accordance with other published SSU-based phylogenies, the NJM 0034 isolate did not group with other known Haliphthoros species in our Maximum Likelihood and Bayesian phylogenies. Instead, the strain formed an orphan lineage, diverging before the separation of the Saprolegniales and Pythiales. Based upon 28S large subunit (LSU) phylogeny, our own isolate and the previously unidentified 0034 strain are both identical to the abalone pathogen Halioticida noduliformans. The genus shares morphological similarities with Haliphthoros and Halocrusticida and forms a clade with these in LSU phylogenies. Here, we confirm the first recorded occurrence of H. noduliformans in European lobsters and associate its presence with pathology of the egg mass, likely leading to reduced fecundity.

Selectivity of plant extracts for Trichogramma pretiosum Riley (Hym.: Trichogrammatidae).

We evaluated the selectivity of three plant extracts with potential insecticidal effects for the parasitoid Trichogramma pretiosum Riley, which is commonly used in biological pest control. The plant extracts assayed were an acetone extract of Toona ciliata M. Roem., commercial neem oil, and a nanoencapsulated formulation of neem oil (NC40). The toxicity of the plant extracts to T. pretiosum was evaluated according to the recommendations of the International Organization for Biological Control- IOBC Working Group. We assessed the susceptibility of adults of the maternal and F1 generations and immature stages of T. pretiosum to the extracts. Females exposed to egg cards treated with commercial neem oil parasitized almost 70% fewer eggs than control eggs treated with water; and this extract was therefore classified as slightly harmful. When the eggs were offered to females 24h after treatment with neem oil and aqueous NC40, the parasitism rate also decreased, and the two extracts were classified as slightly harmful. Adult emergence was lower for parasitoids that fed on host eggs offered 24h after the treatment with the T. ciliata extract, which was considered slightly harmful. The emergence of T. pretiosum from eggs, larvae and pupae treated with the different plant extracts, did not decrease compared to development stages treated with the water control. The use of T. pretiosum, combined with the application of an ethanol extract of T. ciliata and a nanoencapsulated formulation of neem, appears to be feasible in view of these low toxicity indices.

Trap Cropping Systems and a Physical Barrier for Suppression of Stink Bugs (Hemiptera: Pentatomidae) in Cotton.

Euschistus servus (Say), Nezara viridula (L.), and Chinavia hilaris (Say) (Hemiptera: Pentatomidae) are economic pests of cotton in the coastal plain of the southeastern United States. The objective of this 2-yr study was to determine the ability of trap cropping systems, pheromone-baited stink bug traps, and a synthetic physical barrier at the peanut-to-cotton interface to manage stink bugs in cotton. The physical barrier was the most effective management tactic. Stink bug density in cotton was lowest for this treatment. In 2010, boll injury was lower for the physical barrier compared to the other treatments except for soybean with stink bug traps. In 2011, boll injury was lower for this treatment compared to the control. Soybean was an effective trap crop, reducing both stink bug density in cotton and boll injury regardless if used alone or in combination with either stink bug traps or buckwheat. Incorporation of buckwheat in soybean enhanced parasitism of E. servus egg masses by Telenomus podisi Ashmead in cotton. The insertion of eyelets in the lid of the insect-collecting device of a stink bug trap allowed adult stink bug parasitoids, but not E. servus, to escape. Stand-alone stink bug traps were not very effective in deterring colonization of cotton by stink bugs or reducing boll injury. The paucity of effective alternative control measures available for stink bug management justifies further full-scale evaluations into these management tactics for control of these pests in crops.

Trans-generational parasite protection associated with paternal diet.

Multiple generations of hosts are often exposed to the same pathogens, favouring the evolution of trans-generational defences. Because females have more opportunities to transfer protective molecules to offspring, many studies have focused on maternally derived protection. However, males of many species can transfer compounds along with sperm, including chemicals that could provide protection. Here, we assess maternally and paternally derived protection in a monarch butterfly-protozoan parasite system where parasite resistance is heavily influenced by secondary plant chemicals, known as cardenolides, present in the larval diet of milkweed plants. We reared monarch butterflies on medicinal and non-medicinal milkweed species and then measured resistance of their offspring to infection. We also measured cardenolide content in adult monarchs reared on the two species, and in the eggs that they produced. We found that offspring were more resistant to infection when their fathers were reared on medicinal milkweed, while maternal diet had less of an effect. We also found that eggs contained the highest levels of cardenolides when both parents were reared on the medicinal species. Moreover, females reared on non-medicinal milkweed produced eggs with significantly higher levels of cardenolides if they mated with males reared on the medicinal milkweed species. However, we found an equivocal relationship between the cardenolides present in eggs and parasite resistance in the offspring. Our results demonstrate that males reared on medicinal plants can transfer protection to their offspring, but the exact mechanism remains unresolved. This suggests that paternal protection from parasitism might be important, particularly when there are environmental sources of parasite resistance and when males transfer spermatophores during mating.

The effect of floral resources on parasitoid and host longevity: prospects for conservation biological control in strawberries.

The strawberry tortricid, Acleris comariana Lienig and Zeller (Lepidoptera: Tortricidae) is an important pest in Danish strawberry production. Its most common parasitoid is Copidosoma aretas (Walker) (Hymenoptera: Chalcidoidea: Encyrtidae). To identify selective flowering plants that could be used to increase functional biodiversity, the longevity of C. aretas and its host A. comariana was assessed on 5 flowering species: buckwheat, Fagopyrum esculentum Moench (Caryophyllales: Polygonaceae); borage, Borago officinalis L. (Boraginaceae); strawberry, Fragaria x ananassa Duchesne (Rosales: Rosaceae); phacelia, Phacelia tanacetifolia Bentham (Boraginaceae); and dill, Anethum graveolens L. (Apiales: Apiaceae). Dill was only tested with C. aretas. Sucrose and pollen served as positive controls, and pure water as a negative control. In a subsequent field experiment, A. comariana larval density was assessed at 1, 6, and 11 m distances from buckwheat flower strips in 3 fields. The proportion of field-collected larvae that were parasitized by C. aretas or fungi was assessed. Among the tested floral diets, buckwheat was superior for C. aretas, increasing its longevity by 1.4 times compared to water. Although buckwheat also increased longevity of A. comariana, its longevity and survival on buckwheat, borage, and strawberry was not significantly different, so buckwheat was chosen for field experiments. A. comariana densities in the 3 fields with sown buckwheat flower strips were 0.5, 4.0, and 8.3 larvae per m per row of strawberry respectively. Of the collected larvae, a total of 1%, 39%, and 65% were parasitized by C. aretas, respectively. The density of A. comariana and the proportion parasitized by C. aretas were highly significantly correlated. Distance from floral strips had no significant effect on either A. comariana larval density or on the proportion of individuals parasitized by C. aretas. Few other parasitoids emerged from collected larvae, and no larvae were infected by entomopathogenic fungi. Still, total A. comariana mortality was significantly affected by distance to flower strips, with the highest mortality near the flower strips. As no effect of buckwheat flower strips on C. aretas parasitism was found, the positive effect they had on A. comariana control stems from unknown mortality factors. As literature indicates that buckwheat for flower strips can augment a more complex suite of natural enemies, one such mortality factor could be a non-consumptive predator and/or parasitoid effect, but this requires further study. If confirmed, buckwheat may be utilized together with a selective food plant, once identified.

Nutritional requirements of Pochonia chlamydospora and ARF18, fungal parasites of nematode eggs.

Twenty carbohydrates (C), 18 nitrogen compounds (N), and 9 vitamins were examined for their effects on the growth and conidiation of the nematode-egg-parasitic fungi Arkansas Fungus 18 (ARF18, isolate 908) and Pochonia chlamydospora var. chlamydospora in solid and liquid cultures. Glycogen was the best, and inulin, D-(+)-galactose, and soluble starch were good C sources for the growth of ARF18 in both solid and liquid cultures. ARF18 could not utilize alpha-cellulose in liquid; alpha-lactose and D-mannitol in solid; and D-(+)-xylose, L-(-)-sorbose, and D-(-)-ribose in both solid and liquid cultures. Casein was the best N source for ARF18 growth in both solid and liquid cultures and L-aspartic acid, DL-glutamic acid, peptone, and L-histidine were good in solid culture. ARF18 could not utilize L-cystine and L-tyrosine in solid culture, and L-cystine, DL-methionine, peptone, L-proline, and ammonium nitrate in liquid culture. Supplement of vitamins appeared to be unnecessary for ARF18 to grow. For P. chlamydospora var. chlamydospora growth, all test C sources, except L-(-)-sorbose, alpha-cellulose, citric acid, and D-(+)-glucose, were good in both solid and liquid cultures. Most N compounds were good for P. chlamydospora var. chlamydospora growth with casein and peptone the best. Vitamins had limited effect on P. chlamydospora var. chlamydospora growth. P. chlamydospora var. chlamydospora conidiation was well supported by D-(-)-ribose, D-(-)-fructose, melibiose, and D-(+)-galactose as C sources and by L-aspartic acid, DL-glutamic acid, and L-arginine as N sources. Excluding myo-inositol from the medium containing all other test vitamins increased P. chlamydospora var. chlamydospora conidiation, while excluding pyridoxine appeared to reduce its conidiation.

Artificial diets for rearing the Colorado potato beetle, Leptinotarsa decemlineata.

Colorado potato beetles have been reared successfully through 12 generations on artificial diets containing either 2.5% potato leaf powder or 2.5% lettuce leaf powder/0.75% potato leaf powder. For all but one of the treatment groups, the mean duration of each of the four larval stages was between 0.8 and 1.5 days longer than the durations exhibited by control beetles that had been fed on potato leaves. Maximum weights of prepupae, newly emerged adults and day 5-9 adults were approximately 78, 80 and 82%, respectively, of the weights for comparable stages of control beetles. Mean percent mortality for 1(st) instars was two to six times higher for artificial diet-fed CPBs than for leaf-fed beetles. However, since pupal mortality was four times higher for control beetles than for beetles reared on artificial diet, mean percent total mortality (newly hatched through the 9 day old adult) was equivalent for leaf-fed beetles and for later generations of potato and Lettuce+Potato diet-fed CPBs. Hemolymph ecdysteroid levels and fluctuations in mature 4th instar larvae and prepupae were similar in controls and experimental groups. Number of hatchlings produced per adult pair per day (fertility) was approximately eight times greater in control beetles than in later generations of artificial diet-fed beetles, primarily because fewer egg masses were laid per day, percent hatch was lower and cannibalism of eggs was higher in these latter groups. Interestingly, the mean percent hatch, although only 68% of the control value, was 1.5 times greater for beetles reared on diet containing lettuce-leaf powder, and a small percentage of potato leaf powder, than on diet containing only potato leaf powder. Percent hatch was equal for beetles fed on diet containing only lettuce-leaf powder and those fed on potato leaves. Finally, it is noteworthy that the quality of eggs, as judged by the ability of the wasp parasitoid, Edovum puttleri, to parasitize and develop in the eggs, was similar for eggs produced by control beetles and for those produced by beetles fed on potato and Lettuce+Potato diets. The diets and rearing system described here will be useful for providing beetles on a year-round basis for experiments designed to evaluate the effects of potential insect control agents, to investigate the mechanism(s) by which insects become resistant to control agents and for other applied and fundamental studies related to the control of this serious pest. The use of lettuce leaf powder in place of most of the potato leaf powder is especially advantageous because of the much reduced cost and greater availability of lettuce as compared to potato leaves.

Soil contamination with geohelminth ova in a tea plantation.

OBJECTIVE: To determine the degree of soil pollution in an area of high prevalence and the risk of exposure to geohelminth infection. DESIGN: A cross-sectional study. SETTING: A division of a large tea plantation in Kandy. SUBJECTS: All persons resident in one division of the tea plantation, above the age of 1 year. MEASUREMENTS: Prevalence of geohelminth infection measured by microscopic examination of faeces of the study group. Soil samples from 38 specific locations, obtained on 3 different occasions, about 6 months apart, analysed for helminth ova. The average temperature, rainfall and the number of rainy days during the sampling period. RESULTS: Of the 99 subjects of the study group, 77.7% were positive for at least one geohelminth infection. The prevalence rates for ascariasis, trichuriasis and hookworm infection were 54%, 28%, and 22% respectively. At the first sampling, 10.5% of the soil samples were positive for helminth ova with 7.8% and 2.6% samples showing Ascaris and Necator ova respectively. At the second sampling of the same locations nearly 40% of the samples were positive for helminth ova with 21.6%, 7.8% and 10.5% of the samples positive for Ascaris, Trichuris and Toxocara ova respectively. At the third sampling, 26.2% of the samples were positive with 21%, 2.6% and 2.6% of the samples positive for Ascaris, Trichuris and Toxocara ova respectively. Necator ova were found only once in 2.6% of the samples of the first sampling. The larger number of samples positive for helminth ova were in the second and the third samplings which coincided with high rainfall in the pre-sampling period. CONCLUSIONS: The prevalence of geohelminthiases among the study group was high. Results of soil analysis confirmed continued contamination of soil in and around the home gardens and the work areas with human and dog faeces. With the knowledge of exact locations of soil pollution it was possible to advice the management on remedial measures.