The Dissemination of Metarhizium brunneum Conidia by Females of the Red Palm Weevil, Rhynchophorus ferrugineus, Suggests a New Mechanism for Prevention Practices.

Direct contact between the conidia of entomopathogenic fungi (EPF) and their host is a prerequisite to successful infection; the host can, therefore, be infected by both direct treatment and by transmission of fungal inoculum from infested surfaces. This unique characteristic makes EPF especially relevant for the control of cryptic insects. In the case of the red palm weevil (RPW) Rhynchophorus ferrugineus, the eggs and larvae are almost inaccessible to direct-contact treatment. The objective of the present study was to investigate the mechanism of conidia transmission from a treated surface to host eggs and larvae. Foam pieces infested with Metarhizium brunneum conidial powder, conidial suspension, or distilled water were used as a laying surface for RPW females. The number of eggs laid was not affected by the EPF treatments and ranged from 2 to 14 eggs per female. However, hatching rate and larval survival were significantly reduced in the conidial powder treatment, resulted in 1.5% hatching and no live larvae. In the conidial suspension treatment, 21% of laid eggs hatched, compared to 72% in the control treatment. In both M. brunneum treatments, females' proboscis, front legs and ovipositor were covered with conidia. The females transferred conidia in both treatments to the laying holes, reaching up to 15 mm in depth. This resulted in reduced egg-hatching rate and significant larval mortality due to fungal infection. The stronger effect on egg and larval survival using dry conidia seemed to result from better conidial adhesion to the female weevil in this formulation. In future studies, this dissemination mechanism will be examined as a prevention strategy in date plantations.

Survival of Nematode Larvae after Treatment with Eugenol, Isoeugenol, Thymol, and Carvacrol.

BACKGROUND: Helminthiases inflict annual losses on the meat and dairy livestock industries. The commonest species of ruminant parasites are the nematodes: Strongyloides papillosus and Haemonchus contortus, which lay eggs in the intestine and enter the feces. There, the eggs develop into larvae, which when voided with the feces crawl onto plants. METHODS: In our experiment, we evaluated the survivability of the noninvasive and invasive (L1-2 and L3, respectively) larvae of S. papillosus, H. contortus (L3), and Muellerius capillaris (L1) in vitro by subjecting each to natural compounds present in the essential oils of many plants. In the experiment, we used aqueous emulsions of eugenol, isoeugenol, thymol, and carvacrol. RESULTS: Administering 1% concentrations of those compounds killed 100% of the nematode larvae following 24 h of exposure. Thymol, eugenol, and isoeugenol at a concentration of 0.1% also caused high larvae mortality (over 96%). CONCLUSIONS: Continuous usage of synthetic anthelmintic drugs in veterinary medicine has led to the parasites developing resistance, thus, a search for novel nematicidal drugs is required. Eugenol, isoeugenol, thymol, and carvacrol are promising compounds against nematodes. However, additional research is required regarding peculiarities in their actions toward the bodies of mammals and parasitic nematodes.

Evaluation of Toxicity with Brine Shrimp Assay.

The in vivo toxicity of new metallodrugs either as Small Bioactive Molecules (SBAMs) or Conjugates of Metals with Drugs (CoMeDs) or their hydrogels such as with hydroxyethyl-methacrylate (HEMA) (pHEMA@SBAMs or pHEMA@CoMeDs) are evaluated by the brine shrimp assay. Thus individuals of Artemia salina larvae are incubated in saline solutions with SBAMs, CoMeDs, pHEMA@SBAMs or pHEMA@CoMeDs or without for 24 h. The toxicity is then determined in terms of the mortality rate of brine shrimp larvae. Brine shrimp assay is a low cost, safe, no required feeding during the assay, while it requiring only a small amount of the tested agent.

Addition of a surfactant to water increases the acaricidal activity of extracts of some plant species used to control ticks by Zimbabwean smallholder farmers.

BACKGROUND: Many studies have revealed that bioactive compounds for different indications are not extracted from plants with water, the only extractant practically available to rural communities. We compared the acaricidal activity of acetone extracts of 13 species used traditionally to protect cattle against ticks. We also investigated if the extraction of biologically active compounds against Rhipicephalus (Boophilus) decoloratus ticks could be enhanced by adding a liquid soap that is locally available to smallholder farmers. METHODS: A total of 13 plant species selected based on reported traditional use in Zimbabwe, were dried and finely ground before extraction with water, or water plus a surfactant, or acetone. The adapted Shaw Larval Immersion Test (SLIT) method was used to determine the activity of acetone and crude water extracts with or without liquid soap against the tick larvae. The activity of four fractions of crude acetone extracts (extracted using solvents of different polarity), of the most active plant species, Maerua edulis (tuber and leaf) was also compared to identify the most active fraction. RESULTS: Aqueous plant extracts were not toxic to ticks, but the addition of 1% liquid soap as a surfactant increased mortality of the R. (B) decoloratus larvae significantly. With the Maerua edulis tuber extract, the efficacy of the 1% liquid soap was comparable to that of the amitraz based commercial synthetic acaricide. The use of acetone as an extractant, also increased the mortality of the tick larvae in all the plant species. With M. edulis (tuber and leaf), Monadenium lugardae and Kleinia sp. acetone extracts, the activity was comparable to that of the positive control (a commercially available amitraz-based synthetic acaricide). The non-polar fractions of the acetone extract of leaf and tuber of M. edulis caused up to 100% mortality. This indicates that non-polar to intermediate polarity compounds are responsible for the acaricidal activity. CONCLUSION: Organic solvents such as acetone extracted active compounds but water did not. By adding commonly available dishwashing soap to water active compounds were extracted leading to a high acaricidal activity of the plant extracts. In some cases, it was as active as non-polar extracts and a synthetic commercial acaricide (positive control). This approach makes it possible for the smallholder farmers and traditional healers to extract biologically active compounds from plants by using water.

Acquirement of CRY8DB Transgenic Tall Fescue (Festuca arundinacea Schreb.) by Agrobacterium tumefaciens to Develop Resistance Against Pentodon idiota Herbest.

Tall fescue plants are widely exposed to white grubs. Regarding the rate of damage caused by the white grubs to tall fescue and difficulty of its ecological and economical control, production of resistant cultivars is a priority. In this experiment, for the first time, we report production of transgenic lines resistant to white grub using CR8DB gene. For this, mature seeds were placed on MS basal medium with 0-15 mg L-1 2,4-D for callogenesis and 0-1.75 mg L-1 BA for regeneration. 'Asterix' (54.11%) in 7.5 and 'Talladega' (52.53%) in 10 mg L-1 2,4-D showed maximum callogenesis. Regeneration percentage was higher in 0.5 mg L-1 BA. Agrobacterium tumefaciens strain LBA4404 harbouring binary vector pCAMBIA 1301 with CRY8DB gene, which contains HPTII gene and uidA and various types MS media were used for transformation of calli. The highest percentage of gus enzyme activity and hygromycin resistance in calli was related to the modified medium type 11. The PCR and RT-PCR analysis was done to confirm the presence and expression of the target gene in transgenic 5 lines in 'Asterix' and 3 lines in 'Talladega'. According to bioassay, larvae mortality of 91.66% was observed in transgenic plants, whereas it was 15.52% in control plants.

Ecotoxicity response of chlorpyrifos in Aedes aegypti larvae and Lactuca sativa seeds after UV/H2O2 and UVC oxidation.

Chlorpyrifos (CP) is an organophosphate pesticide widely used in agriculture known to cause neurological and immunological effects in addition to interfering in the reproduction and development of organisms. In this study, CP degradation by UV/H2O2 process and UVC radiation was investigated, and the ecotoxicity and phytotoxicity was evaluated using bioassays of Aedes aegypti larvae and Lactuca sativa seeds. CP degradation was monitored by HPLC-DAD, and kinetic parameters were calculated for all processes evaluated. Results demonstrated that both processes are efficient, showing a reduction of over 97% of initial CP after 20 and 60 min of UV/H2O2 and UVC radiation, respectively. However, samples treated by UV/H2O2 process demonstrated increase of toxicity, leading to larvae mortality (>90% of organisms) and inhibition effects in seed root growth. The relationship between increased toxicity and the CP byproducts formed was not confirmed due to its low concentration. However, the direct influence of acetonitrile solvent, specifically their toxic byproducts, was observed. This study provides insights into parent compound abatement using oxidative treatment and the changes in toxicity due to the transformation of CP byproducts and complex mixtures (acetonitrile as solvent and hydrogen peroxide).

Efecto de la calidad del agua de criaderos de mosquitos (Diptera: Culicidae) sobre la patogenicidad e infectividad de las zoosporas del hongo Leptolegnia chapmanii (Straminipila: Peronosporomycetes)

Effect of water quality in mosquito breeding sites on the pathogenicity and infectivity of zoospores from the fungus Leptolegnia chapmanii (Straminipila: Peronosporomycetes). The fungus Leptolegnia chapmanii is highly pathogenic to mosquito larvae in Argentina. We studied if physical and chemical characteristics of the water from mosquito breeding sites affect pathogenicity, and the infectivity of zoospores of L. chapmanii. Water samples were taken from pools filled by rains, urban ditches with domestic waste water, pools filled by overflow from Río de la Plata, and flower vases from the Cemetery of La Plata city. Sub-samples of water were analyzed for physical and chemical characteristics, while other sub-samples were used for laboratory bioassays. Containers with 150 ml of water samples, 25 Aedes aegypti larvae, and 2.8 x 105 zoospores of L. chapmanii, were incubated under controlled environment, and larval mortality was recorded after 48 h. There were highly significant differences among mortalities in water from cemetery vases (70.2%), rain pools water (99.5%), and pools with water from Rio de la Plata (95%). There were no significant differences among larval mortalities in water from ditches, rain pools and Río de la Plata pools. Leptolegnia chapmanii was successful as a biological control agent in all kinds of tested water qualities, producing high larval mortality. Rev. Biol. Trop. 57 (1-2): 371-380. Epub 2009 June 30.

Leptolegnia chapmanii es un potente patógeno de larvas de mosquitos. El objetivo del trabajo fue determinar si las características físico-químicas del agua de criaderos de culícidos de importancia sanitaria a nivel mundial afectan la patogenicidad e infectividad de las zoosporas de L. chapmanii. Se tomaron muestras de cuatro tipos de agua en Argentina: de lluvia, de zanja, del Río de La Plata, y de floreros del cementerio local. A una parte del agua se le realizaron estudios físicos-químicos, y otra parte fue utilizada en ensayos. Se colocó 150 ml de cada muestra en recipientes plásticos, con 25 larvas sanas de Aedes aegypti y 2.8 x 105 zoosporas de L. chapmanii. Fueron incubadas a 25 ºC y 12-12 (L-O). La mortalidad larval fue registrada a las 48 horas. Se utilizaron tres recipientes y un control por tratamiento, con tres repeticiones realizadas en distintos días. Se observaron diferencias altamente significativas entre la mortalidad larval en aguas de floreros del cementerio (70.2%), aguas de lluvias (99.53%), y del río de La Plata (95%). No hubo diferencias significativas entre la mortalidad larval en agua de zanja, lluvia y río. Leptolegnia chapmanii resultó exitoso en los cuatro tipos de aguas evaluados, produciendo elevados niveles de mortalidad larval.