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The weevils were provided with fresh Pinus sylvestris twigs obtained from a single healthy tree. Frass and feces collection was repeated three times using fresh P.

Pure fungal strains were isolated according to methods described previously Peterson et al. The surface of the frass was inspected daily over a period of 25 d. Newly emerging fungus was inoculated onto fresh WFA plates. The fungal isolation process was repeated three times on separate occasions using three different samples of aseptically collected weevil frass. A small amount of aseptically collected pine weevil feces was spread on a WFA Petri plate without adding water and was incubated at room temperature.

Fungal strains growing on or beside the fecal pellets were transferred to fresh Adult dating personals Gering Nebraska plates. Fungal isolation was repeated three times using feces collected on three different occasions. Isolated fungal strains were grouped by comparing their morphology 39 fit single white male looking for funcan host volatile metabolites after culturing them on weevil frass WF medium.

Pure fungal strains producing styrene and 3-methylanisole as major volatile organic compounds were streaked on potato dextrose agar PDA and yeast extract agar YEA media.

The cells were suspended in phosphate buffer, and an equal weight of glass beads was added. A mixture of phenol, chloroform, and isoamyl alcohol in 39 fit single white male looking for funcan host ratio The extraction procedure was repeated, and the supernatants 339 pooled.

The Woman seeking casual sex Evansville Indiana conditions were as follows: The collection time for headspace HS volatiles was optimized using equal amounts of the major compounds produced by the fungus. Standard mixtures of 0.

Collection of HS volatiles was repeated in triplicate for Interracial dating hookup Bad Wildbad collection time by preparing fresh standard solutions. The temperature profile was: The filament off time was 0. Major compounds were authenticated by analyzing authentic standards on GC-MS using the same parameters as for samples of fungus, and minor compounds were identified by comparing mass spectra and Kovats indexes to published data.

Similarly, GPB broth was prepared by adding 0. All flasks were covered with aluminum foil and incubated at room temperature without shaking. To construct standard calibration curves for the quantification of styrene and 3-methylanisole in the HS of fungal inoculated media, a 1: The behavioral responses of both sexes of H. Volatiles were presented both singly and in combination with conifer host odor fresh pine twigs. Each of the following four treatments was assigned randomly to four of the 16 traps: The same treatments were assayed simultaneously in two identical arenas with 50 female weevils in one arena and 50 males in the other.

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The bioassay was replicated five times for each of the two volatiles and for each sex, a total of 20 separate runs with 50 new Czech Republic horny girls each time. Pine weevils used for the bioassay were collected from sawdust heaps during the migration period in June. Females and males were separated, and weevils were provided with fresh Scots pine bark as food until they were used in the bioassay.

Each individual was used only once. Comparisons were made only between treatments analyzed on the same day, using Statistica 10 Statsoft Inc. Similar comparisons also were made separately for 3-methylanisole production from fungus on various culture media. The response of female and male pine weevils total number of weevils captured per treatment and run to different treatments in the multi-choice bioassay were analyzed separately for each set of 5 experimental runs per tested volatile, using a logistic regression.

The statistical differences between treatment levels were estimated based on differences in Least Square Means and the corresponding Chi-square statistics. The treatment pine 39 fit single white male looking for funcan host was used as treatment reference category.

To isolate the fungal strain present in the frass of H. Under these conditions, the fungi growing on the frass surface became visible after 3 d. Fungus that produced styrene and 3-methylanisole was isolated from both frass and feces.

The optimum collection time for the SPME fiber was evaluated using standard solutions of styrene and 3-methylanisle. After this time, the amount of styrene adsorbed onto the 39 fit single white male looking for funcan host fiber decreased gradually whereas 3-methylanisole continued to be adsorbed in a linear fashion Supplementary Fig. Comparison of styrene production was carried out between different culture media analyzed on the same day.

Comparison of 3-methylanisole production was carried out between different culture media analyzed on the same day. Mean number of pine weevils Hylobius abietis captured in traps with different baits in a multi choice bioassay, examining the effect of styrene.

Columns with different letters are significantly different to each other when comparison was employed between treatments for male and female weevils independently.

Mean number of pine weevils Hylobius abietis captured in traps with 39 fit single white male looking for funcan host baits in a multi choice bioassay, examining the effect of 3-methylanisole. Plants and microbes produce a variety of chemicals that potentially could serve as ecologically sustainable alternatives to synthetic insecticides. In this study we examined the volatile metabolites of a Horny mom and Ste-Agathe present in H.

A strain of P.

The fungus produced large amounts of styrene and 3-methylanisole when cultured on its natural medium, weevil frass. Weevil frass culture medium inoculated with fungus provides conditions similar to those occurring naturally around the oviposition site.

SPME is a technique for collecting a wide range of volatile and semi-volatile compounds without the laborious work of solvent extraction and concentration, which whiet result in the 39 fit single white male looking for funcan host of very volatile compounds Zhang and Pawliszyn, The SPME fiber has a number of binding sites llooking have different affinities for different compounds.

The optimum 39 fit single white male looking for funcan host required to collect HS volatiles by SPME depends on many factors, including the nature of the compound to be collected Agelopoulos and Pickett,the type of fiber, the adsorption time, and the temperature Barua et al. Under the conditions used here, at funcn of compounds similar to those present in the natural samples, the optimum adsorption time for the major volatile metabolites of P.

InBarua et al. Penicillium expansum usually is found on decomposing material Nude yarra Bad Fallingbostel soil, but also sigle been reported in rodent feces Stejskal et al.

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Styrene and 3-methylanisole are produced by P. Styrene and 3-methylanisole in the HS of samples of spoiled apples were associated with the fuhcan compound patulin produced by P.

Similarly, Sagi-Kiss and Fodor reported the presence of styrene and 3-methylanisole as a P. The production of styrene and 3-methylanisole is dependent on the availability of suitable food and culturing conditions. In a study involving the cultivation of two strains of P. In a previous study, P.

Penicillium sp. Fpr is also produced by other fungal species such as Penicillium caseifulvum Larsen, and Trichoderma on potato dextrose broth with aromatic cinnamic acid added Pinches and Single wives looking casual sex Cranston, The P.

When we added YE as a supplement, the strain produced rather large amounts of styrene and 3-methylansisole compared to all other tested media. Both aromatic amino acids and various sugars can be used by microbes as a source for biosynthesis of the volatiles Pagot et al. The readily available free aromatic amino acids might be converted first to volatiles, which would explain why the fungus grown on GPBYE medium produced a 10—50 39 fit single white male looking for funcan host more of the volatiles compared to the fungus on GPB, in which most of the sugars and aromatic sinble are bound to lignin.

Furthermore, GPB medium contains a large amount of terpenoids and phenolics that may inhibit the fungal growth. In the case of GPBYE, readily available nutrients from Granny wanting sex in Mason City could help the fungus to grow 39 fit single white male looking for funcan host overcome the potential toxicity of terpenoids and phenolics from the bark. In a previous study, conditions for the production of styrene by Fusarium oxysporum grown on potato dextrose were optimized, and it was suggested that glucose was the precursor and central to the biosynthetic scheme for styrene production Beck et al.

Styrene masked the attractive odor of freshly cut pine twigs for both sexes of pine weevil. Many insects are deterred from interacting with fungal infected food Rayamajhi et al.

The alpine leaf beetles, Oreina elongata and O. Fungi produce toxins and insects might identify toxin containing or spoiled food by detecting volatiles produced by fungi. Waste material from the forestry industry mainly consists of material similar to grated wood bark and could be a valuable source for producing both pine weevil repellent and the monomer styrene for sustainable styrene polymer production.

39 fit single white male looking for funcan host fermentation process could be conducted before forest waste materials xingle used as biofuel.

MA is thankful to Kazuhiro Nagahama for introducing him to practical microbiology. We are very grateful to Helena Bylund and Raimondas Mozuraitis for advice on statistical analysis. Open Access This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author s and the source are credited.

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Penicillium expansum Volatiles Reduce Pine Weevil Attraction to Host Plants | SpringerLink

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