Diurnal behaviour of <Emphasis Type="Italic">Cephalcia lariciphila</Emphasis> (Hymenoptera: Pamphiliidae): relation to climatic factors and significance for monitoring

Adults of Cephalcia lariciphila emerge and start their activity at Vetrny Jenikov village (Czech Republic) during mid-April. First specimens were collected from Malaise traps at the same time as from emergence traps. Last specimens flew at the beginning of May or even the beginning of June. Adult occurrence typically lasted about 20 (–30) days. One Malaise trap and five yellow sticky boards were placed on three plots, and sawflies were collected at 1 hour intervals in two two-day periods. The variables with the best explanatory power for activity of imagos were temperature in interaction with humidity. Sawflies were most active at 16°C with 20% air humidity and numbers caught decreased steeply up to 0°C and 80% air humidity. The daily flight activity began at 10 a.m. and ended after 5 p.m. Both methods, the Malaise traps and the yellow sticky boards, should be used in forest practices from the middle of April.     

Detection of organophosphate and pyrethroid resistance alleles in Czech Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) populations by molecular methods

BACKGROUND: Pyrethroids and organophosphates are the most frequently used insecticides for Colorado potato beetle, Leptinotarsa decemlineata (Say), control in the Czech Republic. Based on molecular methods, organophosphate and pyrethroid resistance alleles have been detected in samples from three sites. The accuracy of restriction fragment length polymorphism (RFLP) and bidirectional polymerase chain reaction amplification of specific alleles (Bi‐PASA) for detection of resistance alleles is compared. RESULTS: Leptinotarsa decemlineata from three sites showed higher frequencies of resistance alleles to organophosphates than to pyrethroids. The rates of occurrence of individuals homozygous resistant (RR) to pyrethroids ranged from 20.0 to 22.9%, while the rates of occurrence of individuals RR to organophosphates ranged from 52.9 to 66.7%. The incidences of individuals with resistance alleles to both organophosphates and pyrethroids ranged from 8.6 to 13.6%. No relationship was found between incidence of the pyrethroid resistance allele and site, while incidence of the organophosphate resistance allele differed significantly according to site. CONCLUSION: Both RFLP and Bi‐PASA were suitable for detecting resistance alleles to pyrethroids, and in most cases also for detecting resistance alleles to organophosphates. In contrast to Bi‐PASA, RFLP was also suitable for samples with lower DNA quality when testing for the resistance allele to pyrethroids. On the other hand, RFLP was not as accurate as Bi‐PASA in detection of the organophosphate resistance allele. Copyright © 2010 Society of Chemical Industry     

Transformation of C-labelled lignin and humic substances in forest soil by the saprobic basidiomycetes Gymnopus erythropus and Hypholoma fasciculare

Litter decomposing basidiomycetous fungi produce ligninolytic oxidases and peroxidases which are involved in the transformation of lignin, as well as humic and fulvic acids. The aim of this work was to evaluate their importance in lignin transformation in forest litter. Two litter decomposing basidiomycete species differing in their abilities to degrade lignin - Hypholoma fasciculare, and Gymnopus erythropus - were cultured on sterile or non-sterile oak litter and their transformation of a ¹⁴C-labelled synthetic lignin (dehydrogenation polymer ¹⁴C-DHP) was compared with that of the indigenous litter microflora. Both in sterile and non-sterile litter, colonisation by basidiomycetes led to higher titres of lignocellulose-degrading enzymes, in particular of laccase and Mn-peroxidase (MnP). The titres of the latter were 6 to 40-fold increased in the presence of basidiomycetes compared to non-sterile litter. During 10 weeks, G. erythropus mineralised over 31% of ¹⁴C-DHP in sterile litter and 23% in non-sterile litter compared to 14% in the non-sterile control. Lignin mineralization by H. fasciculare was comparable to the non-sterile control, 12% in sterile litter and 16% in the non-sterile litter. The largest part of ¹⁴C from ¹⁴C-DHP was transformed into humic compounds during litter treatment with both fungi as well as in the control. In addition to the fast lignin mineralization, microcosms containing G. erythropus also showed a lower final content of unaltered lignin and 23-28% of the lignin was converted into water-soluble compounds with relatively low molecular mass (<5 kDa). Both G. erythropus and H. fasciculare were also able to further mineralise humic compounds. During a 10-week fungal treatment of an artificial ¹⁴C-humic acid (¹⁴C-HA) supplemented to the natural humic material of a forest soil, the fungi mineralised 42% and 19% of the labelled material, respectively, under sterile conditions. The ¹⁴C-HA mineralization by introduced basidiomycetes in microcosms containing non-sterile humic material, however, did not significantly differ from that of a non-sterile control and was around 12%. Altogether the results show that saprobic basidiomycetes can considerably differ in their rates of lignin and humic substance conversion. Furthermore, lignin degradation in forest soil can rather slow down by interspecific competition than it is accelerated by cooperation of different microorganisms occupying specific nutritional niches. Therefore, the overall contribution of saprobic basidiomycetes depends on their particular eco-physiological status and the competitive pressure, and may be often lower than initially expected. Significant lignin transformation including partial mineralization is seemingly not exclusively dependent on exceptional high titres of ligninolytic enzymes but also on so far unknown factors. Higher endocellulase production and subsequent weight loss was found in microcosms where saprobic basidiomycetes were combined with indigenous microbes. Potentially, lignin degradation by the basidiomycetes may have increased cellulose availability to the indigenous microbes.     

Methylglyoxal-induced modifications of significant honeybee proteinous components in manuka honey: Possible therapeutic implications

Methylglyoxal (MGO) is a major antibacterial component of manuka honey. Another antibacterial component found in Revamil honey, peptide defensin1, was not identified in manuka honey. The primary aim of the study was to evaluate the content of defensin1 in honeys of different botanical origins and to investigate a presumed effect of reactive MGO on defensin1 and a dominant protein of honey MRJP1 in manuka honey. Immunoblotting of honey samples showed that defensin1 was a regular but quantitatively variable component of honeys. One of the reasons of varying contents of defensin1 in different honeys seems to be constitutive but varying defensin1 expression in individual honeybees in bee populations that we documented on samples of nurse and forager bees by RT-PCR. Comparative analyses of honeys revealed a size modification of defensin1, MRJP1 and probably also α-glucosidase in manuka honey. We further showed that (i) the treatment of purified defensin1 in solution containing high amount of MGO caused a time-dependent loss of its antibacterial activity and (ii) increasing MGO concentrations in a non-manuka honey were connected with a gradual increase in the molecular weight of MRJP1. Obtained results demonstrate that MGO abrogates the antibacterial activity of defensin1 and modifies MRJP1 in manuka honey. We assume that MGO could also have negative effects on the structure and function of other proteins/peptides in manuka honey, including glucose oxidase, generating hydrogen peroxide.     

Characterization of <Emphasis Type="Italic">Agrobacterium</Emphasis> species by capillary isoelectric focusing

Tumorigenic and non-tumorigenic strains of Agrobacterium tumefaciens, A. rhizogenes, A.rubi, and A.vitis were examined using capillary isoelectric focusing, phenotypic determinative tests, PCR and fatty acid analysis. The isoelectric points (pI) of the 40 strains investigated clearly differentiated the strains according to their respective species. The different species were characterized with the following pI values: A. tumefaciens 2.2, A. rhizogenes 4.0, A. rubi 2.15, and A. vitis 2.6. This differentiation corresponded to the phenotypic, PCR and fatty acid characterizations. Strains with the similar chromosomal background but different plasmid content, e.g. A.vitis strain S4, and F2/5 gave the same pI values. Strains of Rhizobium species differed from Agrobacterium strains in their pI values. The advantage of capillary isoelectric focusing over the phenotypic determinative tests, PCR and fatty acid analysis is its speed (15 min), relative simplicity, and the very small amount of chemicals used. This rapid and simple method is a major improvement over the classical methods of separation of Agrobacterium species and should prove useful for rapid characterization of Agrobacterium-like colonies isolated from plant tumours for epidemiological and generic diversity studies.     

Long-term animal impact modifies potential production of N<Subscript>2</Subscript>O from pasture soil

At cattle overwintering areas, inputs of nutrients in animal excrements create conditions favourable for intensive microbial activity in soil. During nitrogen transformations, significant amounts of N₂O are released, which makes overwintering areas important sources of N₂O emission. In previous studies, however, increasing intensity of long-term cattle impact did not always increase emissions of N₂O from the soil: in some cases, N₂O emissions from the soil were lower at the most impacted area than at the moderately impacted one. Thus, the relationships between the level of long-term animal impact and potential production of N₂O from soil by denitrification were investigated in field and laboratory experiments. Field measurements indicated that the production of N₂O after glucose and nitrate amendments was greater in severely and moderately impacted locations than in an unimpacted location, while differences between the severely and moderately impacted locations were not significant. In laboratory experiments, the potential production of N₂O (measured as anaerobic production of N₂O after addition of glucose and nitrate) was highest in the moderately impacted soil. Surprisingly, potential N₂O production was lower in the most impacted than in the moderately impacted soil, and the net N₂O production in the highly impacted soil was further decreased by a significant reduction of N₂O to N₂. The expected stimulating effect of an increasing ratio of glucose C to nitrate N on the reduction of N₂O to N₂ during denitrification was not confirmed. The results show that cattle increase the denitrification potential of the soil but suggest that the denitrification potential does not increase indefinitely with increasing cattle impact.     

Postnatal functional maturation of blood phagocytes in pig

Developmental changes of functional ability of peripheral blood phagocytes from days 1 to 100 of life were investigated. Luminol enhanced chemiluminiscence was used to establish the ability of phagocytes to produce reactive oxygen species (ROS). Simple superoxide anion production was determined by spectrophotometrical measurement of cytochrome c. Activity of surface aminopeptidase N was assessed by spectrophotometrical measurement of l-alanine-p-nitroanilide. Flow cytometric measurements of CD18 and CD45 expression were performed. The ROS production per 0.5microl of blood did not show any trend; however, the values recalculated per 500 granulocytes had a decreasing course. The most noteworthy increase in production of superoxide anion occurred between days 17 and 26. Activity of aminopeptidase N decreased during the first 4 weeks. Expression of CD18 and CD45 intensively increased from days 1 to 14 with gradual decrease by day 100. Natural immunity develops during the early postnatal life and seems to be influenced by exposure of the organism to environmental antigens.     

Food selection and internal processing in Archegozetes longisetosus (Acari: Oribatida)

The nutritional biology of the oribatid mite Archegozetes longisetosus Aoki was examined using a control (starvation) and four types of food: Protococcus sp. algae from tree bark, two fungi—Stachybothrys sp. and Alternaria sp.—and filter paper. Direct observations, histology, enzymology and plating techniques were employed to record contact with food, contents and structure of the alimentary tract, presence and viability of bacterial microorganisms inside the mite body, and chitinase and cellulase activity of mite homogenates. Algae were highly palatable, resulting in high apocrine secretion and guts that were continuously full. Initially there was no evidence of chitinase or cellulase production. Chitinase activity started after 10 days, probably due to consumption of fungi that invaded the algal cover, and correlated with the presence of chitinolytic bacteria (Serratia rubidea) in the mite homogenate. Alternaria was grazed intensively, but cell walls of spores and hyphae remained intact and no chitinase was detected suggesting that only cell contents were enzymatically digested. Stachybothrys sp. was rejected as food. The bacterium Alcaligenes faecalis was dominant in homogenates in each of the treatments, but under starvation Achromobacter xylosoxidans became co-dominant. Cellulase activity was not detected in any treatment, but strong chitinase activity was induced with a filter paper (colonized by invasive fungi) diet. Furthermore, bacteria were common in mesenchyme between the internal organs in the filter paper and starvation treatments.