Biodegradation of the Pyrethroid Pesticide Esfenvalerate by Marine-Derived Fungi

Esfenvalerate biodegradation by marine-derived fungi is reported here. Esfenvalerate (S,S-fenvalerate) and its main metabolites [3-phenoxybenzaldehyde (PBAld), 3-phenoxybenzoic acid (PBAc), 3-phenoxybenzyl alcohol (PBAlc), and 2-(4-chlorophenyl)-3-methylbutyric acid (CLAc)] were quantitatively analyzed by a validated method in triplicate experiments. All the strains (Penicillium raistrickii CBMAI 931, Aspergillus sydowii CBMAI 935, Cladosporium sp. CBMAI 1237, Microsphaeropsis sp. CBMAI 1675, Acremonium sp. CBMAI 1676, Westerdykella sp. CBMAI 1679, and Cladosporium sp. CBMAI 1678) were able to degrade esfenvalerate, however, with different efficiencies. Initially, 100 mg L^?1 esfenvalerate (Sumidan 150SC) was added to each culture in 3 % malt liquid medium. Residual esfenvalerate (64.8–95.2 mg L^?1) and the concentrations of PBAc (0.5–7.4 mg L^?1), ClAc (0.1–7.5 mg L^?1), and PBAlc (0.2 mg L^?1) were determined after 14 days. In experiments after 7, 14, 21, and 28 days of biodegradation with the three most efficient strains, increasing concentrations of the toxic compounds PBAc (2.7–16.6 mg L^?1, after 28 days) and CLAc (6.6–13.4 mg L^?1, after 28 days) were observed. A biodegradation pathway was proposed, based on HPLC-ToF results. The biodegradation pathway includes PBAld, PBAc, PBAlc, ClAc, 2-hydroxy-2-(3-phenoxyphenyl)acetonitrile, 3-(hydroxyphenoxy)benzoic acid, and methyl 3-phenoxy benzoate. Marine-derived fungi were able to biodegrade esfenvalerate in a commercial formulation and showed their potential for future bioremediation studies in contaminated soils and water bodies.     

Steroid Compounds from Far Eastern Starfishes <Emphasis Type="Italic">Henricia aspera</Emphasis> and <Emphasis Type="Italic">H. tumida</Emphasis>

Six new natural compounds were isolated from two Far Eastern starfish species, Henricia aspera and H. tumida, collected in the Sea of Okhotsk. Two new glycosylated steroid polyols were obtained from H. aspera: asperoside A and asperoside B, which were shown to be (20R,24R, 25S)-3-O-(2,3-di-O-methyl-β -D-xylopyranosyl)-24-methyl-5α-cholest-4-ene-3β, 6β,8,15α,16β,26-hexaol and (20R, 24R,25S,22E)-3-O-(2,4-di-O-methyl-β-D-xylopyranosyl)-24-methyl-5α-cholest-22-ene-3β,4β,6β,8,15α,26-hexaol, respectively. Two other glycosylated polyols, tumidoside A, with the structure elucidated as (20R, 22E)-3-O-(2,4-di-O-methyl-β -D-xylopyranosyl)-26,27-dinor-24-methyl-5α-cholest-22-ene-3β,4β,6β,8,15α,25-hexaol, and tumidoside B, whose structure was elucidated as (20R,24S)-3-O-(2,3-di-O-methyl-β-D-xylopyranosyl)-5α-cholestan-3β,4β,6β,8,15α,24-hexaol, were isolated from the two starfish species. (20R, 24S)-5α-Cholestan-3β,6β,15α,24-tetraol and (20R, 24S)-5α-cholestan-3β,6β,8,15α,24-pentaol were identified only in H. tumida. The known monoglycosides henricioside H1 and laeviuscolosides H and G were also identified in both species.     

Multi-locus phylogeny and morphology reveal five new species of <Emphasis Type="Italic">Fomitiporia</Emphasis> (Hymenochaetaceae) from China

Five newly identified species of Fomitiporia (F. alpina, F. gaoligongensis, F. hainaniana, F. subrobusta and F. subtropica) and their morphological and molecular characterisation are described in this paper. Fomitiporia alpina sp. nov. is distinguished by its pileate basidiomata, parallel tramal hyphae and large basidiospores (6.5–8 × 6–8 μm), and by its gymnosperm wood-living habitat. Fomitiporia gaoligongensis sp. nov. is distinct from other species due to its semicircular pileus and subglobose to globose basidiospores (6.5–7.6 × 6–7.4 μm). Fomitiporia hainaniana sp. nov. is marked by its resupinate basidiomata, the presence of setae and small globose basidiospores (4–5 × 3.8–4.4 μm). Fomitiporia subrobusta sp. nov. is characterised by its triquetrous basidiomata, small pores (6–9 per mm) with entire and thick dissepiments, and subglobose to obovoid basidiospores (6.2–6.8 × 5.2–6 μm). Fomitiporia subtropica sp. nov. can be differentiated by its resupinate basidiomata, smaller pores (6–10 per mm) and smaller basidiospores (5.2–6 × 4.4–5 μm). Phylogenetic analysis, based on multi-gene comparison of the internal transcribed spacer regions (ITS), nuclear large subunit ribosomal RNA gene regions (nLSU), the translation elongation factor 1-α gene (tef1α) and the second subunit of RNA polymerase II (rpb2), confirmed affinity with the Fomitiporia species and showed association with similar fungi in the genus.     

Two new steroid glycosides from the Far East starfish <Emphasis Type="Italic">Hippasteria kurilensis</Emphasis>

Two new steroid glycosides were isolated from the Far East starfish Hippasteria kurilensis collected in the Sea of Okhotsk. They were characterized as (22E,24R)-3-O-(2-O-methyl-β-D-xylopyranosyl)-24-O-[2-O-methyl-β-D-xylopyranosyl-(1→5)-α-L-arabinofuranosyl]-5α-cholest-22-ene-3β,4β,6α,7α,8,15β,24-heptaol (kurilensoside I) and (24S)-3-O-(2-O-methyl-β-D-xylopyranosyl)-24-O-(α-L-arabinofuranosyl)-5α-cholestane-3β,4β,6β,15α,24-pentaol (kurilensoside J). In addition, the earlier known glycosides linkosides F and L1, leviusculoside G, forbeside L, desulfated echinasteroside, and granulatoside A were isolated and identified. The structures of the new compounds were established with the help of two-dimentional NMR spectroscopy and mass- spectrometry.     

Intestinal enterobacteria of the hibernating <Emphasis Type="Italic">Apis mellifera mellifera</Emphasis> L. bees

Dynamics of enterobacteria of normal intestinal microflora was studied in Apis mellifera mellifera L. bees hibernating under snow in the Western Urals. The cell numbers (N) of the predominant species Klebsiella oxytoca increased from 10-10⁶ CFU/bee in November 2004 to 10⁴-10⁷ CFU/bee in March 2005; its frequency of occurrence (P) increased from 92 to 100%. Increase of Providencia rettgeri (11.2004: N up to 10⁶, P 25%; 03.2005: N 10²-10⁶, P 80%) was accompanied by the substitution of Morganella morganii (11.2004: N up to 10⁶, P 25%) with Proteus vulgaris (03.2005: N up to 10⁵, P 8%). By spring, Hafnia alvei and Citrobacter sp., which are pathogenic to bees, disappeared (11.2004: N up to 10⁵, P 13 and 10%, respectively). Endophytic species Pantoea agglomerans, Leclecria sp., and other representatives of the “Enterobacter agglomerans” group were present in November and after the first emergence in spring (N up to 10⁵; November: P 15%; April: P 23%). In April, the number of enterobacteria decreased to 10⁵, and P. rettgeri became the predominant species (P 54%) instead of K. oxytoca (P 43%).     

Genetic divergence of sympatric charrs of the genus <Emphasis Type="Italic">Salvelinus</Emphasis> from Nachikinskoe Lake (Kamchatka)

We studied genetic differentiation of two charr species, Dolly Varden Salvelinus malma malma Walbaum and resident lacustrine charr Salvelinus sp., which sympatrically inhabit Nachikinskoe Lake (the Bol’shaya River basin) in southwestern Kamchatka Peninsula. Using restriction analysis (RFLP), three mitochondrial DNA fragments (ND1/ND2, ND5/ND6, and Cyt b/D-loop) amplified in polymerase chain reaction (PCR) were compared. The divergence of the mtDNA sequences between Salvelinus sp. and S. malma malma was 2.8%; Salvelinus sp. and S. taranetzi, 0.36%; Salvelinus sp. and S. krogiusae, 0.21%; Salvelinus sp. and S. alpinus, 3.0%. These results point to reproductive isolation of charrs in Nachikinskoe Lake and support the earlier suggestion on a close relationship between Salvelinus sp., S. taranetzi, and S. krogiusae.     

High level of tetracosapolyenoic fatty acids in the cold-water mollusk <Emphasis Type="Italic">Tochuina</Emphasis><Emphasis Type="Italic">tetraquetra</Emphasis> is a result of the nudibranch feeding on soft corals

Fatty acid (FA) markers are used to trace predator–prey relationships in the marine environment. Soft corals contain tetracosapolyenoic acids (TPA), namely 24:5n-6 and 24:6n-3, which are considered as octocoral FA markers. The nudibranch mollusks are known to feed on soft corals. To check whether TPA are transferred from soft corals to nudibranch mollusks during their nutrition, we determined and compared FA compositions of total lipids of Tochuina tetraquetra and soft corals (Acanella sp., Anthomastus rylovi, Gersemia fruticosa, and Paragorgia arborea) collected together near the Kuril Islands (the depths of 80–550 m). FA compositions of T. tetraquetra, Acanella sp. and A. rylovi were described for the first time. In Acanella sp., G. fruticosa, A. rylovi, and P. arborea, the average contents of TPA were 6.5, 13.4, 5.9, and 12.0 % of total FAs, whereas the 24:5n-6/24:6n-3 ratio was 1.0, 1.4, 5.4, and 2.6, respectively. The high level of TPA (21.7 %) found in T. tetraquetra indicates that 24:5n-6 and 24:6n-3 are transferred from soft corals to the mollusks during their feeding and accumulated in mollusk tissues. The most possible feed source of the mollusk is suggested to be G. fruticosa and/or Acanella sp., because the 24:5n-6/24:6n-3 ratios in T. tetraquetra and these soft corals were similar. Thus, the TPA could be used as FA markers to estimate a proportion of soft corals in feeding of cold-water nudibranch mollusks.     

Bacterial diversity of the outflows of a Polichnitos (Lesvos,Greece) hot spring,laboratory studies of a <Emphasis Type="Italic">Cyanobacterium</Emphasis> sp. strain and potential medical applications

The bacterial diversity of the outflows of Polichnitos (Lesvos, Greece) hot spring has been investigated. Cyanobacteria showing high sequence homologies with Phormidium sp. and Cyanobacterium aponinum were found. Members of the Alphaproteobacteria closely related to Rhodobium sp. Albidovulum sp., Rhodobacter sp., Microvigra sp., Nitratireductor sp. and Phaeobacter sp. Gammaproteobacteria, Betaproteobacteria and Firmicutes were represented by members of Idiomarina sp., Marinobacter sp., Shinella sp., Bacillus sp. and Clostridium sp. with sequence homologies ranging from 92% to 100%. Members of the Bacteroidetes and Planctomycetes were represented by sequences of novel phylogenetic linkages exhibiting 87–90% sequence homology with type strains. When the hot spring consortium was cultivated in bioreactor repeated batch culture under photo-autotrophic growth conditions at temperature < 30 °C, Cyanobacterium sp. dominated over Phormidium sp. Cyanobacterium sp. seems to have biotechnological potential since its extracellular broth exhibited a strong insecticidal activity against larvae of Aedes aegypti (a vector of important human diseases) and significant anti-cancer activity against the PC3 human prostate cancer cell line, while its toxicity against human endothelial cells was relatively low.     

Biochemical and genetic analysis of a unique poly(ADP-ribosyl) glycohydrolase (PARG) of the pathogenic fungus <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis>

The genome sequence of the plant pathogen Fusarium oxysporum f. sp. lycopersici contains a single gene encoding a predicted poly(ADP-ribose) glycohydrolase (FOXG_05947.2, PARG). Here, we assessed whether this gene has a role as a global regulator of DNA repair or in virulence as an ADP ribosylating toxin homologue of bacteria. The PARG protein was purified after expressing its encoding gene in Escherichia coli. Its inhibition by 6,9-diamino-2-ethoxyacridine lactate monohydrate and tannins was similar to its human orthologue that is involved in DNA repair. A deletion strain of F. oxysporum f. sp. lycopersici showed no growth defects and was not affected in pathogenicity. Together, our results indicate that the PARG protein of F. oxysporum f. sp. lycopersici is involved in DNA repair and does not act in pathogenicity as an effector.     

Nivalenol production by<Emphasis Type="Italic">Fusarium poae</Emphasis>

Fusarium sp. were isolated from Swedish nivalenol containing grain and tested for toxin production. OnlyF. poae, 6 of 10 isolates, produced nivalenol. Highest production (44.7 μg/g) was obtained cultured on rice during 4 week at room temperature and under near UV-light. FiveF. poae isolates from other countries did not produce nivalenol but T-2/HT-2 toxin. One Swedish isolate produced both types of trichothecenes. Treatment with fungicides in aF. poae infected experimental field reduced the nivalenol concentration in the harvested grain.     

Isolation,characterization and virulence of bacteria from <Emphasis Type="Italic">Ostrinia nubilalis</Emphasis> (Lepidoptera: Pyralidae)

Isolation, characterization and virulence of the culturable bacteria from entire tissues of larval Ostrinia nubilalis (Hübner) (Lepidoptera: Pyralidae) were studied to obtain new microbes for biological control. A total of 16 bacteria were isolated from living and dead larvae collected from different maize fields in the Eastern Black Sea Region of Turkey. The bacterial microbiota of O. nubilalis were identified as Pseudomonas aeruginosa (On1), Brevundimonas aurantiaca (On2), Chryseobacterium formosense (On3), Acinetobacter sp. (On4), Microbacterium thalassium (On5), Bacillus megaterium (On6), Serratia sp. (On7), Ochrobactrum sp. (On8), Variovorax paradoxus (On9), Corynebacterium glutamicum (On10), Paenibacillus sp. (On11), Alcaligenes faecalis (On12), Microbacterium testaceum (On13), Leucobacter sp. (On14), Leucobacter sp. (On15) and Serratia marcescens (On16) based on their morphological and biochemical characteristics. A partial sequence of the 16S rRNA gene was also determined to confirm strain identification. The highest insecticidal activities were obtained from P. aeruginosa On1 (80%), Serratia sp. On7 (60%), V. paradoxus On9 (50%) and S. marcescens On16 (50%) against larvae 14 days after treatment (p < 0.05). Also, the highest activity from previously isolated Bacillus species was observed from Bacillus thuringiensis subsp. tenebrionis Xd3 with 80% mortality within the same period (p < 0.05). Our results indicate that P. aeruginosa On1, Serratia sp. On7, V. paradoxus On9, S. marcescens On16 and B. thuringiensis subsp. tenebrionis Xd3 show potential for biocontrol of O. nubilalis.     

Myrothecium-like (Ascomycota,Hypocreales) species from tropical areas: <Emphasis Type="Italic">Digitiseta</Emphasis> gen. nov. and additions to <Emphasis Type="Italic">Inaequalispora</Emphasis> and <Emphasis Type="Italic">Parvothecium</Emphasis>

Inaequalispora and Parvothecium are two myrothecium-like, closely related genera of Hypocreales. They are also morphologically similar, sharing sporodochial conidiomata, penicillate conidiophores, fusiform to ellipsoidal conidia accumulating in a green slimy drop, and hypha-like setoid extensions emerging through the conidial mass. During a revision of myrothecium-like isolates originating from rainforest areas of South America (Ecuador, Brazil) and Southeast Asia (Singapore), multilocus phylogenetic inferences (based on DNA sequence data of ITS, partial nuc 28S, and partial tef1a, rpb2 and tub2) and morphological studies concordantly revealed the occurrence of two undescribed species of Inaequalispora (I. longiseta sp. nov. and I. cylindrospora sp. nov.) and one undescribed species of Parvothecium (P. amazonensesp. nov.). Myrothecium setiramosum, M. dimorphum, and two undescribed taxa form the base of a new lineage, sister to the current Parvothecium lineage. This lineage is recognized as Digitiseta gen. nov., typified by D. setiramosa comb. nov. Digitiseta dimorpha comb. nov. is also proposed, and the new species D. parvodigitata sp. nov. and D. multidigitata sp. nov. are described.     

Genetic diversity near the <Emphasis Type="Italic">DGAT1-2</Emphasis> gene for high oleic acid content and kernel trait variation in a maize breeding collection

Previous studies in maize have identified three quantitative trait loci (QTLs) coding for high oleic acid content (HOAC) (QTLs oleic6-1, oleic6-2, and oleic6-3) at bins 6.04–6.05, proximal to the DGAT1-2 gene. The aims of this work were (i) to discover new markers for linkage disequilibrium (LD) and haplotype analysis distal to DGAT1-2, (ii) to develop a new DGAT1-2 PCR probe to detect the allele determining HOAC (F469 insertion), (iii) to conduct cluster analysis for kernel traits, and (iv) to assess genetic diversity, LD, and association analysis for kernel traits with the DGAT1-2 PCR probe and 13 markers previously mapped near DGAT1-2 in the Argentine temperate maize collection of 111 inbred lines. The results showed high haplotype diversity distal to DGAT1-2 and relatedness between the inbred line LP199 (with HOAC) and the Non-Stiff Stalk line W22 (reference genome). The frequency of F469 was low (20%). F469 was clustered with flint-grain type characteristics, whereas HOAC was associated with F469 across linear models. Genetic diversity at bins 6.04–6.05 was high (0.62), whereas LD extent was low (r²?≤?0.45). This low extent of LD indicates a high level of recombination and no LD between DGAT1-2 and markers flanking QTLs oleic6-1 to 3. Nevertheless, the significant LD between markers flanking those QTLs and the cosegregation of F469 with nc009 (markers flanking QTLs oleic6-1 and 2) during inbred line conversion suggest that these QTLs might contribute to HOAC in the breeding collection. However, further studies are needed to precise mapping at bins 6.04–6.05 for breeding purposes.     

New species of the tenebrionid-beetle genera <Emphasis Type="Italic">Tagonoides</Emphasis> Fairm., <Emphasis Type="Italic">Gnaptorina</Emphasis> Rtt. and <Emphasis Type="Italic">Agnaptoria</Emphasis> Rtt. (Coleoptera,Tenebrionidae) from China

Six new species Tagonoides belousovi sp. n., T. yunnana sp. n., Gnaptorina australis sp. n., Agnaptoria elongata sp. n. (Yunnan), A. lecta sp. n., and A. ruida sp. n. (Sichuan), and a new subspecies Gnaptorina potanini minxiana subsp. n. are described from China. Morphological adaptations of tenebrionids of the tribe Blaptini associated with phytophagous habit are discussed.     

New and little known species of the dance-fly subgenus <Emphasis Type="Italic">Xanthempis</Emphasis> Bezzi,genus <Emphasis Type="Italic">Empis</Emphasis> L. (Diptera,Empididae), from the Caucasus

Five new species of the subgenus Xanthempis Bezzi are described from the Caucasus: Empis (Xanthempis) annae sp. n. (Russia: Krasnodar Territory), E. (X.) grichanovi sp. n. (Russia: Krasnodar Territory; Georgia), E. (X.) pseudoconcolor sp. n. (Russia: Krasnodar and Stavropol territories; Georgia: Abkhazia), E. (X.) teberdaensis sp. n. (Russia: Karachay-Cherkessia), and E. (X.) zamotajlovi sp. n. (Russia: Krasnodar Territory and Adygea). The females of E. (X.) alanica Shamshev and E. (X.) kovalevi Shamshev are described for the first time. New data on the distribution of some previously described species are reported. The geographical distribution of Xanthempis is discussed. A key to Xanthempis species from the Caucasus is compiled.     

<Emphasis Type="Italic">Deinococcus sedimenti</Emphasis> sp. nov. isolated from river sediment

A novel Gram-positive, oval-shaped, non-motile bacterium designated strain 16F1L^T was isolated from sediment collected from the Han River in Seoul, Republic of Korea. Based on the 16S rRNA gene sequence (1,448 bp), this strain was identified as a member of the genus Deinococcus that belongs to the class Deinococci. Similarities in the 16S rRNA gene sequence were shown with Deinococcus daejeonensis MJ27^T (99.0%), D. grandis DSM 3963^T (98.1%), D. radiotolerans C1^T (97.5%), and D. caeni Ho-08^T (97.2%). Strain 16F1L^T was classified as a different genomic species from closely related Deinococcus members, based on less than 70% DNA-DNA relatedness. Genomic DNA G+C content of strain 16F1L^T was 67.2 mol%. Strain 16F1L^T was found to grow at temperatures of 10–37°C (optimum 25°C) and pH 7–8 (optimum pH 7) on R2A medium, and was catalase-positive and oxidase-negative. Strain 16F1L^T showed resistance to gamma radiation (D₁₀ > 2 kGy). In addition, this strain had the following chemotaxonomic characteristics: the major fatty acids were C_15:1ω6c and C_16:1ω7c; the polar lipid profile contained phosphoglycolipids, unknown aminophospholipids, an unknown aminoglycolipid, unknown aminolipids, an unknown glycolipid, an unknown phospholipid, and an unknown polar lipid; the major quinone was MK-8. Phylogenetic, genotypic, phenotypic, and chemotaxonomic characteristics indicated that strain 16F1L^T represents a novel species within the genus Deinococcus, for which the name Deinococcus sedimenti sp. nov. is proposed. The type strain is 16F1L^T (=KCTC 33796^T =JCM 31405^T).     

In Situ Enzymatic Conversion of <Emphasis Type="Italic">Nannochloropsis oceanica</Emphasis> IMET1 Biomass into Fatty Acid Methyl Esters

Conventionally, production of methyl ester fuels from microalgae occurs through an energy-intensive two-step chemical extraction and transesterification process. To improve the energy efficiency, we performed in situ enzymatic conversion of whole algae biomass from an oleaginous heterokont microalga Nannochloropsis oceanica IMET1 with the immobilized lipase from Candida antarctica. The fatty acid methyl ester yield reached 107.7% for dry Nannochloropsis biomass at biomass to t-butanol to methanol weight ratio of 1:2:0.5 and a reaction time of 12 h at 25 °C, representing the first report of efficient whole algae biomass conversion into fatty acid methyl esters at room temperature. Different forms of algal biomass including wet Nannochloropsis biomass were tested. The maximum yield of wet biomass was 81.5%. Enzyme activity remained higher than 95% after 55 days of treatment (equal to 110 cycles of reaction) under the conditions optimized for dry algae biomass conversion. The low reaction temperature, high enzyme stability, and high yield from this study indicate in situ enzymatic conversion of dry algae biomass may potentially be used as an energy-efficient method for algal methyl ester fuel production while allowing co-product recovery.     

Moniliformin and the European Corn Borer (<Emphasis Type="Italic">Ostrinia nubilalis</Emphasis>)

A representative survey was made of maize ears of the 1988 and 1989 crop in Austria to establish the influence of corn borer injuries onFusarium species involved in ear fusariosis andFusarium toxin production.TheFusarium species most frequently isolated from rot-damaged ears wereFsacchari var. subglutinans (about 50 %) andF. graminearum (about 30 %). There was a striking difference between theFusarium species of the Liseola and the Discolor section concerning their occurrence on corn borer-damaged ears. More than 80 % of the ears infected withF. sacchari var. subglutinans andF. verticillioides, but less than 15 % of the ears infected withF. graminearum, F. crookwellense andF. culmorum showed corn borer injuries.Toxin analyses of the infected ears corresponded to the known toxigenicity of the respectiveFusarium species. Ears infected withF. sacchari var. subglutinans contained moniliformin (up to 20 mg/kg), those infected withF. verticillioides fumonisin B₁ and B₂ (up to 15 mg/kg). In ears infected withF. graminearum, F. culmorum andF. crookwellense zearalenone (up to 40 mg/kg) and deoxynivalenol (up to 500 mg/kg) or nivalenol (up to 10 mg/kg), respectively, could be detected. Hence measures to combat the European corn borer will mainly reduce moniliformin and fumonisin contamination, but will affect zearalenone, deoxynivalenol and nivalenol contents of the ears to a much lesser extent.