Sampling to Assess a Re-Established Appalachian Forest in Ohio Based on Gelechioid Moths (Lepidoptera: Gelechioidea)

A list of microlepidoptera belonging to the superfamily Gelechioidea was produced from June trap samples from sites within the Wayne National Forest (Lawrence County), an Appalachian forest in Southern Ohio that was once a greatly disturbed area and has since re-established over a period of nearly 100 years. The composition and diversity of Lawrence county is compared to lists of gelechioid moths generated by other surveys in the eastern United States: the Great Smoky Mountains National Park, an Ohio study (Summerville and Crist 2003), and unpublished data from Connecticut (Wagner). From comparison with these studies, we address two questions: (1) How well do passive surveys of Gelechioidea compare to more labor intensive surveys? (2) How does the regenerated Wayne National Forest compare to other well documented areas with respect to gelechioid diversity? Our sample of diversity, though more narrow in time and area, compares favorably to more exhaustive sampling and demonstrates that it may be more efficient to focus on target groups in focal localities when time and resources are limited rather than conduct extensive sampling programs.     

Bacteriocinogenic Potential of Bacillus amyloliquefaciens Isolated from Kimchi,a Traditional Korean Fermented Cabbage

Bacteriocin production is considered a favorable property for various beneficial cultures. In addition to their potential as biopreservatives, bacteriocins are also promising alternatives for the control of multidrug-resistant pathogens and the inhibition of some viruses and cancer cells. The objective of this study was to screen and characterize a bacteriocin-producing strain with the aim of its future application for control of Listeria monocytogenes, an important food-borne pathogen. A total of 22 potentially bacteriocinogenic strains active against L. monocytogenes ATCC15313 were isolated from locally produced kimchi through a three-level approach. Pure cultures were obtained according to good microbiological practices and differentiated through RAPD-PCR using the primers OPL01, OPL09, and OPL11. Altogether, 5 strains were selected for further study. Specific focus was given to strain ST05DL based on its specific inhibitory activity against L. monocytogenes ATCC15313, while not affecting different strains belonging to the genera Lactobacillus, Pediococcus, Leuconostoc, and Weissella, most of which are beneficial microorganisms. The strain ST05DL was identified as Bacillus amyloliquefaciens based on its sugar fermentation profile obtained through API50CHB analysis and 16S rRNA partial sequencing. The antimicrobial compound produced by B. amyloliquefaciens ST05DL was found to be sensitive to pepsin and α-chymotrypsin, evidence of its proteinaceous nature. The presence of skim milk, NaCl, Tween 80, glycerol, and SDS did not affect the antimicrobial activity. The addition of 20% cell-free supernatant (CFS) obtained from a 24-h culture of B. amyloliquefaciens ST05DL to an exponentially growing culture of L. monocytogenes ATCC15313 successfully inhibited the test microorganisms during the monitored 10-h incubation. Optimal bacteriocin production by B. amyloliquefaciens ST05DL was observed during the stationary phase at 12 h (800 AU/mL) and remained stable for the next 15 h. The ratio between live and dead cells during this period was 74.37% and 25.66%, respectively, as determined by flow cytometry. The presence of the virulence genes hblA, hblB, hblC, nheA, nheB, and nheC was not detected in the total DNA of B. amyloliquefaciens ST05DL, and the strain was resistant only to ampicillin out of 10 tested antibiotics. Future evaluation of expressed bacteriocin/s by B. amyloliquefaciens ST05DL (amino acid sequence, molecular mass, cytotoxicity, detailed mode of action, etc.), will be the next step in the characterization and its potential application as biopreservative and/or pharmaceutical product.

     

Assessment of the modulating effects of fructo-oligosaccharides on fecal microbiota using human flora–associated piglets

We first used human flora–associated (HFA) piglets, a significantly improved model for research on human gut microbiota, to study the effects of short-chain fructo-oligosaccharides (scFOS) on the gut bacterial populations. Ten neonatal HFA piglets were assigned to receive basal diets alone or supplemented with scFOS (0.5 g/kg body weight/day) from 1 to 37 days after birth (DAB). The impact of scFOS on the fecal bacterial populations of the piglets before (12 DAB), during (17 DAB), and after (25 and 37 DAB) weaning were monitored by PCR-denaturing gradient gel electrophoresis and real-time quantitative PCR. The Bifidobacterium genus was stimulated consistently, except during weaning, confirming the bifidogenic property of scFOS. At 12 DAB, the Clostridium leptum subgroup was decreased and two unknown Bacteroides-related species were increased; at 25 DAB, the C. leptum subgroup and Subdoligranulum variabile-like species were elevated, whereas one unknown Faecalibacterium-related species was suppressed; and at 37 DAB, the Bacteroides genus was decreased. The results showed that effects of scFOS on non-bifidobacteria varied at different developmental stages of the animals, warranting further investigation into the host-development-related effects of prebiotics on the gut microbiota and the host physiology using the HFA piglets as a model for humans.     

Fusarium species complex and mycotoxins in grain maize from maize hybrid trials and from grower's fields

Aims: To quantify and to compare the occurrence of Fusarium species in maize kernels and stalk pieces, to analyse mycotoxins in kernels and maize crop residues, to evaluate two approaches to obtain kernel samples and to compare two methods for mycotoxin analyses. Methods and Results: The occurrence of Fusarium species in maize kernels and stalk pieces from a three‐year maize hybrid trial and 12 kernel samples from grower’s fields was assessed. Nine to 16 different Fusarium species were detected in maize kernels and stalks. In kernels, F. graminearum, F. verticillioides and F. proliferatum were the most prevalent species whereas in stalks, they were F. equiseti, F. proliferatum and F. verticillioides. In 2006, 68% of the kernel samples exceeded the recommended limit for pig feed for deoxynivalenol (DON) and 42% for zearalenone (ZON), respectively. Similarly, 75% of the samples from grower’s fields exceeded the limits for DON and 50% for ZON. In maize crop residues, toxin concentrations ranged from 2·6 to 15·3 mg kg^?1 for DON and from 0·7 to 7·4 mg kg^?1 for ZON. Both approaches to obtain maize kernel samples were valid, and a strong correlation between mycotoxin analysis using ELISA and LC‐MS/MS was found. Conclusions: The contamination of maize kernels, stalk pieces and remaining crop residues with various mycotoxins could pose a risk not only to animal health but also to the environment. With the hand‐picked sample, the entire Fusarium complex can be estimated, whereas combine harvested samples are more representative for the mycotoxin contents in harvested goods. Significance and Impact of the Study: This is the first multi‐year study investigating mycotoxin contamination in maize kernels as well as in crop residues. The results indicate a high need to identify cropping factors influencing the infection of maize by Fusarium species to establish recommendations for growers.     

Autolysis in vitro of cotton (Gossypium hirsutum) fibre cell walls

Root growth of partly defoliated young peach seedlings [ Prunus persica (L.) Batsch. cv. Lovell] was significantly promoted by application of myo-inositol to the cut surface of the stem. Addition of benzylaminopurine (BA) combined with sucrose enhanced the promotive effect of myo-inositol on root growth, but addition of sucrose alone, suppressed it. Spraying rooted peach cuttings (nectarine cv. Sunred) with myo-inositol and defoliating them after 5 days increased the incorporation of amino acids into proteins in excised roots, obtained from the sprayed plants, as compared with roots from plants sprayed with water, or sucrose, or sucrose + myo-inositol. Myo-inositol applied in combination with kinetin or BA to stems of young peach seedlings (cv. Lovell) or rooted peach cuttings (cv. Almog) promoted the basipctal translocation of the two cytokinins in the stem and acropetally into the small lateral roots. Addition of sucrose voided this effect on the cytokinins. BA, when applied together with myo-inositol, was partly converted into an additional cytokinin-active compound in the roots.
Application of BA to either roots or tops of rooted peach cuttings (cv. Almog) resulted in the accumulation of myo-inositol (supplied through the cut surface of the stem) in the plant part to which BA had been applied.     

The influence of plant spacing on density-dependent versus frequency-dependent spore transmission of the anther smut Microbotryum violaceum

The anther smut fungus Microbotryum violaceum is a pollinator-transmitted plant disease. As for other vector-borne diseases, frequency-dependent transmission patterns are predicted, in contrast to the density-dependent transmission of passively spread diseases. Frequency dependence will, however, only arise if vectors compensate for varying plant spacings. To test this assumption, we set up experimental populations of the host plant, Silene latifolia, with varying disease density (number of diseased plants per plot) and frequency (proportion of plants diseased), and three different plant spacings. We measured spore deposition on healthy flowers in these plots on two dates. Spore deposition decreased considerably from the first to the second census, perhaps related to the concomitant decrease in inflorescence sizes of diseased plants. At our first census, spore deposition rates varied with disease frequency, and the effect of frequency depended on plant spacing. While spore deposition was positively frequency dependent at the 1.5-m inter-plant spacing, no effect of disease frequency was found at a spacings of 0.5 m or 3 m. Nor was there an effect of disease density on spore deposition at the first census. At the later census, on the other hand, spore deposition increased almost significantly with increasing disease density (P = 0.08). This difference in deposition pattern together with a significant decrease in spore receipt indicates changes in pollinator spectrum and/or activity. The correlation of spore numbers among flowers within plants, an indication for intra-plant moves by vectors, was significant at 0.5 m and 1.5 m but not at 3 m. Floral traits and sex of individual plants influenced the number of spores they received. On the first census date, spore deposition increased with increasing inflorescence size in female but not in male plants. On the second census date, neither sex nor number of open flowers had an effect on spore receipt. None of the experimental plants became infected, however, probably because of the unusually hot and dry weather. Received: 14 May 1998 / Accepted: 19 November 1998     

The Living Dead: Bacterial Community Structure of a Cadaver at the Onset and End of the Bloat Stage of Decomposition

Human decomposition is a mosaic system with an intimate association between biotic and abiotic factors. Despite the integral role of bacteria in the decomposition process, few studies have catalogued bacterial biodiversity for terrestrial scenarios. To explore the microbiome of decomposition, two cadavers were placed at the Southeast Texas Applied Forensic Science facility and allowed to decompose under natural conditions. The bloat stage of decomposition, a stage easily identified in taphonomy and readily attributed to microbial physiology, was targeted. Each cadaver was sampled at two time points, at the onset and end of the bloat stage, from various body sites including internal locations. Bacterial samples were analyzed by pyrosequencing of the 16S rRNA gene. Our data show a shift from aerobic bacteria to anaerobic bacteria in all body sites sampled and demonstrate variation in community structure between bodies, between sample sites within a body, and between initial and end points of the bloat stage within a sample site. These data are best not viewed as points of comparison but rather additive data sets. While some species recovered are the same as those observed in culture-based studies, many are novel. Our results are preliminary and add to a larger emerging data set; a more comprehensive study is needed to further dissect the role of bacteria in human decomposition.     

EVIDENCE FOR GENETIC DIFFERENTIATION BETWEEN CHOKE-INDUCING AND ASYMPTOMATIC STRAINS OF THE EPICHLOË GRASS ENDOPHYTE FROM BRACHYPODIUM SYLVATICUM

Life cycle and breeding system variation in Epichloë grass endophytes (choke disease) is tightly linked to the degree of stroma formation. It is not known whether this variation results from differences in host resistance, fungal virulence, or environmental conditions. We found genetic differentiation between 173 asymptomatic (NS) and 93 stromata-forming (S) Epichloë strains isolated from one grass species, Brachypodium sylvaticum, based on 13 presumed allozyme loci, of which six were variable. The fungal strains originated from 10 sites in Switzerland, three sites of which were represented by both NS and S subpopulations. In total, 19 allozyme genotypes, that were nonrandomly distributed among S and NS were detected. Genetic variation measured as G_ST between S and NS strains isolated from the same site ranged from 0.73 to 0.98. Clonality, measured as linkage disequilibrium at one site, was significant in the NS subpopulation (P ? 0.001), but not in the S subpopulation (P = 0.21), implying asexual reproduction by NS strains as well as successful horizontal transmission of S strains. Since all seeds are usually infected vegetatively, horizontal transmission implies the occurrence of multiple host infections. Altogether, these results provide indirect evidence that NS and S strains do not belong to one panmictic population and that differentiation patterns of stroma formation found in nature are due to genetic differences among fungi in associations with their host plants. We discuss the direction of evolution of disease expression in this system. The distribution of genetic variability suggests that the asymptomatic strains were derived from stromata-forming populations.     

ANTHER SMUT DISEASE IN DIANTHUS SILVESTER (CARYOPHYLLACEAE): NATURAL SELECTION ON FLORAL TRAITS

Mating opportunities, pollination intensity, and pollen dispersal ability may vary with variation in floral traits such as color, size, and shape. Where these traits are selected by pollinators for enhanced elaboration, they should evolve toward the equilibrium between selection for further elaboration and selection against this through reduced fecundity or vitality. Here we show that pollinator-borne fungal diseases of plants may be a factor influencing the position of this equilibrium. Populations of the rock pink, Dianthus silvester often contain individuals infected with the anther smut fungus Microbotryum violaceum (= Ustilago violacea). In a naturally infected population in the Alps of eastern Switzerland we investigated how intrapopulation variation in flower size and nectar rewards influenced spore deposition and how floral traits varied with disease status. We found that spore deposition increased with increasing petal size, suggesting that large-flowered plants were at a greater risk of disease. Spore deposition was also higher for plants growing in patches with many or a high proportion of diseased neighbors. Multiple regression analyses showed that petal size or nectar reward influenced spore deposition when the effects of neighborhood disease abundance were controlled statistically. In sequential analyses, after removing the effects of disease density or frequency and plant gender, petal length explained significant variation in spore deposition. Diseased plants had reduced female reproductive organs, but calyx size was intermediate between that of healthy perfect and female flowers of this gynodioecious-gynomonoecious species, and diseased plants bore flowers with the largest petals. This may reflect a symptom of this disease or the cause, if larger-flowered plants are more likely to become infected. We conclude that investment to pollinator attraction may bring an enhanced risk of contracting this sterilizing pollinator-borne disease, so natural selection by the fungus M. violaceum acts to lower attractiveness to pollinators.