Experimental Infection of Nosema sp. Ghose in an Unnatural Host Lesioderma sericorne

ABSTRACT. Various doses of a microsporan parasite, Nosema sp., were fed to third and fourth instar larvae of Lesioderma sericorne that infested different types of stored grains. A spore dose of 3 × 10³ spores/individual resulted in a 39% infection rate, reduction in larval and adult weights, and mean spore concentrations of 1.28 ± 0.2 × 10⁸ spores/larva and 1.1 ± 0.2 × 10⁸ spores/adult. At the above dose, mortality was not well marked (about 35% in larvae and 25% in adults). At 3 × 10⁴ spores/individual, the rate of mortality increases to 80% in larvae and 60% in adults. However, more of the pest population (88% of larvae and 73% of adults) died at a dose of 3 × 10⁵ spores/individual. This dose produced mean spore concentrations of 3.91 ± 0.2 × 10⁸ spores/larva and 2.89 ± 0.2 × 10⁸ spores/adult. Insect death was caused by heavy damage to gut epithelia and fat bodies.     

Attempts to control cabbage root flies Delia radicum L. and Delia floralis (Fall.) (Dipt., Anthomyiidae) with entomopathogenic fungi: laboratory and greenhouse tests

One laboratory and three greenhouse experiments were conducted to study the pathogenicity and efficacy of Finnish isolates of entomopathogenic hyphemycetous fungi against cabbage root flies. In Petri dishes, exposure to 1.5 × 10¹⁰ spores of Metarhizium anisopliae per dish caused 40–50% mortality of undifferentiated second- and third-stage larvae of Delia floralis , and 1 × 10⁷ spores per dish caused 40–50% mortality of Delia radicum larvae. In one greenhouse test, 1 × 10⁸ and 1 × 10⁹ spores of M. anisopliae and Paecilomyces fumosoroseus reduced the root damage of head cabbage by 20–70% compared with untreated controls, although this was not accompanied by significant reductions in the number of pupae. Only M. anisopliae consistently grew out of larvae and pupae of D. floralis during incubation that followed their recovery from the soil at the end of an experiment testing different formulations of M. anisopliae and Beauveria bassiana , but the frequency of the latent infections of the pest by M. anisopliae was not associated with reduced severity of damage to seedlings of head cabbage.     

Laboratory studies on the effects of pollen from Bt-maize on larvae of some butterfly species

Abstract:   Three lepidopteran species were tested to determine their susceptibility against the ingestion of pollen from genetically modified maize plants. To prove the existence of dose–response relations between the applied amount of pollen (Bt-maize) and the damages on the tested larvae, a method was developed which makes it possible to feed caterpillars with defined amounts of pollen. If their food plants were contaminated with pollen of a cultivar of the Bt-176 maize-line Pieris brassicae , Pieris rapae and Plutella xylostella -larvae fed less, grew more slowly and showed a higher mortality than caterpillars of an untreated control group. The 50% lethality (LD₅₀)-values were calculated for P.xylostella (L₄) with 19.2, for P. rapae (L₂) with 39.0 and also for P. brassicae (L₂) with 139.2 pollen of the transgenic maize Pactol CB. Studies with P. brassicae -caterpillars of different larval stages indicated, that older individuals showed a higher tolerance against pollen from Bt-maize than younger ones. It must be stated on the basis of the present studies, that ingestion of non-transgenic maize pollen has neither a positive nor a negative effect on caterpillars. It becomes clear from the information presented here that it is still difficult to make general statements about the endangering of butterflies, arising from cultivation of genetically modified maize lines. Further investigations on this issue are needed. Initially, the LD₅₀-values concerning the larvae of certain butterfly species have to be determined to anticipate the risks, and in addition the distances between habitats with caterpillar host plants and maize fields, and the abundance of these plants have to be considered.     

Effects of elevated CO2-grown loblolly pine needles on the growth, consumption, development, and pupal weight of red-headed pine sawfly larvae reared within open-topped chambers

Seedlings of loblolly pine, Pinus taeda , were grown in open-topped chambers under four levels of CO₂: two ambient and two elevated. Larvae of the red-headed pine sawfly, Neodiprion lecontei , were reared from early instar to pupation, primarily on branches within chambers. Larval growth and mortality were assessed and leaf phytochemistry samples of immature and mature leaves collected weekly. Mature leaves grown under elevated CO₂ had significant reductions in leaf nitrogen and increases in non-structural carbohydrate contents, resulting in foliage being a poorer food source for larvae, i.e. higher carbohydrate:nitrogen ratio. Nutritional constituents of immature needles were unaffected by seedling CO₂ treatment. Volatile mono- and sesquiterpenes were unrelated to plant CO₂ treatments for either leaf age class. Larval consumption of immature needles significantly increased on seedlings grown under CO₂ enrichment, while mature needle consumption was not different between the treatments. The average weight gain per larva significantly declined in late instar larvae consuming elevated CO₂-grown needles. In spite of this reduced growth, neither the days to pupation nor pupal weights were different among the CO₂ treatments. This study suggests that enriched CO₂-induced alterations in pine needle phytochemistry can affect red-headed pine sawfly performance. However, compensatory measures by larvae, such as choosing to consume more nutritious immature needles, apparently helps offset enriched CO₂-induced reductions in the leaf quality of mature needles.     

Effects of Host Cell Density on Cell Infection Level in Antheraea eucalypti (Lepidoptera: Saturniidae) Cell Cultures Persistently Infected with Nosema bombycis (Microsporida: Nosematidae)

ABSTRACT. Spores of Nosema bombycis Y9101, isolated from the beet armyworm, Spodoptera exigua , were primed with an alkaline solution and inoculated into Antheraea eucalypti cell cultures. Infected cells were subcultured every five days at three cell densities (2.5 × 10³, 5.0 × 10³, and 1.0 × 10⁴ cells/cm²). A difference was observed in the spread of N. bombycis Y9101 infection between low-density and higher-density cultures of host cells. The host cell density did not affect the productivity of secondary infective forms of the parasite. The principal factor determining the rate of microsporidian infection in vitro was the number of host cells existing within the reach of extruded short-coiled polar tubes from spores germinated intracellularly.     

Mistletoe viscotoxins induce membrane permeabilization and spore death in phytopathogenic fungi

Viscotoxins (Vts) are basic peptides expressed in mistletoe leaves, seeds and stems which have been shown to be cytotoxic to mammalian cells. The aim of this study was to analyse whether Vts were able to control and/or inhibit the growth of phytopathogenic fungi to obtain a clue to their biological function. Incubation of two Vt isoforms, VtA₃ and VtB, at a final concentration of 10 µ M resulted in a complete blockage of the germination of spores from three different pathogenic fungi. It was also shown that lower concentrations than 10 µ M of VtA₃ and VtB inhibit their mycelial growth in a dose-dependent manner. The protein dose required to inhibit the growth of Fusarium solani and Sclerotinia sclerotiorum to a 50% was between 1.5 and 3.75 µ M , which represents a potent activity. No significant differences in the antifungal potency for each Vt isoform, either VtA₃ and VtB, were observed, although they have been shown to exert differential cytotoxicity on mammalian cells. It was also demonstrated that Vts act as fungicidal compounds. To explore the basis of the antifungal activity the ability of VtA₃ to induce changes in membrane permeability and on the oxidative status of F. solani spores was analysed. By using a specific fluorescent probe on intact spores, it was demonstrated that VtA₃ produces rapid changes in fungal membrane permeability. It also induces H₂O₂ production verified by a histochemical staining. The data presented in this study support a direct role of Vts in the plant defence determined by their lethal effect on fungal pathogens.     

Gas exchange and growth responses of ectomycorrhizal Picea mariana, Picea glauca, and Pinus banksiana seedlings to NaCl and Na2SO4

Abstract: Black spruce (Picea mariana), white spruce (Picea glauca), and jack pine (Pinus banksiana) seedlings were inoculated with Hebeloma crustuliniforme or Laccaria bicolor and subjected to NaCl and Na₂SO₄ treatments. The effects of ectomycorrhizas on salt uptake, growth, gas exchange, and needle necrosis varied depending on the tree and fungal species. In jack pine seedlings, ectomycorrhizal (ECM) fungi reduced shoot and root dry weights and in the ECM white spruce, there was a small increase in dry weights. Sodium chloride treatment reduced net photosynthesis and transpiration rates in the three studied tree species. However, NaCl-treated black spruce and jack pine colonized by H. crustuliniforme maintained relatively high photosynthetic and transpiration rates and needle necrosis of NaCl-treated black spruce seedlings was reduced by the ECM fungi. Higher concentrations of Na+ were found in shoots compared with roots of the three examined conifer species. ECM fungi reduced the concentrations of Na+ mainly in the shoots and this reduction was greater in plants treated with NaCl compared with Na₂SO₄. Shoots contained generally higher concentrations of Cl^- compared with roots. In the NaCl-treated black spruce and white spruce, both ECM species significantly reduced Cl^- concentrations. Our results point to overall greater phytotoxicity of NaCl compared with Na₂SO₄ and support our earlier findings which demonstrated beneficial effects of ECM fungi for woody plants exposed to NaCl stress.     

Susceptibility of Vespula vulgaris (Hymenoptera: vespidae) to generalist entomopathogenic fungi and their potential for wasp control

The pathogenicity of Vespula vulgaris wasp workers and larvae to a range of fungi was determined. All fungi were isolated in New Zealand and included isolates from Vespula, known generalist insect pathogens, and isolates generally nonpathogenic to insects. Workers and larvae were highly susceptible to pathogenic isolates at high spore concentrations (>1.75 x 10(5) cfu/individual). Eight isolates, two of Metarhizium anisopliae, five of Beauveria bassiana, and one of Aspergillus flavus were pathogenic while a single isolate of M. flavouiride var. novazealandicum, Cladosporium sp., and Paecilomyces sp. were not. The transfer of spores between workers, and between workers and larvae, was also investigated using several different application methods. Transfer of spores occurred between treated and untreated individuals, and for some of the application methods sufficient spores were transferred to cause mortality of the nontreated individuals. These findings are related to the potential of fungi for the control of wasps.     

Effect of carbohydrate and nitrogen on hydrogen peroxide formation by wood decay fungi in solid medium

Abstract Hydrogen peroxide (H₂O₂) has been implicated in degradation of wood by both brown-rot and white-rot fungi. This study found that low concentrations of nitrogem and carbohydrates (cellobiose, glucose, xylose and mannose) in an agar medium had little effect on H₂O₂ production by white-rot fungi. However, low concentrations of nitrogen and carbohydrates stimulated H₂O₂ production by brown-rot fungi. Use of the chromogen 2,2'-azino-di(3-ethyl benzthiazoline-6-sulphonic acid) (ABTS) with horseradish peroxidase to detect H₂O₂ by the fungi was slightly better than detection by the chromogen o -dianisidine with horseradish peroxidase. An auxiliary test to check the role of H₂O₂ in wood decay found that hydrogen peroxide-negative isolates of the white-rot fungi Pharnerochaete chrysosporium and Ganoderma applanatum were unable to decay sweetgum and southern pine.     

中国松毛虫研究与治理

<正> 在中国,松毛虫是最严重的森林食叶害虫。远在370年前就有发生成灾的记载,至本世纪90年代每年发生面积达270万公顷左右,给林业生产和环境保护造成严重损害。中国现有松毛虫27种(4亚种)。除宁夏和青海外,分布遍及28个直辖市、省和自治区。其中,陕西省有10种,台湾省有4种。从分布范围和危害程度看,发生严重的有:南方的马尾松毛虫,北方的     

The effect of hydrogen peroxide on spores of Clostridium perfringens

Dithiothreitol (DTT)-treated spores of Clostridium perfringens were much more sensitive to lysis by H₂O₂ in the presence of Cu²⁺ than untreated spores. Lysis was greatly inhibited by hydroxyl radical (.OH) scavengers such as thiourea, dimethylthiourea and dimethylsulfoxide, suggesting that lysis of spores by H₂O₂ involves formation of OH by Cu²⁺-catalysed decomposition of the peroxide. DTT-treated spores took up Cu²⁺ at almost the same rate and extent as did isolated cortical fragments. Hydrogen peroxide caused both the decrease in optical density and the hexosamine solubilization of cortical fragments which bound Cu²⁺.     

Distribution and characterisation of entomopathogenic fungi from Korean soils

We investigated the occurrence of entomopathogenic fungi in 1080 soil samples representing multiple locations and conditions in Korea. Entomopathogenic fungi were isolated from soils using a selective medium containing dodine and antibiotics. Following an initial identification based on morphology, the fungal isolates were more precisely identified by the sequence of their nuclear ribosomal RNA (rRNA) internal transcribed spacer (ITS) regions. As a result, entomopathogenic fungi were found to occur in 32% (342 isolates) of the soil samples studied. The most abundant species were Beauveria spp. (125 isolates) and Metarhizium spp. (82 isolates). Entomopathogenic fungi were more often recovered from natural mountain and riparian soils than from agricultural habitats. The pathogenicity of isolated fungi was evaluated by using wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) larvae. It was determined that 60% (207 isolates) of the isolates were pathogenic using this model. These entomopathogenic fungi may, therefore, have potential use against a variety of agricultural pests. This is the first study of the isolation and distribution of entomopathogenic fungi in representative sampling locations throughout Korea.     

Hydroperoxide inactivation of enzymes within spores of Bacillus megaterium ATCC19213

Abstract Hydroperoxide inactivation of the protoplast enzymes enolase, aldolase and glucose-6-phosphate dehydrogenase in intact spores of Bacillus megaterium ATCC19213 was assessed by first treating the cells with lethal levels of H₂O₂, then germinating them in the presence of chloramphenicol prior to permeabilization and enzyme assays. Glucose-6-phosphate dehydrogenase proved to be more sensitive to H₂O₂than enolase or aldolase, in agreement with findings for isolated enzymes. Average D values (time for 90% inactivation) for spores treated with 0.50% H₂O₂ were 173 min for enolase, 67 min for aldolase and 32 min for glucose-6-phosphate dehydrogenase, compared with a D value of 34 min for spore killing. H₂O₂ killing of spores was found to be conditional in that recoveries of survivors were greater on complex medium than on minimal medium. Overall, it appeared that oxidative inactivation of enzymes may be important for hydroperoxide killing of spores.     

Isolation of ribulose bisphosphate carboxylase-oxygenase from non-hardened and hardened needles of Pinus sylvestris

Ribulose bisphosphate carboxylase-oxygenase, RuBP carboxylase (EC 4.1.1.39), was purified from non-hardened and hardened needles of Pinus sylvestris L. Needles were collected from pine seedlings cultivated in nutrient solution in a climate chamber from seedlings grown outdoors, and from a tree in a natural stand. The enzyme was isolated from crude extracts through quantitative precipitation in polyethylene glycol 4000 and MgCl₂, followed by sucrose gradient centrifugation in a fixed angle rotor. The purified enzyme seemed homogeneous by the criterion of (sodium dodecylsulphate) polyacrylamide gel electrophoresis. Contamination by nucleic acids was negligible. The RuBP carboxylase protein content of the gradient fractions was estimated as A₂₈₀^{1 cm}× 0.61 mg ml⁻¹. Carboxylase activities were determined in a radioactive assay at 25°C. The specific activity of RuBP carboxylase isolated from non-hardened needles was approximately 1 μmol CO₂ (mg protein)⁻¹ min⁻¹. For enzyme isolated from hardened needles collected during winter the specific activity was somewhat lower due to loss of enzyme activity during the preparation. The described two-step procedure provides a means for quantitation of the RuBP carboxylase protein in pine needles during all seasons.     

Increase in respiratory rate during feeding in larvae of the cinnabar moth Tyria jacobaeae

ABSTRACT. This study tested the effect of diet and phase of the feeding cycle on oxygen consumption by fifth-instar larvae of the cinnabar moth Tyria jacobaeae (L.) (Lepidoptera: Arctiidae). There was no significant variation in respiratory rates among larvae fed different diets, which were floral parts, upper, middle and lower leaves of the host plant tansy ragwort, Senecio jacobaea. The respiratory rates ( x ± 95% Cl μlO₂h^-1) of feeding larvae (279.5 ± 30.9) were higher than those of larvae at rest (179.7 ± 12.2), but were not significantly different from those of larvae in the post-ingestive phase (272.8 ± 35.8).
The respiratory rates of feeding larvae increased linearly with the ingestion rate, such that an additional 21.1 μl O₂ were consumed for every mg dry mass of plant material eaten per hour.     

Mesophilic chitin-degrading anaerobes isolated from an estuarine environment

Abstract Eight strains of obligately anaerobic, mesophilic, chitinolytic bacteria were isolated from the sediment of an estuarine environment. The isolates were rod-shaped, Gram-negative, and formed terminal spherical spores that swelled the sporangium. The major products from the fermentation of chitin were: acetate, ethanol, formate, CO₂, H₂ and ammonia. Growth of the isolates was possible at pH values ranging from 5.0–9.0. During the fermentation of chitin, N -acetylglucosamine accumulated in the culture fluids and was not metabolized. No organic compounds other than chitin and its oligomers could be demonstrated to support growth of the isolates. Hydrolysis of chitin proceeded at a relatively low rate and was incomplete. Approximately 65% of the initial amount of chitin was hydrolyzed during a period of 5–10 days. Supplementation of the medium with yeast extract, casamino acids or peptone did not enhance the rate of chitin hydrolysis, but reducing agents such as Na₂S₂O₄ and Ti (III)-NTA markedly stimulated the rate of chitin fermentation.
The ecological implications of the high degree of substrate specialization are discussed.     

Mycotoxin Producing Potential of Fungi Associated with Dry Shrimps

Among 114 fungi isolated from 20 dry shrimp samples, 27 isolates were capable of producing mycotoxins. Aspergillus and Penicillium were-the-predominant fungi. Two of 3 Aspergillus flavus could produce aflatoxin B₁ and G₁ on yeast extract sucrose medium and on surface sterilized dry shrimps. For other toxigenic species, toxicity was found mostly in the chloroform extracts of cultures. Visibly mouldy dry shrimps had significantly higher moisture content than the non-mouldy ones. Most of the fungi were isolated from mouldy samples.     

The effect of age on the thermal resistance of arthroconidia from Chrysosporium inops

J. L. KINDERLERER. 1996. Food-borne members of the genus Chrysosporium have been isolated relatively infrequently. The heat resistance of arthroconidia of the xerophilic fungus, Chrysosporium inops Carmichael, was determined in 0.1% peptone at 66C. The survival curve was sigmoid in shape. The initial lag period was due to the chains of arthroconidia. Thermal inactivation occurred when one viable conidium was left per chain. The presence of chains of arthroconidia was confirmed with the cryo scanning electron microscope. The decimal reduction times were obtained from the regression line of the linear death phase for the heat-sensitive spores. The decimal reduction time (D₆₆) increased with increasing spore age. It was 1.67 min for 3-week-old spores, 1.95 min for 4-week-old spores and 5.49 min for 6-week-old spores. The older spores could recover from thermal death if they were given sufficient time. There was a significant increase in D₆₆ value for 6-week-old spores from 3.97 min to 5.49 when the counts were obtained after 14 d incubation (compared to counts after incubation for 10 d). This effect was not seen for the 3- and 4-week-old spores. There was a small population of heat-resistant spores. The initial population of arthroconidia was greater than log 7 cfu ml^-1. After heating for 1 h at 66C approximately log 2.2 cfu ml^-1 survived. These survivors represented approximately 0.001% of the original population.     

Effects of elevated temperature and carbon dioxide on the nutritional quality of leaves of oak (Quercus robur L.) as food for the Winter Moth (Operophtera brumata L.)

1. Pedunculate Oak trees were grown in ambient and elevated temperatures and CO₂. Leaves were fed to Winter Moth caterpillars reared either in constant conditions or with the trees (caged or on-tree).
2. Caterpillars in constant conditions ate the same mass and produced the same mass of faeces whether fed elevated or ambient temperature leaves. However, less was assimilated from elevated leaves, resulting in lighter pupae and fewer, lighter eggs.
3. Caterpillars in constant conditions ate more and produced more faeces when fed elevated CO₂ leaves than when fed ambient CO₂ leaves, but the mass assimilated and pupal mass were unchanged.
4. Caged caterpillars reared with the trees from which they were fed had constant pupal mass in all treatments, but pupated earlier at elevated temperature. Pupal mass was also unaffected when caterpillars fed on the trees.
5. Nitrogen was reduced in both elevated temperature and elevated CO₂ leaves. Increased fibre in the former prevented increased consumption and resulted in reduced pupal mass and fecundity. Reduced fibre in the latter allowed increased consumption, resulting in pupae of normal mass.
6. Despite the clear effect of nutrient quality, experiments rearing caterpillars and trees together suggest that anticipated climatic change will have no nutritional effect on Winter Moth development.     

Effect of biological and chemical insecticides on Spodoptera species (Lep., Noctuidae) and marketable yields of tomatoes

Various biological and chemical insecticide treatments were evaluated against beet armyworm Spodoptera exigua (Hübner), yellowstriped armyworm Spodoptera ornithogalli (Guenée) and southern armyworm Spodoptera eridania (Cramer) to determine their effects on Spodoptera species, fruit quality and marketable yields of tomatoes Lycopersicon esculentum Mill. Biological insecticides included several Bacillus thuringiensis (Berliner) products including Condor^® OF, Dipel^® 2X, Javelin^® WG, Bactec^® III, Biobit^® FC, Cutlass^® WP and Lepinox^® G. Other biological treatments included a baculovirus, SeNPV isolated from S. exigma , and an entomophagous nematode, Steinernema carpocapsae Weiser. Chemical treatments consisted of several insecticides from various classes including a carbamate (methomyl), pyrethroid (fenpropathrin) and an organophosphate (chlorpyrifos). A natural extract from the neem tree azadirachtin and an untreated control were also included in the evaluation studies. Population densities of S. exigua were below the economic threshold level in control plots and there were no significant differences for small, medium and large larvae. Densities of S. ornithogalli and S. eridania larvae exceeded the threshold level and significant treatment differences were observed in their populations. Fruit injuries were significantly higher in nonefficacious treatments that included S. carpocapsae , SeNPVs and untreated controls. Average weight per fruit was not significantly affected by treatment rates of applications, but total marketable yields were significantly higher in efficacious biological, chemical and combination treatments compared with the control.