The entomopathogenic nematode Steinernema kraussei and Metarhizium anisopliae work synergistically in controlling overwintering larvae of the black vine weevil,Otiorhynchus sulcatus,in strawberry growbags

Previously, the combination of reduced rate of entomopathogenic nematodes (EPN) and fungus caused additive or synergistic mortality to third-instar black vine weevil (BVW), Otiorhynchus sulcatus. In this study, we examined this interaction in unheated glasshouses during winter and compared a combination of commercial formulation of a cold-tolerant EPN, S. kraussei (Nemasys L?) and fungus Metarhizium anisopliae strain V275 against overwintering third-instar BVW. The combination of M. anisopliae with S. kraussei at a rate of 1×10¹⁰ conidia+250,000 nematodes/growbag resulted in additive or synergistic effects, providing 100% control of overwintering larvae.     

Efficacy of soil application of Metarhizium anisopliae and the use of GF-120 spinosad bait spray for suppression of Bactrocera invadens (Diptera: Tephritidae) in mango orchards

Field trials were conducted for two seasons in mango orchards at Nthagaiya, Kenya, to evaluate the efficacy of soil inoculation with Metarhizium anisopliae (Hypocreales: Clavicipitaceae) and GF-120 spinosad bait sprays, applied either alone or in combination, for suppression of the invasive fruit fly, Bactrocera invadens (Diptera: Tephritidae). During the 2006/2007 season, average post-treatment samples showed that B. invadens catches from the control orchards were four times higher than the number of flies captured in the plots receiving M. anisopliae+GF-120. Fruit infestation was 16, 45, 30 and 60% in the M. anisopliae+GF-120, M. anisopliae alone, GF-120 alone and control orchards, respectively. In the 2007/2008 season, average B. invadens post-treatment samples in the control orchards were seven times higher than the treatment with M. anisopliae+GF-120; and fruit infestation was 11, 38, 28, and 52% in the orchards assigned to M. anisopliae+GF-120, M. anisopliae alone, GF-120 alone and control treatments, respectively. Enumeration of conidial densities from soil samples on agar plates showed initial densities of 1.1–2.1×10⁵ colony forming units (cfu)/g of dry weight of soil but decreased to 1.0–1.4×10³ cfu/g at the end of the experimental period. Exposure of laboratory-reared pupariating larvae to soil samples taken from fungus-treated fields showed significant reduction in B. invadens adult emergence (25–36%) compared with the control (80–82%). Our results suggest that the combined use of soil application of M. anisopliae and GF-120 spinosad bait spray is an effective IPM strategy for field suppression of B. invadens on mango.     

Toxicity of non-ionic surfactants and interactions with fungal entomopathogens toward Bemisia tabaci biotype B

Several non-ionic surfactants can be used to enhance insecticidal performance of various bio-based products and agrochemicals. In this study, we describe the toxicity of four commercial non-ionic surfactants against immature Bemisia tabaci (Gennadius) biotype B whiteflies along with their spreading ability. Results revealed that trisiloxane-based surfactants (Break-thru^® and Silwet^® L-77) exhibited the highest toxicity to 1st–2nd (early) and 3rd–4th (late) instar nymphs as well as the greatest wetting performance. We also investigated whether sub-lethal concentrations of Break-thru^® and Silwet^® L-77 improved the insecticidal efficacy of the fungal entomopathogens Beauveria bassiana (Balsamo) Vuillemin (strain CG1229) and Isaria fumosorosea Wise (strain CG1228) conidial suspensions against whiteflies. Germination of hydrophobic conidia of both isolates were unaffected by these surfactants over the range 100–1000 ppm. The combinations of both fungi with trisiloxane carriers significantly increased nymphal mortality with mostly additive and synergistic effects on early and late nymphs, respectively. In screenhouse trials, both fungi (10⁷ conidia ml^?1) mixed with 200 ppm of Silwet^® L-77 significantly improved effectiveness against early nymphs (72–74 % mortality) compared with controls treated with water and Silwet^® L-77 alone. In addition, reduced volume rates of I. fumosorosea in Silwet^® L-77 (equivalent to 100 l ha^?1) were at least as effective against late nymphs as higher volume rates (200 l ha^?1) at equivalent conidial doses. Our findings underline the compatibility and enhanced activity of silicon-based surfactants with B. bassiana and I. fumosorosea for use in integrated whitefly management programs.     

Virulence of the entomopathogenic fungus <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> using soybean oil formulation for control of the cotton stainer bug, <Emphasis Type="Italic">Dysdercus peruvianus</Emphasis>

The cotton stainer bug Dysdercus peruvianus (Hemiptera: Pyrrhocoridae) is an insect pest that causes heavy losses in cotton plantations. The need to reduce the use of insecticides for control of this pest has increased steadily, and Metarhizium anisopliae (Ascomycota: Clavicipitaceae) could be an important biopesticide candidate to control this pest. The effect of M. anisopliae on D. peruvianus nymphs and adults using formulations with soybean oil and Agral^® was evaluated. Formulation using 10% soybean oil added to 10⁸ conidia mL^?1 (grown on used and reused rice) was the most effective for nymph and adult, causing 100% mortality 6 and 7 days after exposure, respectively. The SEM analysis of infected insects showed that M. anisopliae conidia were able to adhere anywhere on the exoskeleton, but were more abundant between the joints. Using the same rice for two batches of growth may be an option for improving commercial conidial production of M. anisopliae and may reduce overall costs. Its effect on D. peruvianus adults opens a new possibility for using this fungus as an alternative to chemical pesticides and the use of M. anisopliae in association with integrate pest management.     

Effect of potting media on the efficacy and dispersal of entomopathogenic nematodes for the control of black vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae)

The effect of five commercial potting media, peat, bark, coir, and peat blended with 10% and 20% compost green waste (CGW) on the virulence of six commercially available entomopathogenic nematodes (EPN), Heterorhabditis bacteriophora UWS1, Heterorhabditis megidis, Heterorhabditis downesi, Steinernema feltiae, Steinernema carpocapsae, and Steinernema kraussei was tested against third-instar black vine weevil (BVW), Otiorhynchus sulcatus. Media type was shown to significantly affect EPN virulence. Heterorhabditis species caused 100% larval mortality in all media whereas Steinernema species caused 100% larval mortality only in the peat blended with 20% CGW. A later experiment investigated the effect of potting media on the virulence of EPN species against BVW by comparing the vertical dispersal of EPN in the presence and absence of BVW larva. Media type significantly influenced EPN dispersal. Dispersal of H. bacteriophora was higher than H. megidis, H. downesi, or S. kraussei in all media, whereas, S. feltiae and S. carpocapsae dispersal was much reduced and restricted to peat blended with 20% CGW and coir, respectively. In the absence of larvae, most of the EPN species remained in the same segment they were applied in, suggesting that the larvae responded to host volatile cues. Greenhouse trials were conducted to evaluate the efficacy of most virulent strain, H. bacteriophora in conditions more representative of those in the field, using 2.5 × 10⁹ infective juveniles/ha. The efficacy of H. bacteriophora UWS1 against third-instar BVW was 100% in peat, and peat blended with 10% and 20% CGW but only 70% in bark and coir, 2 weeks after application. These studies suggest that potting media significantly affects the efficacy and dispersal of EPN for BVW control.     

Covariance in water- and nutrient budgets of Dutch peat polders: what governs nutrient retention?

Water and nutrient budgets were constructed for 13 low-lying peat polders in the Netherlands that varied in elevation relative to sea level (?0.2 to ?2.4 m below sea level), land use (7–70% of the total polder area covered by agriculture; largely dairy farming), and surface water prevalence (6–43%). Water balances were verified with chloride budgets and accepted when both met the criterion (total inflows ? total outflows)/(total inflows) <0.05. Apart from precipitation and evapotranspiration (overall means 913 vs. 600 mm), in- and outlet (171 vs. 420 mm) as well as in- and outward seepage (137 vs. 174 mm) were important items in the water budgets. Nutrient budgets, however, were dominated by terms related to agricultural land use (~60% of all inputs, 90% of N-removal and 80% of P removal) rather than water fluxes (8% and 5% of N and P inputs; 6 and 18% of outputs). After agriculture (200 kg N ha^?1 y^?1), mineralisation of the peat soil and atmospheric deposition appear to be important inputs (about 94 and 21 kg N ha^?1 y^?1). Major output terms were agricultural output (209 kg N ha^?1 y^?1) and denitrification (95 kg N ha^?1 y^?1). The average N budget was in balance (difference ~1 kg N ha^?1 y^?1), whereas P accumulated in most polders, particularly those under agriculture. The mean P surplus (15 kg P ha^?1 y^?1 in the 9 mainly agricultural polders) corresponds well with the accumulated difference observed elsewhere (700 kg P ha^?1 in the upper 50 cm in a nature reserve versus 1400 under agriculture) after over 50 years of dairy farming. Bulk retention of N and P in these polders is taking place in the peat soil, through temporary sorption to the matrix and N is lost through denitrification. In a principal components analysis combining land use, landscape pattern, water balance and nutrient budget terms, the three-first principal components explained 63% of the variability. The first component (PC) correlated strongly with the percentage of land under agriculture (r = 0.82) and negatively with the percentage covered by surface water (r = ?0.74). Most input and output terms of the nitrogen budget also correlated with this PC. The second PC covaried distinctly with the total area of a polder (r = ?0.79) and human population density at municipality level (r = 0.75). Phosphorus loads in inlet and outlet water correlated with this PC. This suggests that the variability in nutrient budgets among polders is largely governed by agricultural land use.     

Additive and synergistic interactions amongst <Emphasis Type="Italic">Orius laevigatus</Emphasis> (Heteroptera: Anthocoridae), entomopathogens and azadirachtin for controlling western flower thrips (Thysanoptera: Thripidae)

This study evaluated the efficacy of the foliage-dwelling predator Orius laevigatus, soil applied entomopathogens and azadirachtin alone and in combinations for controlling western flower thrips (WFT). Evaluated products were Nemastar^® Steinernema carpocapsae (E-nema) and O. laevigatus (Re-natur), Metarhizium anisopliae isolate ICIPE-69 and NeemAzal-T (azadirachtin) (Trifolio). Efficacy against WFT was significantly improved by combined treatments achieving 62–97 % reduction in WFT emergence, compared to 45–74 % in single treatments, and interactions resulted in two synergistic and eight additive responses. Metarhizium-based treatments reduced WFT survival by 93–99.6 % when late mortality by mycosis was considered. Halving the number of released predators did not significantly reduce efficacy (86–96 vs. 76–88 % thrips reduction), and when Orius was introduced to target L1 of WFT, 96–98 % reduction was achieved while only 71–89 % for L2. Early release of O. laevigatus and combination with soil application of NeemAzal-T and/or entomopathogens can be a successful and reliable biocontrol strategy for WFT.     

Effect of entomopathogenic Aspergillus strains against the pea aphid,Acyrthosiphon pisum (Hemiptera: Aphididae)

Aphids (Homoptera: Aphididae) are sap-sucking insect pests that feed on several plants of agronomical importance. Entomopathogenic fungi are valuable tools for potential aphid control. As part of a selection process, laboratory bioassays were carried with five different concentrations of Aspergillus clavatus (Desmazières), Aspergillus flavus (Link) and Metarhizium anisopliae ((Metschnikoff) Sorokin) spores against the pea aphid, Acyrthosiphon pisum (Harris). Aspergillus isolates induced higher mortalities than M. anisopliae, which is a well-known entomopathogen in the literature. Lethal concentrations (LC₅₀ and LC₉₀) were 1.23 × 10³ and 1.34 × 10⁷ spores/ml for A. flavus, 4.95 × 10² and 5.65 × 10⁷ spores/ml for A. clavatus, and 3.67 × 10³ and 9.71 × 10⁷ spores/ml for M. anisopliae 5 days after treatment. Mycelia development and sporulation on adult cadavers were observed 48 h after incubation. The intrinsic growth rate of A. pisum decreased with increased spore concentration for all fungal strains, suggesting an increase in pathogen fitness related to a consumption of host resources. In conclusion, Aspergillus species could be useful in aphid control as pest control agents despite their saprophytic lifestyle. This is also to our knowledge the first report of A. clavatus and A. flavus strains pathogenic to aphids.     

Efficacy of entomopathogenic fungi against soil-dwelling life stages of western flower thrips, Frankliniella occidentalis, in plant-growing media

The potential of several entomopathogenic fungi to control soil‐dwelling stages of western flower thrips, Frankliniella occidentalis Pergande (Thysanoptera: Thripidae), was evaluated in various growing media. Two Metarhizium anisopliae (Metsch) Sorokin strains, V275 and ERL700, were the most effective, causing 85–96% mortality of thrips larvae and pupae (as measured by relative adult emergence rates), 11 days after inoculation. Mortality in other M. anisopliae‐treated media ranged from 51–84%; Beauveria bassiana (Balsamo) Vuillemin strains caused 54–84% mortality, and Paecilomyces fumosoroseus (Wize) 63–75% mortality. In contrast, mortality from the insecticide treatment (fipronil 1 g l^?1 medium) was only 15–54%. Metarhizium anisopliae V275 was selected for more in‐depth evaluation under greenhouse conditions. There was no significant difference in M. anisopliae efficacy in growing media whether it was applied as drench (84–93%) or premixed into the medium as dry conidia (85–92%). The use of M. anisopliae with reduced rates of either fipronil or imidacloprid did not significantly improve control. Overall, our study shows that M. anisopliae V275 is robust and offers much promise for the control of soil‐dwelling stages of thrips as part of an integrated pest‐management programme.     

Impacts of fertilizer practices on environmental risk of nitrate in semiarid farmlands in the Loess Plateau of China

The lack of understanding of nitrate dynamics in soil profiles of semiarid regions hampers the assessment of the environmental risks associated with nitrate. A long-term field experiment established in the Loess Plateau of Northwest China in 1984 was used to investigate the seasonal dynamics of water and nitrate contents in the soil profile (0–300 cm) under bare fallow and continuous winter wheat (Triticum aestivum L.) with various fertilizer treatments. For treatments without mineral N input (i.e., no fertilizer, farmyard manure alone, and with P fertilizer), the amount of nitrate accumulated in the soil profile (52–120 kg N ha^?1, the average for June, August, February and April) was significantly lower than that (292 kg N ha^?1) accumulated in the bare fallow treatment. A large amount of nitrate (1,065 kg N ha^?1) was found accumulated in the soil profile with the treatment applied with mineral N at a rate of 120 kg N ha^?1 year^?1 for 17 years (1984–2001) and this nitrate moved downward during the wet season (from August to February). Clearly, the amount of nitrate accumulated in the soil profiles, and its tendency of downward movement, appears to potentially be an environmental risk as it may reach groundwater. Fertilization as mineral N fertilizers coupled with FYM or P resulted in 50–70% less nitrate accumulation in the soil profiles than that using mineral N fertilizer alone, and therefore the environmental risk was reduced. It is proposed that a “break point” of nitrate distribution existed in the soil profiles, providing an indication of soil depth to which nitrate can transfer.     

Effects of four commercial fungal formulations on mortality and sporulation in house flies (Musca domestica) and stable flies (Stomoxys calcitrans)

The house fly Musca domestica L. (Diptera: Muscidae) and stable fly Stomoxys calcitrans (L.) (Diptera: Muscidae) are major pests of livestock. Biological control is an important tool in an integrated control framework. Increased mortality in filth flies has been documented with entomopathogenic fungi, several strains of which are commercially available. Three strains of Beauveria bassiana (Balsamo‐Crivelli) Vuillemin (Hypocreales: Cordycipitaceae) and one strain of Metarhizium brunneum (Petch) (Hypocreales: Clavicipitaceae) were tested in commercial formulations for pathogenicity against house flies and stable flies. There was a significant increase in mortality of house flies with three of the formulations, BotaniGard^® ES, Mycotrol^® O, and Met52^® EC, during days 4–9 in comparison with balEnce? and the control. In stable flies, mortality rates were highest with Met52^® EC, followed by Mycotrol^® O, BotaniGard^® ES and, finally, balEnce?. There was a significant fungal effect on sporulation in both house flies and stable flies. Product formulation, species differences and fungal strains may be responsible for some of the differences observed. Future testing in field situations is necessary. These commercial biopesticides may represent important tools in integrated fly management programmes.     

Influence of organic and inorganic sources of nutrients on the functional diversity of microbial communities in the vegetable cropping system of the Indo-Gangetic plains

The aim of the present study was to assess the effects of different organic and inorganic fertilizers on the functional diversity of soil microbial community under a vegetable production system. The Biolog^® Eco-plate technique and indices, such as average well-colour development (AWCD), McIntosh and Shannon diversity were employed to study the diversity of soil microorganisms. The AWCD, i.e. overall utilization of carbon sources, suggested that different organic treatments had a significant impact on the metabolic activity of soil microorganisms. After 120 h, the highest AWCD values were observed in poultry manure (2.5 t·ha^?1) + vermicompost (3.5 t·ha^?1) (0.63) and farm yard manure (FYM) (10 t·ha^?1) + vermicompost (3.5 t·ha^?1) (0.61). After 72 h, the highest value of the McIntosh diversity index was recorded in poultry manure (2.5 t·ha^?1) + vermicompost (3.5 t·ha^?1) (3.87), followed by poultry manure (2.5 t·ha^?1) + vermicompost (3.5 t·ha^?1) + biofertilizers (Azotobacter 500 g·ha^?1 applied as seed treatment) (3.12). In the case of the Shannon diversity index, the highest values were noticed in organic treatments; however, there was no significant differences between organic and inorganic treatments. Biplot analysis showed a clear differentiation of organic treatments from the inorganic control. The amino acids, phenolics and polymer utilizing microorganisms were dominant in organic treatments. Inorganic control recorded the lowest values of the microbial diversity indices. Through this study, we have identified the best combination of organic nutrients, i.e. poultry manure (2.5 t·ha^?1) + vermicompost (3.5 t·ha^?1) for the stimulation of metabolically active soil microbial communities.     

Comparative study between Larval Packet Test and Larval Immersion Test to assess the effect of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> on <Emphasis Type="Italic">Rhipicephalus microplus</Emphasis> tick larvae

Entomopathogenic fungi, such as Metarhizium anisopliae, for the control of arthropods, have been studied for more than 20 years. The aim of this study was to determine the best methodology to evaluate the in vitro effect of the fungus M. anisopliae on Rhipicephalus microplus tick larvae. We compared a modified Larval Packet Test (LPT) and a Larval Immersion Test (LIT). For the LPT filter papers were impregnated with 1 mL of M. anisopliae suspension in Triton X-100 at 0.02%, in concentrations of 10⁶, 10⁷ and 10⁸ conidia/mL and subsequently folded to include the larval ticks. LIT was performed by immersing the larvae in M. anisopliae suspensions for 5 min using the same three concentrations, then the larvae were placed on filter paper clips. For LPT, the LT₅₀ values obtained were 134.6, 27.2 and 24.8 days for concentrations of 10⁶, 10⁷ and 10⁸ conidia/mL; and the mortality after 21 days was 17.3, 17.6 and 38%, respectively. The LT₅₀ values of LIT were 24.5, 20 and 9.2 days with mortality after 21 days of 50.5, 64.7 and 98% for 10⁶, 10⁷ and 10⁸ conidia/mL, respectively. For the same conidia concentration, LIT showed a higher mortality in a shorter time interval when compared with LPT. These differences between the methods tested must be taking into account in further screening and effect studies with M. anisopliae. The set of results shown here could optimize the protocol used to identify M. anisopliae strains pathogenic against R. microplus.     

Niche-based assessment of contributions of legumes to the nitrogen economy of Western Kenya smallholder farms

Nitrogen (N) deficiency is a major constraint to the productivity of the African smallholder farming systems. Grain, green manure and forage legumes have the potential to improve the soil N fertility of smallholder farming systems through biological N₂-fixation. The N₂-fixation of bean (Phaseolus vulgaris), soyabean (Glycine max), groundnut (Arachis hypogaea), Lima bean (Phaseolus lunatus), lablab (Lablab purpureus), velvet bean (Mucuna pruriens), crotalaria (Crotalaria ochroleuca), jackbean (Canavalia ensiformis), desmodium (Desmodium uncinatum), stylo (Stylosanthes guianensis) and siratro (Macroptilium atropurpureum) was assessed using the ¹⁵N natural abundance method. The experiments were conducted at three sites in western Kenya, selected on an agro-ecological zone (AEZ) gradient defined by rainfall. On a relative scale, Museno represents high potential AEZ 1, Majengo medium potential AEZ 2 and Ndori low potential AEZ 3. Rainfall in the year of experimentation was highest in AEZ 2, followed by AEZ 1 and AEZ 3. Experimental fields were classified into high, medium and low fertility classes, to assess the influence of soil fertility on N₂-fixation performance. The legumes were planted with triple super phosphate (TSP) at 30 kg P ha^?1, with an extra soyabean plot planted without TSP (soyabean-P), to assess response to P, and no artificial inoculation was done. Legume grain yield, shoot N accumulation, %N derived from N₂-fixation, N₂-fixation and net N inputs differed significantly (P<0.01) with rainfall and soil fertility. Mean grain yield ranged from 0.86 Mg ha^?1, in AEZ 2, to 0.30 Mg ha^?1, in AEZ 3, and from 0.78 Mg ha^?1, in the high fertility field, to 0.48 Mg ha^?1, in the low fertility field. Shoot N accumulation ranged from a maximum of 486 kg N ha^?1 in AEZ 2, to a minimum of 10 kg N ha^?1 in AEZ 3. Based on shoot biomass estimates, the species fixed 25–90% of their N requirements in AEZ 2, 23–90% in AEZ 1, and 7–77% in AEZ 3. Mean N₂-fixation by green manure legumes ranged from 319 kg ha^?1 (velvet bean) in AEZ 2 to 29 kg ha^?1 (jackbean) in AEZ 3. For the forage legumes, mean N₂-fixation ranged from 97 kg N ha^?1 for desmodium in AEZ 2 to 39 kg N ha^?1 for siratro in AEZ 3, while for the grain legumes, the range was from 172 kg N ha^?1 for lablab in AEZ 1 to 3 kg N ha^?1 for soyabean-P in AEZ 3. Lablab and groundnut showed consistently greater N₂-fixation and net N inputs across agro-ecological and soil fertility gradients. The use of maize as reference crop resulted in lower N₂-fixation values than when broad-leaved weed plants were used. The results demonstrate differential contributions of the green manure, forage and grain legume species to soil fertility improvement in different biophysical niches in smallholder farming systems and suggest that appropriate selection is needed to match species with the niches and farmers’ needs.     

Effects of soil-treatment with fungal biopesticides on pupation behaviors,emergence success and fitness of tea geometrid,Ectropis grisescens (Lepidoptera: Geometridae)

Ectropisgrisescens Warren (Lepidoptera: Geometridae) is one of the most severe defoliating pests of tea plants. Synthetic pesticides have been widely applied to control this pest in tea plantations, but pesticide residues may decrease the quality and safety of tea products. In the present study, we hypothesized that soil treatment with the two commercial biopesticides (Shuiguxin^®) based on Metarhizium anisopliae (Metchnikoff) Sorokin and Beauveria bassiana (Balsamo) could reduce the survivorship and fitness of E. grisescens. Wandering larvae of E. grisescens were allowed to pupate in soil treated with each biopesticide, and the concentrations of M. anisopliae (Shuiguxin^®) and B. bassiana (Shuiguxin^®) that produced the 50% mortality values (LC₅₀) were 2.9 × 10⁶ and 1.6 × 10⁷ conidia/g soil, respectively. Artificial burying the pupae using soil treated with M. anisopliae (Shuiguxin^®) and B. bassiana (Shuiguxin^®) (1 × 10⁸ or 1 × 10⁹ conidia/g soil for both biopesticides) also significantly reduced emergence success of E. grisescens. In addition, choice tests showed that soil treated with the high concentration of M. anisopliae (Shuiguxin^®) or B. bassiana (Shuiguxin^®) had repellent effects on pupating E. grisescens. However, sublethal concentrations (LC₂₅ and LC₅₀) of both biopesticides did not significantly affect fecundity, fertility and longevity of post-emerged adults. Our study showed that soil treatment with the two commercial biopesticides caused direct mortality of pupating E. grisescens, but may not effectively suppress E. grisescens populations at sublethal concentrations. The realistic application of the fungal dosage in fields should be determined in future studies     

Evaluation of a liquid formulation of Pseudomonas fluorescens against Fusarium oxysporum f. sp. cubense and Helicotylenchus multicinctus in banana plantation

Plant growth promoting rhizobacteria (PGPR) are eco-friendly alternatives to chemical fungicides to manage plant diseases. We evaluated the efficacy of a Pseudomonas fluorescens formulation against Fusarium oxysporum f. sp. cubense and Helicotylenchus multicinctus at multiple banana plantations. Three field trials were conducted to assess the wilt incidence and the populations of nematode and bacteria in the soil treated with a liquid formulation of P. fluorescens at 2.0, 3.0 and 4.0 l ha^?1 using drip irrigation system at 60, 120, 180 and 240 days after planting. The results showed that the treatment at 4.0 l ha^?1 reduced the wilt incidence by 60 %. It also reduced the overall population of H. multicinctus by 41.3–89.0 % in the treated fields. The presence of P. fluorescens in the treated soil was 5.6 × 10⁶ cfu g^?1 of soil at the time of harvest. The treatment of biocontrol agent P. fluorescens also resulted in an overall yield increase in banana production by 36.6–46.5 % compared to the control.     

Effect of nitrogen and phosphorus fertilization on the composition of rhizobacterial communities of two Chilean Andisol pastures

The effect of nitrogen (N) and phosphorus (P) fertilization on composition of rhizobacterial communities of volcanic soils (Andisols) from southern Chile at molecular level is poorly understood. This paper investigates the composition of rhizobacterial communities of two Andisols under pasture after 1- and 6-year applications of N (urea) and P (triple superphosphate). Soil samples were collected from two previously established sites and the composition of rhizobacterial communities was determined by denaturing gradient gel electrophoresis (PCR–DGGE). The difference in the composition and diversity between rhizobacterial communities was assessed by nonmetric multidimensional scaling (MDS) analysis and the Shannon–Wiener index. In Site 1 (fertilized for 1 year), PCR–DGGE targeting 16S rRNA genes and MDS analysis showed that moderate N application (270 kg N ha^?1 year^?1) without P significantly changed the composition of rhizobacterial communities. However, no significant community changes were observed with P (240 kg P ha^?1 year^?1) and N–P application (270 kg N ha^?1 year^?1 plus 240 kg P ha^?1 year^?1). In Site 2 (fertilized for 6 years with P; 400 kg P ha^?1 year^?1), PCR–DGGE targeting rpoB, nifH, amoA and alkaline phosphatase genes and MDS analysis showed changes in rhizobacterial communities only at the highest rate of N application (600 kg N ha^?1 year^?1). Quantitative PCR targeting 16S rRNA genes also showed higher abundance of bacteria at higher N application. In samples from both sites, the Shannon–Wiener index did not show significant difference in the diversity of rhizobacterial communities. The changes observed in rhizobacterial communities coincide in N fertilized pastures with lower soil pH and higher pasture yields. This study indicates that N–P application affects the soil bacterial populations at molecular level and needs to be considered when developing fertilizer practices for Chilean pastoral Andisols.     

Effects of adjuvant and conidial concentration on the efficacy of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> for the control of the two spotted spider mite, <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Greenhouse experiments were conducted on various crops (cucumber, tomato, eggplant, green bean) to ascertain the effects of Break-thru^® (polyether-polymethylsiloxane-copolymer, a silicone surfactant) and an oil emulsion, on Beauveria bassiana (Balsamo) Vuillemin (Bb) applications for the control of the two spotted spider mite, Tetranychus urticae Koch. The objectives were to compare a) the efficacy of Bb control when applied in aqueous Break-thru^® or an oil emulsion; b) the effects of various concentrations of Bb conidia, as affected by each surfactant; and c) the effects of Break-thru^® on the activity of the fungus. Conidia were suspended either in an aqueous Break-thru^® or an emulsifiable formulation at different conidial concentrations (1.05 × 10⁶, 2.1 × 10⁶ and 4.2 × 10⁶ conidia ml^?1) and sprayed onto leaves 2 weeks after artificial pest inoculation. Two sprays were performed, with an interval of one week from one spray to another, and T. urticae population counts (both motile and egg stages) were made on plant leaves 7 days after each spray. Bb conidia in Break-thru^® were more efficacious than conidia in emulsifiable formulation. With the highest rate of conidia (4.2 × 10⁶ conidia ml^?1), mortality of adult mites ranged from 60 ± 4.2 (mean ± SE) to 85.7 ± 4.3% in the Break-thru^® suspension and 39.4 ± 7 to 61.3 ± 6% in the oil emulsion. Leaf damage index was also substantially reduced from 70% in the unsprayed control to 40% by the application of Bb conidia at the highest rate with Break-thru^®. Break-thru^® can be combined with Bb in the integrated management of T. urticae and Isolate R444 is a promising candidate for the control of the pest.     

Impact of fungicides on Metarhizium anisopliae in the rhizosphere, bulk soil and in vitro

The entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae) is registered in the United States and The Netherlands for black vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae) control in container-grown ornamentals. These studies were conducted to determine the compatibility of M. anisopliae (F52) with a wide range of fungicides commonly applied to container-grown ornamentals for the management of soil-borne plant pathogens. The impact of fungicides on spore germination and mycelial growth were determined in vitro. In addition, M. anisopliae persistence in bulk and rhizosphere soil was determined 30 days following dual application of each fungicide at 7–28 days intervals as prescribed. A number of fungicides (thiophanate-methyl, dimethomorph, captan, triflumizole, triflozystrobin, pyraclostrobin, azoxystrobin) inhibited spore germination in vitro. A larger number of fungicides (fosetyl-AI, thiophanate-methyl, dimethomorph, captan, quintozene, triflumizole, fludioxanil, triflozystrobin, pyraclostrobin, fludiox-mefanox, iprodione, azoxystrobin, phosphorus acid/K-salts) inhibited mycelial growth in vitro. Only three fungicides (etridiazole, propamocard and mafanoxam) had no significant impact in vitro on spore germination or mycelial growth. While a number of fungicides had a detrimental impact in vitro, there was no impact on M. anisopliae populations in bulk soil following dual application of any fungicide. However, the fungicides captan and triflumizolet, which have a short reapplication interval, had a detrimental impact on M. anisopliae populations in the rhizosphere. As researchers develop rhizosphere competence as an alternative management strategy for black vine weevil, the fungicides captan and triflumizole should be avoided.     

The l58Val/Met polymorphism of catechol-O-methyl transferase gene and prostate cancer risk: a meta-analysis

The association between COMT Val158Met polymorphism and prostate cancer has been evaluated. However, the results of these studies on the association remain conflicting. To derive a more precise estimation of the relationship, a meta-analysis was performed. A comprehensive search was conducted to identify the eligible studies of COMT Val158Met polymorphism and prostate cancer risk. Summary odds ratios (OR) and 95 % confidence interval (CI) for COMT Val158Met polymorphism and prostate cancer were calculated. Statistical analysis was performed with the software program Review Manage (Version 5.0) and Stata (Version 12.0). Six case–control studies, totally 4,118 persons including 2,143 cases and 1,975 controls, met the included criteria and thus were selected. Our analysis suggested that Val158Met polymorphism was associated with prostate cancer risk in overall population. Collectively, the results of the present study suggest that significant associations of COMT Val158Met polymorphisms with prostate cancer were observed (for additive model: OR = 1.068, 95 % CI = 1.002–1.138, P _{heterogeneity} = 0.363, P = 0.043; for dominant model: OR = 1.266, 95 % CI = 1.057–1.517, P _{heterogeneity} = 0.000, P = 0.011; for recessive model: OR = 1.050, 95 % CI = 0.961–1.146, P _{heterogeneity} = 0.558, P = 0.279; and Val allele versus Met allele OR = 0.932, 95 % CI = 0.894–0.971, P _{heterogeneity} = 0.272, P = 0.001). In the subgroup analysis, we detected no significant association between the COMT 158 Val/Met genotype and prostate cancer risk in Caucasian and Asian populations, while the contrary result for additive model (OR = 2.43, 95 % CI = 1.08–5.43, P _{heterogeneity} = 0.04, P = 0.03) in Asian populations. The result of this meta-analysis suggests that COMT l58Val/Met polymorphism might be contributed to the overall prostate cancer risk.