Distribution of entomopathogenic nematodes in Southern Cameroon

A first survey of entomopathogenic nematodes (EPN) was conducted in three agro-ecological zones of Southern Cameroon in 2007 and 2008. Entomopathogenic nematodes were recovered from 26 of 251 soil samples (10.4%). Three species, Heterorhabditis baujardi, Steinernema sp. A and Steinernema sp. B were found. The two steinernematids were considered unidentified species. Among the positive samples, 23 samples contained only H. baujardi (88.5%), two contained Steinernema sp. A co-occurring with H. baujardi (7.7%), and one sample contained Steinernema sp. B (3.9%). H. baujardi was frequent in forest and fruit crop (cocoa and oil palm plantations). Steinernema sp. A was found in a tree plantation of teak, Steinernema sp. B in a forest habitat. Nematodes were mostly present in acidic soils with pH ranging from 3.7 to 7.0. The highest EPN presence was recorded in sandy loam, sandy clay loam, sandy clay and clay soils. EPNs were not recovered in sand, loamy sand and clay loam soils. Using principal component analysis for elucidating the major variation patterns among sampling sites, four factors explaining for 73.64% of the overall variance were extracted. Factors were a combination of geographical (latitude, longitude, altitude), soil (pH, contents of sand, silt and clay, organic carbon, texture), and moisture (wilting point, field capacity) parameters as well as climatic parameters (mean annual rainfall, mean air temperature). Logistic regression and redundancy analyses (RDA) revealed that soil pH, longitude, available water and altitude were associated with presence and absence of EPN. Both logistic regression and RDA indicated that, increasing soil pH and longitude, associated with decreasing altitude, led to higher percentages of samples containing entomopathogenic nematodes.     

Entrapment of biological control agents applied to entomopathogenic nematodes

Summary Hydrogels of alginate, phospho guar gum, carboxymethyl guar gum, k-carrageenan and cellulose sulphate, respectively were tested to find easily redissolvable gels. The entomopathogenic nematode, Heterorhabditis sp., was entrapped in calcium alginate beads, calcium alginate hollow spheres and foils made from different hydrogels. Emigration from calcium alginate beads after 7 days of storage was 100 % at room temperature and was lowered to 6 % at 6 °C, whereas no emigration from calcium alginate hollow spheres was found at either temperature. Highly concentrated polymer foils produced on gauze showed reduced emigration with a survival of 80 % after 24 h compared to foils produced on glass slides. Calcium alginate beads can be used for a controlled release of the nematode into the environment, while hollow spheres and foils are suitable for storage.Dedicated to Prof. Dr. F. Wagner on the occasion of his 65th birthday     

Nitrification inhibition in drip irrigation systems

The water flow pattern and distribution in drip irrigated soil are different than those for conventional irrigation systems, to an extent that could drastically effect reactions of nutrient ions in the soil.Ammonium applied through the irrigation system was not nitrified in the saturated zone below the emitter. Nitrification occurred in the unsaturated zone surrounding the central cylinder. Nitrate levels and number of nitrifying bacteria are low below the emitter and higher father away.The residual nitrogen in the soil was found to be higher than that found for conventional irrigation plots. It does seem that application of ammonical fertilizer through a drip irrigation system is potentially a controlled release fertilizer system.     

昆虫病原线虫的耐寒性

昆虫病原线虫可开发成生物农药,广泛应用于多种地下及钻蛀害虫的安全防治。但昆虫病原线虫货架期较短,对寒冷等极端环境的耐受性较差,影响了其在生物防治方面的商业开发。本文介绍了寒区的昆虫病原线虫资源,总结了昆虫病原线虫耐寒性的测定方法及增强方法、耐寒性差异的研究进展,并对其耐寒的生理生化机制及分子机理进行了综述。研究昆虫病原线虫的耐寒性,对于解释种群动态,指导昆虫病原线虫的低温保存,以及拓展其在生物防治方面的应用具有重要意义。     

Distribution of the entomopathogenic nematodes from La Rioja (Northern Spain)

Entomopathogenic nematodes (EPNs) distribution in natural areas and crop field edges in La Rioja (Northern Spain) has been studied taking into account environmental and physical-chemical soil factors. Five hundred soil samples from 100 sites of the most representative habitats were assayed for the presence of EPNs. The occurrence of EPNs statistically fitted to a negative binomial distribution, which pointed out that the natural distribution of these nematodes in La Rioja was in aggregates. There were no statistical differences (p < or = 0.05) in the abundance of EPNs to environmental and physical-chemical variables, although, there were statistical differences in the altitude, annual mean air temperature and rainfall, potential vegetation series and moisture percentage recovery frequency. Twenty-seven samples from 14 sites were positive for EPNs. From these samples, twenty isolates were identified to a species level and fifteen strains were selected: 11 Steinernema feltiae, two S. carpocapsae and two S. kraussei strains. S. kraussei was isolated from humid soils of cool and high altitude habitats and S. carpocapsae was found to occur in heavy soils of dry and temperate habitats. S. feltiae was the most common species with a wide range of altitude, temperature, rainfall, pH and soil moisture, although this species preferred sandy soils. The virulence of nematode strains were assessed using G. mellonella as insect host, recording the larval mortality percentage and the time to insect die, as well as the number of infective juveniles produced to evaluate the reproductive potential and the time tooks to leave the insect cadaver to determinate the infection cycle length. The ecological trends and biological results are discussed in relationship with their future use as biological control.     

Antagonism between entomopathogenic fungi and bacterial symbionts of entomopathogenic nematodes

Mutual effects between the symbiotic bacteria of entomopathogenic nematodes, Photorhabdus luminescens and Xenorhabdus poinarii, and entomopathogenic fungi were investigated in vitro. A dual culture assay on nutrient agar supplemented with bromothymol blue and triphenyltetrazolium chloride (NBTA) medium revealed that P. luminescens is antagonistic to Metarhizium anisopliae, Beauveria bassiana, B. brongniartii and Paecilomyces fumosoroseus by inhibiting their growth and conidial production; the fungal growth was not inhibited by X. poinarii. In a second laboratory experiment, crude extract produced by M. anisopliae was tested for its activity against P. luminescens and X. poinarii. Crude extract from M. anisopliae was antibacterial to P. luminescens and X. poinarii at 1000 g/ml and inhibited their growth on NBTA, but had no effect at 100 or 10 g/ml. The influence of the crude extract of M. anisopliae on the dispersal of infective juveniles (IJs) of Heterorhabditis megidis and Steinernema glaseri was assayed on Sabouraud Dextrose Agar (SDA) plates. Results showed that the crude extract of M. anisopliae had no toxic effects even at highest concentration (1000 g/ml).     

Compatibility of entomopathogenic nematodes with fipronil

The survival and infectivity of infective juveniles (IJs) of three species of entomopathogenic nematodes, Steinernema carpocapsae Weiser, S. arenarium (Artyukhovsky) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae), were determined after exposure to different concentrations (250, 500, 1000 and 2000 ppm) of fipronil, an insecticide acting on the GABA receptors to block the chloride channel. Heterorhabditis bacteriophora was very tolerant to all concentrations of fipronil, with the highest mortality of 17% being observed at 2000 ppm of fipronil after 72 h exposure. Steinernema carpocapsae showed a similar response, with the highest mortality of 11.25% of IJs being observed after 72 h exposure to 2000 ppm of fipronil. Steinernema arenarium was, however, more sensitive to fipronil, and at 2000 ppm mortality rates of 94.6% and 100% were observed after 24 and 72 h, respectively. Fipronil had negligible effects on the infectivity of the three nematode species tested. The IJs which survive exposure to all concentrations of fipronil tested can infect and reproduce in Galleria larvae. The moderate effects on entomopathogenic nematodes of a lower fipronil concentration (250 ppm) and the field rates (12-60 ppm) of fipronil used as insecticide, suggest that direct mixing of entomopathogenic nematodes and fipronil at field rates is a viable integrated pest management option.     

Factors limiting short-term persistence of entomopathogenic nematodes

Abstract:  Entomopathogenic nematodes are commonly applied to soil and crops for biocontrol of insects. Typically, when nematodes are applied, recoverable numbers decline quickly. Because ultra-violet light is known to kill entomopathogenic nematodes in the laboratory, many researchers blame poor field persistence on the action of ultra-violet light. We conducted a field experiment to test this hypothesis, using Heterorhabditis bacteriophora applied to turf as a model system. We compared persistence of surface-applied nematodes with subsurface-applied nematodes and found no difference in persistence. Numbers of potential nematode antagonists (mites and collembola) were also monitored. Poor persistence was positively correlated with numbers of mites and collembola in plots where nematodes were surface-applied, but not in plots where they were subsurface-applied.     

The safety of thiamethoxam to pollinating bumble bees (Bombus terrestris L.) when applied to tomato plants through drip irrigation

Thiamethoxam, mainly sold under the trademark of Actara, is a neonicotinoid widely used in covered vegetables for the control of aphids and whiteflies. In these crops, and particularly in covered tomatoes, bumble-bees are used for cross-pollination as an alternative to labour intensive manual techniques. In this study, made on tomatoes grown in separated greenhouse plots in Murcia, Southern Spain, thiamethoxam was applied through drip irrigation at a rate of 200 g ai/ha, and as a split application of the same rate, to evaluate the effects on pollinating bumble bees compared to a foliar application of a toxic standard. The results showed that the toxic foliar standard had a clear effect on the pollination of tomato flowers, declining to zero pollination two weeks after application, whereas both the single and split drip irrigation applications of Actara had no effect on pollination when compared to the control plots. The count of dead adults and larvae did not show any differences between the treatments, whereas the measurement of sugar water consumption was shown to correlate well with pollination. The consumption of sugar water declined in the toxic standard plots by 69% with respect to the control, whilst the decline in lower dose drip irrigation application was only 3%. In regard to hive weight, and number of adults and brood after destructive sampling; there were no statistical differences between the treatments but a negative effect of the foliar treatment was observed. Based on these results we can conclude that a split application of Actara applied in drip irrigation to the soil/substrate has no effect on the bumble-bees used in tomatoes for pollination.     

Spatial patterns of the red palm weevil and applied entomopathogenic nematode Heterorhabditis bacteriophora

The distribution and abundance of red palm weevil (Rhynchophorus ferrugineus) within palm tree-infected stems at Ismaelia governorate, Egypt, were investigated. Taylor's power law was fit (P ≤ 0.01) to its larvae, pupae and/or adults. The slope value of this power law indicated the clumped distribution of these insect stages. Sample size optimisation needed to achieve a predetermined level of sampling error for the insect stages was calculated. Also, soil samples were taken after uniform application of entomopathogenic nematode Heterorhabditis bacteriophora strain EG₁ to the soil under citrus tree canopies at the rate of 10⁴ infective juveniles/400 cm² plots, and assayed for the nematode using the Galleria bait method. At the first and second sampling dates, the nematode displayed contagious distribution and attained mean insect mortality of 15.3 and 4, respectively. At the third date of sampling, the nematode showed random distribution according to chi-squared test and caused 1.8 mean insect mortality. Evaluation of H. bacteriophora EG₁ as a biocontrol measure for the weevil was discussed based on their investigated dispersion indices.     

昆虫病原线虫对黄地老虎致病力的研究

为了探索应用昆虫病原线虫防治黄地老虎Agrotis segetum的可行性,室内生测比较了9个昆虫病原线虫品系对黄地老虎4龄幼虫的致病力。生测结果表明,9个不同昆虫病原线虫品系对黄地老虎4龄幼虫的致病力差别很大,其中格氏线虫Steinernema glaseri ib(Sgib)是对黄地老虎幼虫致病力最强的昆虫病原线虫品系,侵染24、36和72 h黄地老虎幼虫的校正死亡率均高于其他线虫品系。此外,还测定了不同温度、土壤湿度以及线虫剂量对格氏线虫Sgib致病力的影响,结果表明,在21℃-27℃、20%土壤湿度以及侵染期线虫与黄地老虎幼虫数量比不少于10∶1的条件下,格氏线虫Sgib对黄地老虎幼虫的致病效果最好。田间小区实验表明,格氏线虫Sgib对黄地老虎幼虫的防效达到96.67%。     

Mass production of entomopathogenic nematodes for plant protection

Entomopathogenic nematodes of the genera Heterorhabditis and Steinernema are commercially used to control pest insects. They are symbiotically associated with bacteria of the genera Photorhabdus and Xenorhabdus, respectively, which are the major food source for the nematodes. The biology of the nematode-bacterium complex is described, a historical review of the development of in vitro cultivation techniques is given and the current use in agriculture is summarised. Cultures of the complex are pre-incubated with the symbiotic bacteria before the nematodes are inoculated. Whereas the inoculum preparation and preservation of bacterial stocks follow standard rules, nematodes need special treatment. Media development is mainly directed towards cost reduction, as the bacteria are able to metabolise a variety of protein sources to provide optimal conditions for nematode reproduction. The process technology is described, discussing the influence of bioreactor design and process parameters required to obtain high nematode yields. As two organisms are grown in one vessel and one of them is a multicellular organism, the population dynamics and symbiotic interactions need to be understood in order to improve process management. Major problems can originate from the delayed or slow development of the nematode inoculum and from phase variants of the symbiotic bacteria that have negative effects on nematode development and reproduction. Recent scientific progress has helped to understand the biological and technical parameters that influence the process, thus enabling transfer to an industrial scale. As a consequence, costs for nematode-based products could be significantly reduced.     

Dwarf pea response to gibberellic acid applied to soil through a drip irrigation system,and gibberellic acid biodegradation in soil

Gibberellic acid (29 or 290 M) injected into drip irrigation lines significantly stimulated internode elongation of dwarf peas, and the 290-M soil treatment produced significantly taller plants than did the 29-M treatment. GA₃ uptake may limit GA-induced internode elongation when GA₃ is applied to soil, in contrast to results obtained for hydroponically grown plants, where uptake initially appeared to exceed the rate of hormone metabolism (andersonet al.). It is likely that biodegradation or chemical inactivation limited the plant-availability of GA₃ in the soil. Degradation of moderate GA₃ concentrations in a moist, aerobic loamy fine sand was nearly complete within five days, indicating that the inefficiency of soil applications may outweight the benefits provided by reducing labor costs associated with foliar-spray applications.     

Prevalence of potentially thermophilic microorganisms in biofilms from greenhouse-enclosed drip irrigation systems

Drip irrigation systems using reclaimed water often present clogging events of biological origin. Microbial communities in biofilms from microirrigation systems of an experimental greenhouse in Almería, SE Spain, which used two different qualities of water (treated wastewater and reclaimed water), were analyzed by denaturing gradient gel electrophoresis and subsequent sequencing of amplified 16S rRNA gene bands. The most remarkable feature of all biofilms was that regardless of water origin, sequences belonging to Firmicutes were prevalent (53.5 % of total mean band intensity) and that almost all sequences recovered had some similarity (between 80.2 and 97 %) to thermophilic microorganisms. Mainly, sequences were closely related to potentially spore-forming organisms, suggesting that microbial communities able to grow at high temperatures were selected from the microbiota present in the incoming water. These pioneer results may contribute to improve management strategies to minimize the problems associated to biofouling in irrigation systems.     

Harmful effects of mustard bio-fumigants on entomopathogenic nematodes

Mustard (Brassica and Sinapis spp.) green manures tilled into the soil preceding potato crops act as bio-fumigants that are toxic to plant–parasitic nematodes, providing an alternative to synthetic soil fumigants. However, it is not known whether mustard green manures also kill beneficial entomopathogenic nematodes (EPNs) that contribute to the control of pest insects. We used sentinel insect prey (Galleria mellonella larvae) to measure EPN infectivity in Washington State (USA) potato fields that did or did not utilize mustard green manures. We found a trend toward lower rates of EPN infection in fields, where mustard green manures were applied, compared to those not receiving this cultural control method. In a series of bioassays we then tested whether the application of two mustard (Brassica juncea) cultivars, differing in glucosinolate levels, disrupted the abilities of a diverse group of EPN species to infect insect hosts. Mustard-exposure trials were conducted first in laboratory arenas where EPNs were exposed to mustard extracts suspended in water, and then in larger microcosms in the greenhouse where EPNs were exposed to green manure grown, chopped, and incorporated into field soil. In all trials we used G. mellonella larvae as hosts and included multiple EPN species in the genera Steinernema (Steinernema carpocapsae, Steinernema feltiae, Steinernema glaseri, and Steinernema riobrave) and Heterorhabditis (Heterorhabditis bacteriophora, Heterorhabditis marelatus, and Heterorhabditis megidis). In the laboratory, EPN infection rates were lower in arenas receiving mustard extracts than the control (water), and lower still when EPNs were exposed to extracts from plants with high versus low glucosinolate levels. Results were nearly identical when mustard foliage was soil-incorporated into greenhouse microcosms, except that the negative effects of mustards on EPNs developed more slowly in soil. Significantly, in arenas of both types one EPN species, S. feltiae, appeared to be relatively unaffected by mustard exposure. Together, our results suggest that the use of mustard bio-fumigants for the control of plant–parasitic nematodes has the potential to interfere with the biocontrol of insect pests using EPNs. Thus, it may be difficult to combine these two approaches in integrated pest management programs.     

昆虫病原线虫对烟苗的保护效果

小地老虎Agrotis ypsilon是危害烟草的重要害虫.为了实现该害虫的绿色防控,本研究测定了小地老虎幼虫对不同食物(小白菜、蕹菜、人工饲料和麸皮)取食的偏好性,评价添加鲜叶类诱食剂时昆虫病原线虫Steinernema carpocapsae All粉剂、S.feltiae SN虫尸剂、绿僵菌Metarhizium anisopliae和化学农药敌杀死(溴氰菊酯)对小地老虎的防控而实现对移栽期烟苗的保护效果.结果表明,尽管不同时间不同食物吸引的小地老虎幼虫数量有差异,但是这些幼虫对鲜叶类蔬菜(小白菜和蕹菜)具有偏好性.当每株烟苗存在2头小地老虎4龄幼虫时,未添加幼虫诱食剂的情况下,昆虫病原线虫粉剂处理后的烟苗死亡率为56.7％,显著低于绿僵菌处理的74.2％、昆虫病原线虫虫尸剂处理70.0％和对照78.3％,但高于敌杀死(溴氰菊酯)处理的烟苗死亡率37.5％;添加诱食剂后,昆虫病原线虫粉剂处理后烟苗的死亡率为40.8％,显著低于绿僵菌62.5％和昆虫病原线虫虫尸剂67.6％处理和对照62.5％,但与敌杀死(溴氰菊酯)处理的烟苗死亡率39.2％,差异不显著.当每株烟苗存在3头小地老虎4龄幼虫时,不论是否添加小地老虎幼虫诱食剂,昆虫病原线虫粉剂处理后的烟苗死亡率显著低于绿僵菌、昆虫病原线虫虫尸剂处理和对照,但与敌杀死(溴氰菊酯)处理的烟苗死亡率差异不显著.同样,添加幼虫诱食剂后,昆虫病原线虫粉剂处理的烟苗死亡率为36.7％,显著低于未加入小地老虎幼虫诱食剂的处理71.7％.因此,烟苗周围存在2头或3头小地老虎4龄幼虫的情况下,S.carpocapsae All线虫粉剂对刚移栽烟苗具有明显保护作用,在烟苗周围添加小地老虎幼虫诱食剂能显著降低烟苗的被危害率,为烟田小地老虎幼虫防控提供了有效的方法.     

Root distribution and water uptake patterns of corn under surface and subsurface drip irrigation

Information on root distribution and uptake patterns is useful to better understand crop responses to irrigation and fertigation, especially with the limited wetted soil volumes which develop under drip irrigation. Plant water uptake patterns play an important role in the success of drip irrigation system design and management. Here the root systems of corn were characterized by their length density (RLD) and root water uptake (RWU). Comparisons were made between the spatial patterns of corn RWU and RLD under surface and subsurface drip irrigation in a silt loam soil, considering a drip line on a crop row and between crop rows. Water uptake distribution was measured with an array of TDR probes at high spatial and temporal resolution. Root length density was measured by sampling soil cores on a grid centered on crop row. Roots were separated and an estimation of root geometrical attributes was made using two different image analysis programs. Comparisons of these programs yielded nearly identical estimates of RLD. The spatial patterns of RWU and RLD distributions, respectively normalized to the total uptake and root length, were generally similar only for drip line on a crop row, but with some local variations between the two measures. Both RLD and RWU were adequately fitted with parametric models based on semi-lognormal and normal Gaussian bivariate density functions (Coelho and Or, 1996; Soil Sci. Soc. Am. J. 60, 1039–1049).     

Distribution of entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) in natural habitats in California, USA

A total of 270 soil samples from 30 different habitats in 10 geographic regions of California were evaluated for the presence of rhabditid entomopathogenic nematodes. Nematodes were isolated from 26.3% of the samples. The recovered isolates were identified as Steinernema carpocapsae, S. feltiae, S. kraussei, S. longicaudum, S. oregonense, Heterorhabditis marelatus and H.bacteriophora. Among the steinernematids, S. kraussei and S. feltiae were the most commonly encountered species, generally occurring in acidic soils high in organic matter. Among the heterorhabditids, H. bacteriophora was isolated along the southern coast, whereas H. marelatus was recovered along the northern coast of California. Steinernematids were recovered from coniferous forests, oak woodlands and grasslands whereas heterorhabditids were isolated from coastal marshes.