Development of Beauveria bassiana dry formulation for vectoring by honey bees Apis mellifera (Hymenoptera: Apidae) to the flowers of crops for pest control

Using bee pollinators as a means for the dissemination of microbial control agents, such as Beauveria bassiana, against insect pests of agricultural crops is a novel and interesting approach to biological control. In four laboratory trials, one in Canada and three in Jordan, factors affecting the acquisition of B. bassiana by honey bees were evaluated using hive-mounted inoculum dispensers. The numbers of conidia carried by bees emerging from the dispensers differed according to the type of carrier used. Bees that passed through corn flour acquired more inoculum than did those that walked through wheat flour, durum semolina, corn meal, potato starch, potato flakes, oat flour or barley flour. The numbers of conidia acquired by the bees increased with decreasing particle size and moisture content of the carrier, and with increasing density of B. bassiana conidia in the formulation. Time required for a bee to pass through the dispenser did not significantly affect the acquisition of conidia. This study indicated that honeybees (Apis mellifera carnica) have a great potential for vectoring B. bassiana in crop systems. It also opens more avenues for studies on bee delivery of other microbial biological control agents.     

Feeding enrichment methods for Pacific walrus calves

In the wild, walrus calves are occupied with many behaviors necessary for survival. In captivity there is usually less to occupy them. Therefore it is necessary to develop other ways to occupy the animals to prevent negative behavior. In the present study, food in four different types of dispensers were tested on two walrus calves: fish in ice, fish in a nine‐hole container, fish in a two‐hole container, and fish in ice in a one‐hole container. All four methods of offering fish had an effect on the animals' behavior. There were differences in the way the two animals responded to each of the four dispensers. The dispensers occupied one animal on average between 32–95% of the 90‐min test periods, and the other animal for between 14–57% of the test periods. Due to the effect of learning and rapid development of the calves, which were shifting from a 100% formula diet to a 100% fish diet during the study period, the four methods cannot be compared. Besides the time feeding methods occupy animals, the practicality of a dispenser determines how often it will be used by the keepers. Of the four dispenser types tested in the present study, the nine‐hole container was the most practical and was still used frequently by the keepers years after the study. Zoo Biol 0:1–12, 2007. © 2007 Wiley‐Liss, Inc.     

Sex pheromone traps for monitoring the codling moth: effect of dispenser type, field aging of dispenser, pheromone dose and type of trap on male captures

The effects of dispenser type, dispenser aging in the field, pheromone dose in the dispenser and trap type on trapping efficiency of codling moth,Cydia pomonella (L.), males were investigated in Israel. An Israeli-manufactured rubber septum was significantly better than the CM Pherocon cap in attracting males to a Pherocon 1C cap trap or an IPS trap. The effect of aging of the dispenser in the field on trapping efficiency was significant in CM Pherocon caps and the Israeli septa. Captures in traps were negatively correlated with aging of septa. The effect of aging of dispensers was more marked during summer than during spring. The fairly rapid loss of attractancy indicates that the lures should be replaced after 2 weeks at the most. The release rate of the pheromone from the dispensers was measured in a flow system. The emission from the Israeli septa and Pherocon caps decreased sharply after 2 weeks and then was almost constant, which explains the lower attractancy of aged septa. However, there was a marked difference in the release profiles of the pheromone from the two types of dispensers, which may explain the different performance of the two dispensers. Within the range of 0.1 to 100 μg pheromone per dispenser, male response increased positively with the pheromone dose. Pheromone loadings of 100 or 1000 μg per dispenser did not differ significantly in their attractiveness for males. A load of 5000 μg per dispenser was significantly less attractive to males than was 100 or 1000 μg per dispenser. The non-sticky IPS trap was significantly better in capturing codling moth males than was the sticky, commonly used Pherocon 1C trap, provided it was baited with the Israeli dispenser. The two traps were equally effective when baited with the CM Pherocon caps. The possibility of using the non-sticky, nonsaturating and easy-to-handle IPS traps for monitoring codling moth is of great importance. Contribution from the Agricultural Research Organization, The Volcani Center, Bet Dagan 50-250, Israel. No 3702-E, 1992 series.     

Evaluation of traps and lures for mass trapping of Mediterranean fruit fly in citrus groves

Mass trapping has proven to be a powerful weapon in the control of Ceratitis capitata (Wiedemann), and its application in Mediterranean countries has currently increased notably as a control method. In this study, the efficacy of newly developed traps and dispensers of attractants were assessed with the aim of finding the best trap and set the lifetime of the dispensers, thus improving the total efficacy of mass trapping. Efficacy trials with six different types of traps and six different types of female dispensers were carried out. Moreover, the lifetime of three female dispensers, including a new attractant composition dispenser with n-methyl pyrrolidine, were studied. Results show significant differences among the trap types using female attractants, with an advantage of nearly 3 times more catches in best trap. Tested female dispensers showed no significant differences in efficacy between trimethylamine and putrescine attractants regard n-methyl pyrrolidine, however we observed differences in lifetime between dispensers. Thus, there are significant differences among different types of traps and dispensers in efficacy, and the appropriate selection of the trap and dispenser will improve the mass trapping results.     

苹果蠹蛾性信息素缓释剂的控害效果

2010年在甘肃、宁夏、黑龙江3地对苹果蠹蛾Cydia pomonella(L.)性信息素缓释剂(购自澳大利亚Bioglobal公司;性信息素含量为0.16g/根)对苹果蠹蛾的迷向防治效果进行了试验;观察了不同生态区设置不同缓释剂密度的防治效果和持效期。结果表明,在低虫口密度下,在不同生态区每公顷悬挂660、990和1320根性信息素迷向丝,均能取得较好的防治效果;在越冬代活动前处理,基本上可以控制整个生长期苹果蠹蛾对果实的为害。在新侵入的低密度果园,通过诱蛾监测、调查蛀果和树干绑诱虫带等验证方法,初步证明使用该性信息素缓释剂处理1年可以达到铲除的效果。依据本研究结果,结合防治成本,在苹果蠹蛾发生地区,采用660根/hm2的悬挂密度使用进口性信息素缓释剂处理果园可以达到良好的防治效果。     

A novel method for the quantitative assessment of the percolation of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> conidia through horticultural growing media

Incorporation of fungal biological control agents (BCAs) into plant growing media has considerable ergonomic and economic benefits for growers. These agents usually give prophylactic control of target pests and diseases. However, their efficacy is dose dependent and loss of inoculum through leaching could influence the degree of protection they provide. At present there are no protocols to determine the loss of inoculum in the disparate growing media used in horticulture. We describe a method based on a nutrient leaching column to quantify leaching of conidia of the insect-pathogenic fungus Metarhizium anisopliae in a range of growing media. Conidia of this biocontrol agent were applied as a drench or premixed into the medium. Both the application method and growth medium influenced conidial leaching. Inoculum losses were greater following drench application than premixing (95% vs. 15%) irrespective of media type. Comparatively more inoculum was lost from bark and coir following drench application whereas losses were relatively high in peat following premixed application. The leaching column assay provided a simple and accurate method to quantify inoculum loss in real time. This assay could help determine leaching of other fungal BCAs in growing media. It could help in improving pest and disease control by optimizing the rate and frequency of conidial application as well in the design of more efficacious formulations.     

黄瓜青枯病内生拮抗菌株的分离及ARDRA分析

在黄瓜生长的不同阶段从根系分离内生细菌共469株。通过青枯菌平板拮抗试验,从中筛选到具明显拮抗作用的菌株59株。将内生拮抗菌纯培养物扩增近全长的16SrDNA并用限制性内切酶AluⅠ对PCR产物进行ARDRA(amplifiedrDNArestrictionanalysis)多态性分析,共得到5种不同的操作分类单元(OperationalTaxonomicUnit,OTU)。其中属于OTU1共有39株分离物,占内生拮抗菌总数的66%,为优势种群。进一步通过ERIC-PCR指纹图的方法在菌株水平上分析OTU1类群。结果表明,OTU1可分为12种不同的菌株,其中菌株HE-1和HE-2在黄瓜生长的5个不同阶段均可分离到。通过标记天然不具有利福平抗性的HE-1和HE-2菌株,获得抗利福平突变体菌株,回收检测结果表明,在栽培的不同时期,黄瓜植株根内均有HE-1和HE-2菌株的定殖。经防病效果的盆栽试验,发现HE-1和HE-2的浸种处理能有效降低黄瓜青枯病的病发率,与对照比较差异显著。因此确定HE-1和HE-2为黄瓜青枯病生物防治的优良菌株。     

Sex pheromone of the citrus mealybug Planococcus citri: synthesis and optimization of trap parameters

A simple synthesis of the pheromone of the citrus mealybug, Planococcus citri (Risso) (Hemiptera: Pseudococcidae), has been developed. Various factors affecting capture of males have been assessed to optimize the trap design and to develop a lure with high efficacy and longevity. Male capture was the same with the racemic and chiral pheromone; technical pheromone (85% purity) was statistically as attractive as pure pheromone (97%). A special formulation was used to determine the actual release rate of the pheromone under field conditions as related to male capture. Generally, plate traps caught more males than delta traps, and large traps caught more than small ones. The effects of aging on the performance of three types of rubber dispensers were evaluated. It was found that the American dispenser displayed the most consistent trapping performance and could be used for monitoring for at least 16 wk with a load of 200 microg of pheromone. The dose-response of the males to sex pheromone was tested within the range of 25-1,600 microg.     

Effectiveness of honey bees in delivering the biocontrol agent Bacillus subtilis to blueberry flowers to suppress mummy berry disease

Honey bees are important pollinators of commercial blueberries in the southeastern United States, and blueberry producers often use supplemental bees to achieve adequate fruit set. However, honey bees also vector the plant pathogenic fungus Monilinia vaccinii-corymbosi which infects open blueberry flowers through the gynoecial pathway causing mummy berry disease. Here, we report the results of a 3-year field study to test the hypothesis that using bee hives equipped with dispensers containing the biocontrol product Serenade, a commercial formulation of the bacterium Bacillus subtilis which has shown activity against flower infection by M. vaccinii-corymbosi in laboratory experiments, can reduce mummy berry disease incidence when honey bees are used as pollinators in blueberries. Individual honey bees carried 5.1–6.4 × 10⁵ colony-forming units (CFU) of B. subtilis when exiting hive-mounted dispensers with Serenade. On caged rabbiteye blueberry bushes in the field, population densities of B. subtilis vectored by honey bees reached a carrying capacity of <10³ CFU per flower stigma within 2 days of exposure, and there was a highly significant non-linear relationship between B. subtilis populations per stigma and bee activity, expressed as number of legitimate flower visits per time interval per cage (R = 0.6928, P < 0.0001, n = 32). Honey bee density (1600 or 6400 individuals per 5.8-m³ cage) and Serenade treatment (presence or absence of the product in hive-mounted dispensers) significantly (P < 0.05) affected the incidence of fruit mummification on caged bushes, whereby increasing bee density increased disease incidence and application of Serenade reduced disease levels. Taken together, results of this study suggest that use of a hive-dispersed biocontrol product such as Serenade as a supplement during pollination can reduce the risk of mummy berry disease. This may be a prudent practice that optimizes the benefits to pollination of high bee densities while reducing the associated disease-vectoring risk.     

Comparative Statistical Study of the Cornwall, LKB, and Micromedic Dispensers

The working precisions of the 5.0-ml-capacity Cornwall, the LKB 2075, and the Micromedic model 25000 dispensers were compared, and the bias of the LKB dispenser was contrasted with that of the Micromedic dispenser. Three technologists used six Cornwall dispensers, three LKB units with four different pumps, and three Micromedic units with six different pumps to dispense (in 1.0-ml amounts) sucrose solution adjusted to match the normal viscosity of serum. Under these conditions, the LKB dispenser was approximately 30% more precise than the Micromedic dispenser, which was approximately four times as precise as the Cornwall dispenser. Although the left pump site of the Micromedic was slightly more precise than the LKB, overall the Micromedic was less precise than the LKB. Moreover, the LKB was easier to use than the Micromedic.     

Evaluation of extended-season mating disruption of the Oriental fruit moth Grapholita molesta (Busck) (Lep., Tortricidae) in apples

Abstract:  Oriental fruit moth Grapholita molesta (Busck) (Lep., Tortricidae) has recently become a key pest of apples throughout the eastern USA. Pheromone-mediated mating disruption of Oriental fruit moth was successfully used in North Carolina apple orchards in the past few years. However, low levels of late-season fruit damage occurred in some orchards treated in late May with hand-applied pheromone dispensers because of inadequate dispenser longevity. To investigate alternative pheromone application schedules for extended mating disruption control, the following pheromone treatments were compared with conventional insecticides in Henderson County (NC) in 2002: late May application of hand-applied dispensers; late June application of hand-applied dispensers; late May application of hand-applied dispensers supplemented with a late August application of sprayable pheromone dispensers; late May application of hand-applied dispensers which have a longer activity period; and conventional insecticides as a control. All treatments were sprayed with an insecticide at petal fall in late April for thinning and for control of the first generation Oriental fruit moth adults. Pheromone trap catches were significantly reduced in all mating disruption blocks compared with conventional insecticide blocks. Among pheromone treatments, the highest trap captures were recorded in the delayed hand-applied dispenser treatment in June before treatment. However, the mean percentage fruit damage did not vary with timing of application of hand-applied dispensers and the type of pheromone dispenser used. Clearly, the combination of each mating disruption treatment with insecticide application against first generation Oriental fruit moth was as effective as the conventional insecticide treatment under moderate population pressure.     

Mating disruption of codling moth Cydia pomonella with high densities of Ecodian sex pheromone dispensers

A mating disruption approach using high densities of pheromone dispensers, has recently been proposed for controlling codling moth, Cydia pomonella (L.) (Lep., Tortricidae). Ecodian® CP dispensers, made of low‐cost biodegradable material and easy to apply, were formulated with 10 mg of codlemone and placed at a rate of 1400–2000 dispensers/ha. Seasonal dispenser performance was evaluated using different methods. The release rate of new Ecodian dispensers, measured directly by solid‐phase micro‐extraction (SPME), was significantly higher than that of the standard monitoring lure. The release rate of field‐aged dispensers decreased over time; it was comparable with that of the standard monitoring lure after 55–90 days of field exposure and significantly lower beyond. Electroantennographic (EAG) recordings showed that field‐aged dispensers strongly stimulated virgin male moths’ antennae. Dispensers elicited close‐range approaches in wind tunnel irrespective of their age. Traps baited with aged Ecodian CP dispensers captured males throughout the season. Our results demonstrate that Ecodian dispensers achieved a good level of activity and longevity over the season. Field trials carried out from 2002 to 2004 confirmed the efficacy of Ecodian CP dispensers for codling moth control, regardless of the size of the treated area. The potential of this strategy for the control of C. pomonella is discussed.     

Dual pheromone dispenser for combined control of codling moth Cydia pomonella L. and oriental fruit moth Grapholita molesta (Busck) (Lep., Tortricidae) in pears

Codling moth (CM) and oriental fruit moth (OFM) are very important orchard pests worldwide, and particularly in Victoria, Australia, where both species damage pome fruit. Individually CM and OFM have been controlled successfully by pheromone‐mediated mating disruption, but treatment of pome fruit with full registered rates of two individual hand‐applied dispensers for CM and OFM could be uneconomical for growers. Field trials conducted over three seasons in plots sprayed with insecticides consistently demonstrated that dual Isomate C/OFM TT dispensers, designed to disrupt both CM and OFM, were as effective as Isomate CTT and Isomate OFM Rosso dispensers applied individually in pears for control of CM and OFM respectively. The dual‐ and individual‐species dispensers reduced moth catches and fruit damage to a similar degree. The results suggest that combined control of CM and OFM in pears by applying dual Isomate C/OFM TT dispensers at the full‐recommended rate of 500 dispensers per hectare will be more economical than use of individual species dispensers, because the price and application cost of dual dispenser is about half that for individual dispensers.     

Evaluating dispensers loaded with codlemone and pear ester for disruption of codling moth (Lepidoptera: Tortricidae)

Polyvinyl chloride polymer (PVC) dispensers loaded with ethyl (E,Z)-2,4-decadienoate (pear ester) plus the sex pheromone, (E,E)-8,10-dodecadien-1-ol (codlemone) of codling moth, Cydia pomonella (L.), were compared with PVC dispensers and a commercial dispenser (Isomate-C Plus) loaded with codlemone. Evaluations were conducted in replicated plots (0.1-0.2 ha) in apple, Malus domestica (Borkhausen) during both generations of codling moth from 2007 to 2009. Dispensers were applied at 1,000 ha(-1). Male captures in traps baited with virgin female moths and codlemone lures were recorded. Residual analysis of field-aged dispensers over both moth generations was conducted. Dispensers exhibited linear declines in release rates of both attractants, and pear ester was released at a significantly higher rate than codlemone during both time periods. The proportion of virgin female-baited traps catching males was significantly lower with combo dispenser TRE24 (45/110, mg codlemone/mg pear ester) during the second generation in 2007 and the combo dispensers TRE144 (45/75) and TRE145 (75/45) during the first generation in 2008 compared with Isomate-C Plus. Similarly, male catches in female-baited traps in plots treated with the combo dispensers TRE144 during the first generation in 2008 and TRE23 (75/110) during the second generation, in 2007 were significantly lower than in plots treated with Isomate-C Plus. No significant differences were found for male catches in codlemone-baited traps in plots treated with Isomate-C Plus and any of the combo dispensers. However, male catches were significantly lower in plots treated with Cidetrak CM (codlemone-only dispenser) than the combo TRE144 dispenser during both generations in 2009.     

Success evaluation of the biological control of Fusarium wilts of cucumber,banana, and tomato since 2000 and future research strategies

The Fusarium wilt caused by Fusarium oxysporum strains is the most devastating disease of cucumber, banana, and tomato. The biological control of this disease has become an attractive alternative to the chemical fungicides and other conventional control methods. In this review, the research trends and biological control efficiencies (BCE) of different microbial strains since 2000 are reviewed in detail, considering types of microbial genera, inoculum application methods, plant growth medium and conditions, inoculum application with amendments, and co-inoculation of different microbial strains and how those affect the BCE of Fusarium wilt. The data evaluation showed that the BCE of biocontrol agents was higher against the Fusarium wilt of cucumber compared to the Fusarium wilts of banana and tomato. Several biocontrol agents mainly Bacillus, Trichoderma, Pseudomonas, nonpathogenic Fusarium, and Penicillium strains were evaluated to control Fusarium wilt, but still this lethal disease could not be controlled completely. We have discussed different reasons of inconsistent results and recommendations for the betterment of BCE in the future. This review provides knowledge of the biotechnology of biological control of Fusarium wilt of cucumber, banana, and tomato in a nut shell that will provide researchers a beginning line to start and to organize and plan research for the future studies.     

A new long-life trimedlure dispenser for Mediterranean fruit fly

New agricultural techniques are attempting to reduce the application of synthesized pesticides and replace them with new environmentally friendly methods such as mass trapping, mating disruption, or chemosterilization techniques. All these methods are based on the release of a lure for insect attraction or confusion. The success of the chosen method depends on the quality of the attractant emission from the dispenser. Currently, used dispensers with a polymeric matrix and new dispensers with mesoporous inorganic materials were evaluated to obtain more efficient emission kinetics. In this study, the selected pest was the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) and the lure used was trimedlure (TML). The dispensers were validated by means of a field study comparing insect catches with attractant release values. As a result, we have demonstrated that mesoporous dispensers have a clearly longer lifetime than the polymeric plug. Furthermore, the attractant release rate is less dependent on temperature in mesoporous than in polymeric dispensers.     

Application of synthetic herbivore‐induced plant volatiles causes increased parasitism of herbivores in the field

We previously reported that Cotesia vestalis (Hymenoptera, Braconidae), a parasitoid of diamondback moth (DBM) (Plutella xylostella; Lepidoptera, Plutellidae) larvae, was attracted to volatiles from crucifer plants infested by moth larvae kept in a desktop acrylic box, and that a blend of four DBM‐induced plant volatiles was responsible for this attraction. In this study, using a specially designed dispenser to release the four compounds, we demonstrated that the wasp was attracted to intact komatsuna plants (Brassica rapa var. perviridis). The experiments were performed in a climate‐controlled room, which was approximately 1000 times larger than the acrylic box used previously. Similarly, using the dispenser in the field, C. vestalis females were attracted to intact komatsuna plants with the dispenser from a distance of three metres. We also examined the effect of the volatile blend on the incidence of parasitism of DBM larvae in the field. Three small containers containing DBM‐infested komatsuna plants with dispensers, and three control containers containing only infested plants (control) were arranged in two lines running perpendicular to a komatsuna field in which both DBM larvae and C. vestalis populations were maintained, at distances of 12, 30 and 70 m. The results showed that the incidence of DBM parasitism was significantly higher in containers containing dispensers than in the control containers, suggesting that the blend could potentially be applied to DBM control in agroecosystems.     

Enhanced biocontrol activity of Trichoderma virens transformants constitutively coexpressing β-1,3- and β-1,6-glucanase genes

Evidence for the role of chitinases, proteases and β-1,3- and β-1,6-glucanases in mycoparasitism by Trichoderma species has been well documented. Moreover, constitutive over-expression of genes encoding individual cell-wall-degrading enzymes (CWDEs) has been shown to improve the potential of biological agents. In this study, we generated transformants of T. virens in which β-1,3- and β-1,6-glucanase genes, TvBgn2 and TvBgn3 , respectively, were constitutively coexpressed in the same genetic T. virens Gv29.8 wild-type background. The double over-expression transformants (dOEs) grow and sporulate slower than the wild-type (WT). However, the reduction in growth did not seem to affect their mycoparasitic and biocontrol capabilities, as dOEs displayed much higher levels of total β-1,3- and β-1,6-glucanase activity than the WT. This higher enzymatic activity of dOEs positively correlated with observed in vitro inhibition of Pythium ultimum and Rhizoctonia solani mycelia, and with enhanced bioprotection of cotton seedlings against P. ultimum , R. solani and Rhizopus oryzae . Besides effective biocontrol of all pathogens at an original inoculum level, the performance of dOEs was highly enhanced (up to 312% of WT performance) when pathogen pressure was greater (i.e. concentration of inoculum was higher or pathogens applied in combination). These results demonstrate that the strategy of introducing multiple lytic enzyme-encoding genes through transformation of a given biocontrol strain can be successfully used to achieve better biocontrol.     

A comparison of various mating disruption technologies for control of two internally feeding Lepidoptera in apples

Sex pheromone mating disruption (MD) is an approach used to control several moth pest species of pome fruit by disrupting the ability of the males to find females and consequently prevent mating. The following experiments were performed to determine the effectiveness of several new and experimental sex pheromone MD technologies, and dispenser densities for simultaneous control of the codling moth (CM), Cydia pomonella (L.), and the oriental fruit moth (OFM), Grapholita molesta (Busck) (both Lepidoptera: Tortricidae), in Pennsylvania apple orchards. In one study, three MD approaches to control CM and oriental fruit moth – CM and OFM Disrupt Micro‐Flakes, Isomate CM/OFM TT, and both a CideTrak OFM and a CideTrak CM dispenser containing both codlemone and pear ester – and an insecticides‐only treatment were compared over the course of 2 years. In the other studies, the efficacy of several CheckMate Duel dispenser densities (i.e., 250, 375, 425, and 500 dispensers ha^?1) were compared against Isomate CM/OFM TT, and an insecticides‐only treatment. The CideTrak CM/pear ester combination and Isomate CM/OFM TT treatments both substantially reduced CM captures in traps in 2007 and 2008. Meanwhile, OFM trap shutdown was highest in the CheckMate Duel densities of 375 (99.9 ± 0.08%) and 500 dispensers ha^?1 (98.9 ± 0.07%) and the Isomate CM/OFM TT treatment (98.0 ± 1.13%), and lowest in the 250 dispensers ha^?1 density treatment (94.3 ± 3.23%). In orchards where OFM is the dominant pest species, a CheckMate Duel dispenser density of 375 ha^?1 is necessary for effective control, whereas higher densities are needed to control CM.