Efficacy of entomopathogenic nematodes against the tomato leafminer <Emphasis Type="Italic">Tuta absoluta</Emphasis> in laboratory and greenhouse conditions

The tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is an important pest of tomato crops in South America and it has recently arrived in Europe affecting tomato plantations. The susceptibility of T. absoluta larvae and pupae to three species of entomopathogenic nematodes (Steinernema carpocapsae, Steinernema feltiae and Heterorhabditis bacteriophora) was examined under laboratory conditions. Leaf bioassays were conducted to evaluate the nematode’s capability to reach the larvae and kill them within the galleries. The efficacy of the three nematode species after foliar application to potted tomato plants was evaluated under greenhouse conditions. High larval mortality (78.6–100%) and low pupae mortality (<10%) was determined in laboratory experiments. In the leaf bioassay a high level of larval parasitation (77.1–91.7%) was recorded revealing the nematode’s capacity to kill the larvae inside the galleries. In the pot experiments nematode treatment reduced insect infection of tomato plants by 87–95%. The results demonstrate the suitability of entomopathogenic nematodes for controlling T. absoluta.     

Efficacy of some biorational insecticides against Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) under laboratory and greenhouse conditions in Kuwait

The tomato leafminer, Tuta absoluta, is one of the most destructive pests of tomato worldwide. Management of the pest is mainly based on chemical insecticides. Reliance on insecticides is difficult to sustain because of unintended long‐term adverse effects on the environment and human health. Consequently, there is a need to develop pest management strategies that ensure the production of high‐quality products, while at the same time ensuring environmental sustainability and maximum consumer protection. We evaluated the efficacy the biopesticides: Azadirachtin, Bacillus thuringiensis, Steinernema feltiae and Beauveria bassiana individually and in combination against T. absoluta under laboratory and greenhouse conditions. When second instar larvae were exposed to tomato leaf discs treated with Azadirachtin (3 g / L), B. thuringiensis (0.5 g/L) or B. bassiana (1.5 g/L), 70%–86%, 55%–65%, and 45.5%–58.5% mortality was observed, respectively. Steinernema feltiae (1,000 IJs/ml) was the least effective biopesticide, with 26%–42% mortality. In the greenhouse trials on tomato, pest infestation (mines/10 leaves/plant) and fruits damaged were significantly lower on plants treated with Azadirachtin  +  B. thuringiensis or Azadirachtin  +  B. bassiana compared to plants treated with Azadirachtin, B. thuringiensi, B. bassiana or S. feltiae alone. Azadirachtin  +  B. thuringiensis and Azadirachtin  +  B. bassiana resulted in 90% and 81% reduction in fruits damaged in the summer experiments, respectively, and 96% and 91% in winter. The most severe pest infestation was observed on plants treated with S. feltia. The results indicate that the biopesticides, except S. feltia, can contribute to T. absoluta control in greenhouse tomato crops. In particular, the combined use of Azadirachtin with B. thuringiensis or B. bassiana provided the highest level of control of the pest. The potential for including these biopesticides in an overall sustainable integrated pest management programme for T. absoluta is discussed.     

Different host plant species modifies the susceptibility of Bactericera cockerelli to the entomopathogenic fungus Beauveria bassiana

Bactericera cockerelli (Sulc.) is a serious pest of solanaceous crops and a vector of the plant pathogen Candidatus Liberibacter psyllaurous. Entomopathogenic fungi are the most important biological control alternatives for this pest. Host plant species, however, can modify the outcomes of insect–pathogen interactions. We conducted laboratory experiments to quantify the virulence of two isolates of the entomopathogenic fungus Beauveria bassiana (Bals. [Vuill.]), BB40 and BB42, against third instar B. cockerelli nymphs maintained on chilli pepper plants. Owing to the lack of difference in virulence against B. cockerelli nymphs on chilli pepper between the two B. bassiana isolates, only BB42 was used to: compare virulence against nymphs maintained on either chilli pepper, potato or tomato; and in vivo conidia production from nymphs maintained on different host plants. Virulence of the two B. bassiana isolates against B. cockerelli nymphs was similar. Bactericera cockerelli nymphs maintained on tomato were more susceptible to B. bassiana than nymphs maintained on potato or chilli peppers. Infected nymphs maintained on chilli peppers produced the greatest number of conidia followed by infected nymphs maintained on tomato and potato. Host plant affected the susceptibility of B. cockerelli to B. bassiana isolate BB42 and subsequent conidia production. The implications of our results for microbial control of B. cockerelli by B. bassiana are discussed.     

First record of an entomopathogenic fungus of tomato leafminer,Tuta absoluta (Meyrick) in Tanzania

Tuta absoluta (Meyrick) is a destructive pest causing adverse environmental and economic effects globally. Management of T. absoluta by synthetic chemicals is of diminutive achievement due to resistance trick of the pest. Harnessing of biocontrols is currently fascinating and efficient for management of the tremendous pests. This study isolated an entomopathogenic fungus from dead larvae of T. absoluta on tomato leaves from farmer’s field Tanzania. Two Aspergillus fungal isolates were isolated and tested against larvae and adults of T. absoluta under conditions of 19.5°C and 50%RH, 30.4°C and 70% RH, respectively, in the laboratory. Sequence analysis of Aspergillus isolates that we referred as A-Tz1 and A-Tz2 were analysed with the BLAST program available at the National Center for Biological Information (NCBI). The two isolates (A-Tz1 and A-Tz2) showed 100% similarity with Aspergillus oryzae of GeneBank accession numbers EF121337.1 and MG519722.1. Such similarity implied that both A-Tz1 and A-Tz2 isolates were A. oryzae. To confirm pathogenicity, bioassays were conducted against the larvae, pupa and adults of T. absoluta. Results showed that at 1.0?×?10⁸?conidia/mL, fungal isolates caused up to 70% larval mortality 3 days post inoculation and consequently inhibited pupation by 84.5% and adult emergence by 74.4%. The identified A. oryzae reduced the life span of adult T. absoluta up to 5 days post inoculation at 1.0?×?10⁸?conidia/mL whereas in control T. absoluta survived up to 25 days. This study found that isolates of Aspegillus species have the potential to be used as active ingrediennt for the formulation of a new bioinjections for application by farmers to control the pest in fields.     

Establishment of Beauveria bassiana as a fungal endophyte in potato plants and its virulence against potato tuber moth,Phthorimaea operculella (Lepidoptera: Gelechiidae)

The potato tuber moth, Phthorimaea operculella, is the most damaging potato pest in the world and is difficult to control as the larvae are internal feeders in the foliage and tubers. Entomopathogenic fungi that colonize plants as endophytes have lethal and sublethal pathological effects on insect pests. We show that Beauveria bassiana colonizes the aerial parts of potato plants endophytically after inoculation through soil drenching. Endophytic B. bassiana persisted in potato foliage for more than 50 days postinoculation. Bioassays indicated that foliage of B. bassiana-inoculated potato plants were pathogenic against larvae of P. operculella. Sublethal experiments indicated that B. bassiana negatively affected the growth, development, and reproduction of P. operculella. Development experiments showed that the weight of P. operculella pupae reared on B. bassiana-colonized potato plants (4.25 mg) was significantly less than that of those reared on uninoculated control plants (8.89 mg). Compared with newly eclosed larvae fed on control plants, those fed on B. bassiana-inoculated plants had significantly lower survivorship, with only 17.8% developing to the adult stage. Oviposition of P. operculella females reared on B. bassiana endophytically colonized plants was significantly lower (35 eggs/female) than of those reared on uninoculated plants (115 eggs/female). This study demonstrates that endophytic B. bassiana can be a potential biological control agent for the control and management of P. operculella. Comparing pupal weights of P. operculella reared on potato plants inoculated with the B. bassiana strain GZGY-1-3 and on untreated control plants, pupae from the control plants were significantly heavier than those from treated plants.     

Seasonal variation in natural mortality factors of Tuta absoluta (Lepidoptera: Gelechiidae) in open‐field tomato cultivation

The seasonal variation in natural mortality of phytophagous insects is determined by the relative importance of biotic and abiotic factors in agroecosystems. Knowledge regarding these factors throughout the year represents a key concern for IPM programmes. Seasonal population fluctuations of tomato pinworm, Tuta absoluta, led to an investigation of its natural mortality factors during the rainy season when the population level is low and during the dry season when population peaks occur. The aim of this study was to verify the seasonal variation in T. absoluta mortality factors in tomato crops. Immature stages of T. absoluta were obtained from laboratory‐rearing in the laboratory. These were taken to the field and monitored over two years. The mortality causes for each stage of insect development from egg to adult were assessed daily. Multiple biotic and abiotic mortality factors affected the immature T. absoluta stages such as rainfall, physiological disturbances, diseases, parasitoids and predators. The key T. absoluta mortality factor during summer–spring was predation. In addition, larvae predation correlated positively with temperature, wind velocity, photoperiod and rainfall. Nevertheless, during winter–fall, the key mortality factor was parasitism. Therefore, the critical stage for mortality was 3rd‐ and 4th‐instar larvae, being more vulnerable to natural control factors. Finally, the results showed the importance of vertical and horizontal action on natural mortality factors.     

New records of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) predation by Brazilian Hemipteran predatory bugs

The tomato borer Tuta absoluta, native to western South America, is an extremely devastating pest in tomato crops in most of South America, Europe and Africa North of the Sahel, causes yield losses up to 100% and decreases fruit quality in open field and greenhouse crops if control methods are not applied. In Brazil two other important lepidopteran pests – Neoleucinodes elegantalis and Helicoverpa zea – occur in tomato, as well as thrips, whiteflies and aphids. For control of these pests, frequent applications of pesticides of up to 5 times per week are needed, and these resulted in the appearance of resistant populations to a number of active ingredients and decimation of natural enemies. Biological control may offer a better, safer and more sustainable opportunity for pest management. Mirid predatory bugs are currently used with success in southern Europe to control T. absoluta and other pests. In Brazil, four Hemipteran predatory bugs, not yet known to attack T. absoluta, were found to successfully prey on eggs and larvae of this pest. The first results on their predation capacity, development, survival and reproduction on T. absoluta on tomato plants are presented.     

Interrelated responses of tomato plants and the leaf miner Tuta absoluta to nitrogen supply

Plant–insect interactions are strongly modified by environmental factors. This study evaluates the influence of nitrogen fertilisation on the tomato (Solanum lycopersicum L.) cv. Santa clara and the leafminer (Tuta absoluta (Meyrick), Lepidoptera: Gelechiidae). Greenhouse‐grown tomato plants were fed hydroponically on a complete nutrient solution containing either a high nitrogen concentration (HN) sustaining maximum growth or a low nitrogen concentration (LN) limiting plant growth. Insect‐free plants were compared with plants attacked by T. absoluta. Seven and 14 days after artificial oviposition leading to efficacious hatching and larvae development, we measured total carbon, nitrogen and soluble protein as well as defence compounds (phenolics, glycoalkaloids, polyphenol oxidase activity) in the HN versus LN plants. Only in the HN treatment did T. absoluta infestation slightly impair leaf growth and induce polyphenol oxidase (PPO) activity in the foliage. Neither the concentration of phenolic compounds and proteins nor the distribution of nitrogen within the plant was affected by T. absoluta infestation. In contrast, LN nutrition impaired T. absoluta‐induced PPO activity. It decreased protein and total nitrogen concentration of plant organs and enhanced the accumulation of constitutive phenolics and tomatine. Moreover, LN nutrition impaired T. absoluta development by notably decreasing pupal weight and lengthening the development period from egg to adult. Adjusting the level of nitrogen nutrition may thus be a means of altering the life cycle of T. absoluta. This study provides a comprehensive dataset concerning interrelated responses of tomato plants and T. absoluta to nitrogen nutrition.     

Target-oriented dissemination of Beauveria bassiana conidia by the predators,Harmonia axyridis (Coleoptera: Coccinellidae) and Chrysoperla carnea (Neuroptera: Chrysopidae) for biocontrol of Myzus persicae

Dissemination of microbial biocontrol agents via predators may have advantages for safe spore dispersal to targeted pests with the added benefit of predation. A laboratory study was conducted to test the target-oriented dissemination of conidia of Beauveria bassiana using larvae of both the multicoloured Asian lady beetle (Harmonia axyridis) and common green lacewing (Chrysoperla carnea) for control of aphids. Maximum dry conidial attachment occurred within approximately 7 min after exposure. After release of the treated predators on leaves of Chinese cabbage, within 12 hours lacewing larvae dispersed 89% of the attached conidia while Asian lady beetles dispersed 93%. Both predators dispersed conidia up to 2.4 m from the release site. Leaf disk bioassays were conducted to compare two application methods; the dissemination of conidia of B. bassiana by predators and the direct application of conidial suspensions. Mortality in sprayed aphids was 91±2.1% compared to 88±2.1 and 84±4.2%, respectively, when conidia were disseminated by lacewings and lady beetles. Predation was not affected in treated lacewing larvae whereas there was a 20% reduction in predation by lady beetle larvae. It appears that B. bassiana can be effectively delivered using certain insect predators.     

Reaction of some tomato cultivars to tomato leaf curl virus and evaluation of the endophytic colonisation with Beauveria bassiana on the disease incidence and its vector,Bemisia tabaci

Tomato line LA1478 and Pusa Ruby were resistant to tomato leaf curl virus (TLCV) disease. They registered higher plant height, number of branches, total phenol content and yield per plant than the other cultivars. Variety Peto 86 was tolerant to the disease while the other popular tomato cultivars, i.e. Ace, Early Pack, Money Maker, Prichard and Strain B were highly susceptible to the disease. Plant height and number of branches per plant revealed significantly positive association with fruit yield per plant. The disease index of TLCV exhibited significant negative correlations with plant height, total phenol content and fruit yield per plant – 0–4 and 5–25 adult whiteflies were observed on resistant susceptible cultivars. In the case of epiphytically colonisation by Beauveria bassiana conidia, not all developing hyphae on the leaf surface penetrated the whitefly cuticle. Many of the germ tubes elongated to a short distance before terminating its growth. On the other hand, the rapid staining of tomato tissues injected with B. bassiana conidial suspension indicates that the entomopathogenic fungus was established inside tomato tissues until the end time of the trial. The direct injection with the spore suspension yielded high post-colonisation, where the fungus was recovered from sites distant from the point of inoculation. This indicates that the fungus has the potential to move throughout the plant tissues. Laboratory bioassay of tomato whitefly feeding on tomato tissues containing B. bassiana conidial spores indicates that plant endophytic colonisation with entomopathogenic fungi may reduce insect survival on these plants. LT₅₀ values of the test diet were between three and four days. The mortality of Bemisia tabaci was high in the case of endophytically colonisation compared to epiphytically one (90.0% compared to 10.0% during three days) for whiteflies fed tomato tissues containing 1.5 × 10⁷ B. bassiana spores/ml. Application of B. bassiana as an artificial endophyte inside tomato plants can be an important component in the integrated control of tomato whiteflies. The endophytic colonising can achieve biocontrol effect based on induced disease resistance in plant tissues. According the available references, this is the first report on B. tabaci controlling by plant endophytic treatment.     

Can colonization by an endophytic fungus transform a plant into a challenging host for insect herbivores?

Endophytic growth of arthropod pathogenic fungi can parasitize insect herbivores without causing damage to the crop. However, studies addressing this tritrophic interaction are absent. Here, the endophytic arthropod pathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordyciptaceae), the polyphagous two-spotted spider mite Tetranychus urticae Koch (Trombidiformes: Tetranychidae), and its preferred plant host Phaseolus vulgaris L. (Fabales: Fabaceae) were selected to study the multi-kingdom interactions among plants, arthropods, and entomopathogenic fungi. Real-Time PCR analysis of nine defense-related genes revealed that a broad range of plant defense mechanisms is activated in response to the endophytic growth of B. bassiana. Moreover, we studied the molecular mechanism adapted by the two-spotted spider mite that underlies resistance. The analysis of 41 detoxification genes revealed that relatively moderate, high, and few numbers of genes were changed in the adults, nymphs, and eggs stages of T. urticae, respectively, after inoculation on colonized tissues of P. vulgaris. The endophytic growth of B. bassiana can have a negative effect on the growth and performance of the pest, in a developmental stage-dependent manner, by priming plant defense pathways. In parallel, the herbivore induces a broad range of detoxification genes that could potentially be involved in adaptation to endophytically colonized plant tissues.     

二种交配型球孢白僵菌对亚洲玉米螟的生态控制作用

利用生物种间互做关系抑制农业害虫的暴发是生物防治的重要手段。为探讨二种交配型内共生球孢白僵菌与玉米之间的互惠关系及其形成的共生体在亚洲玉米螟控制中的生态效应,以玉米为宿主植物,以球孢白僵菌孢子悬浮液进行灌根,在温室内构建了二种交配型(MAT1-1-1型,B5;MAT1-2-1型,B2)球孢白僵菌-玉米共生体,并研究了共生体对玉米的生长、对亚洲玉米螟的产卵选择和幼虫发育及其对球孢白僵菌生物学特性的影响。结果显示:通过叶片离体培养、ITS基因和交配型基因MAT检测,均能检测到白僵菌的内生定殖;MAT1-2-1型B2菌株定殖检出率高,MAT1-1-1型B5菌株在混合型接种中定殖有优势。回收后的球孢白僵菌菌落直径和毒力无显著性变化,但其产孢量都显著提高其中回收B5处理组来源菌株的产孢量提高最显著。接种过球孢白僵菌的玉米植株地上部生长速度、生物量和地下根系生物量均优于对照组,其中根系干重明显增加,而地上植株干重也相对增加。MAT1-1-1型菌株B5对共生体玉米植株地上高度促生长贡献明显;MAT1-2-1型菌株B2对共生体玉米植株地下干重增加贡献明显。总体上球孢白僵菌内生定殖对玉米地下根系生物量影响大于对地上植株生物量的影响。在产卵选择性试验中,各处理组亚洲玉米螟的产卵量显著少于对照组。共生体对亚洲玉米螟产卵具有明显的趋避作用,MAT1-2-1型菌株B2对产卵的趋避作用明显,而MAT1-2-1型菌株B5的趋避作用较弱。在人工接种幼虫的试验中,处理组回收的亚洲玉米螟幼虫存活率均显著低于对照组,其中,B5组回收幼虫的存活率最低,仅为38.33%;处理组的化蛹率与对照组差异不显著,但B5组的回收幼虫化蛹率显著低于B2组和对照组,仅为34.77%,这说明MAT1-1-1型B5菌株对玉米螟幼虫发育抑制最明显。上述结果表明,不同交配型球孢白僵菌内生定殖效率有差异,在经过内生定殖后在产孢量方面有显著性提高,两个交配型菌株在联合应用时具有协同增效作用;两个交配型菌株均能够通过内生定殖与玉米形成共生体并促进玉米植株的生长,这显示球孢白僵菌和玉米之间已经建立具有互惠关系的共生体。这种共生体通过趋避亚洲玉米螟产卵、抑制幼虫存活和降低化蛹率等方面的潜力虽然不一样,但都有助于对亚洲玉米螟的可持续生态防治,也证明了共生体的建成有效提高了玉米的生态适应性,为利用球孢白僵菌内共生性实施亚洲玉米螟防控提供了新思路。     

Efficacy of <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> (Berliner) in controlling the tomato borer, <Emphasis Type="Italic">Tuta absoluta</Emphasis> (Meyrick) (Lepidoptera: Gelechiidae)

The tomato borer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is considered to be one of the most devastating pests affecting tomato crops in South America, where crop losses range from 60 to 100%. After its detection in the Spanish tomato-growing area at the end of 2006, it spread quickly to other European and northern African countries. Currently, T. absoluta management in these countries is mainly based on chemical treatments. Nonetheless, special emphasis is being placed on implementing environmentally safe strategies. Commercial formulates based on Bacillus thuringiensis may be a good alternative, as they have been used to control other insect pests successfully. The laboratory, greenhouse, and open-field experiments presented in this work are evidence that B. thuringiensis is highly efficient in controlling T. absoluta. First instar larvae were the most susceptible, while susceptibility was lower in second and third instar larvae. Our results have shown that the impact of T. absoluta can be greatly reduced by spraying only B. thuringiensis-based formulates, with no need for chemical insecticides. Furthermore, the integration of this technology with other biological control methods focused on T. absoluta eggs, such as the use of mirid predators or parasitoids, could reduce the number of B. thuringiensis treatments and the use of chemicals, with the consequent reduction of residues on fruits.     

Endophytic blastospores of Beauveria bassiana provide high resistance against plant disease caused by Botrytis cinerea

The entomopathogenic fungus Beauveria bassiana is widely used for insect pest control and can produce three distinct infective unit types under different nutritional and environmental conditions: aerial conidia, blastospores, and submerged conidia. Here, we identified endophytic colonization ability and existing forms of the three types of B. bassiana infective units after inoculating Arabidopsis plants via soil drenching, and tested their effects on their presence mold disease caused by Botrytis cinerea. We found that all B. bassiana infective unit types colonized Arabidopsis leaves, with germinating and producing hyphae by hydrophilic blastopores and submerged conidia; further, we showed that blastospores were more effective in defending against B. cinerea, compared with aerial conidia. These findings suggest that in addition to aerial conidia, the colonization of other two types of entomopathogenic fungal infective units could also have important impacts on plant resistance. This study contributes to better understanding on the function of B. bassiana as fungal endophytes, which could lead to a new paradigm for how to successfully use these organisms in biological control against plant diseases.     

The impact of arbuscular mycorrhizal fungi on tomato plant resistance against Tuta absoluta (Meyrick) in greenhouse conditions

The symbiotic relationship between plants and arbuscular mycorrhizal fungi (AMF) improves plant growth and increases its resistance to pests and diseases. Mycorrhizal fungi are among the specialized fungi associated with the rhizosphere and are completely dependent on plant organic carbon. In this research tomato, Solanum lycopersicum L. was used as the host plant to evaluate the interaction effects between inoculation of tomato plant with AMF and feeding of tomato leaf miner, Tuta absoluta (Meyrick). In addition, plant growth parameters and growth rate of insect were assessed. The mycorrhizal treatment included a mixture of four fungal species (Funneliformis mosseae, Rhizophagus intraradices, R. irregularis and Glomus iranicus). The results of the experiment showed that tomato plant roots were well colonized (66.29%) by AMF and there was a significant mutual relationship between the insects feeding on the plants and the fungi. Feeding by the insects on plants inoculated with the fungus increased percentage of colonization by AMF in plants infested with the insect as compared to the control plants. The results also indicated that growth parameters and phosphorus content of the plants inoculated with fungi significantly increased compared to the control group. Moreover, significantly lower growth rate and consumption index observed in the T. absoluta larvae were fed on the leaves of plants treated with AMF compared to leaves of plants not inoculated with AMF.     

Virulence potential of some fungal isolates and their control-promise against the Egyptian cotton leaf worm,Spodoptera littoralis

A preliminary virulence test of four fungal isolates, Beauveria bassiana IMI 382302, Beauveria bassiana IMI 386701, Trichoderma harzianum T24 and Aspergillus flavus Link against larvae of Spodoptera littoralis was performed. The most effective isolates against larvae of S. littoralis were B. bassiana 302 and T. harzianum T24, which also showed the lower percentage of pupation compared with the other two isolates under the same conditions of treatments. Three concentrations (1 × 10⁶, 1 × 10⁷ and 1 × 10⁸ ml^?1) of the aqueous conidial suspension of the four tested isolates were carried out against both larval and pupal stages of S. littoralis within five days post-treatment. T. harzianum T24 showed 80% larval mortality only when applied at the highest conidial concentration, while A. flavus showed 100% pupal mortality only, at all of its conidial concentrations. However, B. bassiana IMI 382302 showed relatively high dose-dependant larval and pupal mortalities, while strain IMI 386701 of B. bassiana showed a very weak mortality against pupae at higher concentrations, and no virulence against larvae was recorded. Enzymatic and antibiosis bioassays of the four fungal isolates showed relatively high activities against Fusarium spp. for most of the tested isolates. Clear zone of enzyme activity on agar plates proportionally increased with increasing the concentration of enzyme substrate and prolongation of the incubation period. Mtabolites produced in the agar culture inhibited the growth of Fusarium spp. and the productivity differed greatly among isolates or strains of the same isolate. Volatile and non-volatile compounds produced by A. flavus Link showed a higher inhibition activity against Fusarium spp. compared with the other fungal isolates. The humoral antifungal response of insect host is relatively high compared to the anti-bacterial one. Injection of larvae with the immune sensitive bacteria Micrococcus luteus (5 × 10³ bacteria/larva) showed a detectable humoral response by 2 h, peaked around 12 h and became hardly detectable by 24 h post-injection. Injection of larvae with conidial suspension (5 × 10³ conidia/larva) from each of the fungal isolates showed humoral antifungal activity against B. bassiana IMI 386701 and A. flavus only. This activity was detectable by 12 h, peaked around 36 h and became hardly detectable by 48 h post-injection. Although the humoral antifungal response was started slowly compared to the antibacterial one, it lasted for longer and enabled larvae to withstand the infection with these immune-sensitive fungal strains. No humoral activity was detected against B. bassiana IMI 382302, although however, weak activity was detected against T. harzianum T24 only at the low conidial concentration but not at the higher one (1 × 10⁸ ml^?1). Thus, this study concludes that larvae of S. littoralis showed immune-dependant sensitivity to T. harzianum T24 and B. bassiana IMI 382302. Therefore, this study may recommend these two fungal isolates as mycoinsecticides in the battle against cotton leaf worm in Egypt. Hence, they have been selected for future comprehensive bioassays in the laboratory under conditions similar to that in the field. This, in fact, may help for developing effective mycoinsecticides against this pest. Penetration mechanims of insect cuticle by entomopathogenic fungi will be discussed.     

Isolation of Beauveria species from Lebanon and evaluation of its efficacy against the cedar web-spinning sawfly, Cephalcia tannourinensis

Larvae of the cedar web-spinning sawfly, Cephalcia tannourinensis Chevin (Hymenoptera: Pamphiliidae), infected with a white fungus were collected from the Tannourine-Hadath El-Jebbeh cedar forest. Macro- and micro-morphological data based on the examination of colonies, conidiophores, and conidial shape of the fungus suggested a Beauveria species. Sequence analysis of the internal transcribed spacer regions of the isolated fungus showed that it is most closely related to isolates of B. bassiana Clade C. The present study showed that the isolated B. bassiana is a naturally occurring entomopathogenic fungus parasitizing the larvae of C. tannourinensis in Lebanon. Laboratory bioassays showed that B. bassiana caused high mortality of eggs and larvae. The infected eggs turned brownish in color, while larvae of the first instar ceased feeding and showed immobility and rigidity within 5 days and before sporulating conidial mat appeared on their cuticle. Second and third larval instars took longer time to show fungal sporulation: mortalities ranged between 85 and 100% within 7 days when treated with different conidial concentrations. The efficacy of control of C. tannourinensis using B. bassiana was higher or equal to the reference insect growth regulator, diflubenzuron, suggesting the possibility of its success as a biological control agent.     

Potential of betabaculoviruses to control the tomato leafminer Tuta absoluta (Meyrick)

The tomato leafminer Tuta absoluta (Meyrick 1917) (Lepidoptera:Gelechiidae) is a devastating pest, causing losses of up to 100%. An interesting tool for its control is the use of the insect specific viruses of Baculoviridae family. Despite its high potential, its use on T. absoluta larvae has been poorly studied. In this work, Colombian granuloviruses VG013 and VG003 isolated from T. absoluta and Tecia solanivora (Lepidoptera:Gelechiidae) larvae sampled in tomato and potato crops, respectively, were morphologically, biologically and molecularly characterized. Occlusion bodies showed ovoid shape containing one nucleocapsid. Restriction endonuclease analysis revealed a pattern similar to Phthorimaea operculella granulovirus and bioinformatics studies showed that both isolates are variants of that baculovirus specie. Similar mean lethal concentrations (LC₅₀) on T. absoluta larvae were estimated for both viruses, although VG013 exhibited shorter mean time to death than VG003. The latter reached a higher OBs yield in comparison with VG013. These results demonstrated an interesting potential of evaluated betabaculoviruses to control T. absoluta populations and pointed key features to its use under field conditions.     

Pathogenicity of Beauveria bassiana ANU1 to the red imported fire ant,Solenopsis invicta workers in Korea

The red imported fire ant (RIFA), Solenopsis invicta, is one of the globalized invasive pests. This study focused on pathogenicity and virulence of entomopathogenic fungi as one of the biological control agents to RIFA workers under different temperatures. The fungal pathogen, Beauveria bassiana ANU1 was isolated from Korea in 2015 and showed the pathogenicity to RIFA workers. A conidial suspension (1 × 10⁷ conidia/ml) induced a low mortality from day 2 after treatment and reached to 100% mortality at day 7 and day 8 after treatment for major and minor workers, respectively. The median lethal concentrations of B. bassiana ANU1 were calculated as 3.9 × 10³ for major and 4.6 × 10³ for minor workers at day 7 after treatment. Low temperatures decreased a virulence of B. bassiana ANU1 (1 × 10⁷ conidia/ml) to RIFA and showed mortality of 26.6% for major and 20% for minor workers. Based on bioassay results, this study provides one of possibilities of effective and successful strategy for controlling RIFA by entomopathogenic fungi.     

Laboratory studies on the competition for insect haemocoel between Beauveria bassiana and Steinernema ichnusae recovered in the same ecological niche

Laboratory assays evaluated the combined action of the entomopathogenic nematode Steinernema ichnusae and the entomopathogenic fungus Beauveria bassiana isolated from the same ecological niche, an oak forest in Sardinia (Italy). Galleria mellonella larvae were used as the test insect with the aim of understanding what happens in host haemocoel during a simultaneous infection with two different entomopathogens. Larval mortality assays were performed using nematodes and fungi both alone and together, at the same and different times, and in different concentrations. No additive or synergistic effects were observed, but there was a clear antagonism and competition for survival space in the haemocoel. Moreover, mutual effects between the symbiotic bacteria of the entomopathogenic nematode Xenorhabdus bovienii and entomopathogenic fungi were investigated. In laboratory experiments, X. bovienii crude extracts were tested for their activity against fungal growth. Compounds produced by B. bassiana were tested for their activity against the growth of bacteria, revealing that X. bovienii and B. bassiana are antagonistic to each other.