Future prospects for application of insect pathogens as a component of integrated pest management in tropical root crops

Insect pests and phytophagous mites cause a considerable loss to tropical root crops in the field. Major pests include the sweet potato weevil Cylas puncticollis, cassava mealybug Phenacoccus manihoti, cassava green spider mite Mononychellus tanajoa, yam beetle Heteroligus meles, and taro hornworm Hippotion celerio. Field and laboratory evaluation experiments indicate that entomopathogenic microorganisms may be adequately used in the management of insect and mite pests in root crops. The highest promise probably lies with fungal pathogens (Beauvaria bassiana, Hirsutella thompsonii, Metarhizium anisopliae, Nomuraea rileyi, Entomophthora thaxteriana, and E. parvispora), but bacterial (Bacillus thuringiensis), microsporidian (Nosema locustae) nematode (Steinernema feltiae) and even viral (Baculoviruses) pathogens may be exploited in an integrated pest management programme of tropical root crop pests.     

Pathogenicity of entomopathogenic fungi to different developmental stages of Rhipicephalus sanguineus (Acari: Ixodidae).

The pathogenicity of 4 species of entomopathogenic fungi (Hyphomycetes species: Beauveria bassiana, Metarhizium anisopliae, Metarhizium flavoviride, and Paecilomyces fumosoroseus) to various developmental stages of Rhipicephalus sanguineus (Acari: Ixodidae) ticks was compared under laboratory conditions. The most virulent isolate, M. anisopliae-108, caused 92-96% mortality to unfed larvae and nymphs on day 7 postinfection (PI) and 100% mortality to unfed adults and engorged females on day 21 PI. The pathogenicity of M. anisopliae-108 to engorged larvae and nymphs was lower--82.6 and 60%, respectively. All tested B. bassiana, M. flavoviride, and P. fumosoroseus isolates were significantly less virulent (P < 0.05) or avirulent toward most life stages of R. sanguineus. The M. anisopliae and M. flavoviride isolates also prevented or reduced the ability of the ticks to lay eggs several days before their deaths. Female ticks infected by the fungi achieved only 11.3-60.8% of their egg-laying capacity compared with the controls.     

Potential of fungi as biocontrol agents of polyphagotarsonemus latus (Acari: Tarsonemidae)

Infection of broad mites, Polyphagotarsonemus latus, by conidia of Beauveria bassiana, Hirsutella thompsonii, and Paecilomyces fumosoroseus conidia was investigated in the laboratory under controlled temperature and moisture conditions. Infection of P. latus by the fungus (LC₅₀) was related positively to the 1.16 × 10⁶ B. bassiana conidia per ml, 2.39 × 10³ H. thompsonii conidia per ml, and 1.29 × 10⁵ P. fumosoroseus conidia per ml. Mortality caused by B. bassiana occurred the fastest among densities fluctuating between 65 and 125 mites per leaf. The pathogens B. bassiana and P. fumosoroseus, adjuvants (oil and molasses), and the acaricide ABG6364, a microbial insecticide containing the Beta-exotoxin of Bacillus thuringiensis were evaluated for efficacy against P. latus in a greenhouse test. The B. bassiana treated plants had the greatest and most persistent effect on percentage mortality of total broad mites present per leaf (88 %) followed by the acaricide ABG6364. Use of adjuvants (oil and molasses) did not increase infection of P. latus.     

Mycopathogens of Mites in Poland - A Review

Studies on diseases of mites caused by entomopathogenic fungi have been undertaken in Poland about a half a century later than in West European countries. Nevertheless, during the last 30 years almost 40 species of arthropod pathogenic fungi have been identified, among them 23 species of Entomophthorales, 12 species of Hyphomycetales and four species of ectoparasitic Laboulbeniales. The most common are representatives of the genus Hirsutella , found both on phytophagous and parasitic or predaceous mites. Percentage infection by H. thompsonii in populations of some tarsonemid and eriophyid mites increases slowly from the end of spring reaching a maximum of 30-60% in August-September. Neozygites floridana is a common entomophthoralean species causing epizootics in spider mite populations; infection also peaks in late summer and autumn. The most numerous fungal taxa have been identified from the resting spores produced internally within the body of the host and show features of the genus Tarichium . From the total number of the taxa recorded within these studies, 22 were described as species new to science. Current research on invertebrate pathology in Poland includes many groups of mites and among their pathogens fungal diseases seem to be the most common.     

Diversity of acaropathogenic fungi in Poland and other European countries

The occurrence, species diversity and some aspects of taxonomical affinity and host selectivity of acaropathogenic fungi associated with phytophagous, saprotrophic and predacious mites in Poland and other European countries were investigated on wild and cultivated plants, in insect feeding sites under the bark and in decayed wood. From among 33 species of fungi affecting mites only five species of Entomophthorales were separated and the most numerous were Neozygites floridana mostly on Tetranychus urticae, N. abacaridis on a few eriophyid species, and Conidiobolus coronatus attacking gamasid mites most frequently of the genus Dendrolaelaps. The most frequent mite pathogens occurring in mite communities on plants and in wood infested by insects were of the genus Hirsutella. Until now 13 of their form-species have been recognized in these habitats, but only H. kirchneri, H. necatrix and H. thompsonii (including its variety synnematosa) can be treated as exclusive oligophagous pathogens of phytophagous mites, though their potential host range seems to embrace only selected eriophyid or tarsonemid mites. Taxonomical differentiation of fungal strains was based on close morphological observations and molecular analysis of ITS region sequences. Two new species of acaropathogenic fungi were described in these studies. Hirsutella danubiensis sp. nov. was found in the tetranychid T. urticae, whereas H. vandergeesti sp. nov. affected phytoseiid mites of the genera Amblyseius, Neoseiulus, Seiulus and Typhlodromus, and the tarsonemid Tarsonemus lacustris.     

Hirsutella thompsonii and Metarhizium anisopliae as potential microbial control agents of Varroa destructor,a honey bee parasite

The potential of Hirsutella thompsonii Fisher and Metarhizium anisopliae (Metschinkoff) as biological control agents of the parasitic mite, Varroa destructor Anderson and Trueman was evaluated in the laboratory and in observation hives. In the laboratory, time required for 90% cumulative mortality of mites (LT(90)) was 4.16 (3.98-4.42) days for H. thompsonii and 5.85 (5.48-7.43) days for M. anisopliae at 1.1 x 10(3) conidia mm(-2). At a temperature (34+/-1 degrees C) similar to that of the broodnest in a honey bee colony, Apis mellifera L., H. thompsonii [LC(90)=9.90 x 10(1) (5.86-19.35) conidia mm(-2) at Day 7] and M. anisopliae [LC(90)=7.13 x 10(3) (2.80-23.45) conidia mm(-2) at Day 7] both showed significant virulence against V. destructor. The applications of H. thompsonii to observation hives resulted in significant mortality of mites, and reduction of the number of mites per bee 21 and 42 days post-treatments. The treatments did not significantly affect the mite population in sealed brood. However, the fungus must have persisted because infected mites were still observed [82.97+/-(0.6)%] 42 days post-treatment. In addition, the fungus was found to sporulate on the host. A small percentage [2.86+/-(0.2)%] of dead mites found in the control hives also showed fungal infection, suggesting that adult bees drifted between hives and disseminated the fungus. H. thompsonii was harmless to the honey bees at the concentrations applied and did not have any deleterious effects on the fecundity of the queens. Microbial control with fungal pathogens provides promising new avenues for control of V. destructor and could be a useful component of an integrated pest management program for the honey bee industry.     

The Susceptibility of Boophilus annulatus (Ixodidae) Ticks to Entomopathogenic Fungi

The pathogenicity of five species of entomopathogenic fungi (Deuteromycetes, species: Beauveria bassiana, Metarhizium anisopliae, Metarhizium flavoviride, Paecilomyces fumosoroseus and Verticillium lecanii ) to the various developmental stages of Boophilus annulatus ticks was compared under laboratory conditions. M. anisopliae and B. bassiana strains were most virulent to engorged females and caused 85-100% mortality within 7-10 days post-inoculation (PI). The highest mortality of engorged females caused by other fungi reached only 25-60%. All tested fungi prevented or reduced the egg laying capability of the ticks several days before their death. Females surviving after treatment with the most virulent M. anisopliae strain (Ma-7) reached only 7-8% of their egg laying capacity as compared with the control. Other fungi caused a reduction of the weight of laid eggs by 35.4-80.8% as compared with untreated females. Only M. anisopliae and B. bassiana strains caused 70-98% mortality of the treated eggs. Unfed larvae of Boophilus annulatus were sensitive to M. anisopliae and M. flavoviride strains. The Ma-7 strain was most virulent to unfed larvae, with a mortality rate of 80.4% at a concentration of 1 ×107 spores ml -1 and 100% mortality at a concentration of 1 ×108 spores ml -1 .     

Susceptibility of adult Aethina tumida (Coleoptera: Nitidulidae) to entomopathogenic fungi

Aethina tumida Murray (Coleoptera: Nitidulidae) is an invasive parasite species in populations of honey bees, Apis mellifera L. Aiming toward substitution of chemical control, we here identified a naturally occurring fungal pathogen of adult A. tumida from its endemic range in South Africa [Metarhizium anisopliae (Metschnikoff) Sorokin variety anisopliae strain FI-203]. The susceptibility of adult beetles (n = 400) to this fungus and to three other generalist entomopathogenic fungal isolates [Metarhizium anisopliae, Beauveria bassiana (Balsamo) Vuillemin, and Hirsutella illustris Minter & Brady] was assessed using spore suspension bioassays. The data revealed significantly increased mortality in the B. bassiana (74.00 +/- 8.94%) and M. anisopliae variety anisopliae (28.00 +/- 16.43%) tests but not in the H. illustris (2.00 +/- 4.47%) and M. anisopliae (12.00 +/- 8.37%) groups. The results indicate a potential for entomopathogenic fungi as an alternative control of A. tumida.     

Perspectives for the biological control of Cameraria ohridella

The horse chestnut leaf-miner, Cameraria ohridella Deschka et Dimic (Lepidoptera, Gracillariidae) is a serious invasive pest of Aesculus hippocastanum in Europe. The larvae of this species feed on leaf parenchyma and can reduce the tree growth. We studied the impact of parasitoids on C. ohridella in the Czech Republic and also searched for entomopathogenic fungi associated with this pest. The results showed that the rate of parasitism varied between 5% and 15% in most cases. The most parasitized stages of C. ohridella were spinning stages and especially pupae. The most abundant parasitoid species were Minotetrastichus frontalis, Pnigalio sp. and Pediobius saulius (Hymenoptera, Eulophidae). All species are polyphagous. Using the Galleria-bait method we isolated many strains of entomopathogenic fungi. Dominant species were Paecilomyces fumosoroseus, Paecilomyces farinosus and Beauveria bassiana. The perspectives of fungal bioagents in control of C. ohridella is discussed.     

利用黄粉虫分离土壤昆虫病原真菌

利用黄粉虫Tenebrio molitorL.作为寄主从土壤中诱感并分离昆虫病原真菌。结果显示,从吉林市的18个土样中分离出球孢白僵菌(Beauveria bassiana)、金龟子绿僵菌(Metarhiziumanisopliae)和玫烟色拟青霉(Paecilomyces fumosoroseus)。土样中的昆虫病原真菌检出率为77.78%。这表明利用黄粉虫分离土壤中的昆虫病原真菌是一种简单、有效的方法。     

Pathogenicity of three formulations of entomopathogenic fungi for control of adult Haematobia irritans (Diptera: Muscidae)

Powder formulations of three species of entomopathogenic fungi were evaluated for their pathogenic effect upon adult horn flies, Hematobia irritans (L.) (Diptera: Muscidae). Flies were treated with conidia and blastospores of the entomopathogenic fungi Beauveria bassiana (Bals.) Vuill. (strain GHA), Metarhizium anisopliae (Metschnikoff) Sorokin (strain ESCI), and Paecilomyces fimosoroseus (Wize) Brown & Smith (strain ARSEF 3581) in the laboratory. At 4 d postexposure, flies treated with B. bassiana had an average of 98.4% mortality versus 43.5% from treatment with M. anisopliae and 13.0% from treatment with P. fiimosoroseus. At 7 d postexposure, flies treated with B. bassiana had an average of 100.0% mortality compared with 73.0% from treatment with M. anisopliae and 33.3% from treatment with P.fumosoroseus. Mean lethal time (LT50) was 2.70, 4.98, and 7.97 d for B. bassiana, M. anisopliae, and P. fiumosoroseus, respectively. Entomopathogenic fungi such as B. bassiana and M. anisopliae may have the potential for controlling populations of horn flies. These studies indicate that B. bassiana and M. anisopliae were not only pathogenic to adult horn flies, but they caused mortality in a short time.     

几种虫生真菌对斜纹夜蛾的致病性

用布氏白僵菌、球孢白僵菌、玫烟色拟青霉、绿僵菌和莱氏野村菌5种真菌的固体培养物,对斜纹夜蛾2、3龄幼虫进行了毒力测定.结果表明：布氏白僵菌和莱氏野村菌两种菌株对斜纹夜蛾幼虫有明显的致病效果,对2龄幼虫的致死中时(LT₅₀)分别为2.95 d和4.10 d,累计校正死亡率分别为100%和95.2%;对3龄幼虫的致病力低于2龄,致死中时(LT₅₀)分别为19.67 d和19.63 d,累计校正死亡率分别为56.6%和52.2%.玫烟色拟青霉、球孢白僵菌两菌株也有一定的致病力,对2龄幼虫的致死中时(LT₅₀)分别为4.89 d 和6.34 d,累计校正死亡率分别为85.7%和71.4%.     

Potential of a Dried Rice/Mycelium Formulation of Entomopathogenic Fungi to Suppress Subterranean Pests in Small Fruits

The potential of a dried rice/mycelium formulation of three species of entomopathogenic fungi ( Metarhizium anisopliae, Beauveria bassiana and Paecilomyces farinosus ) was assessed according to five criteria: storage life, field efficacy against two subterranean pests of cranberry, persistence and efficacy against first-instar black vine weevil larvae in potted strawberry and field persistence in a cranberry bog. The number of conidia sporulated from the formulation did not decline between 4 and 20 days. In small plot field trials, the numbers of black vine weevil, Otiorhynchus sulcatus (F.), were significantly lower in plots treated with 1000 gm - 2 of formulated M. anisopliae than in untreated plots. In similar trials, the numbers of adult cranberry girdler, Chrysoteuchia topiaria (Zeller), inside emergence cages were significantly lower on plots treated with the formulated M. anisopliae than on untreated plots. The formulation suppressed first-instar black vine weevil larvae in potted strawberry regardless of the species of entomopathogenic fungus or the post-inoculation interval. M. anisopliae was periodically isolated for up to 637 days from plots in cranberry bogs treated with the formulation. In general, the dried rice/mycelium formulation may be a useful fungal mycopesticide for the management of subterranean pests of small fruits, especially if applied at high rates to areas of serious pest infestation.     

A Proposed Role for the Cuticular Fatty Amides of Liposcelis bostrychophila (Psocoptera: Liposcelidae) in Preventing Adhesion of Entomopathogenic Fungi with Dry-conidia

Maximum challenge exposure of Liposcelis bostrychophila to Beauveria bassiana, Paecilomyces fumosoroseus, Aspergillus parasiticus or Metarhizium anisopliae resulted in no more than 16% mortality. We investigated several of L. bostrychophila's cuticular lipids for possible contributions to its tolerance for entomopathogenic fungi. Saturated C14 and C16 fatty acids did not reduce the germination rates of B. bassiana or M. anisopliae conidia. Saturated C6 to C12 fatty acids that have not been identified in L. bostrychophila cuticular extracts significantly reduced germination, but the reduction was mitigated by the presence of stearamide. Cis-6-hexadecenal did not affect germination rates. Mycelial growth of either fungal species did not occur in the presence of caprylic acid, was reduced by the presence of lauric acid, and was not significantly affected by palmitic acid. Liposcelis bostrychophila is the only insect for which fatty acid amides have been identified as cuticular components. Stearamide, its major fatty amide, did not reduce germination of B. bassiana or M. anisopliae conidia or growth of their mycelia. Adhesion of conidia to stearamide preparations did not differ significantly from adhesion to the cuticle of L. bostrychophila. Pretreatment of a beetle known to be fungus-susceptible, larval Oryzaephilus surinamensis, with stearamide significantly decreased adhesion of B. bassiana or M. anisopliae conidia to their cuticles. This evidence indicates that cuticular fatty amides may contribute to L. bostrychophila's tolerance for entomopathogenic fungi by decreasing hydrophobicity and static charge, thereby reducing conidial adhesion.     

球孢白僵菌和金龟子绿僵菌不同菌株对黑足角胸叶甲成虫的致病力评价

本研究在测定不同球孢白僵菌Beauveria bassiana和金龟子绿僵菌Metarhizium anisopliae菌株的生长速率与产孢量的基础上, 采用孢悬液浸渍法进行了对油茶新害虫--黑足角胸叶甲Basilepta melanopus成虫的生物测定, 旨在筛选出感染该成虫的高致病力菌株, 为防治该虫提供新的生物资源。结果表明： 不同菌株生长速率和产孢量间存在显著差异, MaYTTR-04, BbFZ-17, MaZPTR-01和BbTK-01生长速率和产孢量均显著高于其他菌株。接种后, 叶甲成虫累积死亡率随时间的增加而逐渐增高, 接种白僵菌7 d后, 成虫校正死亡率全部达到100%; 接种MaZPTR-01和MaYTTR-04两个绿僵菌菌株14 d后, 成虫死亡率分别为80.3%和78.8%。而且接种白僵菌后, 叶甲成虫的僵虫率显著较绿僵菌高, 尤其以BbTK-01和BbFZ-17两个菌株较好, 分别达到85.7%和75.8%。BbXJ-01, BbFZ-17和BbTK-01 3个白僵菌菌株的LT₅₀最小, 分别为3.0, 3.3和3.4 d; MaYTTR-04和MaZPTR-01两个绿僵菌的LT₅₀分别为6.0和6.2 d。结果说明, 白僵菌对叶甲成虫的致病力较强, 尤其是BbTK-01和BbFZ-17两个菌株, 不仅致死率高, 且致死速度快, 僵虫率高, 同时这2个菌株生长速度快、 产孢量大, 具有优良的生产特性, 在黑足角胸叶甲的生物防治中将有重要的应用价值。     

大别山地区虫生真菌群落结构与生态分布

本文对安徽大别山地区虫生真菌的群落结构和生态分布规律进行了研究。在天堂寨和鹞落坪2个自然保护区共设样方278个,先后采集罹病标本1986个,共计50种,隶属于4目、4科、16属。优势种依次为粉拟青霉、细脚拟青霉、下垂虫草、球孢白僵菌和金龟子绿僵菌。在垂直分布上,种的丰度、多度和物种多样性指数都以海拔1100-1150m最高,随着海拔升高或降低,各多样性指标基本上表现递减的趋势。季节分布上,夏季是虫生真菌物种分布最丰富的季节,随着温度下降和雨量的减少,种的丰度和多度也明显下降。另外,该区虫生真菌群落呈现出明显的水平地带性分布规律。     

Effects of nutrient medium composition and temperature on the germination of conidia and the entomopathogenic activity of the fungi

The effects of nutrient medium composition and temperature on the germination of conidia of the fungi Beauveria bassiana (strain AlG) and Metarhizium anisopliae (strain M-99) and their entomopathogenic activity have been studied. It was demonstrated that the presence of carbohydrates alone was sufficient for the spores of M. anisopliae M-99 to germinate, whereas the germination of B. bassiana AlG spores was inhibited by carbohydrates. Addition of KJ, ZnSO4, or KBr into the Czapek medium increased the entomopathogenic activity of B. bassiana. The optimum temperature for spore germination was 20-35 degrees C in both fungal species.     

Seasonal and Habitat Variability in the Fungal Pathogens, Neozygites cf. floridana and Hirsutella thompsonii, Associated with Cassava Mites in Benin, West Africa

A survey of the pathogenic fungi associated with mites on cassava in Benin, West Africa, revealed both geographical and seasonal variation in the presence of Neozygites cf. floridana (Weiser and Muma) and Hirsutella thompsonii Fisher on Mononychellus tanajoa (Bondar) and Oligonychus gossypii (Zacher). Few dead and infected mites were found during the dry season, regardless of vegetation zone. In three of 30 surveyed sites, N. floridana was found infecting 1% of the dead M. tanajoa and 2% of the dead O. gossypii, while H. thompsonii was observed infecting 20% of the dead M. tanajoa in a single site. The frequency of sites having infected mites during the wet season was 3.5 times greater than that seen during the dry season. N. floridana infected 10% of the dead M. tanajoa and 19% of the dead O. gossypii on young leaves. Mites infected with N. floridana were found either in the coastal Southern Forest Mosaic (SFM) or in the Northern Guinea Savanna vegetation zones. N. floridana was rare in the low mite densities associated with mature leaves. H. thompsonii was found on 19% and 29% of the dead M. tanajoa on young and mature leaves respectively. All M. tanajoa infected with H. thompsonii on young leaves and mature leaves (75%) were found in the SFM. A single M. tanajoa was the only infected mite found in the Southern Guinea Savanna. Relatively few O. gossypii were infected with H. thompsonii. N. floridana and H. thompsonii were found together in three sites, but never on the same host. Phytoseiids were never found infected with either pathogen. In a regression analysis, the number of dead mites was significantly estimated from the total number of mites for both species, regardless of leaf age. The numbers of dead M. tanajoa on mature leaves were also estimated from the proportion infected with H. thompsonii. The numbers of infected mites on young leaves were estimated from their association with the SFM for M. tanajoa infected with H. thompsonii, and from total mites for O. gossypii infected with N. floridana. On mature leaves, infected mite numbers were estimated from the numbers of dead M. tanajoa infected with H. thompsonii. The merit of introducing more virulent or better adapted isolates of N. floridana to control M. tanajoa in Africa is discussed.     

Pests and Diseases of Betelvine Piper Betle and Their Natural Enemies in India

Betelvine (Piper betle) is an important cash crop in India. The cultivation of this crop is seriously threatened by several pest insects, for example the homopterans Aleurocanthus rugosa, Dialeuroides pallida, Aphis gossypii and Tricentrus gibbosulus and the thysanopterans Membrothrips indicus, Zaniothrips ricini, Mymarothrips garuda and Aroidothrips longistylus and by the plant pathogenic fungi Curvularia lunata, Phytophthora parasitica, Colletotrichum piperis and Fusarium moniliforme. The pest and pathogen species are attacked by natural enemies belonging to the Coleoptera, Diptera, Heteroptera, Hymenoptera, Aranea and Acari. While occurrence of non-acarine enemies is confined to certain seasons, the acarine enemies are associated with the insect pests and fungal pathogens throughout the year. The predatory mites Walzia indiana and Proctolaelaps pygmaeus and the mycophagous mites Unguizetes clavatus and Acarus sp. seem to have potential to control the insect pests and fungal pathogens, respectively.