A novel two-stage cultivation method to optimize carbon concentration and carbon-to-nitrogen ratio for sporulation of biocontrol fungi

A two-stage solid cultivation method was used to determine the precise requirements of carbon concentration (1–16 g/L) and C: N ratio (0.625: 1 to 80: 1) for the sporulation of six biocontrol fungi. The C concentration and C: N ratio producing the highest conidia yield were 1 g/L and 5: 1 for Paecilomyces lilacinus IPC-P; 2 g/L and 10: 1 for P. lilacinus M-14; 16 g/L and 80: 1 for Metarhizium anisopliae SQZ-1-21; 4 g/L and 5: 1 for M. anisopliae RS-4-1 and Lecanicillium lecanii CA-1-G; and 2 g/L and 10: 1 for Trichoderma viride TV-1. Sporulation was more affected by C: N ratio than by C and N concentration per se. More spores per colony were produced by the two-stage method than by a conventional, single-stage cultivation method. These results should be useful for improving the mass production of these biocontrol agents.     

Effects of carbon concentration and carbon to nitrogen ratio on the growth and sporulation of several biocontrol fungi

Effects of carbon concentration and carbon to nitrogen (C:N) ratio on six biocontrol fungal strains are reported in this paper. All fungal strains had extensive growth on the media supplemented with 6–12 g l⁻¹ carbon and C:N ratios from 10:1 to 80:1, and differed in nutrient requirements for sporulation. Except for the two strains of Paecilomyces lilacinus, all selected fungi attained the highest spore yields at a C:N ratio of 160:1 when the carbon concentration was 12 g l⁻¹ for Metarhizium anisopliae SQZ-1-21, 6 g l⁻¹ for M. anisopliae RS-4-1 and Trichoderma viride TV-1, and 8 g l⁻¹ for Lecanicillium lecanii CA-1-G. The optimal conditions for P. lilacinus sporulation were 8 g l⁻¹ carbon with a C:N ratio of 10:1 for M-14 and 12 g l⁻¹ carbon with a C:N ratio of 20:1 for IPC-P, respectively. The results indicated that the influence of carbon concentration and C:N ratio on fungal growth and sporulation is strain dependent; therefore, consideration for the complexity of nutrient requirements is essential for improving yields of fungal biocontrol agents.     

Carbon Requirements of Some Nematophagous, Entomopathogenic and Mycoparasitic Hyphomycetes as Fungal Biocontrol Agents

Thirty-three carbon sources were evaluated for their effects on spore germination, hyphal growth and sporulation of 11 fungal biocontrol agents, i.e. the nematophagous fungi Paecilomyces lilacinus, Pochonia chlamydosporia, Hirsutella rhossiliensis, H. minnesotensis and Arkansas Fungus 18, the entomopathogenic fungi Lecanicillium lecanii, Beauveria bassiana and Metarhizium anisopliae, and the mycoparasitic fungus Trichoderma viride. Variations in carbon requirements were found among the fungal species or strains tested. All strains studied except for T. viride grew on most carbon sources, although B. bassiana had more fastidious requirements for spore germination. Monosaccharides and disaccharides were suitable for fungal growth. For most isolates, d-glucose, d-mannose, sucrose and trehalose were superior to pectin and soluble starch among the polysaccharides and lactic acid among the organic acids. Both ethanol and methanol could accelerate growth of most isolates but not biomass. d-mannose, d-fructose and d-xylose were excellent carbon sources for sporulation, while d-glucose, sucrose, cellobiose, trehalose, chitin, dextrin, gelatin and lactic acid were better for some isolates. Neither sorbic acid nor linoleic acid could be utilized as a single carbon source. These findings provided a better understanding of the nutritional requirements of different fungal biocontrol agents that can benefit the mass production process.     

Effect of storage conditions on the survival of two potential biocontrol agents of nematodes,the fungi Paecilomyces lilacinus and Pochonia chlamydosporia

The nematophagous fungi Paecilomyces lilacinus and Pochonia chlamydosporia have been extensively studied as biological control agents for plant-parasitic nematodes. This study describes the formulation of alginate pellets containing mycelia of these fungi and also describes the effect of storage conditions on shelf-life of the pellets. The shelf-lives of P. lilacinus and P. chlamydosporia, which were measured monthly for 6 months, were significantly improved at low temperatures and low water activity (a _w) values (<0.33). Vacuum did not affect the viability of the formulated P. lilacinus but increased the viability of P. chlamydosporia. Carbon dioxide reduced the activity of P. lilacinus as compared to ambient air but increased the activity of P. chlamydosporia. Nitrogen, however, significantly improved the viability of both fungi. The optimal parameters of each factor for our formulation of P. lilacinus and P. chlamydosporia included a temperature range of 4 to ?20°C, a _w=0.12, and a nitrogen-filled atmosphere.     

Enzymatic analysis and in vitro ovicidal effect of Pochonia chlamydosporia and Paecilomyces lilacinus on Oxyuris equi eggs of horses

The aim of this work was to analyse the enzymatic activity and in vitro ovicidal effect of Pochonia chlamydosporia (VC1 and VC4) and Paecilomyces lilacinus (PL1) on Oxyuris equi eggs of horses. The growth of isolates and their enzymatic production were evaluated on agar media supplemented with gelatin (GA), casein (CA), olive oil (OOA) or starch (SSA). The ovicidal effect was evaluated according to morphological alterations. Following, the P. chlamydosporia crude extract production and proteolytic content was evaluated (VC1 and VC4) in liquid medium at the interval of 15, 30, 45 and 60 min during incubation of P. chlamydosporia and P. lilacinus which grew and showed enzymatic activity on agar media (GA, CA, OOA and SSA). At the 15th day, VC1, VC4 and PL1 showed results on GA, CA, OOA and SSA media, for Type 3 effect of (78, 54, 52 and 68%), (72, 50, 58 and 78%) and (62, 57, 65 and 63%). Pochonia chlamydosporia was able to grow in liquid medium (gelatin) and at Day 5 showed proteolytic activity. The results of the present work suggest that P. chlamydosporia and P. lilacinus can be used in the biological control of O. equi eggs.     

Effect of plant growth promoting rhizobacteria,nematode parasitic fungi and root-nodule bacterium on root-knot nematodes Meloidogyne javanica and growth of chickpea

Effects of plant growth promoting rhizobacteria (Pseudomonas putida MTCC No. 3604 and Pseudomonas alcaligenes MTCC No. 493) and parasitic fungi (Pochonia chlamydosporia KIA and Paecilomyces lilacinus KIA) were studied, alone and together with Rhizobium sp. (charcoal commercial culture) on the growth of chickpea and multiplication of Meloidogyne javanica. Individually, P. putida 3604, P. alcaligenes 493 and Rhizobium caused a significant increase in the growth of chickpea in both nematode inoculated and uninoculated plants. Inoculation of Rhizobium with a parasitic fungus or with plant growth promoting rhizobaterium caused a greater increase in the growth of plants inoculated with nematodes than caused by either of them singly. Individually, P. lilacinus KIA caused a greater increase in the growth of nematode inoculated plants than caused by P. putida 3604 or P. alcaligenes 493. P. lilacinus KIA caused a greater reduction in galling and nematode multiplication followed by P. chlamydosporia KIA, P. putida 3604 and P. alcaligenes 493. Combined use of P. lilacinus KIA with Rhizobium was better in reducing galling and nematode multiplication than any other treatment. P. putida 3604 caused a greater colonization of root than P. alcaligenes 493 while P. lilacinus KIA was isolated from more nematodes than P. chlamydosporia KIA.     

Effects of Carbon Concentrations and Carbon to Nitrogen ratios on Sporulation of Two Biological Control Fungi as Determined by Different Culture Methods

The nematophagous fungus Pochonia chlamydosporia (Clavicipitaceae) and entomopathogenic fungus Beauveria bassiana (Cordycipitaceae) have great potential for biological control. However, a significant barrier to their commercial development as mycopesticides is the high costs associated with production. Carbon (C) concentration and C to nitrogen ratio (C:N ratio) greatly affect fungal growth and sporulation. Effects of C concentration and C:N ratio differed when the fungi were cultivated using two different methods: the conventional (continuous cultivation) method and a novel “two-stage” method. Sporulation of P. chlamydosporia (HSY-12-14) was the highest when the media contained 6 g l⁻¹ C and a C:N ratio of 40:1 or 8 g l⁻¹ C and C:N ratios of 20:1 or 40:1 for the conventional method but 8 g l⁻¹ C and a C:N ratio of only 10:1 with the novel “two-stage” method. Sporulation of B. bassiana (IBC1201) was the highest when the media contained 12 g l⁻¹ C and a C:N ratio of 40:1 with the conventional method but only 4 g l⁻¹ C and a C:N ratio of 5:1 with the novel “two-stage” method. In addition, the nutritional requirements as determined by the conventional method differed for mycelial growth and sporulation. Understanding the effects of nutrition on sporulation can help programs seeking to use these organisms as biological control agents and is essential for their mass production and commercialization.     

Isolation of nematophagous fungi from eggs and females of Meloidogyne spp. and evaluation of their biological control potential

Fungi were isolated from Meloidogyne spp. eggs and females on 102 field-collected root samples in China. Of the 235 fungi isolated (representing 18 genera and 26 species), the predominant fungi were Fusarium spp. (42.1% of the isolates collected), Fusarium oxysporum (13.2%), Paecilomyces lilacinus (12.8%), and Pochonia chlamydosporia (8.5%). The isolates were screened for their ability to parasitise Meloidogyne incognita eggs in 24-well tissue culture plates in two different tests. The percentage of eggs parasitised by the fungi, the numbers of unhatched eggs and alive and dead juveniles were counted at 4 and 7 days after inoculation. The most promising fungi included five Paecilomyces isolates, 10 Fusarium isolates, 10 Pochonia isolates and one Acremonium isolate in test 1 or test 2. Paecilomyces lilacinus YES-2 and P. chlamydosporia HDZ-9 selected from the in vitro tests were formulated in alginate pellets and evaluated for M. incognita control on tomato in a greenhouse by adding them into a soil with sand mixture at rates of 0.2, 0.4, 0.8 and 1.6% (w/w). P. lilacinus pellets at the highest rate (1.6%) reduced root galling by 66.7%. P. chlamydosporia pellets at the highest rate reduced the final nematode density by 90%. The results indicate that P. lilacinus and P. chlamydosporia as pellet formulation can effectively control root-knot nematodes.     

Biochemical properties of different entomopathogenic fungi and their virulence against Chilo suppressalis (Lepidoptera: Crambidae) larvae

We determined the virulence of Beauveria bassiana, Metarhizium anisopliae, Isaria fumosoroseus and Lecanicilium lecanii against larvae of Chilo suppressalis Walker by bioassay and evaluated several enzymatic and non-enzymatic components. LC₅₀ values of the entomopathogenic fungi revealed 90, 32, 45,000, 4600, 42,000 and 1,540,000 spores/larva for isolates BB1–BB3 of B. bassiana, I. fumosoroseus, M. anisopliae and L. lecanii, respectively. Isolate BB3 and I. fumosoroseus had the highest amounts of total protein and hydrophobin and isolates BB3 and M. anisopliae showed the highest activities of lipases and chitinases. In case of proteases, the highest activities were observed for Pr1 of BB1 and Pr2 of L. lecanii. The highest general esterase activities were obtained in I. fumosoroseus and BB1 when 1-naphtyl acetate and 2-naphtyl acetate were used as substrates. The highest activity of glutathione S-transferase (GST) was observed in I. fumosoroseus by using both reagents but BB1 demonstrated the highest activities of alkaline phosphatase and acid phosphatase. Clustering of the fungi using biochemical enzymes revealed BB2 and BB3 as a separate group of entomopathogenic fungi. In another group, I. fumosoroseus and L. lecanii had the most similarity and were separated from BB1 and M. anisopliae. The fungi exhibited different virulence on larvae of C. suppressalis by producing adhering protein and extracellular enzymes. Overall, results of the bioassays and clustering based on enzymatic activities revealed that isolate BB2 was the most effective fungus against larvae of C. suppressalis.     

Natural occurrence of soil-borne entomopathogenic fungi in the Moroccan Endemic forest of <Emphasis Type="Italic">Argania spinosa</Emphasis> and their pathogenicity to <Emphasis Type="Italic">Ceratitis capitata</Emphasis>

The occurrence and abundance of entomopathogenic fungi were analysed in 203 soil samples of the Moroccan endemic forests of Argania spinosa, the world main refuge of the Mediterranean fruit fly, Ceratitis capitata. Using the Galleria baiting method and selective media, entomopathogenic fungi were isolated from 186 of the 203 (91.62%) soil samples, with only three species found: Beauveria bassiana (Balsamo) Vuillemin, Metarhizium anisopliae (Metschnikoff) Sorokin and Paecilomyces lilacinus (Thom.) Samson. B. bassiana was the most widespread entomopathogenic fungi (90.64%) in the Argan forest whereas M. anisopliae was less common (15.27%) and P. lilacinus was very rare (1.48%). This is the first report of natural occurrence of M. anisopliae and P. lilacinus in Morocco. Furthermore, 118 Moroccan B. bassiana isolates were studied for their pathogenicity to C. capitata and thermotolerance. Most of these autochtonous B. bassiana isolates were virulent (86.44%) to Medfly pupae and tolerant (55.08%) to temperature stress at 45°C for 2 h. Only 60.17% of Moroccan B. bassiana isolates might be considered as highly entomopathogenic and will serve as a source of potential biological control agents to C. capitata. The percentage of thermotolerant and pathogenic B. bassiana to C. capitata were shown to decrease significantly at winter time characterized by low temperatures and absence of any noticeable medfly in the Argan forest. The occurrence, thermotolerance and virulence of B. bassiana isolates to C. capitata seemed to be related to the sampling periods and location. Our data are discussed with respect to fungal ecology and biocontrol potential of B. bassiana isolates in relation to their habitat.     

In vitro soil receptivity assays to egg-parasitic nematophagous fungi

Soil application of nematophagous fungi for the biological control of plant-parasitic nematodes often fails, and in many cases it has been difficult to reisolate the agent delivered to the soil. A reason for these results could be the inability of the fungi to proliferate in soil. We used a soil–membrane technique to study the capacity of several isolates of the nematophagous fungi Pochonia chlamydosporia and Paecilomyces lilacinus to grow and establish in sterilized and nonsterilized sandy soils from SE Spain and Western Australia. Growth of all fungi tested was inhibited in nonsterilized soil, although there was intraspecific variability in sensitivity among isolates of the same species. With respect to hyphal density, P. chlamydosporia isolate 5 (from Italy) was the least inhibited in nonsterilized soil from both sites. Relative growth analyses confirmed this result for soil from SE Spain, while with this method, P. chlamydosporia isolate 4624 (from Australia) appeared to be least inhibited in the Australian soil. The results indicate that a soil can be more receptive to its indigenous isolates than to nonindigenous isolates. Apparently, soil microbiota can determine the ability of nematophagous fungi to proliferate in soil.     

Biological control of root rot-root knot disease complex of tomato

Efficacy of Pseudomonas aeruginosa alone or in combination with Paecilomyces lilacinus was evaluated in the control of root-knot nematode and root-infecting fungi under laboratory and field conditions. Ethyl acetate extract (1 mg/ml) of P. lilacinus and P. aeruginosa,respectively, caused 100 and 64% mortality of Meloidogyne javanica larvae after 24 h. Ethyl acetate fractions of biocontrol agents were more effective than hexane extracts in the suppression of M. javanica larvae, indicating that active nematicidal compounds are intermediary in polarity. In field experiments, biocontrol fungus and bacterium significantly suppressed soilborne root-infecting fungi including Macrophomina phaseolina, Fusarium oxysporum, Fusarium solani, Rhizoctonia solani and Meloidogyne javanica, the root-knot nematode. P. lilacinus parasitized eggs and female of M. javanica and this parasitism was not significantly influenced in the presence of P. aeruginosa. P. aeruginosa was reisolated from the inner root tissues of tomato, whereas P. lilacinusdid not colonize tomato roots. This revised version was published online in June 2006 with corrections to the Cover Date.     

Occurrence and diversity of entomopathogenic fungi in cultivated and uncultivated soils in Pakistan

The distribution of entomopathogenic fungi in various geographical areas of Punjab, Pakistan, is poorly understood. The present study was planned to explore the occurrence and diversity of entomopathogenic fungi from soils collected from cultivated and non‐cultivated habitats. The detailed survey of different habitats (crop fields, fruits, vegetables, forests) was conducted to collect soil samples and the associated fungi were isolated using Galleria bait method. Among 210 soil samples, 168 fungal isolates were recovered and identified, with 98 from forests, 32 from vegetables, 30 from field crops and 8 from fruits. The major entomopathogenic fungi recovered from these samples were Beauveria bassiana, Metarhizium anisopliae, Paecilomyces lilacinus, B. brongniartii, P. chlamydosporia and Lecanicillium attenuatum. The diversity of entomopathogenic fungi was greater in soil samples from forests compared to crop fields, vegetables and fruits, respectively. The geographical attributes (altitude, longitude, latitude) greatly influenced the occurrence of entomopathogenic fungi with the highest number of isolates found from >600 m altitude, 33°–34′N latitude, and 73°74′E longitude from soil samples. The results of the surveys clearly indicated that the entomopathogenic fungi are distributed in the soils which may be used in successful IPM programs in future.     

In vitro studies of antagonistic fungi against the root-knot nematode,Meloidogyne incognita

The present study was carried out in vitro to determine the efficacy of indigenous fungi isolated from egg masses of root-knot nematode, Meloidogyne incognita on egg parasitism, egg hatching, mobility and mortality against root-knot nematode, M. incognita. The tested fungi were Acremonium strictum, Aspergillus terreus, A. nidulans, A. niger, Chetomium aubense, Chladosporium oxysporum, Fusarium chlamydosporium, F. dimarum, F. oxysporum, F. solani, Paecilomyces lilacinus, Pochonia chlamydosporia, Trichoderma viride and T. harzianum. All tested fungi showed varied effects against the nematodes. Culture filtrates of A. strictum was very effective against the nematode in regards to egg parasitism (53%), egg hatching inhibition (86%) and mortality (68%) compared to controls. A. strictum was found to have an advantage over P. lilacinus, P. chlamydosporia, T. viride and T. harzianum in that it caused greater mortality of the second stage juveniles (J₂). A. terreus did not show egg parasitism but was found to be highly toxic against second stage juveniles (J₂) causing high mortality (around 68%). Thus, A. strictum and A. terreus showed good biocontrol potential against root-knot nematode, M. incognita under in vitro conditions.     

Effects of crop plants on abundance of Pochonia chlamydosporia and other fungal parasites of root‐knot and potato cyst nematodes

The effects of a host plant on reproduction/abundance of fungal populations in relation to soil nutrients released by plants in the rhizosphere were studied. Abundance in the soil and potato rhizosphere of the fungi Paecilomyces lilacinus, Monographella cucumerina (CABI 380408) and Pochonia chlamydosporia var. chlamydosporia (Pc280, potato cyst nematode biotype) and P. chlamydosporia var. catenulata (Pc392, root‐knot nematode biotype) were assessed. The different ability of break crops (oilseed rape, sugarbeet and wheat) in the potato rotation to support Pa. lilacinus, Pochonia isolates Pc280 and Pc392 and abundance of the latter two isolates in soil and rhizosphere of potato plants infected with Meloidogyne incognita were also studied. Potato chits and crop seedlings were planted into boiling tubes containing 5000 chlamydospores or conidia g^?1 in acid washed sand (pH 6) and kept in a growth chamber at 20°C, and 16 h of light for up to 9 weeks. The abundance of the fungi in sand (fallow) differed significantly between fungal species, being in general less abundant in the absence than in the presence of the plant, although there was no interaction between plant species and fungal isolate. There was evidence of a different response to Me. incognita for Pc392 than for Pc280 but there was no significant effect of the presence of the nematode on the rate of increase of the fungus.     

Growth and sporulation of Beauveria bassiana and Metarrhizium anisopliae on media containing various amino acids

Natural isolates of two entomogenous fungi, Beauveria bassiana and Metarrhizium anisopliae, were cultured in liquid culture media containing 24 amino acids and KNO₃ to determine their effect on growth and sporulation. In addition, the growth and medium pH changes for each isolate grown on an asparagine-containing medium were compared. Tryptophan and alanine were most effective for growth and sporulation of B. bassiana, although glutamine and KNO₃ also produced large numbers of regularly shaped spores. Tryptophan, glutamic acid, and histidine were all well utilized for both growth and sporulation of M. anisopliae. Nitrogen sources containing sulfur were poorly utilized for sporulation by M. anisopliae. In general, B. bassiana produces greater mycelial mass and much larger numbers of spores than M. anisopliae. Both fungi attained nearly the same growth maximum on asparagine medium though B. bassiana exhibited an initially more rapid growth rate. In both fungi this rapid growth phase was accompanied by a decline in medium pH followed by a rise in pH during the decline phase of growth.     

Pathogenicity of hyphomycetous fungi against <Emphasis Type="Italic">Cyclocephala signaticollis</Emphasis>

Susceptibility of the white grub Cyclocephala signaticollis Burmeister (Coleoptera: Scarabaeidae: Dynastinae) larvae to seven isolates of Beauveria bassiana (Balsamo) Vuillemin, five of Metarhizium anisopliae (Metschnikoff) Sorokin and two of Paecilomyces lilacinus (Thom) Samson (Deuteromycotina: Hyphomycetes) was investigated. Among 14 fungal isolates screened the most virulent was a B. bassiana isolate (Bb 53) that caused 70% mortality of third instar larvae in 40 days after inoculation at 1 × 10⁸ conida/ml. Strains of M. anisopliae and P. lilacinus showed low efficacy or no virulence to the target host.     

Effects of carbon and nitrogen sources,carbon-to-nitrogen ratio,and initial pH on the growth of nematophagous fungus <Emphasis Type="Italic">Pochonia chlamydosporia</Emphasis> in liquid culture

The effects of carbon and nitrogen sources, carbon-to-nitrogen ratio (C:N) and initial pH value on the growth and sporulation of the nematophagous fungus Pochonia chlamydosporia in liquid culture were examined. Among the 21 carbon sources and 15 nitrogen compounds tested, the optimal carbon and nitrogen sources for mycelial growth were sweet potato and L-tyrosine, and for sporulation were sweet potato and casein peptone. A C:N ratio of 10:1 at pH 3.7 gave the maximum yield of conidia and a C:N ratio of 40:1 at pH 6.8 gave the maximum biomass. The initial pH value had a significant effect on mycelial growth and conidial production, with the optimal ranges being 3.5–4.5 for sporulation and 5–6 for growth. Maximum conidial production was obtained at an initial pH of 4.0 and the maximum biomass at pH 6.0. The results also showed that the final pH after 7 days cultivation was always higher than the initial value. The variability in growth and sporulation of seven strains of P. chlamydosporia in liquid culture was also compared and discussed.     

Preliminary In Vitro Insights into the Use of Natural Fungal Pathogens of Leaf-cutting Ants as Biocontrol Agents

Leaf-cutting ants are one of the main herbivores of the Neotropics, where they represent an important agricultural pest. These ants are particularly difficult to control because of the complex network of microbial symbionts. Leaf-cutting ants have traditionally been controlled through pesticide application, but there is a need for alternative, more environmentally friendly, control methods such as biological control. Potential promising biocontrol candidates include the microfungi Escovopsis spp. (anamorphic Hypocreales), which are specialized pathogens of the fungi the ants cultivate for food. These pathogens are suppressed through ant behaviors and ant-associated antibiotic-producing Actinobacteria. In order to be an effective biocontrol agent, Escovopsis has to overcome these defenses. Here, we evaluate, using microbial in vitro assays, whether defenses in the ant-cultivated fungus strain (Leucoagaricus sp.) and Actinobacteria from the ant pest Acromyrmex lundii have the potential to limit the use of Escovopsis in biocontrol. We also explore, for the first time, possible synergistic biocontrol between Escovopsis and the entomopathogenic fungus Lecanicillium lecanii. All strains of Escovopsis proved to overgrow A. lundii cultivar in less than 7 days, with the Escovopsis strain isolated from a different leaf-cutting ant species being the most efficient. Escovopsis challenged with a Streptomyces strain isolated from A. lundii did not exhibit significant growth inhibition. Both results are encouraging for the use of Escovopsis as a biocontrol agent. Although we found that L. lecanii can suppress the growth of the cultivar, it also had a negative impact on Escovopsis, making the success of simultaneous use of these two fungi for biocontrol of A. lundii questionable.     

Efficiency of Nimbecidine and certain entomopathogenic fungi formulations against bean aphids,Aphis craccivora in broad bean field

This study investigates the effect of certain entomopathogenic fungi formulations (Beauveria bassiana, Verticillium lecanii, Metarhizium anisopliae and Paecilomyces fumosoroseus) compared with a botanical insecticide, Nimbecidine against Aphis craccivora in broad bean field. Bio-Catch (Verticillium lecanii) was the most effective insecticide to achieve 73.3% reduction followed by Nimbecidine (67.7%), Bio-Magic (61.6%), Priority (50.3%) and the least effective was Bio- Power (Beauveria bassiana) which caused 45.5% reduction in individual aphid populations after two sprayings at 15 days interval between the first and the second sprayings. Bio-Catch and Nimbecidine had promise compounds in controlling Aphis craccivora in faba bean fields.