Aptness of Beauveria bassiana and enhanced diatomaceous earth (DEBBM) for control of Rhyzopertha dominica F.

A diatomaceous earth formulation enhanced with bitterbarkomycin (DEBBM) combined with Beauveria bassiana (Balsamo) Vuillemin was evaluated against lesser grain borer Rhyzopertha dominica F. (Coleoptera: Bostrychidae) under laboratory conditions. DEBBM was applied at the rates of 15 and 30 ppm alone as well as in combination with 6.69 × 10⁶, 6.69 × 10⁸ and 6.69 × 10¹⁰ conidia/kg of wheat. Mortality of treated adults was recorded after 5, 10 and 15 days of exposure. Bioassays were carried out at 20, 25 and 30°C with 55 and 75% relative humidity. The emergence of progeny was also assessed 60 days post exposure. The combined use of DEBBM and B. bassiana considerably increased adult mortality especially at increasing temperatures and longer exposure intervals compared with DEBBM and B. bassiana alone. Progeny production was less in wheat treated with high dose rates of DEBBM +B. bassiana. The per cent mycosis in the cadavers was maximum where B. bassiana was applied at low dose rates. The results of the present study indicated that a combination of DEBBM and B. bassiana may provide effective control of R. dominica.     

Entomopathogenic fungus as a biological control agent against Rhyzopertha dominica F. (Coleoptera: Bostrychidae) on stored wheat

To evaluate the pathogenicity of Metarhizium anisopliae (Metschinkoff) Sorokin (Deuteromycotina: Hyphomycetes) a bioassay was designed under laboratory conditions against Rhyzopertha dominica F. (Coleoptera: Bostrychidae) on stored wheat. The fungus was applied at the dose rates of 8 × 10³, 8 × 10⁵, 8 × 10⁷ and 8 × 10⁹ conidia/kg of wheat and the bioassay was conducted at 25°C with 60% relative humidity. The data regarding the mortality was recorded after 7 and 14 days exposure intervals. All the treatments gave the significant mortality of R. dominica and M. anisopliae of 8 × 10⁹ conidia/kg was found to be the most effective after a 14-day exposure interval. There was greater production of progeny when the low rate of M. anisopliae was applied to wheat. Overall, our study showed that M. anisopliae is vigorous when applied at a high dose rate which revealed an effective control of R. dominica and also played a pivotal role in the integrated pest management program (IPM) of stored wheat insect pests.     

Effect of five diatomaceous earth formulations against Tribofium castaneum （Coleoptera： Tenebrionidae）, Oryzaephflus surinamensis （Coleoptera： Silvanidae） and Rhyzopertha dominica （Coleoptera： Bostrychidae）

Laboratory bioassays were conducted to determine the effect of food source on the survival of Tribolium castaneum Herbst, Oryzaephilus surinamensis L. and Rhyzopertha dominica F., after exposure to five diatomaceous earth （DE） formulations： Protect-It, Insecto, Perma-GuardTM, Dryacide and SilicoSec. Adults of these species were exposed to DEs at the rate of 0.5 mg/cm^2 for 1 day on filter paper inside plastic Petri dishes. After exposure, the initial mortality was counted and live individuals of the three species were held for a week in glass vials containing 50 mg wheat flour, rice and whole wheat, respectively. In the second experiment, after 1 day exposure to DEs, beetles were transferred to Petri dishes without food and held for a week to determine if the presence of food source would decrease the mortality of beetles. Experiments were carried out at 27℃ and 55% RH in the dark. The initial mortality in both of the experiments reached 100% for the three species exposed to Protect-It and in the case ofR. dominica and O. surinamensis exposed to Dryacide. In contrast, low level of mortality （〈 10%） was observed for T. castaneum exposed to Perma-GuardTM and Insecto. The mortality after the post-treatment period on food was decreased for the three species exposed to Perma-GuardTM and in the case of T. castaneum and R. dominica exposed to Insecto and SilicoSec. Adults of O. surinamensis were the most susceptible followed by R. dominica and 100% adult mortality was obtained, whereas T. castaneum were the least susceptible beetles to DEs. Protect-It and Dryacide were the most efficient DE formulations and can be used effectively in a stored grain integrated pest management program.     

Evaluation of Isaria fumosorosea (Hypocreales: Cordycipitaceae) for control of the Asian citrus psyllid,Diaphorina citri (Hemiptera: Psyllidae)

A laboratory bioassay was developed to evaluate strains of Isaria fumosorosea Wize, against Diaphorina citri. Up to 100% of adult psyllids were killed at concentrations between 10⁶ and 10⁷ blastospores/ml after 12 days, with derived LC₅₀ values (at 7 days post treatment) between 1.4 × 10⁵ and 2.0 × 10⁶ blastospores/ml for strains ARSEF 3581, FE 9901 and Apopka-97. A significantly higher value (1.5 × 10⁷) was obtained with a conidial formulation of Apopka-97. Average survival times were dosage dependent, i.e. between 10.2 days at 10³ blastospores/ml and 3.5 days at 10⁸ blastospores/ml. Rates of mycosis were also dosage dependent, with up to 100% sporulation on cadavers at 10⁸ blastospores/ml but declining at lower concentrations. The Apopka-97 strain (commercially available as PFR-97) was tested against established D. citri infestations in potted citrus in greenhouse cages. Treatments at label rates reduced psyllid populations by approximately 50% over 3 weeks. The combination of PFR-97 with emulsifiable oils (0.25% v/v) did not increase psyllid mortality compared with either agent alone. Imidacloprid applied as a drench killed 100% of psyllids within 3 weeks. Subsequent greenhouse tests during humid conditions were hampered by natural dissemination of I. fumosorosea to untreated psyllids, suggesting that this fungus is spread by air movement and may be highly effective under very humid conditions. In later tests, a Cladosporium sp. rapidly colonised psyllid cadavers and leaf surfaces, but was not pathogenic in laboratory tests. Our studies confirm the potential of I. fumosorosea to be used in IPM strategies for D. citri that rely on other tactics, such as insecticidal oils and native or introduced biological control agents.     

Using different amounts of vacuum and acrolein in various exposure periods to control two stored products insects

In this research, adults of Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) were exposed to various air pressures solely (first phase) and air pressures together with acrolein (second phase) in various durations and mortality rates were determined. Complete mortality of T. castaneum was obtained at 1, 3 and 6 h after exposure to 10, 30 and 50 mmHg, respectively. R. dominica showed 96.25% and 60% mortality during 3 and 10 h exposed to 10 and 30 mmHg, respectively. Percentage mortality of the insects in second phase was significantly higher than the rate in first phase. T. castaneum was more susceptible than R. dominica in the both phases.     

Conidia production by Isaria fumosorosea on solid substrates and its pathogenicity towards Bemisia tabaci

Large amounts of conidia must be recovered to use Isaria fumosorosea as a biological control agent. Aerial conidia are mass-produced in solid substrate culture. Conidia obtained from growth on different substrates must be similar in quantity and quality (viability, purity and virulence). In the present work, several solid substrates were evaluated: rice, sorghum, sugarcane, corncob, plantain and wheat (derived from the production of Trichogramma sp.) to find alternatives that are economical and available in different regions. The highest conidia production was obtained on rice, plantain and corncob substrates, which yielded 5.33 × 10⁹ conidia/g, 5.24 × 10⁹ conidia/g and 4.8 × 10⁹ conidia/g substrate, respectively. At concentrations of 1 × 10⁷ conidia/mL, the mortality rates obtained with conidia developed on rice, plantain and corncob substrates were 98%, 97% and 88%, respectively. The lethal concentration 50 (LC₅₀₎ values of conidia obtained from rice and plantain were 7.2 × 10⁴ conidia/mL and 5.1 × 10⁴ conidia/mL, respectively.     

Comparison of aerial conidia and blastospores from two entomopathogenic fungi against Diaphorina citri (Hemiptera: Liviidae) under laboratory and greenhouse conditions

This study compared the insecticidal activity of liquid culture-produced blastospores and solid substrate-produced aerial conidia of Beauveria bassiana GHA and Isaria fumosorosea ARSEF3581 strains against Diaphorina citri adults. Insects exposed to 10⁷ propagules/ml in a spray residue contact leaf bioassay died within 6 days at 25°C, with no significant differences between fungal treatments. At higher concentrations (10⁸ propagules/ml), I. fumosorosea conidia killed psyllids faster compared to its blastospore formulation, i.e. 4 versus 5 days, respectively. In greenhouse tests, the same treatments applied to infested citrus plants (2?×?10⁶ spores/ml) all significantly reduced the number of nymphs compared with the untreated controls over 3 weeks; however, only I. fumosorosea blastospores significantly reduced the number of F1 adult psyllids when compared with controls. Similar results were observed in the follow-up greenhouse test, where I. fumosorosea blastospores were the most effective treatment overall, reducing D. citri populations by about 60% after 21 days; by contrast, imidacloprid killed almost 100% of psyllids within a week in both tests. Fewer psyllids exhibited mycosis in the greenhouse (i.e. ≈20 versus?≥?87% in the laboratory). This is the first report comparing both conidial and blastospore formulations of B. bassiana and I. fumosorosea for the control of a psyllid pest. Field testing is required to determine how successful different spore formulations might be under various environmental conditions.     

Dissolved oxygen levels affect dimorphic growth by the entomopathogenic fungus Isaria fumosorosea

The entomopathogenic fungus Isaria fumosorosea is capable of dimorphic growth (hyphal or yeast-like) in submerged culture. Using 250-mL baffled flasks, culture volumes of 50, 100, 150, and 200 mL were grown in a shaker incubator at 350 rpm and 28°C. Dissolved oxygen (DO) was continuously monitored using a non-invasive oxygen monitoring system. Culture volumes of 50 mL maintained DO concentrations above 10% throughout the 3-day growth period and accumulated biomass and produced blastospores more rapidly (1.2×10⁹ blastospores mL^?1 in 2 days) than the other culture volumes tested. Dissolved oxygen was depleted in culture volumes of 100, 150, and 200 mL after 20.5, 16.8, and 13.5 h, respectively. The DO in 150 and 200 mL cultures remained exhausted (<3%) throughout the growth period resulting in significantly lower blastospore yields and increased hyphal growth. These results were used to establish oxygen levels (>20% DO) for I. fumosorosea growth in 100-L bioreactors resulting in blastospore production (1.1×10⁹ blastospores mL^?1 in 2 days) comparable to highly aerated, low volume shake flask cultures. In addition, maintaining higher DO levels resulted in increased blastospore production by cultures of I. fumosorosea grown on low-cost nitrogen sources (cottonseed meal and soy flour) that previously elicited excessive hyphal growth. These studies showed that oxygen availability is essential for significant yeast-like growth by I. fumosorosea cultures and that continuous monitoring of oxygen concentrations in shake flask cultures can be used to establish aeration conditions for bioreactors.     

Selection of entomopathogenic fungi for use in combination with sub-lethal doses of imidacloprid: perspectives for the control of the leaf-cutting ant <Emphasis Type="Italic">Atta sexdens</Emphasis><Emphasis Type="Italic">rubropilosa</Emphasis> Forel (Hymenoptera: Formicidae)

In the first part of this study, four isolates of the fungus Beauveria bassiana (Bals.) Vuillemin (LPP1, LPP2, CG05 and CG24) and one isolate of Metarhizium anisopliae (Metsch.) Sorokin (CG46) were tested against adult foragers of Atta sexdens rubropilosa. Ants were allowed to walk on filter paper discs, inside Petri dishes, previously impregnated with 1 ml of a conidia suspension (2 × 10⁷ conidia ml⁻¹), maintained at 80% RH and 26°C for 24 h and subsequently, transferred to sterile Petri dishes, maintained at 23°C, 80% RH, 24 h dark. The mean values of LT₅₀ for LPP2, LPP1, CG46, CG24 and CG05 were 3.5, 3.7, 3.8, 4.2 and 4.4 days, respectively. Control insects for all tests in this study showed less than 10% mortality. Experiments were carried out to test the toxicity of imidacloprid (IMI) to A. sexdens rubropilosa. Mortality was evaluated 10 days following a 24 h exposure to the insecticide. Percent mortality caused by 500, 200, 100 and 10 ppm IMI was 77.8, 56.7, 45.5 and 5.5 respectively. Insects treated with 10 ppm IMI were observed to have reduced locomotor activity 24 h after exposure to the insecticide. The LC₅₀ of IMI was 154.3 ppm. Subsequent tests were carried out to evaluate the combination of a sub-lethal dose of IMI (10 ppm) and infection by CG24 (1 × 10⁷ conidia ml⁻¹). Mortality due to fungal infection alone was 43.3%. Mortality of insects treated with IMI followed by exposure to the fungus was 64.3%. These results indicate that IMI significantly increases the susceptibility of ants to infection by B. bassiana CG24.     

Population development of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae) on different stored products

The lesser grain borer, Rhyzopertha dominica, is an important pest of various stored products around the world. In this study, the development, survival, reproduction, and life table parameters of R. dominica were investigated on six stored products (angelica, jujube, maize, rice, soybean, and wheat). The developmental time of the immature stage of R. dominica was shortest on wheat (40.20 days) and longest on angelica (67.04 days). The survival rate of the immature stage was highest on wheat (76.33%) and lowest on angelica (41.00%). The fecundity level of R. dominica was highest on wheat (246.05 eggs/female) and lowest on angelica (69.38 eggs/female). The net reproductive rate (R₀) and intrinsic rate of increase (r_m) of R. dominica differed significantly among the six stored products. The highest R₀ of R. dominica was on wheat (68.50), followed by rice (41.28), maize (32.32), soybean (27.17), jujube (23.16), and angelica (20.18); the r_m values showed a similar trend, with values of 0.059, 0.046, 0.042, 0.039, 0.036, and 0.033, respectively. Our results indicate that wheat was the most suitable stored product, whereas angelica was the least suitable, for the feeding, development, and population increase of R. dominica. These findings provide basic information about the occurrence trends and characteristics of R. dominica that will be useful for the control of this pest on different stored products. The physicochemical properties of angelica should be further explored for potential application in the control or integrated management of R. dominica.     

Susceptibility of immature Ixodes scapularis (Acari: Ixodidae) to desiccants and an insecticidal soap

Efficacy of commercially available formulations of desiccants (Drione, Dri-dic, diatomaceous earth) and insecticidal soap (Safer's) comprised of active ingredients were evaluated against Ixodes scapularis Say immatures in petri dish and grass turf bioassays. In petri dish bioassays at label rate, all products, except diatomaceous earth, provided a high degree of control by 24 and 48 h (71–100% mortality). Mortality of larvae exceeded 94% by 4 h after treatment with Drione (10× label rate held at 85% RH), Dri-die (label rate held at both 85% and 98% RH) and Safer's insecticidal soap (10× label rate held at 98% RH). Nymphal mortality was highest by 4 h after treatment with Safer's insecticidal soap (up to 96%). Larvae were more susceptible than nymphs to desiccants, however, nymphs were more susceptible than larvae to Safer's insecticidal soap. Immature ticks treated with Safer's insecticidal soap exhibited sublethal effects with significantly decreased attachment to hosts and no engorgement. In grass turf bioassays, mortality of nymphs was equivalent (85–95%) between treatments of chlorpyrifos (0.6 kg [AI]/ha, Safer's insecticidal soap, and Drione. In the same bioassays, nymphal mortality was moderate after treatment with Dri-die (23–29%) and low following application with diatomaceous earth (16%) and for untreated controls (6%).     

Susceptibility of certain Egyptian and imported wheat cultivars to infestation by Sitophilus oryzae and Rhyzopertha dominica

Stored wheat grains are vulnerable to attack by Sitophilus oryzae and Rhyzopertha dominica, which deteriorate the quality of the infested commodities, depletes nutrition value and makes them unfit for human consumption. The susceptibility of two Egyptian local (MISR1 and SIDS12) and four imported wheat (Hard Australian wheat, American Hard Red Winter, American Soft Red Winter and Romanian wheat) cultivars to infestation by S. oryzae and R. dominica was investigated. The Romanian cultivar was the most susceptible at infestation level of 20 adults/250g. For the other two infestation levels (10 and 15 adults/250g), the Romanian and American (SRW) varieties showed high susceptibility to S. oryzae by 5.13 and 5.52% and 4.32 and 4.76% for both varieties, in respect. On the other hand, the infestation with R. dominica showed the minimum weight loss in Romanian wheat and unexpectedly showed maximum number of emerged adults after three months storage. Maximum means of damaged grains were recorded in Romanian and American (HRW) cultivars caused by S. oryzae and R. dominica infestations, respectively. Moreover, chemical changes were recorded in nutritional composition of the grains when they were subjected to infestation with 20 unsexed adults/250g wheat by S. oryzae and R. dominica for three months storage.     

The culture ofEntomophthora muscae (C) Fres. in carrot flies (Psila rosae F.) and the effect of temperature on the pathology of the fungus

A method for maintaining anin vivo culture ofEntomophthora muscae (C) Fres. on its original host, adult carrot flies (Psila rosae F.), is described. The lethal time for adult carrot flies was greatly influenced by temperature, both for infected and for uninfected flies. In the range 8.2°C–20.2°C the LT₅₀ for infected flies was about 5.4 times shorter than the estimated average life-span for uninfected flies. The discharge of primary spores was also strongly dependent on temperature. The total number of primary spores discharged per fly at 100% RH and in darkness ranged between 1.2×10⁴ and 9.6×10⁴ with a mean of 5.1×10⁴.      

Lethality impact of diatomaceous earth (Sayan®), bran,sawdust and clay on adult of six stored-product insects

The lethality impact of diatomaceous earth (Sayan®), bran, sawdust and clay on six major stored-product insects under laboratory conditions was determined. The lethal effects were investigated in two different methods. In the first method, experimental surface was permeated with test material, and in the second method, the test compounds were mixed with rearing mediums separately. In the first technique, adults of Sitophilus oryzae, Rhyzopertha dominica, Oryzephilus surinamensis, Lasioderma serricorne, Cryptolestes ferrugineus and Callosobruchus maculatus were placed in a Petri dish, which was contaminated with a known amount of Sayan®, bran, sawdust or clay. Mortality rate was recorded at 24, 48 and 72 h after commence of each exposure. In the second method, the lethality of test compounds which were mixed with wheat was investigated against S. oryzae, R. dominica, O. surinamensis, L. serricorne and C. ferrugineus for 1, 7 and 14 days of exposure. In the first technique, the results indicated that bran and sawdust caused more mortality rate on S. oryzae compared to Sayan®. In the second method, the results demonstrated a higher mortality caused by bran and sawdust compared to Sayan® on S. oryzae. For L. serricorne, mortality rate caused by bran and sawdust was similar, whereas, in C. maculates case, mortality rate caused by bran was more than the other treatments. The protective effects of test compounds on life span of S. oryzae and R. dominica showed significant differences among the emerged adult of insects in different treatments. In the bran treatment, S. oryzae adult's emergence was the lowest, while, in the R. dominica case, a non-significant difference among resultant number of adult insects was detected.     

Infection and mortality of Microtheca ochroloma (Coleoptera: Chrysomelidae) by Isaria fumosorosea (Hypocreales: Cordycipitaceae) under laboratory conditions

Microtheca ochroloma Stål, the yellowmargined leaf beetle, is a pest in crucifer crops during the late fall and winter months in Florida. On organic farms, it is difficult to control due to the restricted use of insecticides, in addition to the lack of specific natural enemies. The objective of this study was to evaluate a blastospore-formulated product of Isaria fumosorosea (PFR-97^TM 20% WDG) against this beetle. In the first experiment, four of the beetle's life stages were treated with a suspension of 3 × 10⁷ blastospores/ml. Mean corrected mortality of treated insects was significantly higher in 1st and 3rd instars than in the egg, pupal, and adult stages. Larvae infected by I. fumosorosea exhibited reduced growth and unsuccessful molting. The second experiment quantified mortality of first instars of M. ochroloma by four concentrations of PFR-97^TM. Mean corrected larval infection/treatment was significantly (2.6 times) higher with a concentration of 4?g of product per 100?ml of water compared to concentrations of 1–3?g per 100?ml of water. Different factors that might have affected the pathogenicity of I. fumosorosea against M. ochroloma are discussed.     

Insecticidal efficacy of three vegetable oils as post-harvest grain protectants of stored wheat against Rhyzopertha dominica （F.） （Coleoptera： Bostrychidae）

The lesser grain borer, Rhyzopertha dominica is a major insect pests of stored grain in the tropics. Vegetable oils （chamomile, sweet almond and coconut） at 2.5, 3.5, 5.0, 7.0 and 10.0 mL/kg were tested against Rhyzopertha dominica （F.） in wheat grain. All bioassays were conductr, d at 30℃ and 65% ＋ 2% RH. Treatments with vegetable oils at high dose （10.0 mL/kg） achieved over 95% control within 24 h of exposure to freshly treated grain, There was little difference between the three oils in their effect. Persistence of oils in grains was tested at short-term storage intervals （48, 72 and 96 h） and intermediate-term intervals （10, 20 and 30 days） after treatments. The activity of all products decreased with storage period. Seed viability was reduced by the high dose rate （10.0 mL/kg） of oil treatments. The potential use of vegetable oils as supplementary or alternative grain protectants against insect damage in traditional grain storage in developing countries is discussed.     

Biochemical analysis and pathogenicity of entomopathogenic fungi to Diaphorina citri Kuwayama (Hemiptera: Liviidae)

Asiatic citrus psylla, Diaphorina citri, Kuwayama (Hemiptera: Liviidae) is an economic pest of citrus groves and a vector of the bacterium, Candidatus Liberibacter spp., one of the causative agents of citrus greening. In order to estimate the infectivity of six different isolates of Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea, fungal bioassay was performed on the adults of D. citri. Adults of D. citri were treated individually with 1 × 10⁵, 1 × 10⁶, 1 × 10⁷, 1 × 10⁸, 2 × 10⁸ spores/mL fungal concentrations by the immersion method. Subsequent to fungal bioassay, treated D. citri were used to determine the levels of esterase and glutathione S‐transferases (GST) enzymes over a period of 3–7 days. The mortality results suggest that I. fumosorosea isolates (If‐02) caused 82.2% mortality on the seventh day of treatment. However, B. bassiana isolate (Bb‐08) with lowest LC₅₀ (1.4 × 10⁷ spores/mL) proved to be highest potential isolate against D. citri. Biochemical determination of esterase and GST activity assay showed significant differences in activities after infection of fungi. Significantly high activity of esterase was observed by Bb‐01 (27.0 unit per mg protein) on the seventh day, while Ma‐11.1 and If‐2.3 (16.9 and 36.3 unit per mg protein) on the third day post treatment. However, maximum GST's activity was showen by isolates Bb‐08,Ma‐M₂ and If‐2.3 (37.6, 1.40 and 10.9 unit per mg protein) on the third day. The current investigation will help to explore the relations between the insect defense system and entomopathogenic fungi. Moreover, the determination of enzymatic activities will be useful for selecting the most pathogenic isolates.     

Insecticidal effect of the combined application of spinosad,Beauveria bassiana and diatomaceous earth for the control of Tribolium confusum

Laboratory bioassays were carried out to evaluate the insecticidal effect of Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) germinated conidia, a commercial formulation of diatomaceous earth and a liquid formulation of spinosad, applied alone or in all possible combinations on wheat, against adults of the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae). Diatomaceous earth was applied at one (200 ppm), the fungus at two (100 and 300 ppm) and spinosad at three (0.01, 0.1 and 0.5 ppm) dose rates, whereas all combinations of two or three of the insecticide concentrations were also tested. Insect mortality was determined after 2, 7 and 14 d of exposure. The simultaneous use of B. bassiana germinated conidia with diatomaceous earth notably increased mortality of T. confusum adults by sixfold 7 d after exposure, as compared with either B. bassiana or diatomaceous earth alone. When diatomaceous earth and spinosad were combined, diatomaceous earth did not increase the efficacy of spinosad. In general, the combination of all three agents did not differ from either the application of the agents tested alone, or their binary combinations, in most treatments examined.     

Time-dose-mortality modelling and virulence indices for six strains of Verticillium lecanii against sweetpotato whitefly, Bemisia tabaci (Gennadius)

Bioassays of six strains of Verticillium lecanii (Zimmermann) Viégas were conducted with the sweet potato whitefly, Bemisia tabaci. For inoculation, batches of third‐instar whitefly nymphs on sweetpotato seedlings were immersed in conidial suspensions of five dosages at concentrations from 10³ to 10⁷ conidia/ml. Each dosage was used to inoculate 150–250 nymphs. The nymphs were maintained at 25°C, under 95% RH, photo phase of 16 : 8 (L : D) and observed daily for mortality. The resulting data were analysed over 8 days by a complementary log‐log (CLL) time‐dose‐mortality model, based on the Hosmer–Lemeshow test, analysing the time‐dose trends for five concentrations of six strains of V. lecanii simultaneously. The parameters from the model were used to estimate the virulence indices (the values of LC₅₀) of six strains against B. tabaci. Based on the time‐dose‐mortality relationships fitted and the virulence indices, the virulence of the six strains of V. lecanii for B. tabai was compared. Results indicated that the strain Vl6063 imported from Canada and the domestic strains V3450 and Vp28 derived from B. tabaci and a scale insect, respectively, were more virulent than the others with LC₅₀ values of 2.57 × 10⁵, 6.03 × 10⁵ and 6.03 × 10⁵, respectively.     

Evaluation of Pochonia chlamydosporia var. chlamydosporia as a control agent of Meloidogyne javanica on pistachio

The potential of isolates of Pochonia chlamydosporia var. chlamydosporia as biocontrol agents for root-knot nematodes was investigated in vitro and on pistachio plants. On potato dextrose agar, growth of all isolates started at temperatures above 10°C, reached maximum between 25 and 28°C and slowed down at 33°C. On water agar, all isolates parasitized more than 85% of the eggs of Meloidogyne javanica at 18°C after 3 weeks. Filtrates of isolates grown on malt extract broth did not cause more than 5% mortality on second-stage juveniles of M. javanica after 48 h of incubation. A single application of 10×10³ chlamydospores (produced on sand–barley medium) g^–1 soil, was applied to unsterilised soil planted with pistachio cv. Kalehghochi, and plants were inoculated with 3000 nematode eggs. After 120 days in the glasshouse, nematode multiplication and damage were measured. Ability of fungus isolates to survive in the soil and to grow on roots were estimated by counting colony forming units (cfu) on semi-selective medium. Fungal abundance in soil increased nearly 3-fold and 10×10³ and 20×10³ cfu g^–1 root of pistachio were estimated in pots treated with isolates 40 and 50, respectively. Strain 50 was more abundant in soil and on the roots, infected more eggs (40%) on the roots and controlled 56% of total population of M. javanica on pistachio roots, whereas isolate 40 parasitized 15% of the eggs on the roots and controlled ca. 36% of the final nematode population.