Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR

The objective of this work was to evaluate the survival of two Trichoderma harzianum co-transformants, TE 10 and TE 41, carrying genes for green fluorescent protein (egfp) and for resistance to benomyl, during four weeks in a contained soil microcosm. Selective culture media were used to detect viable fungal material, whose identity was confirmed by the observation of the fluorescent phenotype by direct epifluorence microscopy. PCR using two nested primer pairs specific to the egfp gene was also used to detect the transformed fungi. Although it was not possible to reliably detect the egfp gene directly from soil extracts, an enrichment step involving selective culture of soil samples in liquid medium prior to DNA extraction enabled the consistent detection of the T. harzianum co-transformants by nested PCR for the duration of the incubation period.

Pachycondyla obscuricornis as natural enemy of the spittlebug Deois flavopicta

The objective of this work was to evaluate the potential control of the ant Pachycondyla obscuricornis Emery (Hymenoptera Formicidae) on populations of nymphs of the spittlebug, Deois flavopicta Stal (Hemiptera Cercopidae). Foraging behavior and prey consumption rate of P. obscuricornis were evaluated. Field data revealed that P. obscuricornis does not show aggressive behavior against individuals of the same species, when they are not carrying a prey; they can patrol distances larger than 10 m searching for prey, and they can build their nest as close as 1 m from each other. The ant has a solitary patrolling habit, there is no recruitment behavior, and individuals dislocate fast, browsing on soil and vegetation for prey. Predation rate on spittlebug nymphs increased relative to the spittlebug abundance, reaching 93.8% of captured prey. Pachycondyla obscuricornis is a voracious predator and may control the population of spittlebugs in cultivated pastures.

Emergence, longevity and fecundity of Trissolcus basalis and Telenomus podisi after cold storage in the pupal stage

Pupae of Trissolcus basalis (Wollaston) and Telenomus podisi Ashmead (Hymenoptera: Scelionidae) were stored at 12ºC and 15ºC for 120-210 days, after different periods of parasitism at 18ºC in order to evaluate adult emergence, longevity and ovipositional capacity. There was no emergence of adults at 12ºC. The rate of emergence of parasitoids transferred to 15ºC at the beginning of the pupal stage was 1.5% and 26.3%, for T. basalis and T. podisi respectively, whereas those parasitoids transferred one day before the expected date of emergence at 18ºC showed 86.4% of emergence for T. basalis and 59.9% for T. podisi. Mean adult longevity was also significantly lower when pupae were transferred to 15ºC at the beginning of the pupal stage. Females emerged after storage and maintained for 120 to 210 days at 15ºC parasitized host eggs after transference to 25ºC; however, fecundity of T. podisi was reduced in about 80% after cold storage.

Parasitism by Campoletis flavicincta on Spodoptera frugiperda in corn

Parasitism by Campoletis flavicincta (Ashmead) (Hymenoptera: Ichneumonidae) on Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) and consequent reduction of production losses were evaluated on caged corn plants in the field. Treatments consisted of plots infested with 0 (control), 15 and 30 pairs of C. flavicincta with egg masses per cage and plot infested without cage and liberation of the parasitoid. Parasitoid release was done when S. frugiperda larvae were three-day-old. Fifty corn plants (40%) per plot were collected seven days after infestation and S. frugiperda larvae present were reared in glass cups on an artificial diet. Number of S. frugiperda larvae was reduced by C. flavicincta but mortality of the pest and parasitoid sex ratio in laboratory were similar among treatments. Total progeny and female production from collected larvae were similar among densities of released parasitoid. Parasitism rate was higher on 30 than on 15 pairs of C. flavicincta. Damage on corn plants at seven and 14 days after S. frugiperda infestation had similar grades at 0, 15 or 30 C. flavicincta pairs and higher values than the plots without cage. Damage by S. frugiperda was lower at 30 C. flavicincta pairs after 21 days of infestation. Final stand, stand reduction by plant death and corn productivity were similar among treatments.

Parasitism of the mite Acarophenax lacunatus on Tribolium castaneum

The objective of this work was to assess the parasitism potential of Acarophenax lacunatus (Cross & Krantz) on Tribolium castaneum (Herbst). The experimental units encompassed Petri dishes containing 25 g of wheat infested with 20 adults of this beetle species. After seven days of infestation, different mite densities (0, 2, 4, 6, 8 and 10 females) were inoculated into the Petri dishes. The increase in mite density led to a linear increase in parasitism, and a consequent reduction in populations of this insect. It is possible to use this mite species in the biological control of T. castaneum.

Induction of systemic resistance in tomato by the autochthonous phylloplane resident Bacillus cereus

The objective of this work was to verify if the induced resistance mechanism is responsible for the capacity of a phylloplane resident bacteria (Bacillus cereus), isolated from healthy tomato plants, to control several diseases of this crop. A strain of Pseudomonas syringae pv. tomato was used as the challenging pathogen. The absence of direct antibiosis of the antagonist against the pathogen, the significant increase in peroxidases activity in tomato plants exposed to the antagonist and then inoculated with the challenging pathogen, as well as the character of the protection, are evidences wich suggest that biocontrol efficiency presented by the antagonist in previous works might be due to induced systemic resistance (ISR).

Scanning electron microscopy of the interaction between Cryptococcus magnus and Colletotrichum gloeosporioides on papaya fruit

The objective of this work was to investigate possible modes of action of the yeast Cryptococcus magnus in controlling anthracnose (Colletotrichum gloeosporioides) on post harvested papaya fruits. Scanning electron microscopy was used to analyze the effect of the yeast on inoculations done after harvest. Results showed that C. magnus is able to colonize wound surfaces much faster than the pathogen, outcompeting the later for space and probably for nutrients. In addition, C. magnus produces a flocculent matrix, which affects hyphae integrity. The competition for space and the production of substances that affect hyphae integrity are among the most important modes of action of this yeast.

Euschistus heros mass rearing technique for the multiplication of Telenomus podisi

The objective of this work was to improve the mass rearing technique of Euschistus heros in laboratory. Nymphs and adults were reared at densities 100, 200, 300 and 400 eggs per Petri dish (9 cm diameter), and at 50, 100, 150 and 200 couples per rearing cages (900 mL), respectively. Survival rate of immature stages and survivorship and reproduction of adults were determinated. Survivorship of nymph to adult was the highest (89%) at density 100 eggs per dish. Adult survivorship was independent of density, and 100 couples per cage were the best to improve quality of the produced progeny. In these conditions, fecundity was 160.8±9.28 eggs per female, and a total of 8,950±456 eggs per cage per month was produced. Two hundred couples per cage showed a negative effect on reproduction, which decreased to 65%. With this technique, a colony of 35 cages with 100 couples per cage yields about 313.3 thousands eggs per month, which is enough to supply the egg parasitoid Telenomus podisi to colonize about 35 ha of soybean field.

New species of Rhinoleucophenga, a potential predator of pineapple mealybugs

The objective of this work was to describe a new species of Rhinoleucophenga (Diptera: Drosophilidae). Rhinoleucophenga capixabensis sp. nov. is described based on specimens collected from pineapple (Ananas comosus var. comosus) infested with Dysmicoccus brevipes (Hemiptera: Pseudococcidae) in the state of Espírito Santo, Brazil. Distinguishing characters of R. capixabensis sp. nov. include hyaline wings, a strong seta on the second antennal segment, body length of about 2-3 mm, uniformly dark brown scutellum, one pair of strong prescutellar setae, ventral epandrial lobes with about 17-18 teeth, and female cerci with approximately 20 long setae.

Biology and morphometrics of the immature stages of Epinotia aporema on artificial diet

The objective of this work was to establish a life table for the immature stages of Epinotia aporema, as part of a wider investigation on its biological control. Insects were reared on an artificial diet at 25±1ºC and a 16:8 (light:dark) hour photoperiod. For the identification of larval instars for the study of pathogen-insect interactions under laboratory conditions, head capsule widths (HCWs) were also determined. The egg incubation period was 4.13±0.30 days, larval stage took 11.64±0.49 days, and the development time of the pupal phase was sex-dependent with 8.51±0.69 days for the females and 9.41±0.65 days for the males. Five larval instars were identified.

Induced-feeding bioassays for detection of Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera)

The objective of this work was to establish and test the induced-feeding bioassay in order to detect Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera: Coccinellidae). Larvae were induced to swallow high concentrations of spore-crystal suspensions of seven exotic and 30 Argentine B. thuringiensis strains. The great majority of strains showed no toxicity to E. paenulata larvae, and observed mortality was lower than 30%. Induced-feeding bioassay is feasible, and should be used for prospecting strains that produce right combinations of Cry proteins needed to an efficient pest control.

Egg parasitoid wasps as natural enemies of the neotropical stink bug Dichelops melacanthus

The objective of this work was to determine the potential of five species of Scelionidae wasps - Telenomus podisi, Trissolcus basalis, Trissolcus urichi, Trissolcus teretis and Trissolcus brochymenae - as natural enemies of the neotropical stink bug Dichelops melacanthus, and to determine if the presence of eggs of other stink bug species influences the parasitism and development of the parasitoids. Two kinds of experiments were done in laboratory: without choice of hosts (eggs of D. melacanthus) and with choice (eggs of D. melacanthus and of Euschistus heros). Biological parameters, including proportion of parasitism, immature survivorship, progeny sex ratio, immature stage development period, and host preference were recorded. All the evaluated parasitoids can parasitize and develop on D. melacanthus eggs. The first choice of eggs did not influence the proportion of D. melacanthus eggs parasitized by Tr. basalis, Tr. teretis or Tr. brochymenae. However, D. melacanthus eggs as the first choice of Te. podisi and Tr. urichi increased, respectively, 9 and 14 times the chance for parasitism on eggs of this species. Behavioral and ecological aspects of parasitoids should be considered prior to their use in biological control programs.

Bacterial spot and early blight biocontrol by epiphytic bacteria in tomato plants

The objective of this work was to evaluate in vitro and in vivo biocontrol of bacterial spot (Xanthomonas vesicatoria) and early blight (Alternaria solani) by the epiphytic bacteria Paenibacillus macerans and Bacillus pumilus. Tomato plants were previously sprayed with epiphytic bacteria, benzalkonium chloride and PBS buffer and, after four days, they were inoculated with A. solani and X. vesicatoria. To determine the phytopathogenic bacteria population, leaflet samples were collected from each treatment every 24 hours, for seven days, and plated on semi-selective medium. The effect of epiphytic bacteria over phytopathogens was performed by the antibiosis test and antagonistic activity measured by inhibition zone diameter. The epiphytic and benzalkonium chloride drastically reduced the severity of early blight and bacterial spot in comparison to the control (PBS). In detached leaflets, the epiphytic bacteria reduced in 70% the number of phytopathogenic bacteria cells in the phylloplane. The antibiosis test showed that the epiphytic bacteria efficiently inhibit the phytopathogens growth. In all the bioassays, the epiphytic bacteria protect tomato plants against the phytopathogens

Biology and parasitic potential of Doryctobracon areolatuson Anastrepha fraterculus larvae

The objective of this work was to study the biology and parasitic potential of Doryctobracon areolatus on larvae of Anastrepha fraterculus. The egg-adult period, the sex ratio, the longevity, the pupal viability, and the parasitism rate of D. areolatus were determined in laboratory, using A. fraterculus as host. The parasitoid development from oviposition to adult emergence required 25.00±1.70 days, the sex ratio was 0.62±0.09, and the mean longevity was 16.36±3.62 days for males and 10.24±1.71 days for females. The mean parasitism rate was 53.50±8.93%, varying from 41.60 to 68.60%, which shows the potential of this parasitoid for biological control of A. fraterculus.

Biocontrol of seed pathogens and growth promotion of common bean seedlings by Trichoderma harzianum

The objective of this work was to evaluate isolates of Trichoderma harzianum regarding biocontrol of common bean seed-borne pathogens, plant growth promotion, and rhizosphere competence. Five isolates of T. harzianum were evaluated and compared with commercial isolate (Ecotrich), Carboxin+Thiram, and an absolute control. Bean seeds of the cultivar Jalo Precoce, contaminated with Aspergillus, Cladosporium, and Sclerotinia sclerotiorum, were microbiolized with antagonists, and seed health tests were carried out. Isolates were evaluated on autoclaved substrate and in field conditions. Ten days after sowing (DAS), plant length was measured. To test rhizosphere competence, isolates were applied in boxes containing autoclaved washed sand, and root colonization was evaluated at 10 DAS, using five plants per box. The most effective isolates in the seed health tests were: CEN287 and CEN289 to control Aspergillus; the commercial isolate to control Cladosporium; and CEN287 and CEN316 to control S. sclerotiorum. Isolates CEN289 and CEN290 promoted bean growth in greenhouse and field. Seed treatment with T. harzianum reduces the incidence of Aspergillus, Cladosporium, and S. sclerotiorum in 'Jalo Precoce' common bean seeds.