Evaluation of the efficacy of entomopathogenic nematodes on Ctenocephalides felis felis larvae (Siphonaptera: Pulicidae)

Abstract Ctenocephalides felis felis is a relevant and widely distributed ectoparasite that acts as a vector of disease-causing pathogens. Moreover, it is responsible for economic losses due the use of harmful chemicals to the environment and that favor the emergence of insecticide-resistant populations. Research on entomopathogenic nematodes may open up an alternative route to the insect chemical control. The present study aimed to evaluate the killing efficacy of Heterorhabditis bacteriophora (HP88) against C. felis larvae in 400 μL, 600 μL and 1000 μL of suspension containing 120, 160 and 200 infective juveniles/larva and 600 μL of suspension containing the same concentrations of Heterorhabditis indica (LPP30), divided into two groups (absence and presence of diet) and a control group with three replications containing only distilled water. In the bioassay with H. bacteriophora, the groups in 600 μL of suspension showed higher mortality rates than those in the other tested volumes, which were above 80% at all concentrations. On the other hand, H. indica achieved mortality rates above 70% at all concentrations used. Results indicate that flea larvae are susceptible to in vitro infection by H. bacteriophora and H. indica

Ultrastructural and Cytotoxic Effects of Metarhizium robertsii Infection on Rhipicephalus microplus Hemocytes

Metarhizium is an entomopathogenic fungus widely employed in the biological control of arthropods. Hemocytes present in the hemolymph of invertebrates are the cells involved in the immune response of arthropods. Despite this, knowledge about Rhipicephalus microplus hemocytes morphological aspects as well as their role in response to the fungal infection is scarce. The present study aimed to analyze the hemocytes of R. microplus females after Metarhizium robertsii infection, using light and electron microscopy approaches associated with the cytotoxicity evaluation. Five types of hemocytes (prohemocytes, spherulocytes, plasmatocytes, granulocytes, and oenocytoids) were described in the hemolymph of uninfected ticks, while only prohemocytes, granulocytes, and plasmatocytes were observed in fungus-infected tick females. Twenty-four hours after the fungal infection, only granulocytes and plasmatocytes were detected in the transmission electron microscopy analysis. Hemocytes from fungus-infected tick females showed several cytoplasmic vacuoles with different electron densities, and lipid droplets in close contact to low electron density vacuoles, as well as the formation of autophagosomes and subcellular material in different stages of degradation could also be observed. M. robertsii propagules were more toxic to tick hemocytes in the highest concentration tested (1.0 × 108 conidia mL−1). Interestingly, the lowest fungus concentration did not affect significantly the cell viability. Microanalysis showed that cells granules from fungus-infected and uninfected ticks had similar composition. This study addressed the first report of fungal cytotoxicity analyzing ultrastructural effects on hemocytes of R. microplus infected with entomopathogenic fungi. These results open new perspectives for the comprehension of ticks physiology and pathology, allowing the identification of new targets for the biological control

Antifungal activity of Stenotrophomonas maltophilia in Stomoxys calcitranslarvae

The microbiota present in Stomoxys calcitrans larvae may assist their survival in contaminated environments through production of inhibitory substances. Bacteriological identification methods, the polymerase chain reaction (PCR) and scanning electron microscopy (SEM) were used to detect a bacterium naturally present in mucus and maceratedS. calcitrans larvae. The antifungal activity was determined based on the results from disk diffusion tests on an artificial solid medium. The bacterium was identified as Stenotrophomonas maltophilia and presented antifungal activity againstBeauveria bassiana sensu lato isolates CG 138, CG 228 and ESALQ 986. This result suggests that the larval microbiota is a factor that can compromise the use of B. bassiana s.l. fungus for biological control of S. calcitrans larvae

Isolation of fungi in Musca domestica Linnaeus, 1758 (Diptera: Muscidae) captured at two natural breeding grounds in the municipality of Seropédica, Rio de Janeiro, Brazil

The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica. The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus (23.8%), A. niger var. niger (14.4%), Penicillium corylophilum (21.4%), P. fellutanum (11.9%), Cladosporium cladosporoides (4.7%), Fusarium sp. (4.7%), Alternaria alternata (11.9%), Curvularia brachyspora (2.4%), Mycelia sterilia (2.4%) and the Mucorales order (2.4%)

Efeito dos fungos entomopatogenicos Metarhizium anisopliae e Beauveria bassiana em ovos de Rhipicephalus sanguineus (acarlixodidae) Effect of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana on eggs of Rhipicephalus sanguineus (acarlixodidae)

Este experimento foi conduzido para conhecer os efeitos de duas espécies de fungos, Beauveria bassiana e Metarhizium anisopliae em ovos do carrapato Rhipicephalus sanguineus sob temperatura e umidade constantes (27 &plusmn; 1&deg; C e > 80% de umidade relativa (UR)) e variáveis (25 &plusmn;5&deg; C e > 60% UR). Ovos de R. sanguineus foram imersos em quatro concentrações (10(8), 10(7), 10(6) e 10(5) conídios/ml) de dois isolados de B. bassiana (Bb 986 e Bb 747) e três isolados de M. anisopliae ( Ma 959, Ma E9 e Ma 319). Os períodos de incubação e eclosão das larvas de todos os ovos tratados foram estatisticamente não significantes (P>0,05). Entretanto, a porcentagem de eclosão das larvas, oriundas dos ovos tratados, foi inversamente proporcional à concentração de conídios/ml. A porcentagem de eclosão variou entre 0 à 26,66% para a concentração 10(8) conídios/ml, e 6,66 à 100% para a concentração 10(5) conídios/ml. A porcentagem de eclosão das larvas de ovos mantidos em temperatura e umidade variáveis foi menor quando comparada com a das larvas de ovos mantidos em temperatura de 27&deg;C e umidade > 80% UR (PThis experiment was conducted to evaluate the effects of two species of fungi Beauveria bassiana and Metarhizium anisopliae on the eggs of the tick Rhipicephalus sanguineus at constant (27 &plusmn; 1&deg; C and > 80% RH) and variable (25 &plusmn; 5&deg; C and > 60% RH), temperatures and humidities. Eggs of R. sanguineus were immersed in four concentrations (10(8), 10(7), 10(6), 10(5) conidia/ml) of two isolated of B. bassiana (Bb 986 and Bb 747) and three isolated of M. anisopliae ( Ma 959, Ma E9 and Ma 319). The incubation and hatch periods of all treated eggs and controls were statistically not significant (P> 0.05). Otherwi- se, the percentage of hatch of larvae in all treated eggs was inversely proportional to the concentration of conidia/ml. The percentage of hatch ranged from 0 to 26.66% for the 10(8) concentration and from 6.66 to 100% for the 10(5) concentration. The percentage of eclosion of eggs maintened at variable temperatures and humiditis were lower as compared with eggs maintened at 27 &plusmn; l&deg; C and > 80% RH (P< 0.05). The DL 50 and 90 for inhibicion of eclosion of larvae were also lower at variable temperatures and humidities (P<0.05)

Isolation of Fungi in Musca domestica Linnaeus, 1758 (Diptera: Muscidae) Captured at Two Natural Breeding Grounds in the Municipality of Seropédica, Rio de Janeiro, Brazil

The objective of this study was to isolate and identify fungal species found in natural association with adults of Musca domestica   . The adult insects were collected from two natural breeding grounds: hog pens and an urban sanitary landfill. The isolated fungi were identified as: Aspergillus flavus   (23.8%), A. niger var. niger (14.4%), Penicillium corylophilum   (21.4%), P. fellutanum (11.9%), Cladosporium cladosporoides   (4.7%), Fusarium   sp. (4.7%), Alternaria alternata   (11.9%), Curvularia brachyspora   (2.4%), Mycelia sterilia   (2.4%) and the Mucorales order (2.4%)

Antimicrobial activity of Stomoxys calcitransagainst Beauveria bassiana sensu lato isolates

This study had the aims of evaluating the antimicrobial characteristics of Stomoxys calcitrans (Diptera: Muscidae) larvae against the fungal isolates CG138, CG228 and ESALQ986 of Beauveria bassiana sensu lato (Balsamo-Crivelli) Vuillemin, 1912 (Hypocreales: Cordycipitaceae). S. calcitrans eggs, larvae and pupae were exposed to these same isolates. Statistical analysis showed that the immature stages of S. calcitrans were not susceptible to the fungal isolates used, regardless of the exposure method. Diffusion test on solid culture medium reveled that macerated S. calcitrans larvae exposed to isolate CG138 reduced CG138 fungal development. The analysis of the chromatographic profiles indicated that the macerate or mucus of larvae of the control group and the groups exposed to the isolate CG138 presented different profiles. Reduced development of the isolate CG138 on the larvae cuticle was observed by means of scanning electron microscopy