Comparison of Methods for Isolating Entomopathogenic Fungi from Soil Samples.

The goal of the present study is to compare the effectiveness of using insect baits versus artificial selective medium for isolating entomopathogenic fungi (EPF) from soil samples. The soil is a rich habitat for microorganisms, including EPF particularly belonging to the genera Metarhizium and Beauveria, which can regulate arthropod pests. Biological products based on fungi are available in the market mainly for agricultural arthropod pest control. Nevertheless, despite the high endemic biodiversity, only a few strains are used in commercial bioproducts worldwide. In the present study, 524 soil samples were cultured on potato dextrose agar enriched with yeast extract supplemented with chloramphenicol, thiabendazole, and cycloheximide (CTC medium). The growth of fungal colonies was observed for 3 weeks. All Metarhizium and Beauveria EPF were morphologically identified at the genus level. Additionally, some isolates were molecularly identified at the species level. Twenty-four out of these 524 soil samples were also surveyed for EPF occurrence using the insect bait method (Galleria mellonella and Tenebrio molitor). A total of 51 EPF strains were isolated (41 Metarhizium spp. and 10 Beauveria spp.) from the 524 soil samples. All fungal strains were isolated either from croplands or grasslands. Of the 24 samples selected for comparison, 91.7% were positive for EPF using Galleria bait, 62.5% using Tenebrio bait, and 41.7% using CTC. Our results suggested that using insect baits to isolate the EPF from the soil is more efficient than using the CTC medium. The comparison of isolation methods in addition to the identification and conservation of EPF has a positive impact on the knowledge about biodiversity. The improvement of EPF collection supports scientific development and technological innovation.

Acute toxicity of Bisphenol A (BPA) to tropical marine and estuarine species from different trophic groups.

BPA is chemical pollutant of very high concern due to its toxicity to the environment and risks for human health. Environmental concern consists in BPA entrance into aquatic ecosystems due to acute and chronic toxicity to invertebrates and vertebrates. This study aimed to determine acute BPA toxicity to tropical estuarine-marine species of four trophic levels and integrate BPA toxicity values using species sensitivity distribution (SSD) analysis. Our hypothesis is that BPA toxicity increases towards higher trophic levels. Microalga (Tetraselmis sp.), zooplanktonic grazer (Artemia salina), deposit-feeder invertebrate (Heleobia australis), and omnivorous fish (Poecilia vivipara) were chosen as experimental models. Tetraselmis sp. showed the highest BPA tolerance, without a concentration-dependent response. Species sensitivity have increased from A. salina (LC50,96h = 107.2 mg L-1), followed by H. australis (LC50,96h = 11.53.5 mg L-1), to P. vivipara (LC50,96h = 3.5 mg L-1). Despite the toxicity hierarchy towards trophic levels, which partially supported our hypothesis, SSD did not evidence a clear pattern among estuarine-marine trophic groups. Our study disclosed the sensitivity of not yet investigated species to BPA and, in an integrative way, highlighted BPA toxic effects at different trophic levels. Although estimated acute hazardous concentration (HC5 = 1.18 mg L-1) for estuarine and marine species was higher than environmentally relevant concentrations, sublethal adverse effects induced by BPA exposure may lead to unbalances in population levels and consequently affect the ecological functioning of tropical coastal systems.

Anti-helminthic activity of Momordica charantia L. against Fasciola hepatica eggs after twelve days of incubation in vitro.

Fasciolosis, a parasitic disease caused by the trematode Fasciola hepatica underreported is expanding both in human and animal population, throughout the world. The constant use of synthetic drugs to treat this condition has led to the natural selection of resistant strains of the parasite. Hence, there is a growing focus on the potential anti-helminthic properties of medicinal plants and phytopharmaceuticals. The current study assessed the potential anti-fasciolicide action of Momordica charantia leaf extracts and fractions on the eggs of F. hepatica parasites. The lyophilized crude extract (CE) of M. charantia leaves and its sub-fractions, obtained from liquid-liquid partitioning with organic solvents, were analysed by High Performance Liquid Chromatography (HPLC), suspended in 1% DMSO and used in in vitro tests. Quadruplicates of 50F. hepatica eggs were incubated at 23°C with M. charantia leaf CE in different concentrations. After 12days no larvae were formed in eggs incubated with CE concentrations above 12.5mg/mL. Eggs incubated with CE sub-fractions at concentrations of 1000, 100, 10, 1, 0.1, 0.01µg/mL affected embryonic development, with n-butanol presenting the strongest inhibition of miracidia formation. In contrast, on the 12th day, 90% of the miracidia hatched in the control experiments using 0.03% DMSO whereas embryogenesis was completely abolished with any concentration of albendazole sulphoxide ABZ(SO). Chemical analysis of the CE and sub-fractions revealed a prominent presence of flavonoids. HPLC-MS confirmed Quercetin to be one of the main flavonoids present in the CE and the n-butanol subfraction. This is the first study to analyse the potential anti-fasciolicide action of M. charantia leaf CE and subfractions.