[Identification and antifungal activity of halophilic bacteria isolated from saline soils in Campeche, México]. / Identificación y actividad antifúngica de bacterias halófilas aisladas de suelos salinos en Campeche, México.

Phytopathogenic fungi Alternaria alternata and Colletotrichum gloeosporioides cause diseases in plant tissues as well as significant postharvest losses. The use of chemical fungicides for their control has negative effects on health and the environment. Secondary metabolites from halophilic bacteria are a promising alternative for new antifungal compounds. In the present study, halophilic bacteria were isolated and characterized from two sites with saline soils called branquizales in Campeche, Mexico. A total of 64 bacteria were isolated. Agrobacterium, Bacillus, Inquilinus, Gracilibacillus, Metabacillus, Neobacillus, Paenibacillus, Priestia, Staphylococcus, Streptomyces and Virgibacillus were among the identified genera. The antifungal potential of the culture supernatant (CS) of 39 halophilic bacteria was investigated against C. gloeosporioides and A. alternata. The bacteria showing the greatest inhibition of mycelial growth corresponded to Bacillus subtilis CPO 4292, Metabacillus sp. CPO 4266, Bacillus sp. CPO 4295 and Bacillus sp. CPO 4279. The CS of Bacillus sp. CPO 4279 exhibited the highest activity and its ethyl acetate extract (AcOEt) inhibited the germination of C. gloeosporioides, with IC50 values of 8,630µg/ml and IC90 of 10,720µg/ml. The organic partition of the AcOEt extract led to three fractions, with acetonitrile (FAcB9) showing the highest antifungal activity, with values exceeding 66%. Halophilic bacteria from 'blanquizales' soils of the genus Bacillus sp. produce metabolites with antifungal properties that inhibit the phytopathogenic fungus C. gloeosporioides.

Plant Extracts from the Yucatan Peninsula in the In Vitro Control of Curvularia lunata and Antifungal Effect of Mosannona depressa and Piper neesianum Extracts on Postharvest Fruits of Habanero Pepper.

Plant extracts are a valuable alternative for the control of phytopathogenic fungi in horticultural crops. In the present work, the in vitro antifungal effect of ethanol and aqueous extracts from different vegetative parts of 40 native plants of the Yucatan Peninsula on Curvularia lunata ITC26, a pathogen of habanero pepper (Capsicum chinense), and effects of the most active extracts on postharvest fruits were investigated. Among these, the ethanol extracts of Mosannona depressa (bark from stems and roots) and Piper neesianum (leaves) inhibited 100% of the mycelial growth of C. lunata. The three extracts were partitioned between acetonitrile and n-hexane. The acetonitrile fraction from M. depressa stem bark showed the lowest mean inhibitory concentration (IC50) of 188 µg/mL against C. lunata. The application of this extract and its active principle α-asarone in the postharvest fruits of C. chinense (500 µg/mL) was shown to inhibit 100% of the severity of the infection caused by C. lunata after 11 days of contact. Both samples caused the distortion and collapse of the conidia of the phytopathogen when observed using electron microscopy at 96 h. The spectrum of M. depressa enriched antifungal action is a potential candidate to be a botanical fungicide in the control of C. lunata in cultivating habanero pepper.

Activity of Aqueous Extracts from Native Plants of the Yucatan Peninsula against Fungal Pathogens of Tomato In Vitro and from Croton chichenensis against Corynespora cassiicola on Tomato.

Plant extracts are a valuable alternative to control pathogens of horticultural crops. In the present study, four species of pathogenic fungi were isolated from leaf spots on Solanum lycopersicum and identified by traditional and molecular techniques as Alternaria alternata ITC24, Corynespora cassiicola ITC23, Curvularia lunata ITC22, and Fusarium equiseti ITC32. When 11 aqueous extracts from eight native plants of the Yucatan Peninsula were tested against the four fungi in vitro, the extract from Croton chichenensis roots was most active, inhibiting mycelial growth (79-100%), sporulation (100%), and conidial germination (71-100%) at 3% (w/v). A logarithmic-diagrammatic scale of the pathosystem C. cassiicola-S. lycopersicum was established and used to assess disease severity on inoculated tomato plants in a greenhouse after treatment with the aqueous extract from C. chichenensis roots at 12% (w/v). After 21 days, the disease severity was 57% lower than on the control without extract applied. This dose of the extract was not phytotoxic to tomato leaves and was compatible with the beneficial organisms Bacillus subtilis CBCK47 and Trichodema asperellum Ta13-17. The antifungal efficacy of C. chichenensis is highly promising for incorporation into integrated disease management of tomato crops.

Nematicidal Screening of Aqueous Extracts from Plants of the Yucatan Peninsula and Ecotoxicity.

Active metabolites from plants are considered safer than synthetic chemicals for the control of plant-parasitic nematodes of the genus Meloidogyne. In the present work, 75 aqueous extracts (AEs) from different vegetative parts of 34 native plant species of the Yucatan Peninsula were evaluated against second-stage juveniles (J2s) of Meloidogyne incognita and M. javanica in microdilution assays. The highest mortality (M) against both Meloidogyne species was produced by the foliar AE from Alseis yucatanensis (M ≥ 94%) and Helicteres baruensis (M ≥ 77%) at 3% w/v after 72 h. Other active AEs at 3% were from the leaves of Croton itzaeus and stems of H. baruensis (M: 87-90%) on M. javanica and the stems of Annona primigenia and the leaves of Morella cerifera on M. incognita (M: 92-97%). The AEs from A. yucatanensis had the lowest LD50 against M. incognita (0.36% w/v), and against M. javanica (3.80% w/v). In an acute ecotoxicity assay of the most promising AEs using non-target earthworms (Eisenia fetida), the AE of A. yucatanensis had slight acute toxicity (LD50: 2.80% w/v), and the rest of the most active AEs were not ecotoxic. These tropical plants are potential candidates for further studies as biorational agents for controlling Meloidogyne species.

In Vitro Assessment of Organic and Residual Fractions of Nematicidal Culture Filtrates from Thirteen Tropical Trichoderma Strains and Metabolic Profiles of Most-Active.

The nematicidal properties of Trichoderma species have potential for developing safer biocontrol agents. In the present study, 13 native Trichoderma strains from T. citrinoviride, T. ghanense (2 strains), T. harzianum (4), T. koningiopsis, T. simmonsii, and T. virens (4) with nematicidal activity were selected and cultured in potato dextrose broth to obtain a culture filtrate (CF) for each. Each CF was partitioned with ethyl acetate to obtain organic (EA) and residual filtrate (RF) fractions, which were then tested on second-stage juveniles (J2s) of the nematodes Meloidogyne javanica and M. incognita in a microdilution assay. The most lethal strains were T. harzianum Th43-14, T. koningiopsis Th41-11, T. ghanense Th02-04, and T. virens Th32-09, which caused 51-100% mortality (%M) of J2s of both nematodes, mainly due to their RF fractions. Liquid chromatography-diode array detector-electrospray-high resolution mass spectrometry analysis of the most-active fractions revealed sesquiterpene and polyketide-like metabolites produced by the four active strains. These native Trichoderma strains have a high potential to develop safer natural products for the biocontrol of Meloidogyne species.

Anthelmintic Activity of Extracts and Active Compounds From Diospyros anisandra on Ancylostoma caninum, Haemonchus placei and Cyathostomins.

The present study aimed to evaluate the anthelmintic activity of leaf and bark extracts of Diospyros anisandra collected during different seasons and their major constituents on eggs of Ancylostoma caninum, Haemonchus placei, and cyathostomins. Specifically, the eclosion inhibition of the methanolic extracts of the leaves and bark of D. anisandra collected during the dry and rainy seasons (600-37.5 µg/ml) were evaluated in addition to the fractions, sub-fractions (300-37.5 µg/ml) and active major constituents (150-2.3 µg/ml). The rainy season bark extract had the highest percentage of eclosion inhibition (PEI) against the evaluated nematodes (≥ 90% at 75 µg/ml) along with high ovicidal activity (90.0 to 93.4% at 75 µg/ml). The purification of the rainy season bark extract showed that its biological activity came from the non-polar n-hexane fraction (≥ 93% at 75 µg/ml). The bioguided fractionation pointed to sub-fraction 5 as having the highest anthelmintic activity against the three evaluated genera of nematodes (PEI ≥ 93% at 37.5 µg/ml). Gas chromatography and mass spectrometry revealed that the major constituent in sub-fraction 5 was plumbagin. Upon evaluation, plumbagin was confirmed to be responsible for the anthelmintic activity of D. anisandra, with a PEI ≥ 90% at 2.3 µg/ml on the three evaluated nematodes. Additionally, the compounds betulin and lupeol in the bark of D. anisandra were evaluated but presented low anthelmintic activity (PEI ≤ 5.3% at 2.3 µg/ml). In conclusion, the rainy season bark extract of D. anisandra exerts a high ovicidal activity against the eggs of the three studied nematodes. Plumbagin is the active compound responsible for this activity and represents a potential alternative for the control of different genera of gastrointestinal nematodes given the current scenario of anthelmintic resistance.

Extracts from Six Native Plants of the Yucatán Peninsula Hinder Mycelial Growth of Fusarium equiseti and F. oxysporum, Pathogens of Capsicum chinense.

Fusarium equiseti strain FCHE and Fusarium oxysporum strain FCHJ were isolated from the roots of wilting habanero pepper (Capsicum chinense Jacq.) seedlings with root rot. Toward developing a biorational control of these serious phytopathogenic strains, ethanolic (EE) and aqueous (AE) extracts of different vegetative parts of 40 tropical native plants of the Yucatán Peninsula were screened for antifungal activity. Extracts of six out of 40 assayed plants were effective, and the most inhibitory extracts were studied further. EEs from Mosannona depressa (bark from stems and roots), Parathesis cubana (roots), and Piper neesianum (leaves) inhibited mycelial growth of both strains. Each active EE was then partitioned between hexane and acetonitrile. The acetonitrile fraction from M. depressa stem bark (MDT-b) had the lowest minimum inhibitory concentration of 1000 µg/mL against both pathogens and moderate inhibitory concentration (IC50) of 462 against F. equiseti and 472 µg/mL against F. oxysporum. After 96 h treatment with EE from M. depressa stem bark, both strains had distorted hyphae and conidia and collapsed conidia in scanning electron micrographs. Liquid chromatography-ultraviolet-high resolution mass spectrometry analysis revealed that the major component of the fraction was α-asarone. Its antifungal effect was verified using a commercial standard, which had an IC50 of 236 µg/mL against F. equiseti and >500 µg/mL against F. oxysporum. Furthermore, the P. cubana hexane fraction and P. neesianum acetonitrile fraction had antifungal activity against both Fusarium pathogens. These compounds provide new options for biorational products to control phytopathogenic fungi.

Identification of Insect-Deterrent Metabolites from Acremonium masseei strain CICY026, a Saprophytic Fungus from a Sinkhole in Yucatán.

Micromycetes from unexplored sources represent an opportunity to discover novel natural products to control insect pests. With this aim, a strain of Acremonium masseei CICY026 isolated from a tropical sinkhole was identified, cultured on fermented rice, and its ethyl acetate extract (EAE) was evaluated against three serious phytophagous insects (Bemisia tabaci, Myzus persicae, and Rhopalosiphum padi). DNA from A. masseei CICY026 was used to confirm its identity. EAE caused settling inhibition (SI) of M. persicae and R. padi (67.5% and 75.3%, respectively). Bioassay-guided fractionation of the active EAE led to the isolation of a novel metabolite, named hexahydroacremonintriol (1), and of acremonin A glucoside (2). The structures of 1 and 2 were determined using IR, one- and two-dimensional NMR, HRMS, and confirmed by theoretical data. The aphid M. persicae was noticeably sensitive to 1 and 2 (SI: 55.6% and 67.2%, respectively), whereas R. padi was only slightly affected by 1 (SI: 59%). This new knowledge about mycobiota from these special sinkhole ecosystems will inform the development of new biorational pesticides.

Antitubercular Activity of the Fungus Gliocladium sp. MR41 Strain.

Tuberculosis (TB) is a leading cause of death worldwide from infectious diseases and its inadequate treatment has led to emergence of resistant strains. The emergence of these strains renders the search for new drugs for the treatment of TB. The aim of this study was the evaluation of the anti-TB activity of the extract from fungus Gliocladium sp. MR41, and bioassay-guided fractionation and identification of majority compounds was carried out. Fungal strain culture was lyophilized and extracted by maceration in Ethyl Acetate (EtOAc). This extract was fractionated by liquid-liquid partitioning and chromatographic techniques, and the compounds were identified by their spectroscopic data. Furthermore, the EtOAc extract, fractions, and pure compounds were tested on Mycobacterium tuberculosis using the Microplate Alamar Blue Assay. From the bioactive AcetoNitrile Fraction (AcNF; MIC = 3.13 µg/mL) of the EtOAc extract, four compounds were isolated: ergosterol (1), ergosterol-5, 8-peroxide (2), 1, 6-di-O-acetyl-2,3,4,5-tetrahydroxy-hexane (3), and allitol (4). Only 2 exhibited potent activities against M. tuberculosis (MIC = 0.78 µg/mL). Additionally, this is the first report, to our knowledge, of polyols 3 and 4 from this fungus.

Molecular Identification of Selected Streptomyces Strains Isolated from Mexican Tropical Soils and their Anti-Candida Activity.

The increasing incidence of Candida albicans infections and resistance to current antifungal therapies has led to the search for new and more effective antifungal compounds. Actinobacterial species from the Streptomyces genus are recognized as some of the major producers of antimicrobial compounds. Therefore, the aims of this study were: (1) the identification of Streptomyces strains isolated from Mexican tropical acidic soils, (2) the evaluation of their antifungal activity on C. albicans, and (3) the exploration of the presence of polyketide synthase genes in their genome and antifungal secondary metabolites in their extracts. Four actinobacterial strains, isolated from previously unexplored soils with antibacterial antecedents, were selected. These strains were identified as Streptomycesangustmyceticus S6A-03, Streptomyces manipurensis S3A-05 and S3A-09, and Streptomyces parvisporogenes S2A-04, according to their molecular analyses. The ethanol extract of the lyophilized supernatant of S. parvisporogenes displayed the most interesting antifungal activity against C. albicans, with a minimum inhibitory concentration (MIC) of 0.5 mg/mL. Type I polyketide synthase (PKS-I) and non-ribosomal peptide synthase (NRPS) genes were detected in all strains. In addition, type II PKS genes (PKS-II) were also found in S.manipurensis S3A-05 and S. parvisporogenes. LC-UV-HRMS analysis of the active organic extract of S. parvisporogenes indicated the presence of the known antifungal compound carbazomycin G as the major component.

Medium-Chain Fatty Acids from Eugenia winzerlingii Leaves Causing Insect Settling Deterrent, Nematicidal, and Phytotoxic Effects.

Eugenia winzerlingii (Myrtaceae) is an endemic plant from the Yucatan peninsula. Its organic extracts and fractions from leaves have been tested on two phloem-feeding insects, Bemisia tabaci and Myzus persicae, on two plant parasitic nematodes, Meloidogyne incognita and Meloidogyne javanica, and phytotoxicity on Lolium perenne and Solanum lycopersicum. Results showed that both the hexane extract and the ethyl acetate extract, as well as the fractions, have strong antifeedant and nematicidal effects. Gas chromatography-mass spectrometry analyses of methylated active fractions revealed the presence of a mixture of fatty acids. Authentic standards of detected fatty acids and methyl and ethyl derivatives were tested on target organisms. The most active compounds were decanoic, undecanoic, and dodecanoic acids. Methyl and ethyl ester derivatives had lower effects in comparison with free fatty acids. Dose-response experiments showed that undecanoic acid was the most potent compound with EC50 values of 21 and 6 nmol/cm2 for M. persicae and B. tabaci, respectively, and 192 and 64 nmol for M. incognita and M. javanica, respectively. In a phytotoxicity assay, medium-chain fatty acids caused a decrease of 38-52% in root length and 50-60% in leaf length of L. perenne, but no effects were observed on S. lycopersicum. This study highlights the importance of the genus Eugenia as a source of bioactive metabolites for plant pest management.

Insecticidal and Nematicidal Contributions of Mexican Flora in the Search for Safer Biopesticides.

Plant metabolites have been used for many years to control pests in animals and to protect crops. Here, we reviewed the available literature, looking for the species of Mexican flora for which extracts and metabolites have shown activity against pest insects and parasitic nematodes of agricultural importance, as well as against nematodes that parasitize domestic cattle. From 1996 to 2018, the search for novel and eco-friendly biopesticides has resulted in the identification of 114 species belonging to 36 botanical families of Mexican plants with reported biological effects on 20 insect species and seven nematode species. Most plant species with detected pesticide properties belong to the families Asteraceae, Fabaceae, and Lamiaceae. Eighty-six metabolites have been identified as pesticidal active principles, and most have been terpenoids. Therefore, the continuation and intensification of this area of research is very important to contribute to the generation of new products that will provide alternatives to conventional pesticide agents. In addition, future studies will contribute to the recognition and dissemination of the importance of propagating plant species for their conservation and sustainable use.

In Vitro Lethal Activity of the Nematophagous Fungus Clonostachys rosea (Ascomycota: Hypocreales) against Nematodes of Five Different Taxa.

This study was aimed to evaluate the in vitro lethal activity of the nematophagous fungi Clonostachys rosea against 5 nematodes species belonging to different taxa. Two groups of 35 Petri dishes (PD) each were divided into 5 series of 7 (PD). Group 1 (series 1, 2, 3, 4, and 5) contained only water agar; meanwhile group 2 plates (series 6, 7, 8, 9, and 10) contained C. rosea cultures growth on water agar. Every plate from the two groups was added with 500 nematodes corresponding to the following genera/specie: Haemonchus contortus, Caenorhabditis elegans, Rhabditis sp., Panagrellus redivivus, and Butlerius sp. After 5-day incubation at room temperature, free (nontrapped) larvae were recovered from plates using the Baermann funnel technique. Recovered nematodes were counted and compared with their proper controls. Results shown an important reduction percentage of the nematode population attributed to the fungal lethal activity as follows: H. contortus (L3) 87.7%; C. elegans 94.7%; Rhabditis sp. 71.9%; P. redivivus 92.7%; and Butlerius sp. 100% (p ≤ 0.05). The activity showed by C. rosea against the H. contortus can be crucial for further studies focused to the biological control of sheep haemonchosis, although the environmental impact against beneficial nematodes should be evaluated.

Population fluctuation of Schistocerca piceifrons piceifrons (Orthoptera: Acrididae) in the Yucatán Península and its relation with the environmental conditions / Fluctuación poblacional de Schistocerca piceifrons piceifrons (Orthoptera: Acrididae) en la Península de Yucatán y su relación con las condiciones climáticas

Abstract The Central American Locust Schistocerca piceifrons piceifrons is one of the most damaging plant pest in Mexico and Central America. The present work was carried out to evaluate the seasonal population fluctuation of S. p. piceifrons and vegetation diversity and their association with weather factors and edaphic conditions in the gregarious zone of the Yucatán Península. The study was performed in seven sites during three seasons: North-wind (December 2013), rainy (June 2014) and dry (April 2014). The locust density was sampled in transect of 100 m2, as well as the vegetation in 16 m2: plant species richness (PSR) and relative species density (RSD), and analyzed by generalized linear models. Additionally, soil samples were obtained at 10 cm depth into a 4 × 6 m quadrat, land use in the sites was classified and temperature, precipitation and evaporation of each site were obtained from the database and they were analyzed with multiple factor analysis. The population density of S. p. piceifrons was higher in the sites Panaba, Tizimin, Tunkas and Cenotillo (F= 74.3, P < 0.0001). Characterization of vegetation showed that PSR and RSD were higher during the rainy season relative to those in the dry season (F= 50.4, P < 0.0001). RSD was identified as the most important group associated with locust density (0.86), followed by isotherm/isohyets (0.63), maximum precipitation and temperature (0.60), as well as the land use (0.65); no correlation between locust density and soil characteristics was found. Locust density was positively correlated with the abundance of the grass Panicum maximum (Sr2= 0.85, PC5= 0.87). This work shows that the population of S. p. piceifrons is high in the rainy season and influenced primarily by the abundance of the grass P. maximum and the precipitation. The results indicate that surveys for early detection and control of the locust on the Yucatán Península can focus on areas with the grass P. maximum to predict risk areas and target survey efforts. Rev. Biol. Trop. 66(1): 403-414. Epub 2018 March 01.

Resumen La langosta Centroamericana Schistocerca piceifrons piceifrons es una de las plagas más dañinas en México y Centroamérica. El presente trabajo se realizó para evaluar la fluctuación estacional de poblaciones de S. p. piceifrons y su asociación con la diversidad de vegetación, factores climáticos y edáficos en la zona gregarígena de la Península de Yucatán. Se seleccionaron siete sitios y se muestrearon a lo largo de tres estaciones: nortes (diciembre 2013), lluvias (junio 2014) y sequías (abril 2014). La densidad poblacional de la langosta fue muestreada en transectos de 100 m2, así como la vegetación en 16 m2 obteniendo la riqueza de especies vegetales (REV) y la densidad relativa de especies (DRE), y fueron analizadas por modelos lineales generalizados. Adicionalmente se obtuvieron muestras de suelo de 10 cm de profundidad en un área de 4 × 6 m, se clasificó el uso del suelo de cada sitio y se obtuvieron en una base de datos las condiciones de temperatura, precipitación y evaporación para cada sitio, estos datos se analizaron con análisis de factores múltiples. La densidad poblacional de S. p. piceifrons fue mayor en los sitios de Panaba, Tizimin, Tunkas y Cenotillo (F= 74.3, P < 0.0001). La caracterización de la vegetación mostró que la REV y la DRE fueron mayores en la estación de lluvias que en la de sequías (F= 50.4, P < 0.0001). La DRE fue el grupo más importante asociado a la densidad de la langosta (0.86), seguido por las isotermas/isoyetas (0.63) y la precipitación-temperatura máxima (0.60), así como el uso del suelo (0.65); no hubo correlación con las características del suelo. La densidad de la langosta fue correlacionada positivamente con la abundancia del pasto Panicum maximum (Sr2= 0.85, PC5= 0.87). El estudio mostró que las poblaciones de S. p. piceifrons fueron mayores en la estación lluviosa e influenciadas principalmente por la abundancia del pasto P. maximum y la precipitación. Los resultados indican que las exploraciones, la detección temprana y el control de la langosta en la Península de Yucatán puede enfocarse sobre áreas cultivadas con P. maximum para predecir áreas con riesgo y eficientar los recursos.

Fungal Screening on Olive Oil for Extracellular Triacylglycerol Lipases: Selection of a Trichoderma harzianum Strain and Genome Wide Search for the Genes.

A lipolytic screening with fungal strains isolated from lignocellulosic waste collected in banana plantation dumps was carried out. A Trichoderma harzianum strain (B13-1) showed good extracellular lipolytic activity (205 UmL-1). Subsequently, functional screening of the lipolytic activity on Rhodamine B enriched with olive oil as the only carbon source was performed. The successful growth of the strain allows us to suggest that a true lipase is responsible for the lipolytic activity in the B13-1 strain. In order to identify the gene(s) encoding the protein responsible for the lipolytic activity, in silico identification and characterization of triacylglycerol lipases from T. harzianum is reported for the first time. A survey in the genome of this fungus retrieved 50 lipases; however, bioinformatic analyses and putative functional descriptions in different databases allowed us to choose seven lipases as candidates. Suitability of the bioinformatic screening to select the candidates was confirmed by reverse transcription polymerase chain reaction (RT-PCR). The gene codifying 526309 was expressed when the fungus grew in a medium with olive oil as carbon source. This protein shares homology with commercial lipases, making it a candidate for further applications. The success in identifying a lipase gene inducible with olive oil and the suitability of the functional screening and bioinformatic survey carried out herein, support the premise that the strategy can be used in other microorganisms with sequenced genomes to search for true lipases, or other enzymes belonging to large protein families.

Insect deterrent and nematicidal screening of microfungi from Mexico and anti-aphid compounds from Gliomastix masseei.

Fungal metabolites are promising alternatives for the development of biorational pesticides. In this sense, microfungi from tropical regions are valuable sources of natural compounds for pest management. With the aim of broadening the search for new eco-friendly products to manage plant pests, this study was carried out to evaluate the biological activity of 23 tropical fungal extracts on three species of phytophagous insects and a plant parasitic nematode. In addition, the active principles of the most effective extract were identified. The insect deterrent activity of fungal extracts was evaluated on the settling of aphids Myzus persicae and Rhopalosiphum padi, and on the feeding of lepidoptera larva Spodoptera littoralis; the nematostatic activity was evaluated on the mobility of Meloidogyne javanica. Active metabolites from Gliomastix masseei were identified by GC-MS techniques and by comparison with commercial standards. Results showed seven extracts with strong effect on the settling of M. persicae and R. padi (settling inhibition >80%). The calculated median of effective concentration (EC50) values ranged from 8 to 38µg/cm2 for the extracts of Clonostachys rosea and G. masseei, respectively. Bioassay-guided separation of the ethyl acetate extract of G. masseei revealed the presence of fatty acids and their derivatives, where methyl 9-octadecenoate was the most active compound with EC50 values of 16µg and 35µg/cm2 for M. persicae and R. padi, respectively. Extracts of C. rosea and G. masseei could be a promising option in the control of pest aphids in agriculture.

Insect deterrent and nematicidal screening of microfungi from Mexico and anti-aphid compounds from Gliomastix masseei / Evaluación de la actividad disuasiva de insectos y nematotóxica de microhongos de México y compuestos anti-áfidos de Gliomastix masseei

Fungal metabolites are promising alternatives for the development of biorational pesticides. In this sense, microfungi from tropical regions are valuable sources of natural compounds for pest management. With the aim of broadening the search for new eco-friendly products to manage plant pests, this study was carried out to evaluate the biological activity of 23 tropical fungal extracts on three species of phytophagous insects and a plant parasitic nematode. In addition, the active principles of the most effective extract were identified. The insect deterrent activity of fungal extracts was evaluated on the settling of aphids Myzus persicae and Rhopalosiphum padi, and on the feeding of lepidoptera larva Spodoptera littoralis; the nematostatic activity was evaluated on the mobility of Meloidogyne javanica. Active metabolites from Gliomastix masseei were identified by GC-MS techniques and by comparison with commercial standards. Results showed seven extracts with strong effect on the settling of M. persicae and R. padi (settling inhibition >80%). The calculated median of effective concentration (EC50) values ranged from 8 to 38 µg/cm² for the extracts of Clonostachys rosea and G. masseei, respectively. Bioassay-guided separation of the ethyl acetate extract of G. masseei revealed the presence of fatty acids and their derivatives, where methyl 9-octadecenoate was the most active compound with EC50 values of 16 µg and 35 µg/cm² for M. persicae and R. padi, respectively. Extracts of C. rosea and G. masseei could be a promising option in the control of pest aphids in agriculture.

Los metabolitos fúngicos son agentes prometedores para el desarrollo de plaguicidas biorracionales. En este sentido, los hongos microscópicos de zonas tropicales representan una valiosa fuente de compuestos naturales para el manejo de plagas. Con la finalidad de ampliar la investigación en productos amigables con el medio ambiente, en este estudio se evaluó la actividad biológica de 23 extractos de hongos sobre 3 especies de insectos fitófagos y un nematodo fitoparásito; además se identificaron los componentes del extracto más activo. El efecto disuasivo de los extractos fúngicos se evaluó en el asentamiento de Myzus persicae y Rhopalosiphum padi, así como en la alimentación de Spodoptera littoralis; la actividad nematostática se evaluó sobre la movilidad de Meloidogyne javanica. Los metabolitos activos de Gliomastix masseei se identificaron por cromatografía de gases-espectrometría de masas y por comparación con muestras comerciales. Los resultados mostraron 7 extractos con fuerte efecto en la inhibición del asentamiento (> 80%) de M. persicae y R. padi. Los valores de la concentración efectiva media (CE50) estuvieron en el rango de 8 a 38 µg/cm² para los extractos de Clonostachys rosea y G. masseei, respectivamente. El extracto de acetato de etilo de G. masseei se fraccionó por un proceso biodirigido y reveló la presencia de ácidos grasos y sus derivados, donde el 9-octadecenoato de metilo fue el más activo, con una CE50 de 16 µg/cm² para M. persicae y 35 µg/cm² para R. padi. Los extractos fúngicos de C. rosea y G. masseei pueden ser una alternativa promisoria en el control de áfidos que son plaga en la agricultura.

Insecticidal effects of plant extracts on immature whitefly Bemisia tabaci Genn. (Hemiptera: Aleyroideae)

Background: The whitefly (Bemisia tabaci Genn.) is a widely distributed and highly harmful plant pest species. The management of B. tabaci has been typically carried out by chemical pesticides. In the last decade however, there has been an increasing interest in natural products, particularly those of plant origin, to control this pest species. In the present work, aqueous and ethanolic extracts of native plants from the flora of the Yucatán peninsula (Acalypha gaumeri, Annona squamosa, Carlowrightia myriantha, Petiveria alliaceae and Trichilia arborea) and the introduced plant Azadirachta indica were collected and evaluated for insecticidal activity against eggs and nymphs Bemisia tabaci. Results: Most of the aqueous and ethanolic extracts showed high insecticidal effects on B. tabaci eggs. The lowest LC50 values were recorded in the aqueous extracts of A. gaumeri (0.39% w/v), A. squamosa (0.36% w/v), P. alliaceae (0.42% w/v) and A. indica (0.30% /v), as well as in the ethanolic extracts of P. alliaceae (2.09 mg mL-1) and T. arborea (2.14 mg mL-1). On the other hand, B. tabaci nymphs were not affected by the aqueous extracts, but were highly sensitive to the ethanolic extracts of the tested plants. The lowest LC50 values were recorded in the ethanolic extracts of P. alliaceae (1.27 mg ml-1) and T. arborea (1.61 mg mL-1). The GC-MS analysis showed that phytol was the major component of the ethanolic extract of P. alliaceae and fatty acids were the major components of ethanolic extract of T. arborea. Conclusions: Overall, results suggest that ethanolic extracts of P. alliaceae and T. arborea leaves showed the highest insecticidal effects on eggs and nymphs B. tabaci. The extracts from P. alliaceae and T. arborea are good candidates to be developed as sources of natural insecticides for the management of immature B. tabaci since their effects were comparable with that showed by the extracts of A. indica, a well-known plant species for its insecticidal activity.

Antimycobacterial and antileishmanial effects of microfungi isolated from tropical regions in México.

A total of 82 fungal extracts were selected and screened against Mycobacterium tuberculosis and promastigotes of Leishmania mexicana strains. Results showed inhibitory activity in 29 % of the fungal strains against at least one of the targets tested. The most significant antituberculosis (antiTB) effects were presented by Cylindrocarpon sp. XH9B, Fusarium sp. TA54, Fusarium XH1Ga, Gliocladium penicillioides TH04 and TH21, Gliocladium sp. TH16, Kutilakesa sp. MR46, and Verticillium sp. TH28 strains (minimal inhibition concentration (MIC) = 1.56-25 µg/ml). Mortality of L. mexicana promastigotes was displayed by only four strains, Fusarium sp. TA50, Fusarium sp. TA54, Verticillium sp. TH28, and the unidentified 2TA2 strain (IC(50) = 14.23-100 µg/ml and IC(100) = 50-100 µg/ml). Seven of these most active strains were defatted and their corresponding fractions evaluated again. The results showed the best antiTB activity in Gliocladium sp. TH16 (MIC = 1.56 µg/ml) and the highest leishmanicidal potential in Fusarium sp. TA54 (IC(50) = 6.36 µg/ml). These results show that fungi living in the tropical regions of México have the ability to produce bioactive metabolites that could be used in the near future as natural products to control neglected tropical diseases.