Abstract Background: Members of the genus Cupiennius Simon, 1891 are categorized as wandering spiders and are part of the family Trechaleidae. The genomics and proteomics of Cupiennius spiders from North America remain uncharacterized. The present study explores for the first time molecular data from the endemic species Cupiennius chiapanensis Medina, 2006, and also presents new data for Cupiennius salei (Keyserling, 1878), both collected in southern Mexico. Methods: In total, 88 Cupiennius specimens were collected from southern Mexico and morphologically identified. DNA was extracted and the mitochondrial COI fragment was amplified. COI sequences were analyzed, and a phylogenetic tree was inferred for species from the Americas. Genetic diversity was analyzed using haplotype networks and gene distances. Venom was obtained from C. chiapanensis and C. salei by electrostimulation. The venom was separated by HPLC, visualized using SDS-PAGE, and quantified for use in toxicity bioassays in mice and insects. Results: Analysis of COI sequences from C. chiapanensis showed 94% identity with C. salei, while C. salei exhibited 94-97% identity with sequences from Central and South American conspecifics. The venom from C. chiapanensis exhibited toxic activity against crickets. Venoms from C. chiapanensis and C. salei caused death in Anastrepha obliqua flies. Analysis of venom fractions from C. salei and C. chiapanensis revealed molecular masses of a similar size as some previously reported toxins and neurotoxic components. We determined the amino acid sequences of ChiaTx1 and ChiaTx2, toxins that are reported here for the first time and which showed toxicity against mice and insects. Conclusion: Our work is the first to report COI-based DNA barcoding sequences from southern Mexican Cupiennius spiders. Compounds with toxic activity were identified in venom from both species. Background Simon 189 Trechaleidae uncharacterized Medina 2006 Keyserling, Keyserling (Keyserling 1878, 1878 , 1878) Methods total 8 amplified Americas distances C electrostimulation HPLC SDSPAGE, SDSPAGE SDS PAGE, PAGE SDS-PAGE insects Results 94 9497% 9497 97% 97 94-97 conspecifics crickets flies components ChiaTx ChiaTx2 Conclusion COIbased based 18 200 187 9 949 94-9 1 20 94- 2

Abstract Background: Phonotimpus pennimani (Araneae, Phrurolithidae) is a small-sized (3-5 mm) spider endemic to the Tacaná volcano in Chiapas, Mexico, where it is found in soil litter of cloud forests and coffee plantations. Its venom composition has so far not been investigated, partly because it is not a species of medical significance. However, it does have an important impact on the arthropod populations of its natural habitat. Methods: Specimens were collected in Southeastern Mexico (Chiapas) and identified taxonomically by morphological characteristics. A partial sequence from the mitochondrial gene coxI was amplified. Sequencing on the Illumina platform of a transcriptome library constructed from 12 adult specimens revealed 25 toxin or toxin-like genes. Transcripts were validated (RT-qPCR) by assessing the differential expression of the toxin-like PpenTox1 transcript and normalising with housekeeping genes. Results: Analysis of the coxI-gene revealed a similarity to other species of the family Phrurolithidae. Transcriptome analysis also revealed similarity with venom components of species from the families Ctenidae, Lycosidae, and Sicariidae. Expression of the toxin-like PpenTox1 gene was different for each developmental stage (juvenile or adult) and also for both sexes (female or male). Additionally, a partial sequence was obtained for the toxin-like PpenTox1 from DNA. Conclusion: Data from the amplification of the mitochondrial coxI gene confirmed that P. pennimani belongs to the family Phrurolithidae. New genes and transcripts coding for venom components were identified. Background Araneae, Araneae (Araneae Phrurolithidae smallsized small sized 35 3 5 (3- mm Chiapas plantations investigated significance However habitat Methods (Chiapas characteristics amplified 1 2 toxinlike like RTqPCR RT qPCR (RT-qPCR PpenTox Results coxIgene Ctenidae Lycosidae Sicariidae juvenile female male. male . male) Additionally DNA Conclusion P (3 (

Resumen Guarianthe skinneri está sujeta a protección debido a la disminución de sus poblaciones silvestres, y la germinación asimbiótica in vitro de semillas es una alternativa viable para la conservación y restauración de poblaciones. Este proceso de laboratorio también facilita el estudio detallado de las fases de germinación, ahora, en el caso de las orquídeas, repartido en dos etapas, la pregerminación y la germinación, como contribución al diseño de una estrategia para la restauración de poblaciones de esta orquídea. Con este fin, se evaluó el efecto de cuatro medios de cultivo (Yasuda 10% + 0.112 mg L-1 6-bencilaminopurina (BAP), Yasuda 100% + 1.125 mg L-1 BAP, Dalla Rosa & Laneri modificado, Murashige & Skoog) y dos condiciones de escotoperiodo (12 y 24 h) sobre las diferentes fases de la pregerminación y germinación de semillas. Los medios Yasuda y Dalla Rosa promovieron el desarrollo y terminación de las fases de germinación, mientras que en el medio MS los porcentajes fueron bajos en el desarrollo de las Fases 4 y 5, ocasionados por los altos porcentajes de oxidación en Fases 2 y 3. Las condiciones de escotoperiodo no afectaron el proceso de germinación de las semillas.

Abstract Guarianthe skinneri is subject to protection due to the loss of natural, wild populations, and the in vitro symbiotic germination of seeds is a viable alternative for the conservation and restoration of such populations. This laboratory process also facilitates the detailed study of the phases of germination, that, in the case of orchids, we now divide into two stages, pre-germination and germination, as a contribution towards the design of a strategy for the restoration of populations of this orchid. To that end, we evaluated the effect of four culture media, (Yasuda 10% + 0.112 mg/L 6-benzylaminopurine (BAP), Yasuda 100% + 1.125 mg/L BAP, modified Dalla Rosa & Laneri, Murashige & Skoog) and two skotoperiods (12 and 24 h), observing the phases of pre-germination and germination of the seeds in all treatments. The treatments with the culture media Dalla Rosa and Yasuda promoted the development and completion of the germination phases. The MS medium gave low percentages of development for phases 4 and 5, due to high percentages of oxidation in phases 2 and 3. The two skotoperiods did not affect the germination process.

The goal of this study was to isolate, select and characterize bacteria with cellulolytic activity from two different coffee residue composting piles, one of which had an internal temperature of 57 |#9702;C and pH 5.5 and the other, a temperature of 61 |#9702;C, and pH 9.3. Culture media were manipulated with carboxymethylcellulose and crystalline cellulose as sole carbon sources. The enzyme activity was assessed by hydrolysis halo formation, reducing sugar production and zymograms. Three out of twenty isolated strains showed higher enzymatic activity and were identified as Bacillus subtilis according to their morphological, physiological, biochemical characteristics and based on the sequence analysis of 16S rDNA regions. The enzymatic extracts of the three selected strains showed exocellulase and endocellulase maximum activity of 0.254 and 0.519 U/ml, respectively; the activity of these enzymes was maintained even in acid pH (4.8) and basic (9.3) and at temperatures of up to 60°C. The enzymatic activities observed in this study are within the highest reported for cellulose produced by bacteria of the genus Bacillus. Endocellulase activity was shown in the zymograms from 24 h until 144 h of incubation. Furthermore, the pH effect on the endocellulase activity is reported for the first time by zymograms. The findings in this study entail the possibility to use these enzymes in the procurement of fermentable substrates for the production of energy from the large amount of residues generated by the coffee agroindustry.

El objetivo de este estudio fue aislar, seleccionary caracterizar bacterias con actividad celulolítica a partir de 2 diferentes pilas de compostaje de residuos de café, una con temperatura interna de 57°C y pH 5,5; la otra con temperatura interna de 61 °C y pH 9,3. Se utilizaron medios de cultivo con carboximetilcelulosa y celulosa cristalina como únicas fuentes de carbono. La actividad enzimàtica fue evaluada por formación de halos de hidrólisis, producción de azúcares reductores y zimogramas. De 20 cepas aisladas, 3 presentaron mayor actividad enzimàtica y fueron identificadas como Bacillus subtilis sobre la base de sus características morfológicas, fisiológicas y bioquímicas y del análisis de las secuencias de la región 16S del ADNr. Los extractos enzimáticos de las 3 cepas seleccionadas presentaron actividad de exocelulasa y de endocelulasa, con máximos de 0,254 y 0,519 U/ml, respectivamente; la actividad de estas enzimas se mantuvo incluso a pH ácido (4,8) o básico (9,3) y a temperaturas de hasta 60 °C. Las actividades enzimáticas halladas en este estudio se ubican dentro de las más altas reportadas para celulasas producidas por bacterias del género Bacillus. En los zimogramas se demostró actividad de endocelulasa desde las 24h hasta las 144h de incubación. Asimismo, se reporta por primera vez el efecto del pH sobre la actividad de endocelulasa observado por zimogramas. Los resultados de este estudio abren la posibilidad de hacer uso de estas enzimas en la obtención de sustratos fermentables para la producción de energía a partir de los residuos generados en grandes cantidades por la agroindustria del café.

El paraquat es un herbicida utilizado ampliamente en la agricultura. Debido a su gran distribución y uso inadecuado, representa un problema grave de contaminación del suelo y el agua. Se ha encontrado que los hongos de la podredumbre blanca son capaces de degradar compuestos contaminantes que poseen estructuras similares a la lignina, como es el caso del paraquat. En el presente trabajo se evaluó la degradación de este herbicida y su efecto en la producción de enzimas ligninolíticas por parte de algunos hongos de la podredumbre blanca aislados del sur de México. Seis cepas fúngicas mostraron tolerancia al herbicida durante el cultivo en medio sólido. Tres de las 6 cepas evaluadas, correspondientes a las especies Polyporus tricholoma, Cilindrobasidium laeve y Deconica citrispora, mostraron niveles de degradación del 32, el 26 y el 47%, en ese orden, a los 12 días de cultivo en presencia del xenobiótico. Se detectó un incremento en las actividades de las enzimas lacasa y Mn-peroxidasa en las cepas que presentaron el mayor porcentaje de degradación, probablemente asociado a la disminución del herbicida. Adicionalmente, se realizaron ensayos con extractos enzimáticos procedentes del medio de cultivo extracelular de las 2 cepas que presentaron mayor degradación. Después de 24 h de incubación, se obtuvo una degradación del 49% del paraquat inicial con los extractos de D. citrispora. Los resultados obtenidos indican que la degradación del herbicida estaría asociada a la presencia de enzimas extracelulares en los hongos de la podredumbre blanca. En este trabajo se muestran las primeras evidencias del potencial de biodegradación de diferentes especies de hongos de la pudrición blanca.