Outstanding impact of soil tillage on the abundance of soil hydrolases revealed by a metagenomic approach

ABSTRACT The soil represents the main source of novel biocatalysts and biomolecules of industrial relevance. We searched for hydrolases in silico in four shotgun metagenomes (4,079,223 sequences) obtained in a 13-year field trial carried out in southern Brazil, under the no-tillage (NT), or conventional tillage (CT) managements, with crop succession (CS, soybean/wheat), or crop rotation (CR, soybean/maize/wheat/lupine/oat). We identified 42,631 hydrolases belonging to five classes by comparing with the KEGG database, and 44,928 sequences by comparing with the NCBI-NR database. The abundance followed the order: lipases > laccases > cellulases > proteases > amylases > pectinases. Statistically significant differences were attributed to the tillage system, with the NT showing about five times more hydrolases than the CT system. The outstanding differences can be attributed to the management of crop residues, left on the soil surface in the NT, and mechanically broken and incorporated into the soil in the CT. Differences between the CS and the CR were slighter, 10% higher for the CS, but not statistically different. Most of the sequences belonged to fungi (Verticillium, and Colletotrichum for lipases and laccases, and Aspergillus for proteases), and to the archaea Sulfolobus acidocaldarius for amylases. Our results indicate that agricultural soils under conservative managements may represent a hotspot for bioprospection of hydrolases.

Classic and molecular study of Giardia duodenalis in children from a daycare center in the region of Presidente Prudente, São Paulo, Brazil / Estudo clássico e molecular de Giardia duodenalis em crianças atendidas em uma creche na região de Presidente Prudente, São Paulo, Brasil

Epidemiological studies on giardiasis by using molecular techniques such as RAPD (Randomly Amplified Polymorphic DNA) may give information on factors related to the transmission of Giardia duodenalis. The aim of this work was to assess the epidemiology of G. duodenalis in 101 children attended at a daycare center in Presidente Bernardes, SP, Brazil. After parasitological examinations in feces samples, 15 children presented cysts of G. duodenalis. Their respective parents, brothers and pets, besides the daycare center workers, also had their feces submitted to parasitological analysis. Seven mothers and nine brothers also presented G. duodenalis cysts, while fathers, daycare workers and pets (dogs) did not presented the parasite. Besides the 15 cases with G. duodenalis, other 23 children presented other enteroparasites (Entamoeba coli, Endolimax nana, Enterobius vermicularis, Ascaris lumbricoides and Trichuris trichiura). Samples of G. duodenalis cysts from children and their relatives were submitted to molecular typing by RAPD after genomic DNA extraction and amplification of a fragment of the 18S rDNA region by PCR. After examining 31 isolates of G. duodenalis (children and their respective mothers and brothers), it was concluded that the parasite transmission occurred in children, probably during daily cohabitation at the daycare center, but not at home among their relatives or pets.

Estudos epidemiológicos sobre giardíase por técnicas moleculares como a RAPD (Amplificação aleatória do DNA polimórfico) podem contribuir para o entendimento de fatores relacionados à transmissão de Giardia duodenalis. O objetivo desse trabalho foi estudar a epidemiologia de G. duodenalis em 101 crianças atendidas em uma creche em Presidente Bernardes, SP, Brasil. Após exames parasitológicos em amostras de fezes, 15 crianças apresentaram cistos de G. duodenalis. Seus respectivos pais, irmãos e animais domésticos, além dos funcionários da creche, foram submetidos a exames parasitológicos de fezes. Destes, sete mães e nove irmãos apresentaram G. duodenalis, enquanto os pais, funcionários da creche e animais de estimação (cães) não apresentaram o parasita. Além dos 15 casos com G. duodenalis, outras 23 crianças apresentaram outros enteroparasitas (Entamoeba coli, Endolimax nana, Enterobius vermicularis, Ascaris lumbricoides e Trichuris trichiura). As amostras contendo cistos de G. duodenalis das crianças e de seus parentes foram submetidas à análise molecular por RAPD após extração do DNA genômico e amplificação de um fragmento de uma região do rDNA 18S por PCR. Entre os 31 isolados de G. duodenalis (crianças e suas respectivas mães e irmãos), concluiu-se que a transmissão dos parasitas ocorreu entre as crianças, provavelmente durante seu convívio na creche, mas não foi decorrente do convívio familiar ou animais de estimação.