Enhancement of Trichoderma harzianum CFAM-422 for cellulase and hemicellulase production by deletion of the carbon catabolite repressor gene cre1.

Carbon catabolite repression (CCR) is a mechanism by which microorganisms can utilize preferably highly energetic compounds over those of difficult degradation. For Trichoderma reesei, the protein that acts as repressor in the presence of glucose is CRE1. In this project, we aim to delete cre1 gene in Trichoderma harzianum CFAM-422 and obtain mutants with enhanced production of biomass degrading enzymes. Disruption of cre1 in T. harzianum CFAM-422 was performed by gene replacement of cre1 for hph (hygromycin B phosphotransferase) via homologous recombination. Hygromycin resistant mutants and parental strains enzyme production was evaluated in both inductive and repressive conditions in four different carbon sources. Enzymatic indexes (EI) were determined and compared. All genetically stable transformants showed increased enzymatic index under inductive conditions and modest inhibition under repressive conditions for most carbon sources, indicating that the deletion of cre1 in T. harzianum can be beneficial to cellulase and hemicellulase production with reduced product inhibition.SINAFERM; SHEB. 3 a 6 de setembro. Seção Trabalhos. Ref. 59019

Sensitive and specific serodiagnosis of Leishmania infantum infection in dogs by using peptides selected from hypothetical proteins identified by an immunoproteomic approach

In Brazil, the percentage of infected dogs living in areas where canine visceral leishmaniasis (CVL) is endemic ranges from 10 to 62%; however, the prevalence of infection in dogs is probably higher than figures reported from serological studies. In addition, problems with the occurrence of false-positive or false-negative results in the serodiagnosis of CVL have been reported. The present work analyzed the potential of synthetic peptides mapped from hypothetical proteins for improvement of the serodiagnosis of Leishmania infantum infection in dogs. From 26 identified leishmanial proteins, eight were selected, considering that no homologies between these proteins and others from trypanosomatide sequence databases were encountered. The sequences of these proteins were mapped to identify linear B-cell epitopes, and 17 peptides were synthesized and tested in enzyme-linked immunosorbent assays (ELISAs) for the serodiagnosis of L. infantum infection in dogs. Of these, three exhibited sensitivity and specificity values higher than 75% and 90%, respectively, to differentiate L. infantum-infected animals from Trypanosoma cruziinfected animals and healthy animals. Soluble Leishmania antigen (SLA) showed poor sensitivity (4%) and specificity (36%) to differentiate L. infantum-infected dogs from healthy and T. cruzi-infected dogs. Lastly, the three selected peptides were combined in different mixtures and higher sensitivity and specificity values were obtained, even when sera from T. cruzi-infected dogs were used. The study’s findings suggest that these three peptides can constitute a potential tool for more sensitive and specific serodiagnosis of L. infantum infection in dogsThis work was supported by grants from the Pró-Reitoria de Pesquisa from UFMG (Edital 07/2012), Instituto Nacional de Ciência e Tecnologia em Nano-biofarmacêutica (INCT-NANOBIOFAR, Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (CBB-APQ-02364-08, CBB-APQ-00356-10, CBB-APQ-00496-11, and CBB-APQ-00819-12), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (APQ-472090/2011-9), and the Instituto Nacional de Ciência e Tecnologia em Vacinas (INCT-V). E.A.F.C. and A.P.F. are CNPq grant recipients. M.A.C.-F. is a FAPEMIG/CAPES grant recipient. This study was supported in Spain, in part, by grants from the Ministerio de Ciencia e Innovación (FIS/PI1100095)

Recent updates and perspectives on approaches for the development of vaccines against visceral leishmaniasis

All rights reserved. Visceral leishmaniasis (VL) is one of the most important tropical diseases worldwide. Although chemotherapy has been widely used to treat this disease, problems related to the development of parasite resistance and side effects associated with the compounds used have been noted. Hence, alternative approaches for VL control are desirable. Some methods, such as vector control and culling of infected dogs, are insufficiently effective, with the latter not ethically recommended. The development of vaccines to prevent VL is a feasible and desirable measure for disease control, for example, some vaccines designed to protect dogs against VL have recently been brought to market. These vaccines are based on the combination of parasite fractions or recombinant proteins with adjuvants that are able to induce cellular immune responses, however, their partial efficacy and the absence of a vaccine to protect against human leishmaniasis underline the need for characterization of new vaccine candidates. This review presents recent advances in control measures for VL based on vaccine development, describing extensively studied antigens, as well as new antigenic proteins recently identified using immuno-proteomic techniquesThis work was supported by grants from Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica, Rede Nanobiotec/Brasil-Universidade Federal de Uberlândia/CAPES, PRONEX-FAPEMIG (APQ-01019-09), FAPEMIG (CBB-APQ-00819-12 and CBB-APQ-01778-2014), and CNPq (APQ-482976/2012-8, APQ-488237/2013-0, and APQ-467640/2014-9). EAFC and LRG are recipients of the grant from CNPq. MACF is the recipient of grants from FAPEMIG/CAPE

Development of RAPD-AFLP map of eggplant and improvement of tolerance to Verticillium wilt

Verticillium wilt is one of the most serious diseases in eggplant and one of the main goals of eggplant breeding is the achievement of Verticillium wilt tolerant hybrids. Solanum sodomeum L. has shown a mechanism of partial resistance against Verticillium wilt. Interspecific hybrids were obtained by using an accession of S. sodomeum from Sicily and the eggplant variety Buia. The results of the screenings for eggplant genotypes tolerant/resistant to Verticillium wilt by using backcrosses are reported. In a naturally infected field, from 1998 to 2001, the resistance to V dahliae of backcrossed progenies was increased by about 60%. The interspecific hybrids were both selfed and backcrossed using different types of eggplant. An integrated linkage map of eggplant (Solanum melongena L.) has been obtained by using 48 plants of an F-2 population. Starting from a genetic RAPDs map with similar to 100 markers mapped on 13 linkage groups, 4 AFLPs primer combinations were screened in order to improve the genetic map with the aim to achieve markers linked to Verticillium tolerance. Mendelian segregation of loci was verified by chi-square tests of the expected 3:1 and 1:1 ratios. Marker order was determined and all the data were combined to construct the most likely map. The integrated analysis of markers resulted in the construction of a map consisting of 273 loci and 12/13 linkage groups spanning 736 cM in a total. Linkage between different AFLP markers and the tolerance to Verticillium is discussed