Genetic diversity and efficiency of DNA microsatellites for genealogic control in Nelore breed

The genetic variability and paternity testing values in Nelore breed were estimated using 11 ISAG/FAO microsatellites. The markers were organized into 4 amplification groups for semi-automated fluorescence genotyping. All markers were highly polymorphic, with an average of 8.2 alleles per locus. With a mean value of 0.48, the observed heterozygosity was lower than the expected for 10 of the loci. A significant deficit of heterozygotes was observed for 9 loci, resulting in a lack of Hardy-Weinberg equilibrium in the studied population. Polymorphism information content values exceeded 0.5 for 10 loci. The power of discrimination was >0.999 and paternity exclusion probabilities when a mother, her offspring and a putative sire are compared or when one or other parental genotype is unavailable for the combined set of markers were, respectively, >0.999 and >0.989. The set of 11 microsatellite markers proved to be an efficient tool for paternity testing in Nelore cattle.Foram estimados na raça Nelore a variabilidade genética e os valores de determinação de paternidade usando-se 11 marcadores microssatélites do painel ISAG/FAO. Estes foram organizados em quatro conjuntos de amplificação para genotipagem semi-automática por fluorescência. Todos os marcadores apresentaram-se altamente polimórficos, com média de 8,2 alelos por loco. A heterozigosidade observada, com média de 0,48, foi menor que a esperada em 10 locos. Foram observadas deficiências de heterozigotos em nove locos, o que resultou no desequilíbrio de Hardy-Weinberg para a população estudada. O conteúdo polimórfico informativo foi superior a 0,5 em 10 locos. O poder de discriminação foi >0,999 e as probabilidades de exclusão de paternidade quando são conhecidos os genótipos de um bezerro, sua mãe e um pai alegado, ou quando um ou outro genótipo parental não está disponível, para o conjunto de marcadores foram >0,999 e >0,989, respectivamente. O conjunto de 11 marcadores constitui método eficiente para a determinação de paternidade na raça Nelore

Insights Into Limnothrix sp. Metabolism Based on Comparative Genomics

Currently only four genome sequences for Limnothrix spp. are publicly available, and information on the genetic properties of cyanobacteria belonging to this genus is limited. In this study, we report the draft genome of Limnothrix sp. CACIAM 69d, isolated from the reservoir of a hydroelectric dam located in the Amazon ecosystem, from where cyanobacterial genomic data are still scarce. Comparative genomic analysis of Limnothrix revealed the presence of key enzymes in the cyanobacterial central carbon metabolism and how it is well equipped for environmental sulfur and nitrogen acquisition. Additionally, this work covered the analysis of Limnothrix CRISPR-Cas systems, pathways related to biosynthesis of secondary metabolites and assembly of extracellular polymeric substances and their exportation. A trans-AT PKS gene cluster was identified in two strains, possibly related to the novel toxin Limnothrixin biosynthesis. Overall, the draft genome of Limnothrix sp. CACIAM 69d adds new data to the small Limnothrix genome library and contributes to a growing representativeness of cyanobacterial genomes from the Amazon region. The comparative genomic analysis of Limnothrix made it possible to highlight unique genes for each strain and understand the overall features of their metabolism

Isolation And Characterization Of 11 Microsatellite Loci In The Mangrove Crab, Ucides Cordatus (ocypodidae: Ucidinae).

Eleven polymorphic microsatellite loci were described for the mangrove crab, Ucides cordatus, an important fishery resource on the Brazilian coast. The number of alleles observed at each locus varied between eight and 23. Observed and expected mean heterozygosities were 0.791 and 0.893 respectively. Amplification of all loci was highly successful, under the same polymerase chain reaction conditions. With the exception of P2D3, all loci adhered to the assumptions of the Hardy-Weinberg equilibrium and did not present deviations reflecting linkage disequilibrium. Given this, these markers will be extremely useful in future management programmes for U. cordatus stocks.91395-

Optimization of a molecular method for the diagnosis of canine babesiosis

Babesiosis is a hemolytic disease caused by protozoans of the genus Babesia (Apicomplexa). This disease occurs worldwide and is transmitted by ticks to a variety of mammals, including humans. The objective of the present study was to optimize a molecular approach for the detection of a fragment of 18S rDNA of Babesia canis, Babesia vogeli, Babesia rossi or Babesia gibsoni based on a single semi-nested Polymerase Chain Reaction (PCR), and compare the efficiency of this approach with that of a simple PCR protocol. To this end, 100 blood samples collected from dogs with suspected hemoparasite infections were analyzed. A comparison of the results of simple PCR and semi-nested PCR indicated a highly significant difference (p value = 0.0000). While only five (5%) of the samples tested positive using the simple protocol, 22 (22%) were positive using the snPCR technique. The results of this study reinforce the findings of previous studies, which have demonstrated the greater sensitivity of tests based on nested or semi-nested PCR. Therefore, to avoid false-negative results due to low levels of parasitemia, we suggest the preferential use of this protocol in epidemiological studies of canine babesiosis, particularly those that require reliable estimates of the prevalence of infection

Virtual screening and repurposing of approved drugs targeting homoserine dehydrogenase from Paracoccidioides brasiliensis

Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Laboratório de Biologia Molecular. Ananindeua, PA, Brasil.Universidade da Amazônia. Ananindeua, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Paracoccidioidomycosis is a systemic mycosis endemic in Latin America, and one of the etiological agents of the disease is Paracoccidioides brasiliensis. Currently, available treatments present adversities, such as duration, side effects, and drug interactions. In search of new therapy possibilities, this study evaluates drugs approved for use against the homoserine dehydrogenase enzyme, by an in silico approach, which performs an important biosynthesis phase for the fungus and is not present in the human body. The three-dimensional structure of the homoserine dehydrogenase enzyme from Paracoccidioides brasiliensis was obtained by homology modeling. The model was validated, and simulations were performed for virtual screening of molecules of drugs approved from the Drugs-libs database by the MTiOpenScreen web server. Molecular dynamics in three replicas were used for four drugs with better results, and in two more molecules as a control, the HS9 with inhibition against enzyme and HON which shows inhibition against mold structure. Based on the results of molecular dynamics and the comparison of binding free energy, the drug that obtained the best result was Bemcentinib. In comparison with the controls, it presented a highly relevant affinity with − 44.63 kcal/mol, in addition to good structural stability and occupation of the active site. Therefore, Bemcentinib is a promising molecule for the inhibition of PbHSD protein (homoserine dehydrogenase of Paracoccidioides brasiliensis) and a therapeutic option to be investigated

Kudoa amazonica n. sp. (Myxozoa; Multivalvulida), a parasite of the esophageal musculature of the freshwater catfish, Hypophthalmus marginatus (Siluriformes: Pimelodidae), from a river of the Amazon region

Federal Rural University of Amazônia. Paragominas, PA, Brazil.Federal Rural University of Amazonia. Carlos Azevedo Research Laboratory. Belém, PA, Brazil.Amapá State University. Laboratory of Morphophysiology and Animal Health. Macapá, AP, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Epidemiologia e Imunologia Aplicada à Leishmaniose. Ananindeua, PA, Brasil.Federal University of Pará. Laboratory of Biomolecular Technology. Belém, PA, Brazil.Federal Rural University of Amazonia. Carlos Azevedo Research Laboratory. Belém, PA, Brazil.The class Myxozoa is a group of spore-producing eukaryote organisms that parasitize both freshwater and marine fish. The multivalvulide myxosporidian parasites of the genus Kudoa infect primarily the musculature of the fish host, some species are producing enzymes (proteases) capable of digesting muscle fibers. In the present study, 50 specimens of the freshwater catfish Hypophthalmus marginatus were collected from the Tocantins River in Pará, Brazil, and were analyzed. Overall, 68% of these specimens presented infections by Kudoa parasite in the esophageal musculature. The morphology of these parasite was examined under light microscopy and nucleotide sequences of the SSU rDNA gene were obtained for phylogenetic analyses. The species formed numerous whitish pseudocysts containing square spores with rounded extremities in the apical view, and four polar capsules of equal size. In the phylogenetic analyses, Kudoa amazonica n. sp. was characterized as a sister taxon of another freshwater species, Kudoa orbicularis. The combination of morphological, morphometric, and molecular data obtained in the present study provided a conclusive diagnosis of Kudoa amazonica n. sp., which is clearly distinct from all other Kudoa taxa described to date

Genomic screening of new putative antiviral lectins from Amazonian cyanobacteria based on a bioinformatics approach

Fundação Amazônia Paraense de Amparo à Pesquisa. Grant Number: ICAAF 099/2014; Conselho Nacional de Desenvolvimento Científico e Tecnológico. Grant Number: 311686/2015‐0Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Naturais. Laboratórios de Investigação Sistemática em Biotecnologia e Biodiversidade Molecular. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Lectins are proteins of nonimmune origin, which are capable of recognizing and binding to glycoconjugate moieties. Some of them can block the interaction of viral glycoproteins to the host cell receptors acting as antiviral agents. Although cyanobacterial lectins have presented broad biotechnological potential, little research has been directed to Amazonian Cyanobacterial diversity. In order to identify new antiviral lectins, we performed genomic analysis in seven cyanobacterial strains from Coleção Amazônica de Cianobactérias e Microalgas (CACIAM). We found 75 unique CDS presenting one or more lectin domains. Since almost all were annotated as hypothetical proteins, we used homology modeling and molecular dynamics simulations to evaluate the structural and functional properties of three CDS that were more similar to known antiviral lectins. Nostoc sp. CACIAM 19 as well as Tolypothrix sp. CACIAM 22 strains presented cyanovirin‐N homologues whose function was confirmed by binding free energy calculations. Asn, Glu, Thr, Lys, Leu, and Gly, which were described as binding residues for cyanovirin, were also observed on those structures. As for other known cyanovirins, those residues in both our models also made favorable interactions with dimannose. Finally, Alkalinema sp. CACIAM 70d presented one CDS, which was identified as a seven‐bladed beta‐propeller structure with binding sites predicted for sialic acid and N‐acetylglucosamine. Despite its singular structure, our analysis suggested this molecule as a new putative antiviral lectin. Overall, the identification and the characterization of new lectins and their homologues are a promising area in antiviral research, and Amazonian cyanobacteria present biotechnological potential to be explored in this regard

A molecular survey of three tick-borne pathogens in dogs from Algodoal village/Maiandeua island on the northeast coast of Pará, Brazil

Ehrlichia spp., Anaplasma spp. and Babesia spp. are obligate intracellular parasitic microorganisms found in the blood of domestic animals. Until then, there were no reports of these hemoparasites in dogs in the northeast of the State of Pará. The aim of this study was to record cases of natural infection by Ehrlichia sp., Anaplasma sp. and Babesia sp. in dogs from Ilha de Algodoal/ Maiandeua, State of Pará, Brazil, through the detection of DNA from these agents. Whole blood samples were collected from 52 animals, without considering breed, sex or age. Include results for different species of animals and parasites. Molecular analysis data showed 50% co-infection for Ehrlichia sp. and Anaplasma sp. This study allowed the detection of Ehrlichia canis and Anaplasma platys in domestic dogs on Algodoal Island, an important ecological tourism site in northern Brazil

Anti-dengue virus activity of scytovirin and evaluation of point mutation effects by molecular dynamics and binding free energy calculations

Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Naturais. Laboratórios de Investigação Sistemática em Biotecnologia e Biodiversidade Molecular. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Tecnologia Biomolecular. Belém, PA, Brazil.The absence of a specific treatment against DENV has led to intensive research into developing strategies for curing the infection. One lectin with high antiviral activity is scytovirin, which was isolated from the cyanobacterium Scytonema varium and has proven activity against HIV and Zaire Ebola Virus. To achieve the results presented here, we tested the affinity of full-length scytovirin, SD1 and SD2 separately, and six SD1 mutants for DENV glycoprotein E carbohydrate by Molecular Dynamics (MD) simulations and binding free energy calculations. It was possible to identify the key residues for protein-ligand interaction such as Glu10, Ala11, Pro17, Ans18, Arg30, Thr41, Ser42 and Arg43, which also has importance action against HIV. All binding free energy calculations showed negative values to ΔGbind of protein-DENV carbohydrate complexation. Additionally, these results are similar to the values of scytovirin and HIV gp120 carbohydrate complexation (-32.20 kcal/mol). Furthermore, we found that SD1 individually has more affinity to the carbohydrate and the Asn9, Glu10, Asn18, Arg30 and Arg43 demonstrated an important role in this matter. We also found that mutant G48R has better affinity (-34.10 kcal/mol) for the DENV carbohydrate than the wild type protein (-27.15 kcal/mol)