A comparative genomics study of the microbiome and freshwater resistome in Southern Pantanal.

This study explores the resistome and bacterial diversity of two small lakes in the Southern Pantanal, one in Aquidauana sub-region, close to a farm, and one in Abobral sub-region, an environmentally preserved area. Shotgun metagenomic sequencing data from water column samples collected near and far from the floating macrophyte Eichhornia crassipes were used. The Abobral small lake exhibited the highest diversity and abundance of antibiotic resistance genes (ARGs), antibiotic resistance classes (ARGCs), phylum, and genus. RPOB2 and its resistance class, multidrug resistance, were the most abundant ARG and ARGC, respectively. Pseudomonadota was the dominant phylum across all sites, and Streptomyces was the most abundant genus considering all sites.

SARS-CoV-2 and dialysis: humoral response, clinical and laboratory impacts before vaccination.

BACKGROUND: Patients with kidney disease on Hemodialysis (HD) are susceptible to Coronavirus Disease (COVID-19) due to multiple risk factors. AIM: This study aims to report the prevalence of antibodies against SARS-CoV-2 among patients on hemodialysis before vaccination in Brazil and to compare with clinical, demographic, and laboratory data. METHODS: Blood samples from 398 Chronic Kidney Disease (CKD) patients treated in three different private institutions in Rio de Janeiro State, Brazil were submitted to the total anti-SARS-CoV-2 testing. Kidney, liver, and hematological markers were also determined. Respiratory samples were tested by real-time PCR for SARS-CoV-2 RNA and positive samples were subjected to high-throughput sequencing on the MinION device. RESULTS: Overall, anti-SARS-CoV-2 prevalence was 54.5 % (217/398) and two individuals had SARS-CoV-2 RNA with variant B.1.1. High anti-SARS-CoV-2 seroprevalence was found in male gender and those with hospital admission in the last 3-months before the inclusion in the study. Lower red blood cell count was observed in the anti-SARS-CoV-2 seropositive group. High levels of anti-SARS-CoV-2 were found in those who reported symptoms, had low levels of eosinophils and low hematocrit, and who practiced physical activity. CONCLUSION: High prevalence of anti-SARS-CoV-2 was found in CKD patients before the universal immunization in Brazil suggesting that dialysis patients were highly exposed to SARS-CoV-2.

Metalloproteinases in Restorative Dentistry: An In Silico Study toward an Ideal Animal Model.

In dentistry, various animal models are used to evaluate adhesive systems, dental caries and periodontal diseases. Metalloproteinases (MMPs) are enzymes that degrade collagen in the dentin matrix and are categorized in over 20 different classes. Collagenases and gelatinases are intrinsic constituents of the human dentin organic matrix fibrillar network and are the most abundant MMPs in this tissue. Understanding such enzymes' action on dentin is important in the development of approaches that could reduce dentin degradation and provide restorative procedures with extended longevity. This in silico study is based on dentistry's most used animal models and intends to search for the most suitable, evolutionarily close to Homo sapiens. We were able to retrieve 176,077 mammalian MMP sequences from the UniProt database. These sequences were manually curated through a three-step process. After such, the remaining 3178 sequences were aligned in a multifasta file and phylogenetically reconstructed using the maximum likelihood method. Our study inferred that the animal models most evolutionarily related to Homo sapiens were Orcytolagus cuniculus (MMP-1 and MMP-8), Canis lupus (MMP-13), Rattus norvegicus (MMP-2) and Orcytolagus cuniculus (MMP-9). Further research will be needed for the biological validation of our findings.

Evolution of Toll, Spatzle and MyD88 in insects: the problem of the Diptera bias.

BACKGROUND: Arthropoda, the most numerous and diverse metazoan phylum, has species in many habitats where they encounter various microorganisms and, as a result, mechanisms for pathogen recognition and elimination have evolved. The Toll pathway, involved in the innate immune system, was first described as part of the developmental pathway for dorsal-ventral differentiation in Drosophila. Its later discovery in vertebrates suggested that this system was extremely conserved. However, there is variation in presence/absence, copy number and sequence divergence in various genes along the pathway. As most studies have only focused on Diptera, for a comprehensive and accurate homology-based approach it is important to understand gene function in a number of different species and, in a group as diverse as insects, the use of species belonging to different taxonomic groups is essential. RESULTS: We evaluated the diversity of Toll pathway gene families in 39 Arthropod genomes, encompassing 13 different Insect Orders. Through computational methods, we shed some light into the evolution and functional annotation of protein families involved in the Toll pathway innate immune response. Our data indicates that: 1) intracellular proteins of the Toll pathway show mostly species-specific expansions; 2) the different Toll subfamilies seem to have distinct evolutionary backgrounds; 3) patterns of gene expansion observed in the Toll phylogenetic tree indicate that homology based methods of functional inference might not be accurate for some subfamilies; 4) Spatzle subfamilies are highly divergent and also pose a problem for homology based inference; 5) Spatzle subfamilies should not be analyzed together in the same phylogenetic framework; 6) network analyses seem to be a good first step in inferring functional groups in these cases. We specifically show that understanding Drosophila's Toll functions might not indicate the same function in other species. CONCLUSIONS: Our results show the importance of using species representing the different orders to better understand insect gene content, origin and evolution. More specifically, in intracellular Toll pathway gene families the presence of orthologues has important implications for homology based functional inference. Also, the different evolutionary backgrounds of Toll gene subfamilies should be taken into consideration when functional studies are performed, especially for TOLL9, TOLL, TOLL2_7, and the new TOLL10 clade. The presence of Diptera specific clades or the ones lacking Diptera species show the importance of overcoming the Diptera bias when performing functional characterization of Toll pathways.

Core of the saliva microbiome: an analysis of the MG-RAST data.

BACKGROUND: Oral microbiota is considered as the second most complex in the human body and its dysbiosis can be responsible for oral diseases. Interactions between the microorganism communities and the host allow establishing the microbiological proles. Identifying the core microbiome is essential to predicting diseases and changes in environmental behavior from microorganisms. METHODS: Projects containing the term "SALIVA", deposited between 2014 and 2019 were recovered on the MG-RAST portal. Quality (Failed), taxonomic prediction (Unknown and Predicted), species richness (Rarefaction), and species diversity (Alpha) were analyzed according to sequencing approaches (Amplicon sequencing and Shotgun metagenomics). All data were checked for normality and homoscedasticity. Metagenomic projects were compared using the Mann-Whitney U test and Spearman's correlation. Microbiome cores were inferred by Principal Component Analysis. For all statistical tests, p < 0.05 was used. RESULTS: The study was performed with 3 projects, involving 245 Amplicon and 164 Shotgun metagenome datasets. All comparisons of variables, according to the type of sequencing, showed significant differences, except for the Predicted. In Shotgun metagenomics datasets the highest correlation was between Rarefaction and Failed (r = - 0.78) and the lowest between Alpha and Unknown (r = - 0.12). In Amplicon sequencing datasets, the variables Rarefaction and Unknown (r = 0.63) had the highest correlation and the lowest was between Alpha and Predicted (r = - 0.03). Shotgun metagenomics datasets showed a greater number of genera than Amplicon. Propionibacterium, Lactobacillus, and Prevotella were the most representative genera in Amplicon sequencing. In Shotgun metagenomics, the most representative genera were Escherichia, Chitinophaga, and Acinetobacter. CONCLUSIONS: Core of the salivary microbiome and genera diversity are dependent on the sequencing approaches. Available data suggest that Shotgun metagenomics and Amplicon sequencing have similar sensitivities to detect the taxonomic level investigated, although Shotgun metagenomics allows a deeper analysis of the microorganism diversity. Microbiome studies must consider characteristics and limitations of the sequencing approaches. Were identified 20 genera in the core of saliva microbiome, regardless of the health condition of the host. Some bacteria of the core need further study to better understand their role in the oral cavity.

Events linked to Geosmin and 2-methylisoborneol (2-MIB) in a Water Supply Source in the State of Rio de Janeiro, Brazil: a case study / Eventos atrelados a Geosmina e 2-metilisoborneol (2-MIB) em Manancial de Abastecimento do Estado do Rio de Janeiro, Brasil: estudo de caso

Tastes and odors in tap water are problems faced by water companies all over the world, with complaints from consumers especially during the summer when cyanobacterial blooms occur that produce compounds such as geosmin and 2-methylisoborneol (2-MIB).  We analyzed data on the intensity of taste and odor, and total concentration of the substances geosmin and 2-MIB, present in drinking water and raw water collected by the sanitation company that supplies the metropolitan region of the state of RJ/Brazil, during water crises in the year 2020, and in the new event of 2021, despite the previous warnings made by experts. Statistical and correlation analysis of the public data, and metagenomic analysis of the raw water captured from the Guandu basin in the year 2020 were performed. Organoleptic data allowed us to signal the presence of these compounds in drinking water, the values of the intensity of taste were more times above the MPV of the Brazilian legislation, with the average of the data for 2020 being 37.5 and in the following year this average was 5 times lower, indicating that the measures to remove the compounds were more effective, but did not eliminate the problem. For the year 2020 there was a linear correlation of 0.97 between the taste organoleptic standard and the total concentration of the compounds. The metagenomic data of the raw water of the year 2020, related to the genes mtf, mic and glys indicated that the substance responsible for the taste and odor was 2-MIB, since the read rate for it was higher, both in the first visit and only in the second visit, when there was still perception of intensity of taste and odor. Modifications in the surveillance system of the quality of the water taken and consumed need to be adopted to circumvent the problems of cyanobacterial proliferation in the Guandu basin, since conditions favorable to blooms will occur as long as the sanitation problems in this watershed are not solved.

Gostos e odores na água da torneira são problemas enfrentados por empresas produtoras de água em todo o mundo, com reclamações dos consumidores principalmente durante o verão, quando ocorrerem florações de cianobactérias produtoras dos compostos como geosmina e o 2-metilisoborneol (2-MIB).  Foram analisados dados de intensidade do gosto e do odor, e concentração total das substâncias geosmina e 2-MIB, presente na água  potável e na água bruta captada pela empresa de saneamento, que abastece a região metropolitana do estado do RJ/Brazil,  durante  crises hídricas do ano 2020, e no novo evento de 2021, a despeito dos alertas prévios feitos por especialistas. Foram realizadas análises estatísticas e de correlação dos dados públicos, e analise metagenômica da água bruta captada no manancial da Bacia do Guandu, no ano de 2020. Dados organolépticos permitiram sinalizar a presença desses compostos na água de consumo, os valores da intensidade do gosto estiveram maior número de vezes acima do VMP da legislação brasileira, sendo a  média dos dados de 2020 de 37,5 e no ano seguinte esta média foi 5 vezes menor, indicando que as medidas para retirar dos compostos foram mais eficazes, mas não eliminaram o problema. Para o ano de 2020 houve correlação linear de 0,97 entre o padrão organoléptico gosto e a concentração total dos compostos. Os dados metagenômicos da água bruta do ano 2020, relacionados com os genes mtf, mic e glys indicaram que a substância responsável pelo gosto e odor foi o 2-MIB, visto o índice de reads para ele ter sido maior, tanto na primeira visita e único na segunda, quando ainda havia percepção de intensidade de gosto e odor. Modificações no sistema de vigilância da qualidade da água captada e de consumo precisam ser adotadas para contornar os problemas de proliferação de cianobactérias na bacia do Guandu, pois condições favoráveis às florações irão acontecer enquanto não forem resolvidos os problemas de saneamento dessa bacia hidrográfica.

Multidrug-resistant Klebsiella quasipneumoniae subsp. similipneumoniae carrying blaNDM-blaCTX-M15 isolated from flies in Rio de Janeiro, Brazil.

OBJECTIVES: Flies have been implicated in the dispersal of medically important bacteria including members of the genus Klebsiella between different environmental compartments. The aim of this study was to retrieve and characterize antibiotic-resistant bacteria from flies collected near to hospitals. METHODS: Flies were collected in the vicinity of medical facilities and examined for bacteria demonstrating phenotypic resistance to ceftriaxone, followed by determination of phenotypic and genotypic resistance profiles. In addition, whole genome sequencing followed by phylogenetic analysis and resistance genotyping were performed with the multidrug-resistant (MDR) strain Lemef23, identified as Klebsiella quasipneumoniae subsp. similipneumoniae. RESULTS: The strain Lemef23, classified by multiple locus sequence typing as novel ST 3397, harboured numerous resistance genes. The blaNDM was located on a Tn3000 element, a common genetic platform for the carriage of this gene in Brazil. Inference of phylogenetic orthology of strain Lemef23 and other clinical isolates suggested an anthropogenic origin. CONCLUSIONS: The findings of this study support the role of flies as vectors of MDR bacteria of clinical importance and provide the first record of blaNDM-1 and blaCTXM-15 in a Brazilian isolate of K. quasipneumoniae subsp. similipneumoniae, demonstrating the value of surveying insects as reservoirs of antibiotic resistance.

Água de reúso: uma alternativa sustentável para o Brasil / Water reuse: a sustainable alternative for Brazil

RESUMO As motivações para a busca de soluções e alternativas que podem diminuir o estresse hídrico são evidentemente importantes. Mesmo que renováveis, os recursos hídricos de qualidade e potabilidade são escassos. Como alternativa, o reúso pode ajudar a diminuir o uso de águas potáveis para atividades que não necessitam de potabilidade. Este trabalho teve como objetivo realizar uma revisão da literatura sobre água de reúso a partir da definição de alternativa sustentável e racional para o uso da água. Realizou-se um levantamento bibliográfico do período de julho a dezembro de 2017 nas bases de dados Medical Literature Analysis and Retrievel System Online (MedLine), Literatura Latino-americana e do Caribe em Ciências da Saúde (LILACS), Embase e Institute for Scientific Information (ISI). Com a revisão bibliográfica foi possível identificar que o tema "reúso" vem sendo debatido para a gestão, a padronização e os riscos associados à utilização. No Brasil, não há uma legislação a nível nacional que regule o uso e determine os padrões de qualidade da água de reúso. No entanto, nas quatro regiões brasileiras foi observado que há sete estados com legislações e normas sobre o tema. Legislações internacionais apresentam uma relação abrangente de parâmetros que podem ser utilizados no debate para a legislação brasileira. Trabalhos produzidos pela comunidade acadêmica fundamentam os riscos sobre esse tipo de água. Publicações científicas sobre água de reúso devem ser estimuladas, assim como legislações mais detalhadas descrevendo os tipos de água de reúso, assim como os padrões de riscos relacionados.

ABSTRACT The motivations for seeking solutions and alternatives that can reduce water stress is clearly important. Even though renewable, water resources of quality and potability are scarce. As an alternative, reuse can help reduce the use of drinking water for activities that do not require drinking. The objective was to carry out a review of the literature on reuse water based on the definition of a sustainable and rational alternative for water use. A bibliographical survey was carried out from July to December 2017 in the MedLine, Lilacs, Embase, and ISI databases. The bibliographic review showed that the reused topic has been discussed with notes to the management, standardization, and risks associated with its use. There is no national Brazilian legislation that regulates the use and quality standards of reuse water. However, in the four Brazilian regions, the literature points to only seven Brazilian states with legislation and norms on the subject. International legislations present a comprehensive list of parameters that can be used in the discussion of Brazilian national legislation. Works have been produced by the academic community, basing the risks of this type of water. Scientific publications on reuse water should be encouraged, as well as more detailed legislation describing the types of reuse water and related risk patterns.

Multilocus Analysis Resolves the European Finch Epidemic Strain of Trichomonas gallinae and Suggests Introgression from Divergent Trichomonads.

In Europe, Trichomonas gallinae recently emerged as a cause of epidemic disease in songbirds. A clonal strain of the parasite, first found in the United Kingdom, has become the predominant strain there and spread to continental Europe. Discriminating this epidemic strain of T. gallinae from other strains necessitated development of multilocus sequence typing (MLST). Development of the MLST was facilitated by the assembly and annotation of a 54.7 Mb draft genome of a cloned stabilate of the A1 European finch epidemic strain (isolated from Greenfinch, Chloris chloris, XT-1081/07 in 2007) containing 21,924 protein coding genes. This enabled construction of a robust 19 locus MLST based on existing typing loci for Trichomonas vaginalis and T. gallinae. Our MLST has the sensitivity to discriminate strains within existing genotypes confidently, and resolves the American finch A1 genotype from the European finch epidemic A1 genotype. Interestingly, one isolate we obtained from a captive black-naped fruit dove Ptilinopsus melanospilus, was not truly T. gallinae but a hybrid of T. gallinae with a distant trichomonad lineage. Phylogenetic analysis of the individual loci in this fruit dove provides evidence of gene flow between distant trichomonad lineages at 2 of the 19 loci examined and may provide precedence for the emergence of other hybrid trichomonad genomes including T. vaginalis.

Aporé virus, a novel mammarenavirus (Bunyavirales: Arenaviridae) related to highly pathogenic virus from South America.

Here, we report the complete genome sequence of the Aporé virus (Bunyavirales: Arenaviridae), obtained from a wild rodent Oligoryzomys mattogrossae captured in Mato Grosso do Sul state, Brazil. The genome of this virus showed strong similarity to highly pathogenic mammarenavirus from South America.

Reproductive performance of Nellore heifers raised in extensive system undergoing different vaccination protocols in fixed-time artificial insemination (FTAI) / Desempenho reprodutivo de novilhas Nelore criadas em sistema extensivo no Pantanal, submetidas a diferentes protocolos vacinais na Inseminação Artificial em tempo Fixo (IATF)

ABSTRACT: This study evaluated the reproductive performance of Nellore heifers without previous vaccination undergoing fixed-time artificial insemination (FTAI) and subsequent natural breeding under different vaccination protocols against infectious bovine herpesvirus 1 (BHV-1), bovine viral diarrhea virus (BVDV), and bovine leptospirosis (BL). A total of 763 animals were included; the group assignment was as follows: T1, control without vaccination; T2, vaccination against BL; T3, vaccination against BHV-1and BVDV; T4,vaccination against combination of BL + BHV-1and BVDV. Vaccination was started 30 days before FTAI (D-40), and a booster dose was administered at D-10. Serological tests were used to verify the occurrence rate of natural infection by BL, BHV-1, and BVDV. Initial pregnancy was diagnosed at D30, D60, and D120 after FTAI. Final pregnancy rates were evaluated by group, weight, body-condition score, occurrence of estrus, inseminator, bull, and occurrence of abortions or embryonic death. In all groups, results indicated seropositivity of 100% for leptospirosis, 77.0-94.0% for BHV-1, and 64.0-87.0% for BVDV. The vaccination protocol was not a significant influencing factor of pregnancy rate and embryonic or fetal mortality; whereas, animal weight at the beginning of the breeding season and presence of estrus at FTAI were significant influencing factors of pregnancy rate. The vaccination protocol had no interference with the reproductive performance of the experimental groups.

RESUMO: O estudo foi conduzido para avaliar o desempenho reprodutivo de novilhas Nelore, não imunizadas para herpesvirus bovino tipo 1 (BHV-1), vírus da diarreia viral bovina (BVDV) e leptospirose bovina (LB), submetidas a diferentes protocolos vacinais em manejo de inseminação artificial em tempo fixo (IATF) e repasse com touro em monta natural. Um total de 763 animais foram distribuídos em quatro grupos: T1 - controle sem vacinação; T2 - vacinação contra LB; T3 - vacinação contra BHV-1 e BVDV T4 - vacinação com LB+ BHV-1 e BVDV. A vacinação foi iniciada 30 dias antes da IATF (D-40) com a aplicação da segunda dose em D-10. Verificou-se a taxa de ocorrência de infecção natural por LB, BHV-1 e BVDV realizando-se exames sorológicos. O diagnóstico de gestação inicial foi feito aos 30, 60 e 120 dias. Avaliou-se as taxas de prenhez em função dos grupos, do peso, escore de condição corporal, presença ou ausência de cio, inseminador, touro, ocorrência de abortos ou morte embrionária. Os resultados obtidos revelaram que soropositividade para leptospirose foi de 100% para todos os grupos, variando de 77,0% a 94,0% para BHV-1 e 64,0% a 87,0% para BVDV. O protocolo vacinal não interferiu na taxa de prenhez, de mortalidade embrionária ou fetal. Houve diferença na taxa de prenhez em relação ao peso dos animais no início da estação reprodutiva e a presença de cio no momento da IATF. Concluiu-se que o protocolo vacinal não interferiu no desempenho reprodutivo dos grupos experimentais.

Aporé virus, a novel mammarenavirus (Bunyavirales: Arenaviridae) related to highly pathogenic virus from South America

Here, we report the complete genome sequence of the Aporé virus (Bunyavirales: Arenaviridae), obtained from a wild rodent Oligoryzomys mattogrossae captured in Mato Grosso do Sul state, Brazil. The genome of this virus showed strong similarity to highly pathogenic mammarenavirus from South America.

Systematic Identification and Classification of ß-Lactamases Based on Sequence Similarity Criteria: ß-Lactamase Annotation.

ß-lactamases, the enzymes responsible for resistance to ß-lactam antibiotics, are widespread among prokaryotic genera. However, current ß-lactamase classification schemes do not represent their present diversity. Here, we propose a workflow to identify and classify ß-lactamases. Initially, a set of curated sequences was used as a model for the construction of profiles Hidden Markov Models (HMM), specific for each ß-lactamase class. An extensive, nonredundant set of ß-lactamase sequences was constructed from 7 different resistance proteins databases to test the methodology. The profiles HMM were improved for their specificity and sensitivity and then applied to fully assembled genomes. Five hierarchical classification levels are described, and a new class of ß-lactamases with fused domains is proposed. Our profiles HMM provide a better annotation of ß-lactamases, with classes and subclasses defined by objective criteria such as sequence similarity. This classification offers a solid base to the elaboration of studies on the diversity, dispersion, prevalence, and evolution of the different classes and subclasses of this critical enzymatic activity.

Homology Inference Based on a Reconciliation Approach for the Comparative Genomics of Protozoa.

Protozoa parasites are responsible for several diseases in tropical countries, such as malaria, sleeping sickness, Chagas disease, leishmaniasis, amebiasis, and giardiasis, which together threaten millions of people around the world. In addition, most of the classic parasitic diseases due to protozoa are zoonotic. Understanding the biology of these organisms plays a relevant role in combating these diseases. Using homology inference and comparative genomics, this study targeted 3 protozoan species from different Phyla: Cryptosporidium muris (Apicomplexa), Entamoeba invadens (Amoebozoa), and Trypanosoma grayi (Euglenozoa). In this study, we propose a new approach for the identification of homologs, based on the reconciliation of the results of 2 different homology inference software programs. Our results showed that 46.1% (59/128) of the groups inferred by our reconciliation approach could be validated using this methodology. These validated groups are here called homologous Supergroups and were compared with SUPERFAMILY and Pfam Clans.

Mining of potential drug targets through the identification of essential and analogous enzymes in the genomes of pathogens of Glycine max, Zea mays and Solanum lycopersicum.

Pesticides are one of the most widely used pest and disease control measures in plant crops and their indiscriminate use poses a direct risk to the health of populations and environment around the world. As a result, there is a great need for the development of new, less toxic molecules to be employed against plant pathogens. In this work, we employed an in silico approach to study the genes coding for enzymes of the genomes of three commercially important plants, soybean (Glycine max), tomato (Solanum lycopersicum) and corn (Zea mays), as well as 15 plant pathogens (4 bacteria and 11 fungi), focusing on revealing a set of essential and non-homologous isofunctional enzymes (NISEs) that could be prioritized as drug targets. By combining sequence and structural data, we obtained an initial set of 568 cases of analogy, of which 97 were validated and further refined, revealing a subset of 29 essential enzymatic activities with a total of 119 different structural forms, most belonging to central metabolic routes, including the carbohydrate metabolism, the metabolism of amino acids, among others. Further, another subset of 26 enzymatic activities possess a tertiary structure specific for the pathogen, not present in plants, men and Apis mellifera, which may be of importance for the development of specific enzymatic inhibitors against plant diseases that are less harmful to humans and the environment.

Morphofunctional diversity of equine of varied genetic compositions raised in the Pantanal biome of Brazil.

Evaluating phenotypic diversity makes it possible to identify discrepancies in aptitudes among animals of different genetic bases, which is an indicator of adaptive or selective differences between populations. The objective of this work was to evaluate the morphofunctional diversity of 452 male and female adult equines (Arabian, Quarter Mile, Pantaneiro, and Criollo breeds, and undefined crossbreeds of horses and mules) raised in the Pantanal biome (Brazil). Linear measurements were performed to estimate conformation indexes. Initially, a discriminant analysis was performed, regardless of the animal's size, followed by factor analysis. The factors were characterized and used as new variables. The diversity among equines and their relationship with the factors were evaluated using multivariate analysis. The factors were classified according to their decreasing importance: balance, rusticity, and robustness for the measurement factors; and load, ability, conformation, and equilibrium for the index factors. The genetic groups of equines have well-defined morphofunctional characteristics. The main differences are based on the rusticity and ability typologies in relation to those based on performance. Equines introduced to the Pantanal biome presented a more robust and compact body with good conformation. As a result, these horses may have superior athletic performance during equestrian activities when compared to the Pantaneiro local breed. However, this biotype may represent less rusticity (less adaptive capacity). Therefore, the regional breed can be equal or better in equestrian activities than breeds introduced to the Pantanal biome. Thus, breeders may cross horses from local breeds as an alternative to those introduced. Undefined crossbred male equines presented a different profile from the Pantaneiro breed, which may indicate little use of crossbreeds in breeding.

ProtozoaDB 2.0: A Trypanosoma Brucei Case Study.

Over the last decade new species of Protozoa have been sequenced and deposited in GenBank. Analyzing large amounts of genomic data, especially using Next Generation Sequencing (NGS), is not a trivial task, considering that researchers used to deal or focus their studies on few genes or gene families or even small genomes. To facilitate the information extraction process from genomic data, we developed a database system called ProtozoaDB that included five genomes of Protozoa in its first version. In the present study, we present a new version of ProtozoaDB called ProtozoaDB 2.0, now with the genomes of 22 pathogenic Protozoa. The system has been fully remodeled to allow for new tools and a more expanded view of data, and now includes a number of analyses such as: (i) similarities with other databases (model organisms, the Conserved Domains Database, and the Protein Data Bank); (ii) visualization of KEGG metabolic pathways; (iii) the protein structure from PDB; (iv) homology inferences; (v) the search for related publications in PubMed; (vi) superfamily classification; and (vii) phenotype inferences based on comparisons with model organisms. ProtozoaDB 2.0 supports RESTful Web Services to make data access easier. Those services were written in Ruby language using Ruby on Rails (RoR). This new version also allows a more detailed analysis of the object of study, as well as expanding the number of genomes and proteomes available to the scientific community. In our case study, a group of prenyltransferase proteinsalready described in the literature was found to be a good drug target for Trypanosomatids.

The role of amides in seminal cryopreservation of wild silverside, Odontesthes bonariensis.

Amides were tested as internal cryoprotectants for the preservation of wild silverside (Odontesthes bonariensis) sperm. The semen was diluted in modified Mounib's medium and cryopreserved by adding 2, 5, 8 or 11% of dimethyl acetamide (DMA), dimethyl formamide (DMF) or methyl formamide (MF). Dimethyl sulfoxide (DMSO) at a concentration of 10% diluted in modified Mounib's medium was used as a control. The rate motility (17.7 ± 1.9%) and time motility (143.2 ± 9.7 s) (P < 0.05) of the sperm were higher with 2% DMF when compared with the other treatments. Despite the better motility results obtained with 2% DMF, the solution was not able to maintain cellular structure integrity of the cryopreserved sperm. The 10% DMSO and 8% MF treatment allowed for completeness of the plasma membrane (34.8% and 29%), functional mitochondria (19.8% and 16.2%) and plasma membrane fluidity (39.4% and 46.4%); furthermore, rate motility (11.8% and 10%) and time motility (81.4 s and 71.8 s) of the sperm were found to be at suitable levels when compared with 2% DMF. Thus, our evaluation suggests that 10% DMSO and 8% MF provide better cryopreservation of O. bonariensis sperm cells.