Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Application of PolyPRep tools on HIV protease polyproteins using molecular docking

Abstract In recent years, the development of high-throughput technologies for obtaining sequence data leveraged the possibility of analysis of protein data in silico. However, when it comes to viral polyprotein interaction studies, there is a gap in the representation of those proteins, given their size and length. The prepare for studies using state-of-the-art techniques such as Machine Learning, a good representation of such proteins is a must. We present an alternative to this problem, implementing a fragmentation and modeling protocol to prepare those polyproteins in the form of peptide fragments. Such procedure is made by several scripts, implemented together on the workflow we call PolyPRep, a tool written in Python script and available in GitHub. This software is freely available only for noncommercial users.

Resumo Nos últimos anos, o desenvolvimento de tecnologias de alto rendimento para obtenção de dados sequenciais potencializou a possibilidade de análise de dados proteicos in silico. No entanto, quando se trata de estudos de interação de poliproteínas virais, existe uma lacuna na representação dessas proteínas, devido ao seu tamanho e comprimento. Para estudos utilizando técnicas de ponta como o Aprendizado de Máquina, uma boa representação dessas proteínas é imprescindível. Apresentamos uma alternativa para este problema, implementando um protocolo de fragmentação e modelagem para preparar essas poliproteínas na forma de fragmentos de peptídeos. Tal procedimento é feito por diversos scripts, implementados em conjunto no workflow que chamamos de PolyPRep, uma ferramenta escrita em script Python e disponível no GitHub. Este software está disponível gratuitamente apenas para usuários não comerciais.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil / Dieta do cachorro-do-mato (Cerdocyon thous) em duas unidades de conservação da Floresta Amazônica, Brasil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Application of PolyPRep tools on HIV protease polyproteins using molecular docking / Aplicação da ferramenta PolyPRep em poliproteínas de protease de HIV usando docking molecular

Abstract In recent years, the development of high-throughput technologies for obtaining sequence data leveraged the possibility of analysis of protein data in silico. However, when it comes to viral polyprotein interaction studies, there is a gap in the representation of those proteins, given their size and length. The prepare for studies using state-of-the-art techniques such as Machine Learning, a good representation of such proteins is a must. We present an alternative to this problem, implementing a fragmentation and modeling protocol to prepare those polyproteins in the form of peptide fragments. Such procedure is made by several scripts, implemented together on the workflow we call PolyPRep, a tool written in Python script and available in GitHub. This software is freely available only for noncommercial users.

Resumo Nos últimos anos, o desenvolvimento de tecnologias de alto rendimento para obtenção de dados sequenciais potencializou a possibilidade de análise de dados proteicos in silico. No entanto, quando se trata de estudos de interação de poliproteínas virais, existe uma lacuna na representação dessas proteínas, devido ao seu tamanho e comprimento. Para estudos utilizando técnicas de ponta como o Aprendizado de Máquina, uma boa representação dessas proteínas é imprescindível. Apresentamos uma alternativa para este problema, implementando um protocolo de fragmentação e modelagem para preparar essas poliproteínas na forma de fragmentos de peptídeos. Tal procedimento é feito por diversos scripts, implementados em conjunto no workflow que chamamos de PolyPRep, uma ferramenta escrita em script Python e disponível no GitHub. Este software está disponível gratuitamente apenas para usuários não comerciais.

Coleoborers in Bambusa sp. in the Southern Amazon / Coleobrocas em Bambusa sp. na Amazonia Meridional

Bamboo (Bambusa sp.) is a grass species with high potential for exploitation, however, this raw material is easily attacked by xylophagous agents such as Coleoptera. The objective of this study is thus to analyse the presence of wood-boring beetles associated with Bambusa sp. in Alta Floresta, Mato Grosso, Brazil. The study was carried out in a bamboo plantation, from August 2016 to July 2017. Two models of ethanolic traps were used, PET Santa Maria and ESALQ-84. Three families of wood-boring beetles were observed in this study: Bostrichidae, Cerambycidae and Curculionidae. 2,144 individuals were identified, belonging to 19 species, in four subfamilies and 11 tribes, and the species Xyleborus affinis was the most frequently collected. Thus, this study constitutes an important contribution to the knowledge of coleoborers associated with bamboo plantations, with the registration of the occurrence of species, as well as contributing to the knowledge of the entomofauna associated with species cultivated in the Southern Amazon.

O bambu (Bambusa sp.) é uma espécie de gramínea com alto potencial de exploração de fibras, no entanto, essa matéria-prima é facilmente atacada por agentes xilófagos, como os coleópteros. O objetivo deste estudo foi analisar a presença de besouros associados a Bambusa sp. em Alta Floresta, Mato Grosso, Brasil. O estudo foi realizado em um plantio de bambu, no período de agosto de 2016 a julho de 2017. Foram utilizados dois modelos de armadilhas etanólicas, o PET Santa Maria e ESALQ-84. Três famílias de coleobrocas foram observadas neste estudo: Bostrichidae, Cerambycidae e Curculionidae. Foram identificados 2,144 indivíduos, pertencentes a 19 espécies, em quatro subfamílias e 11 tribos, e Xyleborus affinis foi a espécie com maior número de indivíduos coletados. Assim, este estudo constitui-se em uma importante contribuição para o conhecimento de coleobrocas associadas a plantios de bamboo, com o registro de ocorrência de espécies, bem como contribui para o conhecimento da entomofauna associada a espécies cultivadas na Amazônia Meridional.

Coleoborers in Bambusa sp. in the Southern Amazon

Abstract Bamboo (Bambusa sp.) is a grass species with high potential for exploitation, however, this raw material is easily attacked by xylophagous agents such as Coleoptera. The objective of this study is thus to analyse the presence of wood-boring beetles associated with Bambusa sp. in Alta Floresta, Mato Grosso, Brazil. The study was carried out in a bamboo plantation, from August 2016 to July 2017. Two models of ethanolic traps were used, PET Santa Maria and ESALQ-84. Three families of wood-boring beetles were observed in this study: Bostrichidae, Cerambycidae and Curculionidae. 2,144 individuals were identified, belonging to 19 species, in four subfamilies and 11 tribes, and the species Xyleborus affinis was the most frequently collected. Thus, this study constitutes an important contribution to the knowledge of coleoborers associated with bamboo plantations, with the registration of the occurrence of species, as well as contributing to the knowledge of the entomofauna associated with species cultivated in the Southern Amazon.

Resumo O bambu (Bambusa sp.) é uma espécie de gramínea com alto potencial de exploração de fibras, no entanto, essa matéria-prima é facilmente atacada por agentes xilófagos, como os coleópteros. O objetivo deste estudo foi analisar a presença de besouros associados a Bambusa sp. em Alta Floresta, Mato Grosso, Brasil. O estudo foi realizado em um plantio de bambu, no período de agosto de 2016 a julho de 2017. Foram utilizados dois modelos de armadilhas etanólicas, o PET Santa Maria e ESALQ-84. Três famílias de coleobrocas foram observadas neste estudo: Bostrichidae, Cerambycidae e Curculionidae. Foram identificados 2,144 indivíduos, pertencentes a 19 espécies, em quatro subfamílias e 11 tribos, e Xyleborus affinis foi a espécie com maior número de indivíduos coletados. Assim, este estudo constitui-se em uma importante contribuição para o conhecimento de coleobrocas associadas a plantios de bamboo, com o registro de ocorrência de espécies, bem como contribui para o conhecimento da entomofauna associada a espécies cultivadas na Amazônia Meridional.

Coleoborers in Bambusa sp. in the Southern Amazon / Coleobrocas em Bambusa sp. na Amazonia Meridional

Abstract Bamboo (Bambusa sp.) is a grass species with high potential for exploitation, however, this raw material is easily attacked by xylophagous agents such as Coleoptera. The objective of this study is thus to analyse the presence of wood-boring beetles associated with Bambusa sp. in Alta Floresta, Mato Grosso, Brazil. The study was carried out in a bamboo plantation, from August 2016 to July 2017. Two models of ethanolic traps were used, PET Santa Maria and ESALQ-84. Three families of wood-boring beetles were observed in this study: Bostrichidae, Cerambycidae and Curculionidae. 2,144 individuals were identified, belonging to 19 species, in four subfamilies and 11 tribes, and the species Xyleborus affinis was the most frequently collected. Thus, this study constitutes an important contribution to the knowledge of coleoborers associated with bamboo plantations, with the registration of the occurrence of species, as well as contributing to the knowledge of the entomofauna associated with species cultivated in the Southern Amazon.

Resumo O bambu (Bambusa sp.) é uma espécie de gramínea com alto potencial de exploração de fibras, no entanto, essa matéria-prima é facilmente atacada por agentes xilófagos, como os coleópteros. O objetivo deste estudo foi analisar a presença de besouros associados a Bambusa sp. em Alta Floresta, Mato Grosso, Brasil. O estudo foi realizado em um plantio de bambu, no período de agosto de 2016 a julho de 2017. Foram utilizados dois modelos de armadilhas etanólicas, o PET Santa Maria e ESALQ-84. Três famílias de coleobrocas foram observadas neste estudo: Bostrichidae, Cerambycidae e Curculionidae. Foram identificados 2,144 indivíduos, pertencentes a 19 espécies, em quatro subfamílias e 11 tribos, e Xyleborus affinis foi a espécie com maior número de indivíduos coletados. Assim, este estudo constitui-se em uma importante contribuição para o conhecimento de coleobrocas associadas a plantios de bamboo, com o registro de ocorrência de espécies, bem como contribui para o conhecimento da entomofauna associada a espécies cultivadas na Amazônia Meridional.

Caracterización de la displasia broncopulmonar según dos definiciones en prematuros chilenos / Characterization of bronchopulmonary dysplasia in relation with two definitions in chilean preterm infants

Existen controversias en la definición de la displasia broncopulmonar, siendo las más utilizadas el requerimiento de O2 durante 28 días o a las 36 semanas de edad gestacional corregida (EGC). Nuestro objetivo fue determinar la incidencia y características clínicas de los prematuros nacidos antes de las 32 semanas (RNP≤ 32s) con requerimiento de O2 a los 28 días de vida (DBP28d) y a las 36 semanas de EGC (DBP36s) en una unidad neonatal de Santiago, Chile, entre los años 2012 y 2019. Es un estudio descriptivo, retrospectivo con componente analítico. La población estudiada incluyó 535 RNP≤ 32s, vivos a las 36 semanas o dados de alta después de las 34 semanas de EGC. De los 242 prematuros DBP28d, 203 (83,88%) fueron DBP36s; 16 de los 242 (6%) requirió O2 durante menos de 28 días consecutivos, de los cuales 7, aún lo requerían a las 36 semanas. Los predictores de DBP36s fueron: sexo masculino (OR 2,42, IC del 95%: 1,24-4,69), peso al nacer (OR 1, IC del 95%: 0,99-1), edad gestacional (OR 0,75, IC del 95%: 0,57-0,97), APGAR a los 5 min, (OR 0,01, IC del 95%: 0,003-0,05), el requerimiento de presión positiva continua o cánula nasal de alto flujo (OR 1,1, IC del 95%: 1,04-1,17) y días de ventilación mecánica invasiva (OR 1,1,95% IC: 1-1,2). Conclusiones: No encontramos una diferencia significativa en la incidencia de DBP entre las definiciones de DBP28d y DBP36s; y la mayoría de los RNP< 32s con diagnóstico de DBP36s se pudieron identificar a los 28 días de vida.

Bronchopulmonary dysplasia (BPD) is the most common chronic lung disease in premature newborns. It is commonly defined as a need for supplemental O2 for 28 days or at 36 weeks postmenstrual age (PMA) (BPD36w). Our objective was to determine the incidence and clinical characteristics of premature neonates born at less than 32 weeks (PNB< 32w) with O2 requirement at 28 days of life (DBP28d) and DBP36w in a neonatal unit of Santiago, Chile, between 2012 and 2019. This is a descriptive, retrospective study with an analytical component. The study population included 535 PNB< 32w, alive at 36 weeks or discharged after 34 PMA. Of the 242 premature BPD28d, 203 (83,88%) were BPD36w; 16 (6%) required O2 for less than 28 consecutive days, of which 7 still required it at 36 weeks. The predictors of BPD36w were: male (OR 2.42, 95% CI: 1.24-4.69), birth weight (OR 1, 95% CI: 0.99-1), gestational age (OR 0.75, 95% CI: 0.57-0.97), APGAR at 5 min, (OR 0.01, 95% CI: 0.003-0.05), continuous positive pressure or high-flow nasal cannula requirement (OR 1.1, 95% CI: 1.04-1.17) and days of invasive mechanical ventilation (OR 1.1, 95% CI: 1-1.2). Conclusions: We did not find a significant difference in the incidence of BPD between the definitions of BPD28d and BPD36s; and the majority of PNB < 32w diagnosed with BPD36w can be identified at 28 days of life.

Plant stem cell extract from Coffea canephora shows antioxidant, anti-inflammatory, and skin regenerative properties mediated by suppression of nuclear factor-κB

Plant cell cultures have become a promising production platform of bioactive compounds for biomedical and cosmetic uses in the last decades. However, the success so far has been limited. The study aimed to evaluate the effectiveness of this unique biotechnology process to obtain a bioactive stem cell extract of Coffea canephora (SCECC) with antioxidant, anti-inflammatory, and regenerative properties. Total phenolic and flavonoid contents were determined in the SCECC by spectrophotometry. The chemical composition of the extracts was characterized by mass spectrometry. Antioxidant activity was evaluated using the colorimetric methods of free radical scavenging 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and the ferric reducing ability of plasma (FRAP). The anti-inflammatory activity was determined in lipopolysaccharide-stimulated RAW 264.7 macrophages through the production of superoxide anion (O2•-), nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and the activity of nuclear factor kappa B (NF-κB). Moreover, the ability of SCECC to stimulate the proliferation and migration of fibroblasts was assessed. Five compounds were tentatively identified, two flavonoids, two phenolic acids, and one sugar. High phenolic content and antioxidant activity were observed in the SCECC. SCECC promoted the proliferation and migration of fibroblasts and suppressed the pro-inflammatory mediators O2•-, NO, TNF-α, and IL-6 in a dose-dependent manner. Moreover, SCECC inhibited the NF-κB transcription factor. Therefore, we obtained evidence that the extract from C. canephora stem cells can be used as a natural agent against skin damage. Hence, it could be of interest in cosmetics for preventing skin aging.

Antimicrobial resistance of Pseudomonas aeruginosa isolated from patients with pneumonia during the COVID-19 pandemic and pre-pandemic periods in Northeast Brazil

Healthcare-related infections caused by resistant microorganisms are a severe public health problem and are becoming increasingly prevalent in the hospital environment, especially Pseudomonas aeruginosa. This work aimed to evaluate the resistance profile of Pseudomonas aeruginosa to antimicrobials before the COVID-19 pandemic and during the pandemic period. Bacteria strains were obtained from tracheal aspiration, sputum, and bronchoalveolar lavage for diagnosis and phenotypic characterization. Matrix assisted laser-desorption ionization-time of flight mass spectrometry (MALD-TOF MS) was used to identify strains. Automated Phoenix and VITEK® 2 Compact system and the disc diffusion method were performed to determine the antimicrobial susceptibility profile. A total of 41,000 medical reports from adult patients with pneumonia were analyzed. Of these, 951 patients were positive for P. aeruginosa, of which 373 were related to the pre-pandemic period and 578 to the pandemic period. Older men (≥60 years) were more prevalent in both periods. P. aeruginosa strains were resistant to imipenem in both periods: 38.8 and 42.5%, respectively, followed by meropenem (34.2 and 39.2%), ciprofloxacin (33.6 and 36.7%), and levofloxacin (34.9 and 43.5%). Intensive care units had the highest percentage of affected patients (62 and 65%) compared with other sectors, with a prevalence of 71% in the public network before COVID-19 and 59% during the pandemic. Our data showed a prevalence of P. aeruginosa in elderly patients in both the pre-pandemic and pandemic periods. In addition, an increase in P. aeruginosa resistance to beta-lactams, quinolones, carbapenems, and cephalosporins was observed during the COVID-19 pandemic compared with the period before the pandemic, especially in ICUs.

Unlocking the molecular realm: advanced approaches for identifying clinically and environmentally relevant bacteria

Rapid, effective, and specific identification of clinical and environmental bacterial pathogens is of major importance for their control. Traditionally, bacteria have been identified by phenotypic methods based on morphological, biochemical, and metabolic properties. While these methods are very useful in clinical practice, they have limitations including a poor ability to differentiate within and between species and time-consuming workflows. Newly developed molecular methods can greatly improve the accuracy of taxonomic characterization, identifying specific strains of medical or environmental importance. However, due to high costs and the need for trained professionals, these methods are not yet routine in diagnostic laboratories. Thus, disseminating knowledge on advances in molecular identification techniques is pivotal to make these methodologies accessible. The objective of this work was to review and discuss current molecular techniques for bacteria identification aiming to track and monitor microbial agents in clinical and environmental samples.

P2X7 receptor antagonist improves gastrointestinal disorders in spontaneously hypertensive rats

The purinergic system participates in the control of blood pressure. Hypertension promotes the occurrence of gastrointestinal disorders such as intestinal inflammation and gastric emptying delay. This study aimed i) to investigate the participation of the P2X7 receptor blocker Brilliant Blue G (BBG) on gastric emptying of solids and changes in oxidative stress in the gastric fundus, duodenum, and colon of spontaneously hypertensive rats (SHR) and ii) to study the putative relationship of this effect with the renin-angiotensin system. Rats were divided into five groups: Control, SHR, SHR+BBG, SHR+BBG+ATP, and SHR+BBG+ANG II. In the gastrointestinal tract, we assessed gastric emptying (GE) and oxidative stress markers (NOx, MPO, GSH, SOD). We observed a decrease in the GE rate (P<0.05) in SHR vs control rats (21.8±2.0% vs 42.8±3.5%). The decrease in GE was returned (P<0.05) to control levels by BBG in SHR rats (21.8±2.0% vs 41.6±3.2%). Co-administration of ATP or ANG II together with BBG bypassed the effect of the P2X7 antagonist on GE in SHR (P<0.05) (21.9±5.0% vs 25.6±3.0% vs 41.6±3.2%). The MPO activity increased (P<0.05) in the gastric fundus of SHR compared to control rats (6.12±2.26 vs 0.077±0.02 UMPO/mg tissue); this effect was prevented (P<0.05) by BBG (0.55±0.15 vs 6.12±2.26 UMPO/mg tissue). Data demonstrated that blockage of P2X7 receptors with BBG can improve the GE delay and oxidative stress biomarkers in SHR animals. This preventive effect of BBG on GE delay was abrogated by ANG II and ATP, thus prompting crosstalk between renin-angiotensin and the purinergic signaling systems underlying this phenomenon.

Serratia nevei 9rpt1, a Potential Microorganism for Phosphorus Recovery

Aims: Phosphorus (P) is an essential element for life, finite and irreplaceable. Its constant exploration on a global scale has motivated frequent alerts regarding an eventual crisis due to the shortage of this nutrient. However, it is possible to recycle it and reintroduce it into the ecological cycle. One viable alternative is the microbial recovery of phosphate. Study design: This study is based on systematic bioprospection of bacteria in phosphate-deficient Amazon regions. Place and Duration of Study: Bacteria were isolated from black water samples, collected in the Rio Pretinho, located at Serra do Aracá, Barcelos, Amazonas, Brazil, from January to July 2019. Methodology: Microbial isolation was performed in Luria Bertani agar medium. For the genomic study, the isolate with the best performance in the phosphate uptake test was chosen. The WGS was carried out in a Illumina HiSeq 2500 System. The assembly of the draft genome was carried out with the SPAdes and the annotation by the NCBI Prokaryotic Genome Annotation Pipeline (PGAP). Results: Serratia nevei 9rpt1 recovers 90% of the phosphate available in the culture medium. Its draft genome comprises 5.4 MB, the GC content is 59.52% and 4,922 coding sequences were identified, among these, two pst operons: one complete, containing the five pst genes and one missing pstS, pstC and phoU genes. Conclusion: Serratia nevei 9rpt1, isolated from an Amazonian environment poor in phosphate, is very efficient to uptake this nutrient in a Pi starvation condition. The genomic findings revealed this strain has an additional high affinity Pi uptake pst system containing the ATP-binding protein PstB, the canonical permease PstA, a putative permease other than PstC, upstream of the PstA and two essential enzymes in the polyphosphate metabolism: polyphosphate kinase 1 and exopolyphosphatase.

Analysis of the Frequency of the Toll-like 2 Gene Polymorphism in Leprosy

Polymorphisms in genes that are responsible for encoding cytokines and receptors involved in the immune response, such as Toll-like Receptor (TLR) 2 in leprosy, are of great interest for immunogenetic studies. This work aimed to analyze the possible association of single nucleotide polymorphism (SNP), synonymous, rs3804100 of the TLR2 gene with leprosy. The study was conducted in Bacteriology and Mycology section of Evandro Chagas Institute, Brazil between August 2020 and July 2021.The scope of the study consisted of 122 subjects from cities of Goian閟ia, Rondon, Curion髉olis, Altamira, Parauapebas and Reden玢o of the State of Par�, Brazil. Genotyping was performed by conventional PCR and sequencing in the ABI 3130 Genetic Analyzer (Applied Biosystems�) using primer nucleotides designed by the Primer3Plus program from the genomic region 揌omo sapiens toll like receptor 2 (TLR2) transcript variant X6, mRNA�, deposited in GenBank with reference XM_011532216.2. The analyzes were performed based on Fisher's exact test. It was managed in accordance with Helsinki Declaration and the Brazilian National Health Council and with approval of the ethics committee at Evandro Chagas Institute, under opinion number: 3.950.570. No associations between gender and leprosy were possible (P> 0.05). However, associations were observed between age groups, which were significant between those over 46 years old (P=0.004) and the 2nd dose of BCG as a more protective agent between the groups analyzed (P=0.004). For the subjects with the typed genotypes, 68 contacts had T/T genotype and only 4 T/C genotypes, while in multibacillary (MB) group only 1 T/C genotype was found and none in paucibacillary (PB) (P> 0.05). We conclude that there is no association between the TLR2 SNP rs3804100 and leprosy in the Par� population, which still indicates the need for new immunogenetic studies with other genes involved in the immune response and a greater number of polymorphisms.

Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These by products can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-¹. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-¹. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.