Publications by authors named "Tiago Antônio de Oliveira Mendes"

57 Publications

Ceftriaxone causes dysbiosis and changes intestinal structure in adjuvant obesity treatment.

Pharmacol Rep 2021 Nov 10. Epub 2021 Nov 10.

Department of Nutrition and Health, Universidade Federal de Viçosa, Avenue PH Rolfs s/n, Vicosa, MG, 36570-900, Brazil.

Background: Obesity is still a worldwide public health problem, requiring the development of adjuvant therapies to combat it. In this context, modulation of the intestinal microbiota seems prominent, given that the composition of the intestinal microbiota contributes to the outcome of this disease. The aim of this work is to investigate the treatment with an antimicrobial and/or a potential probiotic against overweight.

Methods: Male C57BL/6J mice were subjected to a 12-week overweight induction protocol. After that, 4-week treatment was started, with mice divided into four groups: control, treated with distilled water; potential probiotic, with Lactobacillus gasseri LG-G12; antimicrobial, with ceftriaxone; and antimicrobial + potential probiotic with ceftriaxone in the first 2 weeks and L. gasseri LG-G12 in the subsequent weeks.

Results: The treatment with ceftriaxone in isolated form or in combination with the potential probiotic provided a reduction in body fat. However, such effect is supposed to be a consequence of the negative action of ceftriaxone on the intestinal microbiota composition, and this intestinal dysbiosis may have contributed to the destruction of the intestinal villi structure, which led to a reduction in the absorptive surface. Also, the effects of L. gasseri LG-G12 apparently have been masked by the consumption of the high-fat diet.

Conclusions: The results indicate that the use of a ceftriaxone in the adjuvant treatment of overweight is not recommended due to the potential risk of developing inflammatory bowel disease.
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http://dx.doi.org/10.1007/s43440-021-00336-xDOI Listing
November 2021

Dibenzoylmethane derivative inhibits melanoma cancer in vitro and in vivo through induction of intrinsic and extrinsic apoptotic pathways.

Chem Biol Interact 2021 Nov 4;351:109734. Epub 2021 Nov 4.

Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, Minas Gerais, 36570-900, Brazil. Electronic address:

Malignant melanoma has a low incidence, but is the most lethal type of skin cancer. Studies have shown that dibenzoylmethanes (DBMs) have interesting biological activities, including antineoplastic properties. These findings led us to investigate whether news DBM derivatives exert antitumor effects against skin cancers. In a previous study, we found that 1,3-diphenyl-2-benzyl-1,3-propanedione (DPBP) has high in vitro antineoplastic activity against murine B16F10 melanoma cells, with an IC of 6.25 μg/mL. In the current study, we used transdermal and topical formulations of DPBP to evaluate its activity and molecular mechanism of action in a murine model of melanoma. The compound induces tumor cell death with high selectivity (selectivity index of 41.94) by triggering apoptosis through intrinsic and extrinsic pathways. DPBP treatment reduced tumor volume as well as serum VEGF-A and uric acid levels. Hepatomegaly and nephrotoxicity were not observed at the tested doses. Histopathological analysis of sentinel lymph nodes revealed no evidence of metastases. According to the observed data, the DPBP compound was effective for the topical treatment of melanoma cancer, suggesting that it acts as a chemotherapeutic or chemopreventive agent.
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http://dx.doi.org/10.1016/j.cbi.2021.109734DOI Listing
November 2021

Uncovering the transcriptional response of popcorn (Zea mays L. var. everta) under long-term aluminum toxicity.

Sci Rep 2021 Oct 4;11(1):19644. Epub 2021 Oct 4.

Departamento de Biologia Geral, Universidade Federal de Viçosa, Viçosa, MG, 36570-000, Brazil.

To date, the investigation of genes involved in Al resistance has focused mainly on microarrays and short periods of Al exposure. We investigated genes involved in the global response under Al stress by tracking the expression profile of two inbred popcorn lines with different Al sensitivity during 72 h of Al stress. A total of 1003 differentially expressed genes were identified in the Al-sensitive line, and 1751 were identified in the Al-resistant line, of which 273 were shared in both lines. Genes in the category of "response to abiotic stress" were present in both lines, but there was a higher number in the Al-resistant line. Transcription factors, genes involved in fatty acid biosynthesis, and genes involved in cell wall modifications were also detected. In the Al-resistant line, GST6 was identified as one of the key hub genes by co-expression network analysis, and ABC6 may play a role in the downstream regulation of CASP-like 5. In addition, we suggest a class of SWEET transporters that might be involved in the regulation of vacuolar sugar storage and may serve as mechanisms for Al resistance. The results and conclusions expand our understanding of the complex mechanisms involved in Al toxicity and provide a platform for future functional analyses and genomic studies of Al stress in popcorn.
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http://dx.doi.org/10.1038/s41598-021-99097-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8490451PMC
October 2021

Development of a chimeric protein based on a proteomic approach for the serological diagnosis of human tegumentary leishmaniasis.

Appl Microbiol Biotechnol 2021 Sep 25;105(18):6805-6817. Epub 2021 Aug 25.

Programa de Pós-Graduação Em Ciências da Saúde: Infectologia E Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.

Leishmania braziliensis is responsible for most cases of human tegumentary leishmaniasis (HTL) and has caused a wide range of clinical manifestations, including cutaneous (CL) and mucosal leishmaniasis (ML). The diagnosis is based on criteria that consider epidemiological data, clinical findings, and laboratory tests and is hard to establish. For laboratory tests, none of the assays available can be considered gold standards for disease detection. In addition, the Montenegro skin test, essential to supporting infectologists in the clinical management of the disease, is no longer available in Brazil. Thus, the aim of this study was to develop new targets to be used in diagnostic tests for HTL. In the first step, we carried out two-dimensional gel electrophoresis, followed by mass spectrometry, combined with heat map analysis and immunoproteomics approach, and disclosed eight proteins expressed in the amastigote stage specifically recognized by serum from CL and ML patients. A chimeric protein was designed based on the combination of thirteen linear B-cell epitopes, identified by immunoinformatics analysis, from L. braziliensis proteins. Our results showed that the strategy used in this work was successful in developing an antigen to be used in immunological assays (100.0% sensitivity and specificity) in the detection of HTL cases and in comparison with results obtained from an ELISA using soluble L. braziliensis antigen (SLb-Antigen) and immunofluorescence assay (Bio-Manguinhos/FIOCRUZ). The present technology opens the door for its use in field exams by means of an immunochromatographic test, which will be even more helpful in regions without laboratory structures.Key points• Rational strategy to develop antigens.• Integration between immunoproteomic and immunoinformatics analysis.• Chimeric protein shows high performance in HTL diagnosis.
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http://dx.doi.org/10.1007/s00253-021-11518-1DOI Listing
September 2021

R132 expresses antimicrobial genes and produces metabolites that modulate immune system.

3 Biotech 2021 Sep 4;11(9):396. Epub 2021 Aug 4.

Department of Botany and Ecology, Laboratory of Biotechnology and Microbial Ecology, Institute of Biosciences, Federal University of Mato Grosso, CuiabáMato Grosso, 78060-900 Brazil.

Actinobacteria is a phylum composed of aerobic, Gram-positive, and filamentous bacteria with a broad spectrum of biological activity, including antioxidant, antitumor, and antibiotic. The crude extract of R132 was fractionated on a C18 silica column and the isolated compound was identified by H and C nuclear magnetic resonance as 3-(phenylprop-2-enoic acid), also known as -cinnamic acid. Antimicrobial activity against human pathogens was assayed in vitro (disk-diffusion qualitative test) and in vivo using larvae (RT-qPCR). The methanol fractions 132-F30%, 132-F50%, 132-F70%, and 132-F100% inhibited the (ATCC 25922) and (MRSA) growth in vitro the most effectively. Compared with the untreated control (60-80% of larvae death), the fractions and isolated -cinnamic acid increased the survival rate and modulated the immune system of larvae infected with pathogenic microorganisms. The anti-infection effect of the R132 fermentation product led us to sequence its genome, which was assembled and annotated using the Rast and antiSMASH platforms The assembled genome consisted of 227 scaffolds represented on a linear chromosome of 8.85 Mb and 71.3% of GC. We detected conserved domains typical of enzymes that produce molecules with biological activity, such as polyketides and non-ribosomal and ribosomal peptides, indicating a great potential for obtaining new antibiotics and molecules with biotechnological application.

Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-021-02942-1.
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http://dx.doi.org/10.1007/s13205-021-02942-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8339190PMC
September 2021

Aspergillus sp. A31 and Curvularia geniculata P1 mitigate mercury toxicity to Oryza sativa L.

Arch Microbiol 2021 Nov 13;203(9):5345-5361. Epub 2021 Aug 13.

Department of Botany and Ecology, Laboratory of Biotechnology and Microbial Ecology, Institute of Biosciences, Federal University of Mato Grosso, Cuiabá, Mato Grosso, 78060-900, Brazil.

Aspergillus sp. A31 and Curvularia geniculata P1 are endophytes that colonize the roots of Aeschynomene fluminensis Vell. and Polygonum acuminatum Kunth. in humid environments contaminated with mercury. The two strains mitigated mercury toxicity and promoted Oryza sativa L growth. C. geniculata P1 stood out for increasing the host biomass by fourfold and reducing the negative effects of the metal on photosynthesis. Assembling and annotation of Aspergillus sp. A31 and C. geniculata P1 genomes resulted in 28.60 Mb (CG% 53.1; 10,312 coding DNA sequences) and 32.92 Mb (CG% 50.72; 8,692 coding DNA sequences), respectively. Twelve and 27 genomes of Curvularia/Bipolaris and Aspergillus were selected for phylogenomic analyzes, respectively. Phylogenetic analysis inferred the separation of species from the genus Curvularia and Bipolaris into different clades, and the separation of species from the genus Aspergillus into three clades; the species were distinguished by occupied niche. The genomes had essential gene clusters for the adaptation of microorganisms to high metal concentrations, such as proteins of the phytoquelatin-metal complex (GO: 0090423), metal ion binders (GO: 0046872), ABC transporters (GO: 0042626), ATPase transporters (GO: 0016887), and genes related to response to reactive oxygen species (GO: 0000302) and oxidative stress (GO: 0006979). The results reported here help to understand the unique regulatory mechanisms of mercury tolerance and plant development.
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http://dx.doi.org/10.1007/s00203-021-02481-6DOI Listing
November 2021

Prepubertal arsenic exposure alters phosphoproteins profile, quality, and fertility of epididymal spermatozoa in sexually mature rats.

Toxicology 2021 08 3;460:152886. Epub 2021 Aug 3.

Programa de Pós-Graduação em Biologia Celular e Estrutural, Departamento de Biologia Geral, Universidade Federal de Viçosa, Av. P.H. Rolfs, s/n, Campus Universitário, 36570-900, Viçosa, Minas Gerais, Brazil; Programa de Pós-Graduação em Medicina Veterinária, Departamento de Medicina Veterinária, Universidade Federal de Viçosa, Av. P.H. Rolfs, s/n, Campus Universitário, 36570-900, Viçosa, Minas Gerais, Brazil. Electronic address:

Arsenic intoxication affects male reproductive parameters of prepubertal rats. Besides, morphological and functional alterations in their testis and epididymis may remain after withdrawal of arsenic insult, causing potential impairment in male fertility during adulthood. In this study, we aimed at analyzing the effect of prepubertal arsenic exposure on the fecundity of epididymal sperm from sexually mature Wistar rats, assessing fertility indexes, sperm parameters, and sperm phosphoproteins content. Male pups on postnatal day (PND) 21 received filtered water (controls, n = 10) and 10 mg L arsenite (n = 10) daily for 30 days. From PND52 to PND81, rats from both groups received filtered water. During this period, the males mated with non-exposed females between PND72 and PND75. Our results showed that sexually mature rats presented low sperm production, epididymal sperm count, motility, and quality after prepubertal arsenic exposure. These findings possibly contributed to the low fertility potential and high preimplantation loss. Epididymal sperm proteome detected 268 proteins, which 170 were found in animals from both control and arsenic groups, 27 proteins were detected only in control animals and 71 proteins only in arsenic-exposed rats. In these animals, SPATA 18 and other five proteins were upregulated, whereas keratin type II cytoskeletal 1 was downregulated (q < 0.1). The results of KEGG pathway analysis demonstrated an enrichment of pathways related to dopaminergic response, adrenergic signaling, protein degradation, and oocyte meiosis in arsenic-exposed animals. Moreover, 26 proteins were identified by phosphoproteomic with different phosphorylation pattern in animals from both groups, but SPATA18 was phosphorylated only in arsenic-exposed animals. We concluded that prepubertal exposure to arsenic is deleterious to sperm quality and male fertility, altering the sperm phosphoproteins profile.
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http://dx.doi.org/10.1016/j.tox.2021.152886DOI Listing
August 2021

Assessing the relationship between the rumen microbiota and feed efficiency in Nellore steers.

J Anim Sci Biotechnol 2021 Jul 15;12(1):79. Epub 2021 Jul 15.

Departamento de Microbiologia, Universidade Federal de Viçosa, Viçosa, MG, 36570-900, Brazil.

Background: Ruminants rely upon a complex community of microbes in their rumen to convert host-indigestible feed into nutrients. However, little is known about the association between the rumen microbiota and feed efficiency traits in Nellore (Bos indicus) cattle, a breed of major economic importance to the global beef market. Here, we compare the composition of the bacterial, archaeal and fungal communities in the rumen of Nellore steers with high and low feed efficiency (FE) phenotypes, as measured by residual feed intake (RFI).

Results: The Firmicutes to Bacteroidetes ratio was significantly higher (P < 0.05) in positive-RFI steers (p-RFI, low feed efficiency) than in negative-RFI (n-RFI, high feed efficiency) steers. The differences in bacterial composition from steers with high and low FE were mainly associated with members of the families Lachnospiraceae, Ruminococcaceae and Christensenellaceae, as well as the genus Prevotella. Archaeal community richness was lower (P < 0.05) in p-RFI than in n-RFI steers and the genus Methanobrevibacter was either increased or exclusive of p-RFI steers. The fungal genus Buwchfawromyces was more abundant in the rumen solid fraction of n-RFI steers (P < 0.05) and a highly abundant OTU belonging to the genus Piromyces was also increased in the rumen microbiota of high-efficiency steers. However, analysis of rumen fermentation variables and functional predictions indicated similar metabolic outputs for the microbiota of distinct FE groups.

Conclusions: Our results demonstrate that differences in the ruminal microbiota of high and low FE Nellore steers comprise specific taxa from the bacterial, archaeal and fungal communities. Biomarker OTUs belonging to the genus Piromyces were identified in animals showing high feed efficiency, whereas among archaea, Methanobrevibacter was associated with steers classified as p-RFI. The identification of specific RFI-associated microorganisms in Nellore steers could guide further studies targeting the isolation and functional characterization of rumen microbes potentially important for the energy-harvesting efficiency of ruminants.
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http://dx.doi.org/10.1186/s40104-021-00599-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8281616PMC
July 2021

Proteomic Analysis of Liver from Finishing Beef Cattle Supplemented with a Rumen-Protected B-Vitamin Blend and Hydroxy Trace Minerals.

Animals (Basel) 2021 Jun 29;11(7). Epub 2021 Jun 29.

Department of Animal Science, Universidade Federal de Viçosa, Viçosa 36570-000, Brazil.

Vitamin B and trace minerals are crucial molecular signals involved in many biological pathways; however, their bioavailability is compromised in high-producing ruminant animals. So far, studies have mainly focused on the effects of these micronutrients on animal performance, but their use in a rumen-protected form and their impact on liver metabolism in finishing beef cattle is poorly known. We used a shotgun proteomic approach combined with biological network analyses to assess the effects of a rumen-protected B-vitamin blend, as well as those of hydroxy trace minerals, on the hepatic proteome. A total of 20 non-castrated Nellore males with 353 ± 43 kg of initial body weight were randomly assigned to one of the following treatments: CTRL-inorganic trace minerals without supplementation of a protected vitamin B blend, or SUP-supplementation of hydroxy trace minerals and a protected vitamin B blend. All animals were fed the same amount of the experimental diet for 106 days, and liver biopsies were performed at the end of the experimental period. Supplemented animals showed 37 up-regulated proteins ( < 0.10), and the enrichment analysis revealed that these proteins were involved in protein folding ( = 0.04), mitochondrial respiratory chain complex I ( = 0.01) and IV ( = 0.01), chaperonin-containing T-complex 2 ( = 0.01), glutathione metabolism ( < 0.01), and other aspects linked to oxidative-stress responses. These results indicate that rumen-protected vitamin B and hydroxy trace mineral supplementation during the finishing phase alters the abundance of proteins associated with the electron transport chain and other oxidation-reduction pathways, boosting the production of reactive oxygen species, which appear to modulate proteins linked to oxidative-damage responses to maintain cellular homeostasis.
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http://dx.doi.org/10.3390/ani11071934DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8300412PMC
June 2021

Thermodynamic and kinetic insights into the interactions between functionalized CdTe quantum dots and human serum albumin: A surface plasmon resonance approach.

Int J Biol Macromol 2021 Aug 28;184:990-999. Epub 2021 Jun 28.

Colloidal and Macromolecular Green Chemistry Group (QUIVECOM), Department of Chemistry, Federal University of Viçosa, Av. P. H. Rolfs s/n, 36570900 Viçosa, MG, Brazil. Electronic address:

To explore in vivo application of quantum dots (QDs), it is essential to understand the dynamics and energetics of interactions between QDs and proteins. Here, surface plasmon resonance (SPR) and molecular docking were employed to investigate the kinetics and thermodynamics of interactions between human serum albumin (HSA) and CdTe QDs (~3 nm) functionalized with mercaptopropionic acid (MPA) or thioglycolic acid (TGA). Kinetic analysis showed that HSA-QD interactions involved transition-complex formation. Despite the structural similarities between MPA and TGA, the [[email protected]] formation by association of free HSA and QDs demanded 70% more energy and higher entropic gain (E= 65.10 and T∆S= 28.62 kJ mol) than the formation of [[email protected]] (E = 38.13 and T∆S = 0.53kJ mol). While the [[email protected]] dissociation required higher energy and lower entropy loss (E = 49.96 and T∆S = - 32.18kJ mol) than the [[email protected]] dissociation (E= 30.78 and T∆S= - 51.12 kJ mol). The stability of [HSA-QDs] was independent of the temperature and functionalizing group. However, the enthalpic and entropic components were highly affected by the substitution of MPA (ΔH° = - 11.83 and TΔS° = 32.72 kJ mol) with TGA (ΔH° = 34.31 and TΔS° = 79.73 kJ mol). Furthermore, molecular docking results indicated that the metal site on the QDs contributes to the stabilization of [HSA-QDs]. Therefore, differences in QD functionalization and surface coverage densities can alter the HSA-QD interaction, thus their application.
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http://dx.doi.org/10.1016/j.ijbiomac.2021.06.158DOI Listing
August 2021

Phenotypic, functional and serological aspects of genotypic-specific immune response of experimental T. cruzi infection.

Acta Trop 2021 Oct 20;222:106021. Epub 2021 Jun 20.

Laboratório de Imunologia e Genômica de Parasitos - Departamento de Parasitologia, Instituto de Ciências Biológicas/ICB, Universidade Federal de Minas Gerais, Minas Gerais, Brasil.

The complexity and multifactorial characteristics of Chagas disease pathogenesis hampers the establishment of appropriate experimental/epidemiological sets, and therefore, still represents one of the most challenging fields for novel insights and discovery. In this context, we used a set of attributes including phenotypic, functional and serological markers of immune response as candidates to decode the genotype-specific immune response of experimental T. cruzi infection. In this investigation, we have characterized in C57BL/6 J mice, the early (parasitemia peak) and late (post-parasitemia peak) aspects of the immune response elicited by T. cruzi strains representative of TcI, TcII or TcVI. The results demonstrated earlier parasitemia peak for TcII/Y strain followed by TcVI/CL-Brener and TcI/Colombiana strains. A panoramic overview of phenotypic and functional features of the TCD4, TCD8 and B-cells from splenocytes demonstrated that mice infected with TcI/Colombiana strain exhibited at early stages of infection low levels of most cytokine cells with a slight increase at late stages of infection. Conversely, mice infected with TcII/Y strain presented an early massive increase of cytokine cells, which decreases at late stages. The TcVI/CL-Brener strain showed an intermediate profile at early stages of infection with a slight increase later on at post-peak of parasitemia. The panoramic analysis of immunological connectivity demonstrated that early after infection, the TcI/Colombiana strain trigger immunological network characterized by a small number of connectivity, selectively amongst cytokines that further shade towards the late stages of infection. In contrast, the TcII/Y strain elicited in more imbricate networks early after infection, comprising a robust number of interactions between pro-inflammatory mediators, regulatory cytokines and activation markers that also decrease at late infection. On the other hand, the infection with TcVI/CL-Brener strain demonstrated an intermediate profile with connectivity axes more stable at early and late stages of infection. The analysis of IgG2a reactivity to AMA, TRYPO and EPI antigens revealed that at early stages of infection, the genotype-specific reactivity to AMA, TRYPO and EPI to distinguish was higher for TcI/Colombiana as compared to TcII/Y and TcVI/CL while, at late stages of infection, higher reactivity to AMA was observed in mice infected with TcVI/CL and TcII/Y strains. The novel systems biology approaches and the use of a flow cytometry platform demonstrated that distinct T. cruzi genotypes influenced in the phenotypic and functional features of the host immune response and the genotype-specific serological reactivity during early and late stages of experimental T. cruzi infection.
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http://dx.doi.org/10.1016/j.actatropica.2021.106021DOI Listing
October 2021

Molecular Evidence of Rickettsia felis in Phereoeca sp.

Rev Bras Parasitol Vet 2021 26;30(1):e015620. Epub 2021 Apr 26.

Departamento de Bioquímica e Biologia Molecular, Universidade de Viçosa - UFV, Viçosa. MG, Brasil.

Rickettsia felis is an obligate intracellular bacterium capable of infecting ticks, fleas, lice, and other arthropods. This bacterium is classified as a member of the Transitional Group (TRG) Rickettsia. It is known the evidence of R. felis mutualistic and obligatory relationship with some eukaryote organisms. However, there aren't scientific accounts of R. felis and moths of the order Lepidoptera association. The current work reports the first identification of the bacteria R. felis in Phereoeca sp. For that, a polymerase chain reaction (PCR) assay using gltA, ompA, and ompB genes was used. The nucleotide sequences showed 100% of identity with other Rickettsia felis sequences. The genus-level identification of the moth larvae was performed by morphological taxonomic keys and PCR analysis of the cytochrome oxidase I (COI) gene. The nucleotide sequenced showed 94.94% similarity with the species Phereoeca praecox. However, with the low number of sequences deposited in the databases, the species was classified as Phereoeca sp. The results suggest that R. felis may develop in an organism without blood-feeding behavior (Lepidoptera), as it has been demonstrated for booklice (Psocoptera). Further investigation is necessary in order to confirm pathogenic or mutualistic association with moths.
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http://dx.doi.org/10.1590/S1984-29612021017DOI Listing
May 2021

Antibiotic Followed by a Potential Probiotic Increases Brown Adipose Tissue, Reduces Biometric Measurements, and Changes Intestinal Microbiota Phyla in Obesity.

Probiotics Antimicrob Proteins 2021 Dec 5;13(6):1621-1631. Epub 2021 Apr 5.

Department of Nutrition and Health, Universidade Federal de Viçosa, Avenue PH Rolfs S/N, Viçosa, Minas Gerais, 36570-900, Brazil.

The development of adjuvant therapies for obesity treatment is justified by the high prevalence of this disease worldwide, and the relationship between obesity and intestinal microbiota is a promising target for obesity treatment. Therefore, this study aimed at investigating the adjuvant treatment of obesity through the use of potential probiotics and antibiotics, either separately or sequentially. In the first phase of the experiment, animals had diet-induced obesity with consumption of a high saturated fat diet and a fructose solution. After this period, there was a reduction in caloric supply, that is the conventional treatment of obesity, and the animals were divided into 5 experimental groups: control group (G1), obese group (G2), potential probiotic group (G3), antibiotic group (G4), and antibiotic followed by potential probiotic group (G5). The adjuvant treatments lasted 4 weeks and were administered daily, via gavage: Animals in G1 and G2 received distilled water, the G3 obtained Lactobacillus gasseri LG-G12, and the G4 received ceftriaxone. The G5 received ceftriaxone for 2 weeks, followed by the offer of Lactobacillus gasseri LG-G12 for another 2 weeks. Parameters related to obesity, such as biometric measurements, food consumption, biochemical tests, histological assessments, short-chain fatty acids concentration, and composition of the intestinal microbiota, were analyzed. The treatment with caloric restriction and sequential supply of antibiotics and potential probiotics was able to reduce biometric measures, increase brown adipose tissue, and alter the intestinal microbiota phyla, standing out as a promising treatment for obesity.
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http://dx.doi.org/10.1007/s12602-021-09760-0DOI Listing
December 2021

Enlarging the Toolbox Against Antimicrobial Resistance: Aptamers and CRISPR-Cas.

Front Microbiol 2021 19;12:606360. Epub 2021 Feb 19.

Laboratory of Synthetic Biology and Modelling of Biological Systems, Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, Brazil.

In the post-genomic era, molecular treatments and diagnostics have been envisioned as powerful techniques to tackle the antimicrobial resistance (AMR) crisis. Among the molecular approaches, aptamers and CRISPR-Cas have gained support due to their practicality, sensibility, and flexibility to interact with a variety of extra- and intracellular targets. Those characteristics enabled the development of quick and onsite diagnostic tools as well as alternative treatments for pan-resistant bacterial infections. Even with such potential, more studies are necessary to pave the way for their successful use against AMR. In this review, we highlight those two robust techniques and encourage researchers to refine them toward AMR. Also, we describe how aptamers and CRISPR-Cas can work together with the current diagnostic and treatment toolbox.
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http://dx.doi.org/10.3389/fmicb.2021.606360DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7932999PMC
February 2021

Secretomic insight into the biomass hydrolysis potential of the phytopathogenic fungus Chrysoporthe cubensis.

J Proteomics 2021 03 1;236:104121. Epub 2021 Feb 1.

Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Av. PH Rolfs, s/n, Viçosa, MG 36570-900, Brazil. Electronic address:

The phytopathogenic fungus Chrysoporthe cubensis has a great capacity to produce highly efficient enzymes for the hydrolysis of lignocellulosic biomass. The bioinfosecretome of C. cubensis was identified by computational predictions of secreted proteins combined with protein analysis using 1D-LC-MS/MS. The in silico secretome predicted 562 putative genes capable of encoding secreted proteins, including 273 CAZymes. Proteomics analysis confirmed the existence of 313 proteins, including 137 CAZymes classified as Glycosyl Hydrolases (GH), Polysaccharide Lyases (PL), Carbohydrate Esterases (CE) and Auxiliary Activities enzymes (AA), which indicates the presence of classical and oxidative cellulolytic mechanisms. The enzymes diversity in the extract shows fungal versatility to act in complex biomasses. This study provides an insight into the lignocellulose-degradation mechanisms by C. cubensis and allows the identification of the enzymes that are potentially useful in improving industrial process of bioconversion of lignocellulose. SIGNIFICANCE: Chrysoporthe cubensis is an important deadly canker pathogen of commercially cultivated Eucalyptus species. The effective depolymerisation of the recalcitrant plant cell wall performed by this fungus is closely related to its high potential of lignocellulolytic enzymes secretion. Since the degradation of biomass occurs in nature almost exclusively by enzyme secretion systems, it is reasonable to suggest that the identification of C. cubensis lignocellulolytic enzymes is relevant in contributing to new sustainable alternatives for industrial solutions. As far as we know, this work is the first accurate proteomic evaluation of the enzymes secreted by this species of fungus. The integration of the gel-based proteomic approach, the bioinformatic prediction of the secretome and the analyses of enzymatic activity are powerful tools in the evaluation of biotechnological potential of C. cubensis in producing carbohydrate-active enzymes. In addition, analysis of the C. cubensis secretome grown in wheat bran draws attention to this plant pathogen and its extracellular enzymatic machinery, especially regarding the identification of promising new enzymes for industrial applications. The results from this work allowed for explanation and reinforce previous research that revealed C. cubensis as a strong candidate to produce enzymes to hydrolyse sugarcane bagasse and similar substrates.
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http://dx.doi.org/10.1016/j.jprot.2021.104121DOI Listing
March 2021

Bacillus spp. metabolites are effective in eradicating Aedes aegypti (Diptera: Culicidae) larvae with low toxicity to non-target species.

J Invertebr Pathol 2021 02 28;179:107525. Epub 2020 Dec 28.

Departamento de Botânica e Ecologia, Universidade Federal de Mato Grosso, Av. Fernando Corrêa da Costa 2367, 78060-900 Cuiabá, Brazil. Electronic address:

The growing spread of dengue, chikungunya and Zika viruses demand the development of new and environmentally safe control methods for their vector, the mosquito Aedes aegypti. This study aims to find novel larvicidal agents from mutualistic (endophytic and rhizospheric) or edaphic bacteria that have no action against non-target organisms. Eleven out of the 254 bacterial strains tested were able to kill Ae. aegypti larvae. Larvicidal activity did not depend on presence of cells, since culture supernatants or crude lipopeptide extracts (CLEs) killed the larvae. Bacillus safensis BacI67 and Bacillus paranthracis C21 supernatants were the best performing supernatants, displaying the lowest lethal concentrations (LC50 = 31.11 µL/mL and 45.84 µL/mL, respectively). Bacillus velezensis B64a and Bacillus velezensis B15 produced the best performing CLEs (LC50 = 0.11 mg/mL and 0.12 mg/mL, respectively). Mass spectrometry analysis of CLEs detected a mixture of surfactins, iturins, and fengycins. The samples tested were weakly- or non-toxic to mammalian cells (RAW 264.7 macrophages and VERO cells) and non-target organisms (Caenorhabditis elegans, Galleria mellonella, Scenedesmus obliquus, and Tetrahymena pyriformis) - especially B. velezensis B15 CLE. The biosynthetic gene clusters related to secondary metabolism identified by whole genome sequencing of the four best performing bacteria strains revealed clusters for bacteriocin, beta-lactone, lanthipeptide, non-ribosomal peptide synthetases, polyketide synthases (PKS), siderophores, T3PKS, type 1 PKS-like, terpenes, thiopeptides, and trans-AT-PKS. Purification of lipopeptides may clarify the mechanisms by which these extracts kill Ae. aegypti larvae.
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http://dx.doi.org/10.1016/j.jip.2020.107525DOI Listing
February 2021

Use of the synbiotic VSL#3 and yacon-based concentrate attenuates intestinal damage and reduces the abundance of Candidatus Saccharimonas in a colitis-associated carcinogenesis model.

Food Res Int 2020 11 25;137:109721. Epub 2020 Sep 25.

Nutritional Biochemistry Laboratory, Department of Nutrition and Health, Universidade Federal de Viçosa - UFV, Viçosa, Minas Gerais, Brazil. Electronic address:

Individuals with inflammatory bowel disease are at high risk of developing colitis-associated cancer; thus, strategies to inhibit disease progression should be investigated. The study aimed to explore the role of the synbiotic (probiotic VSL#3® and yacon-based concentrate) in a colitis-associated carcinogenesis model. IL-10 mice were induced to carcinogenesis with 1,2-dimethylhydrazine and divided into two experimental groups: control and synbiotic. Manifestations of colitis, colon histology, expression of antioxidant enzymes, production of organic acids and intestinal microbiota were evaluated. The use of the synbiotic showed benefits, such as the preservation of intestinal architecture, increased expression of antioxidant enzymes and the concentration of organic acids, especially butyrate. It was also observed different microbial community profiles between the groups during the study. Together, these factors contributed to mitigate the manifestations of colitis and improve intestinal integrity, suggesting the potential benefit of the synbiotic in intestinal diseases.
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http://dx.doi.org/10.1016/j.foodres.2020.109721DOI Listing
November 2020

Screening Unveil the Great Potential of Ruminal Bacteria Synthesizing Lasso Peptides.

Front Microbiol 2020 11;11:576738. Epub 2020 Sep 11.

Departamento de Microbiologia, Universidade Federal de Viçosa, Viçosa, Brazil.

Studies of rumen microbial ecology suggest that the capacity to produce antimicrobial peptides could be a useful trait in species competing for ecological niches in the ruminal ecosystem. However, little is known about the synthesis of lasso peptides by ruminal microorganisms. Here we analyzed the distribution and diversity of lasso peptide gene clusters in 425 bacterial genomes from the rumen ecosystem. Genome mining was performed using antiSMASH 5, BAGEL4, and a database of well-known precursor sequences. The genomic context of the biosynthetic clusters was investigated to identify putative genes and protein sequences from enzymes of the biosynthetic machinery were evaluated to identify conserved motifs. Metatranscriptome analysis evaluated the expression of the biosynthetic genes in the rumen microbiome. Several incomplete ( = 23) and complete ( = 11) putative lasso peptide clusters were detected in the genomes of ruminal bacteria. The complete gene clusters were exclusively found within the phylum , mainly (48%) in strains of the genus . The analysis of the genetic organization of complete putative lasso peptide clusters revealed the presence of co-occurring genes, including kinases (85%), transcriptional regulators (49%), and glycosyltransferases (36%). Moreover, a conserved pattern of cluster organization was detected between strains of the same genus/species. The maturation enzymes LasB, LasC, and LasD showed regions highly conserved, including the presence of a transglutaminase core in LasB, an asparagine synthetase domain in LasC, and an ABC-type transporter system in LasD. Phylogenetic trees of the essential biosynthetic proteins revealed that sequences split into monophyletic groups according to their shared single common ancestor. Metatranscriptome analyses indicated the expression of the lasso peptides biosynthetic genes within the active rumen microbiota. Overall, our screening allowed the discovery of novel biosynthetic gene clusters in the genomes of ruminal bacteria and revealed several strains with the genetic potential to synthesize lasso peptides, suggesting that the ruminal microbiota represents a potential source of these promising peptides.
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http://dx.doi.org/10.3389/fmicb.2020.576738DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7533575PMC
September 2020

Study of the differentially abundant proteins among Leishmania amazonensis, L. braziliensis, and L. infantum.

PLoS One 2020 15;15(10):e0240612. Epub 2020 Oct 15.

Departamento de Parasitologia, Laboratório de Leishmanioses, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.

Leishmaniasis has been considered as emerging and re-emerging disease, and its increasing global incidence has raised concerns. The great clinical diversity of the disease is mainly determined by the species. In several American countries, tegumentary leishmaniasis (TL) is associated with both Leishmania amazonensis and L. braziliensis, while visceral leishmaniasis (VL) is associated with L. (L.) infantum. The major molecules that determine the most diverse biological variations are proteins. In the present study, through a DIGE approach, we identified differentially abundant proteins among the species mentioned above. We observed a variety of proteins with differential abundance among the studied species; and the biological networks predicted for each species showed that many of these proteins interacted with each other. The prominent proteins included the heat shock proteins (HSPs) and the protein network involved in oxide reduction process in L. amazonensis, the protein network of ribosomes in L. braziliensis, and the proteins involved in energy metabolism in L. infantum. The important proteins, as revealed by the PPI network results, enrichment categories, and exclusive proteins analysis, were arginase, HSPs, and trypanothione reductase in L. amazonensis; enolase, peroxidoxin, and tryparedoxin1 in L. braziliensis; and succinyl-CoA ligase [GDP -forming] beta-chain and transaldolase in L. infantum.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0240612PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7561129PMC
December 2020

Stimulation of Bovicin HC5 Production and Selection of Improved Bacteriocin-Producing Streptococcus equinus HC5 Variants.

Probiotics Antimicrob Proteins 2021 06;13(3):899-913

Departamento de Microbiologia, Universidade Federal de Viçosa, Vicosa, MG, 36570-900, Brazil.

Bovicin HC5 is a peptide that has inhibitory activity against various pathogenic microorganisms and food spoilage bacteria. Aiming to improve the productivity of this bacteriocin, we evaluated several potential factors that could stimulate the synthesis of bovicin HC5 and selected variants of Streptococcus equinus (Streptococcus bovis) HC5 with enhanced bacteriocin production by adaptive laboratory evolution (ALE). The highest production of the bacteriocin (1.5-fold) was observed when Strep. equinus HC5 was cultivated with lactic acid (100 mmol/L). For the ALE experiment, Strep. equinus HC5 cells were subjected to acid-shock (pH 3.0 for 2 h) and maintained in continuous culture for approximately 140 generations (40 days) in media with lactic acid (100 mmol/L) and pH-controlled at 5.5 ± 0.2. An adapted variant was selected showing a distinct phenotype (sedimentation, pigmentation) compared with the parental strain. Bacteriocin production increased 2-fold in this adapted Strep. equinus HC5 variant, which appears to be associated with changes in the cell envelope of the adapted variant and enhanced bacteriocin release into the culture media. In addition, the adapted variant showed higher levels of expression of all bovicin HC5 biosynthetic genes compared with the parental strain during the early and late stages of growth. Results presented here indicate that ALE is a promising strategy for selecting strains of lactic acid bacteria with increased production of bacteriocins.
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http://dx.doi.org/10.1007/s12602-020-09703-1DOI Listing
June 2021

Temperature Dramatically Shapes Mosquito Gene Expression With Consequences for Mosquito-Zika Virus Interactions.

Front Microbiol 2020 12;11:901. Epub 2020 Jun 12.

Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA, United States.

Vector-borne flaviviruses are emerging threats to human health. For successful transmission, the virus needs to efficiently enter mosquito cells and replicate within and escape several tissue barriers while mosquitoes elicit major transcriptional responses to flavivirus infection. This process will be affected not only by the specific mosquito-pathogen pairing but also by variation in key environmental variables such as temperature. Thus far, few studies have examined the molecular responses triggered by temperature and how these responses modify infection outcomes, despite substantial evidence showing strong relationships between temperature and transmission in a diversity of systems. To define the host transcriptional changes associated with temperature variation during the early infection process, we compared the transcriptome of mosquito midgut samples from mosquitoes exposed to Zika virus (ZIKV) and non-exposed mosquitoes housed at three different temperatures (20, 28, and 36°C). While the high-temperature samples did not show significant changes from those with standard rearing conditions (28°C) 48 h post-exposure, the transcriptome profile of mosquitoes housed at 20°C was dramatically different. The expression of genes most altered by the cooler temperature involved aspects of blood-meal digestion, ROS metabolism, and mosquito innate immunity. Further, we did not find significant differences in the viral RNA copy number between 24 and 48 h post-exposure at 20°C, suggesting that ZIKV replication is limited by cold-induced changes to the mosquito midgut environment. In ZIKV-exposed mosquitoes, vitellogenin, a lipid carrier protein, was most up-regulated at 20°C. Our results provide a deeper understanding of the temperature-triggered transcriptional changes in and can be used to further define the molecular mechanisms driven by environmental temperature variation.
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http://dx.doi.org/10.3389/fmicb.2020.00901DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7303344PMC
June 2020

Exploring the Potential of CRISPR-Cas9 Under Challenging Conditions: Facing High-Copy Plasmids and Counteracting Beta-Lactam Resistance in Clinical Strains of .

Front Microbiol 2020 30;11:578. Epub 2020 Apr 30.

Department of Biochemistry and Molecular Biology, Universidade Federal de Viçosa, Viçosa, Brazil.

The antimicrobial resistance (AMR) crisis urgently requires countermeasures for reducing the dissemination of plasmid-borne resistance genes. Of particular concern are opportunistic pathogens of . One innovative approach is the CRISPR-Cas9 system which has recently been used for plasmid curing in defined strains of . Here we exploited this system further under challenging conditions: by targeting the AMR gene located on a high-copy plasmid (i.e., 100-300 copies/cell) and by directly tackling -positive clinical isolates. Upon CRISPR-Cas9 insertion into a model strain of harboring on the plasmid pSB1A2, the plasmid number and, accordingly, the gene expression decreased but did not become extinct in a subpopulation of CRISPR-Cas9 treated bacteria. Sequence alterations in were observed, likely resulting in a dysfunction of the gene product. As a consequence, a full reversal to an antibiotic sensitive phenotype was achieved, despite plasmid maintenance. In a clinical isolate of , plasmid clearance and simultaneous re-sensitization to five beta-lactams was possible. Reusability of antibiotics could be confirmed by rescuing larvae of infected with CRISPR-Cas9-treated , as opposed to infection with the unmodified clinical isolate. The drug sensitivity levels could also be increased in a clinical isolate of and to a lesser extent in , both of which harbored additional resistance genes affecting beta-lactams. The data show that targeting drug resistance genes is encouraging even when facing high-copy plasmids. In clinical isolates, the simultaneous interference with multiple genes mediating overlapping drug resistance might be the clue for successful phenotype reversal.
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http://dx.doi.org/10.3389/fmicb.2020.00578DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7203346PMC
April 2020

Chimeric Protein Designed by Genome-Scale Immunoinformatics Enhances Serodiagnosis of Bovine Neosporosis.

J Clin Microbiol 2020 06 24;58(7). Epub 2020 Jun 24.

Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil

Neosporosis has become a concern since it is associated with abortion in cattle. Currently, diagnosis is determined through anamnesis, evaluation of the history, and perception of the clinical signs of the herd. There is no practical and noninvasive test adapted to a large number of samples, which represents a gap for the use of new approaches that provide information about infections and the risks of herds. Here, we performed a search in the genome by linear B-cell epitopes using immunoinformatic tools aiming to develop a chimeric protein with high potential to bind specifically to antibodies from infected cattle samples. An enzyme-linked immunosorbent assay with the new chimeric antigen was developed and tested with sera from natural field -infected bovines. The cross-reactivity of the new antigen was also evaluated using sera from bovines infected by other abortive pathogens, including , sp., , and , and enzootic bovine leucosis caused by bovine leukemia virus, as well as with samples of animals infected with The assay using the chimeric protein showed 96.6% ± 3.4% of sensitivity in comparison to healthy animal sera. Meanwhile, in relation to false-positive results provided by cross-reactivity with others pathogens, the specificity value was 97.0% ± 2.9%. In conclusion, immunoinformatic tools provide an efficient platform to build an accurate protein to diagnose bovine neosporosis based on serum samples.
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http://dx.doi.org/10.1128/JCM.01343-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7315025PMC
June 2020

Genome-Scale Characterization of Fungal Phytases and a Comparative Study Between Beta-Propeller Phytases and Histidine Acid Phosphatases.

Appl Biochem Biotechnol 2020 Sep 6;192(1):296-312. Epub 2020 May 6.

Department of Biochemistry and Molecular Biology, Federal University of Viçosa, Av. PH Rolfs, s/n, Viçosa, MG, 36570-900, Brazil.

This work intended to prospect new phytase-producing organisms. In silico genomic analyses allowed the selection of twelve potential phytase-producing fungi. Based on gene sequence, it was possible to identify four well-defined groups of phytate-degrading enzymes: esterase-like, β-propeller phytases (βPP), phosphoglycerate mutase-like, and phytases of the histidine acid phosphatases (HAP) family. Analysis of the predicted genes encoding phytases belonging to the HAP family and βPP phytases and in silico characterization of these enzymes indicated divergence among the catalytic activities. Predicted fungal βPP phytases exhibited higher molecular mass (around 77 kDa) probably due to the epidermal growth factor-like domain. Twelve sequences of phytases contained signal peptides, of which seven were classified as HAP and five as βPP phytases, while ten sequences were predicted as phytases secreted by non-classical pathways. These fungi were grown in liquid or semi-solid medium, and the fungal enzymatic extracts were evaluated for their ability to hydrolyze sodium phytate at 50 °C and pH ranging from 2.0 to 9.0. Seven fungi were identified as phytase producers based on phosphate release under enzyme assay conditions. Results obtained from in silico analyses combining experimental enzymatic activities suggest that some selected fungi could secrete βPP phytases and HAP phytases.
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http://dx.doi.org/10.1007/s12010-020-03309-7DOI Listing
September 2020

Rational design of mimetic peptides based on the interaction between Inga laurina inhibitor and trypsins for Spodoptera cosmioides pest control.

Insect Biochem Mol Biol 2020 07 29;122:103390. Epub 2020 Apr 29.

Departamento de Bioquímica e Biologia molecular, Universidade Federal de Viçosa, Minas Gerais, Brazil; Instituto de Biotecnologia aplicada à Agropecuaria, BIOAGRO-UFV, Viçosa, Minas Gerais, Brazil.

The interaction of Inga laurina Kunitz inhibitor with insect trypsins is an example of protein-protein interaction with potential application for the pest control. However, the crop field application of proteins as inhibitors is limited due to high production cost, the large molecular size and low environmental stability. The use of mimetic peptides that have molecular features associated with the protein inhibitor can result in a product with lower cost and higher efficiency for the agricultural application. Here, we designed mimetic peptides deriving from globular domains of ILTI that are predicted to interact with trypsin enzymes of Lepidoptera pest. Two linear peptides were identified and synthetized from the interface of interaction between trypsin-ILTI complexes. These peptides were derived due to its high-energy contribution for the biding affinity between the enzyme-protein inhibitor. The peptides showed structural stability, propensity to adopt the bound conformation also without the context of the protein, inhibitory activity of digestive trypsins and toxic effects on the S. cosmioides, indicating that they can be used as potential inhibitor for pest control.
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http://dx.doi.org/10.1016/j.ibmb.2020.103390DOI Listing
July 2020

Effects of enrofloxacin treatment on the bacterial microbiota of milk from goats with persistent mastitis.

Sci Rep 2020 03 10;10(1):4421. Epub 2020 Mar 10.

Laboratory of Bacterial Diseases, Sector of Preventive Veterinary Medicine and Public Health, Veterinary Department, Federal University of Viçosa, Viçosa, MG, Brazil.

Antibiotic resistance has become a major concern for human and animal health. As fluoroquinolones have been extensively used in human and veterinary medicine, there has also been the rapid emergence and spread of antimicrobial resistance around the world. Here, we analysed the microbiome of goat milk using samples from healthy goats and those diagnosed with persistent mastitis and treated using the antibiotic enrofloxacin with 16S rRNA amplicon sequencing. We selected a group of 11 goats and 22 samples of milk that did not respond clinically to enrofloxacin treatment. Milk samples were evaluated before and after treatment to verify changes of the microbiota; the three first lactating goats were selected from the healthy control group. The milk samples from the healthy control animals presented a larger abundance of different species of bacteria of the Staphylococcus genus, but a smaller number of different genera, which indicated a more specific niche of resident bacteria. The Firmicutes phylum was predominantly different between the studied groups. Samples from before-treatment animals had a higher number of new species than those from the control group, and after being treated again. These microbiota received new bacteria, increasing the differences in bacteria even more in relation to the control group. Genotypes such as Trueperella and Mannheimia, between other genera, had a high abundance in the samples from animals with persistent mastitis. The dysbiosis in this study, with marked evidence of a complex microbiota in activity in cases of the failure of antimicrobial treatment for persistent chronic mastitis, demonstrates a need to improve the accuracy of pathogen identification and increases concern regarding antibiotic treatments in milk production herds.
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http://dx.doi.org/10.1038/s41598-020-61407-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7064484PMC
March 2020

Physicochemical characterization of Pseudomonas stutzeri UFV5 and analysis of its transcriptome under heterotrophic nitrification/aerobic denitrification pathway induction condition.

Sci Rep 2020 02 10;10(1):2215. Epub 2020 Feb 10.

Department of Microbiology, Federal University of Viçosa, Viçosa, Minas Gerais, 36570-900, Brazil.

Biological ammonium removal via heterotrophic nitrification/aerobic denitrification (HN/AD) presents several advantages in relation to conventional removal processes, but little is known about the microorganisms and metabolic pathways involved in this process. In this study, Pseudomonas stutzeri UFV5 was isolated from an activated sludge sample from oil wastewater treatment station and its ammonium removal via HN/AD was investigated by physicochemical and molecular approaches to better understand this process and optimize the biological ammonium removal in wastewater treatment plants. Results showed that P. stutzeri UFV5 removed all the ammonium in 48-72 hours using pyruvate, acetate, citrate or sodium succinate as carbon sources, C/N ratios 6, 8, 10 and 12, 3-6% salinities, pH 7-9 and temperatures of 20-40 °C. Comparative genomics and PCR revealed that genes encoding the enzymes involved in anaerobic denitrification process are present in P. stutzeri genome, but no gene that encodes enzymes involved in autotrophic nitrification was found. Furthermore, transcriptomics showed that none of the known enzymes of autotrophic nitrification and anaerobic denitrification had their expression differentiated and an upregulation of the biosynthesis machinery and protein translation was observed, besides several genes with unknown function, indicating a non-conventional mechanism involved in HN/AD process.
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http://dx.doi.org/10.1038/s41598-020-59279-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7010759PMC
February 2020

Genomic and gene expression evidence of nonribosomal peptide and polyketide production among ruminal bacteria: a potential role in niche colonization?

FEMS Microbiol Ecol 2020 02;96(2)

Departamento de Microbiologia, Universidade Federal de Viçosa, Av. P.H. Rolfs, s/n, Viçosa-MG, 36570-900, Brazil.

Genomic and transcriptomic analyses were performed to investigate nonribosomal peptide synthetases (NRPS) and polyketide synthases (PKS) in 310 genomes of ruminal/fecal microorganisms. A total of 119 biosynthetic genes potentially encoding distinct nonribosomal peptides (NRPs) and polyketides (PKs) were predicted in the ruminal microbial genomes and functional annotation separated these genes into 19 functional categories. The phylogenetic reconstruction of the 16S rRNA sequences coupled to the distribution of the three 'backbone' genes involved in NRPS and PKS biosyntheses suggested that these genes were not acquired through horizontal gene transfer. Metatranscriptomic analyses revealed that the predominant genes involved in the synthesis of NRPs and PKs were more abundant in sheep rumen datasets. Reads mapping to the NRPS and PKS biosynthetic genes were represented in the active ruminal microbial community, with transcripts being highly expressed in the bacterial community attached to perennial ryegrass, and following the main changes occurring between primary and secondary colonization of the forage incubated with ruminal fluid. This study is the first comprehensive characterization demonstrating the rich genetic capacity for NRPS and PKS biosyntheses within rumen bacterial genomes, which highlights the potential functional roles of secondary metabolites in the rumen ecosystem.
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http://dx.doi.org/10.1093/femsec/fiz198DOI Listing
February 2020

Didelphis albiventris: an overview of unprecedented transcriptome sequencing of the white-eared opossum.

BMC Genomics 2019 Nov 15;20(1):866. Epub 2019 Nov 15.

Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.

Background: The white-eared opossum (Didelphis albiventris) is widely distributed throughout Brazil and South America. It has been used as an animal model for studying different scientific questions ranging from the restoration of degraded green areas to medical aspects of Chagas disease, leishmaniasis and resistance against snake venom. As a marsupial, D. albiventris can also contribute to the understanding of the molecular mechanisms that govern the different stages of organogenesis. Opossum joeys are born after only 13 days, and the final stages of organogenesis occur when the neonates are inside the pouch, depending on lactation. As neither the genome of this opossum species nor its transcriptome has been completely sequenced, the use of D. albiventris as an animal model is limited. In this work, we sequenced the D. albiventris transcriptome by RNA-seq to obtain the first catalogue of differentially expressed (DE) genes and gene ontology (GO) annotations during the neonatal stages of marsupial development.

Results: The D. albiventris transcriptome was obtained from whole neonates harvested at birth (P0), at 5 days of age (P5) and at 10 days of age (P10). The de novo assembly of these transcripts generated 85,338 transcripts. Approximately 30% of these transcripts could be mapped against the amino acid sequences of M. domestica, the evolutionarily closest relative of D. albiventris to be sequenced thus far. Among the expressed transcripts, 2077 were found to be DE between P0 and P5, 13,780 between P0 and P10, and 1453 between P5 and P10. The enriched GO terms were mainly related to the immune system, blood tissue development and differentiation, vision, hearing, digestion, the CNS and limb development.

Conclusions: The elucidation of opossum transcriptomes provides an out-group for better understanding the distinct characteristics associated with the evolution of mammalian species. This study provides the first transcriptome sequences and catalogue of genes for a marsupial species at different neonatal stages, allowing the study of the mechanisms involved in organogenesis.
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http://dx.doi.org/10.1186/s12864-019-6240-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6858782PMC
November 2019

Carbohydrate-independent antibiofilm effect of Bothrops jararacussu lectin BJcuL on Staphylococcus aureus.

Microb Pathog 2019 Dec 11;137:103745. Epub 2019 Sep 11.

Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Minas Gerais, Brazil. Electronic address:

The antivirulence approach to fighting biofilm-based infections caused by Staphylococcus aureus is a promising therapy that has been studied extensively. Here, we compare the antibiofilm activity of a purified lectin from Bothrops jararacussu venom (BJcuL) and commercial lectins obtained from Triticum vulgaris (Wheat Germ Agglutinin, WGA), Bandeiraea simplicifolia BS-II, and Maclura pomifera. Only WGA had antibiofilm activity, although no effect was seen on pre-formed biofilms. The pre-incubation of WGA and BJcuL with their preferential sugars inhibited the biological activity of WGA, but not that of BJcuL, suggesting that biofilm disruption does not involve carbohydrate-recognition domains (CRDs). Quantitative real-time PCR showed that BJcuL promotes modulation of expression of S. aureus genes involved in biofilm formation. Light microscopy revealed cocci and small cell clusters after biofilm formation in the presence of BJcuL, showing that the lectin treatment was unable to completely disrupt biofilm structure. Exposing the free cells to 50 times the minimum inhibitory concentration of gentamicin or ciprofloxacin did not prevent biofilm reestablishment, although inhibition was stronger than in the control (no lectin). This disruption of the biofilm architecture can expose the bacterial cell and may facilitate clearance by the immune system.
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http://dx.doi.org/10.1016/j.micpath.2019.103745DOI Listing
December 2019
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