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1.
Braz J Microbiol ; 2024 Jul 31.
Article in English | MEDLINE | ID: mdl-39083223

ABSTRACT

Enteroaggregative E. coli (EAEC) is one of the most frequent pathogens isolated from diarrheal patients as well as from healthy individuals in Brazil and has recently also been implicated as an extraintestinal pathogenic E. coli (ExPEC) associated with bloodstream and urinary tract infections. In this study, 37 EAEC isolates, obtained from fecal samples of non-diarrheic children, were molecularly and phenotypically characterized to access the pathogenic features of these isolates. The EAEC isolates were assigned into the phylogroups A (54.1%), D (29.7%), B1 (13.5%) and B2 (2.7%); and harbored genes responsible for encoding the major pilin subunit of the aggregative adherence fimbriae (AAFs) or aggregate-forming pili (AFP) adhesins as follows: aggA (24.3%), agg3A (5.4%), agg4A (27.0%), agg5A (32.4%) and afpA (10.8%). The most frequent O:H serotypes were O15:H2 (8.1%), O38:H25 (5.4%) and O86:H2 (5.4%). Twenty-one isolates (56.8%) produce the aggregative adherence (AA) pattern on HeLa cells, and biofilm formation was more efficient among EAEC isolates harboring the aggA and agg5A genes. PFGE analysis showed that 31 (83.8%) of the isolates were classified into 10 distinct clusters, which reinforces the high diversity found among the isolates studied. Of note, 40.5% (15/37) of the EAEC isolates have a genetic profile compatible with E. coli isolates with intrinsic potential to cause extraintestinal infections in healthy individuals, and therefore, classified as EAEC/ExPEC hybrids. In conclusion, we showed the presence of EAEC/ExPEC hybrids in the intestinal microbiota of non-diarrheic children, possibly representing the source of some endogenous extraintestinal infections.

2.
Sci Rep ; 14(1): 17033, 2024 07 24.
Article in English | MEDLINE | ID: mdl-39043862

ABSTRACT

Tritrichomonas foetus is a flagellated and anaerobic parasite able to infect cattle and felines. Despite its prevalence, there is no effective standardized or legal treatment for T. foetus-infected cattle; the vaccination still has limited success in mitigating infections and reducing abortion risk; and nowadays, the diagnosis of T. foetus presents important limitations in terms of sensitivity and specificity in bovines. Here, we characterize the plasma membrane proteome of T. foetus and identify proteins that are represented in different isolates of this protozoan. Additionally, we performed a bioinformatic analysis that revealed the antigenicity potential of some of those proteins. This analysis is the first study to identify common proteins at the plasma membrane of different T. foetus isolates that could be targets for alternative diagnostic or vaccine techniques in the future.


Subject(s)
Proteomics , Protozoan Proteins , Tritrichomonas foetus , Tritrichomonas foetus/isolation & purification , Proteomics/methods , Protozoan Proteins/metabolism , Protozoan Proteins/analysis , Animals , Proteome/analysis , Cell Membrane/metabolism , Cattle , Membrane Proteins/metabolism , Cattle Diseases/parasitology , Protozoan Infections, Animal/parasitology , Protozoan Infections, Animal/diagnosis , Computational Biology/methods
4.
Acta Trop ; 256: 107242, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38782111

ABSTRACT

Leptospirosis is an infectious disease with a worldwide distribution, which represents a major challenge in animal production across developing countries, mainly in tropical areas. Horses are particularly susceptible to the disease, presenting manifestations ranging from subclinical to the development of uveitis that compromises the visual health of the animals. In recent years, serological studies have been carried out in equid populations from America, demonstrating high exposure. For this reason, the aim of this study was to demonstrate microbiologically and molecularly the presence of the members of the genus Leptospira in urine samples from equids in an endemic state of leptospirosis in Mexico, and to detect the serological presence of anti-Leptospira antibodies in the sampled animals. For this reason, blood and urine samples were collected from 28 horses and one mule from three localities in the state of Veracruz, Mexico. Urine samples were inoculated in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium, and the recovered isolates were typed using a short Multi Locus Sequence Typing scheme. Amplifications of the expected size were subjected to sequencing, and the recovered sequences were compared with those of reference deposited in GenBank using the BLAST tool. To identify their phylogenetic position, we performed a phylogenetic reconstruction using the maximum likelihood method. Additionally, Microscopic Agglutination test was performed on the serum samples to identify anti-Leptospira antibodies. We recovered 16 urine isolates which tested positive for the presence of Leptospira DNA. The phylogenetic reconstruction and the MLST analysis confirmed the presence of several genotypes of Leptospira interrogans and Leptospira santarosai. An overall serological frequency of 97.1 % was detected. Our results represent the first record of the presence of Leptospira through bacteriological isolates in equids from Mexico.


Subject(s)
Antibodies, Bacterial , Horse Diseases , Leptospira , Leptospirosis , Phylogeny , Animals , Leptospirosis/veterinary , Leptospirosis/microbiology , Leptospirosis/epidemiology , Mexico/epidemiology , Horses/microbiology , Horse Diseases/microbiology , Leptospira/isolation & purification , Leptospira/genetics , Leptospira/classification , Antibodies, Bacterial/blood , Leptospira interrogans/isolation & purification , Leptospira interrogans/genetics , Leptospira interrogans/classification , Multilocus Sequence Typing , Sequence Analysis, DNA , DNA, Bacterial/genetics
5.
PeerJ ; 12: e17199, 2024.
Article in English | MEDLINE | ID: mdl-38680892

ABSTRACT

Carbapenem-resistant Acinetobacter spp. is associated with nosocomial infections in intensive care unit patients, resulting in high mortality. Although Acinetobacter spp. represent a serious public health problem worldwide, there are a few studies related to the presence of carbapenemases in health care facilities and other environmental settings in Ecuador. The main aim of this study was to characterize the carbapenem-resistant Acinetobacter spp. isolates obtained from four hospitals (52) and from five rivers (27) close to Quito. We used the disc diffusion and EDTA sinergy tests to determine the antimicrobial susceptibility and the production of metallo ß-lactamases, respectively. We carried out a multiplex PCR of gyrB gene and the sequencing of partial rpoB gene to bacterial species identification. We performed molecular screening of nine carbapenem-resistant genes (blaSPM, blaSIM, blaGIM, blaGES, blaOXA-23, blaOXA-24, blaOXA-51, blaOXA-58, and blaOXA-143) by multiplex PCR, followed by identification using sequencing of blaOXA genes. Our findings showed that carbapenem-resistant A. baumannii were the main species found in health care facilities and rivers. Most of the clinical isolates came from respiratory tract samples and harbored blaOXA-23, blaOXA-366, blaOXA-72, blaOXA-65, blaOXA-70, and blaOXA-143-like genes. The river isolates harbored only the blaOXA-51 and probably blaOXA-259 genes. We concluded that the most predominant type of carbapenem genes among isolates were both blaOXA-23 and blaOXA-65 among A. baumannii clinical isolates.


Subject(s)
Acinetobacter Infections , Acinetobacter , Bacterial Proteins , beta-Lactamases , Ecuador/epidemiology , beta-Lactamases/genetics , beta-Lactamases/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Humans , Acinetobacter Infections/microbiology , Acinetobacter Infections/drug therapy , Acinetobacter/genetics , Acinetobacter/isolation & purification , Acinetobacter/drug effects , Acinetobacter/enzymology , Microbial Sensitivity Tests , Cross Infection/microbiology , Anti-Bacterial Agents/pharmacology , Carbapenems/pharmacology , Rivers/microbiology , Acinetobacter baumannii/genetics , Acinetobacter baumannii/drug effects , Acinetobacter baumannii/isolation & purification , Acinetobacter baumannii/enzymology , Multiplex Polymerase Chain Reaction
6.
Fungal Genet Biol ; 172: 103891, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38621582

ABSTRACT

Candida glabrata (Nakaseomyces glabrata) is an emergent and opportunistic fungal pathogen that colonizes and persists in different niches within its human host. In this work, we studied five clinical isolates from one patient (P7), that have a clonal origin, and all of which come from blood cultures except one, P7-3, obtained from a urine culture. We found phenotypic variation such as sensitivity to high temperature, oxidative stress, susceptibility to two classes of antifungal agents, and cell wall porosity. Only isolate P7-3 is highly resistant to the echinocandin caspofungin while the other four isolates from P7 are sensitive. However, this same isolate P7-3, is the only one that displays susceptibility to fluconazole (FLC), while the rest of the isolates are resistant to this antifungal. We sequenced the PDR1 gene which encodes a transcription factor required to induce the expression of several genes involved in the resistance to FLC and found that all the isolates encode for the same Pdr1 amino acid sequence except for the last isolate P7-5, which contains a single amino acid change, G1099C in the putative Pdr1 transactivation domain. Consistent with the resistance to FLC, we found that the CDR1 gene, encoding the main drug efflux pump in C. glabrata, is highly overexpressed in the FLC-resistant isolates, but not in the FLC-sensitive P7-3. In addition, the resistance to FLC observed in these isolates is dependent on the PDR1 gene. Additionally, we found that all P7 isolates have a different proportion of cell wall carbohydrates compared to our standard strains CBS138 and BG14. In P7 isolates, mannan is the most abundant cell wall component, whereas ß-glucan is the most abundant component in our standard strains. Consistently, all P7 isolates have a relatively low cell wall porosity compared to our standard strains. These data show phenotypic and genotypic variability between clonal isolates from different niches within a single host, suggesting microevolution of C. glabrata during an infection.


Subject(s)
Antifungal Agents , Candida glabrata , Drug Resistance, Fungal , Fungal Proteins , Microbial Sensitivity Tests , Candida glabrata/genetics , Candida glabrata/drug effects , Antifungal Agents/pharmacology , Humans , Drug Resistance, Fungal/genetics , Fungal Proteins/genetics , Fungal Proteins/metabolism , Fluconazole/pharmacology , Cell Wall/genetics , Cell Wall/drug effects , Candidiasis/microbiology , Caspofungin/pharmacology , Evolution, Molecular , Oxidative Stress/genetics , Echinocandins/pharmacology , Transcription Factors/genetics
7.
FEBS Lett ; 598(6): 670-683, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38433717

ABSTRACT

Ferredoxin/flavodoxin-NADPH reductases (FPRs) catalyze the reversible electron transfer between NADPH and ferredoxin/flavodoxin. The Acinetobacter sp. Ver3 isolated from high-altitude Andean lakes contains two isoenzymes, FPR1ver3 and FPR2ver3. Absorption spectra of these FPRs revealed typical features of flavoproteins, consistent with the use of FAD as a prosthetic group. Spectral differences indicate distinct electronic arrangements for the flavin in each enzyme. Steady-state kinetic measurements show that the enzymes display catalytic efficiencies in the order of 1-6 µm-1·s-1, although FPR1ver3 exhibited higher kcat values compared to FPR2ver3. When flavodoxinver3 was used as a substrate, both reductases exhibited dissimilar behavior. Moreover, only FPR1ver3 is induced by oxidative stimuli, indicating that the polyextremophile Ver3 has evolved diverse strategies to cope with oxidative environments.


Subject(s)
Ferredoxins , Flavodoxin , Flavodoxin/metabolism , NADP/metabolism , Ferredoxins/metabolism , Ferredoxin-NADP Reductase/chemistry , Ferredoxin-NADP Reductase/metabolism , Protein Isoforms , Kinetics
8.
Front Cell Infect Microbiol ; 14: 1348093, 2024.
Article in English | MEDLINE | ID: mdl-38516533

ABSTRACT

Introduction: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. are microorganisms referred as the ESKAPE group pathogens. These microorganisms have generated great concern in health institutions around the world since most of them have resistance to multiple antibiotics and cause most infections associated with healthcare, as well as community infections. The aim of this study was the analysis of antibiotic resistance in microorganisms of the ESKAPE group, recovered from clinical samples in 11 health institutions from Hermosillo and Ciudad Obregón in the State of Sonora, México, during the period from 2019 to 2020. Methods: A cross-sectional, descriptive, observational, and temporality epidemiological study was carried out. A comparative and statistical analysis of antibiotic resistance was carried out using the chi-square test, and small values were analyzed using Fisher's exact test p ≤ 0.05. Results and discussion: All the ESKAPE group microorganisms showed significant differences in antibiotic resistance percentages between both cities. High resistance percentages for some antibiotics, like cephalosporins and ciprofloxacin were detected for Klebsiella pneumoniae and Acinetobacter baumannii.


Subject(s)
Acinetobacter baumannii , Klebsiella pneumoniae , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Cross-Sectional Studies , Drug Resistance, Multiple, Bacterial , Mexico , Humans
9.
Am J Med Genet A ; 194(5): e63523, 2024 05.
Article in English | MEDLINE | ID: mdl-38164622

ABSTRACT

The FMR1 5' regulation gene region harbors a CGG trinucleotide repeat expansion (CGG-TRE) that causes Fragile X syndrome (FXS) when it expands to more than 200 repetitions. Ricaurte is a small village in southwestern Colombia, with an FXS prevalence of 1 in 38 men and 1 in 100 women (~100 times higher than the worldwide reported prevalence), defining Ricaurte as the largest FXS cluster in the world. In the present study, using next-generation sequencing of whole exome capture, we genotype 55 individuals from Ricaurte (49 with either full mutation or with premutation), four individuals from neighboring villages (with either the full mutation or with the premutation), and one unaffected woman, native of Ricaurte, who did not belong to any of the affected families. With advanced clustering and haplotype reconstruction, we modeled a common haplotype of 33 SNPs spanning 83,567,899 bp and harboring the FMR1 gene. This reconstructed haplotype was found in all the men from Ricaurte who carried the expansion, demonstrating that the genetic conglomerate of FXS in this population is due to a founder effect. The definition of this founder effect and its population outlining will allow a better prediction, follow-up, precise and personalized characterization of epidemiological parameters, better knowledge of the disease's natural history, and confident improvement of the clinical attention, life quality, and health interventions for this community.


Subject(s)
Fragile X Syndrome , Male , Humans , Female , Fragile X Syndrome/epidemiology , Fragile X Syndrome/genetics , Founder Effect , Molecular Epidemiology , Fragile X Mental Retardation Protein/genetics , Trinucleotide Repeat Expansion , Mutation
10.
Braz J Microbiol ; 55(1): 343-355, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38066229

ABSTRACT

Sulfonamide derivatives have numerous pharmaceutical applications having antiviral, antibacterial, antifungal, antimalarial, anticancer, and antidepressant activities. The structural flexibility of sulfonamide derivatives makes them an excellent candidate for the development of new multi-target agents, although long-time exposure to sulfonamide drugs results in many toxic impacts on human health. However, sulfonamides may be functionalized for developing less toxic and more competent drugs. In this work, sulfonamides including Sulfapyridine (a), Sulfathiazole (b), Sulfamethoxazole (c), and Sulfamerazine (d) are used to synthesize Schiff bases of 7-hydroxy-4-methyl-2-oxo-2H-chromene-8-carbalde-hyde (1a-1d). The synthesized compounds were spectroscopically characterized and tested against hospital isolates of three Gram-positive (Methicillin-resistant Staphylococcus aureus PH217, Ampicillin-resistant Coagulase-negative Staphylococcus aureus, multidrug-resistant (MDR) Enterococcus faecalis PH007R) and two Gram-negative bacteria (multidrug-resistant Escherichia coli, and Salmonella enterica serovar Typhi), compared to the quality control strains from ATCC (S. aureus 29213, E. faecalis 25922, E. coli 29212) and MTCC (S. Typhi 734). Two of the four Schiff bases 1a and 1b are found to be more active than their counterpart 1c and 1d; while 1a have showed significant activity by inhibiting MRSA PH217 and MDR isolates of E. coli at the minimum inhibitory concentration (MIC) of 150 µg/mL and 128 µg/mL with MBC of 1024 µg/mL, respectively. On the other hand, the MIC of 1b was 150 µg/mL against both S. aureus ATCC 29213 and Salmonella Typhi MTCC 734, compared to the control antibiotics Ampicillin and Gentamycin. Scanning electron microscopy demonstrated the altered surface structure of bacterial cells as a possible mechanism of action, supported by the in-silico molecular docking analysis.


Subject(s)
Methicillin-Resistant Staphylococcus aureus , Staphylococcus aureus , Humans , Molecular Docking Simulation , Chromones/pharmacology , Escherichia coli , Schiff Bases/pharmacology , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Sulfanilamide , Ampicillin/pharmacology , Sulfonamides/pharmacology , Microbial Sensitivity Tests
11.
Mem. Inst. Oswaldo Cruz ; 119: e240026, 2024. tab, graf
Article in English | LILACS-Express | LILACS | ID: biblio-1564816

ABSTRACT

BACKGROUND In Brazil, Leishmania (Leishmania) infantum is a widely distributed protozoan parasite. The human leishmaniasis caused by this species is often associated with visceral form. Tegumentary leishmaniasis (TL) cases due to L. (L.) infantum in the country are considered rare but may be underestimated. Although probably uncommon, these cases represent a new challenge to the prevention and control of leishmaniasis. OBJECTIVES Here, we describe two distinct cases of TL with atypical clinical presentations caused by L. (L.) infantum. METHODS AND FINDINGS Parasites were isolated from cutaneous lesions of the two patients and typed as L. (L.) infantum after sequencing of the ribosomal DNA internal transcribed spacer. The dermotropic L. (L.) infantum isolates were compared in terms of growth culture patterns, metacyclogenesis and in vitro infectivity in macrophages. MAIN CONCLUSIONS This study addresses the emergence of L. (L.) infantum as a causative agent of cutaneous disease in a visceral leishmaniasis hotspot located in northeast Brazil. The data presented provides novel information about the presence of dermotropic L. (L.) infantum in the country and demonstrates the infectivity potential of theses isolates.

13.
Microbiol Spectr ; 11(3): e0280222, 2023 06 15.
Article in English | MEDLINE | ID: mdl-37052486

ABSTRACT

An integrative approach combining genomics, transcriptomics, and cell biology is presented to address leaf scald disease, a major problem for the sugarcane industry. To gain insight into the biology of the causal agent, the complete genome sequences of four Brazilian Xanthomonas albilineans strains with differing virulence capabilities are presented and compared to the GPEPC73 reference strain and FJ1. Based on the aggressiveness index, different strains were compared: Xa04 and Xa11 are highly aggressive, Xa26 is intermediate, and Xa21 is the least, while, based on genome structure, Xa04 shares most of its genomic features with Xa26, and Xa11 share most of its genomic features with Xa21. In addition to presenting more clustered regularly interspaced short palindromic repeats (CRISPR) clusters, four more novel prophage insertions are present than the previously sequenced GPEPC73 and FJ1 strains. Incorporating the aggressiveness index and in vitro cell biology into these genome features indicates that disease establishment is not a result of a single determinant factor, as in most other Xanthomonas species. The Brazilian strains lack the previously described plasmids but present more prophage regions. In pairs, the most virulent and the least virulent share unique prophages. In vitro transcriptomics shed light on the 54 most highly expressed genes among the 4 strains compared to ribosomal proteins (RPs), of these, 3 outer membrane proteins. Finally, comparative albicidin inhibition rings and in vitro growth curves of the four strains also do not correlate with pathogenicity. In conclusion, the results disclose that leaf scald disease is not associated with a single shared characteristic between the most or the least pathogenic strains. IMPORTANCE An integrative approach is presented which combines genomics, transcriptomics, and cell biology to address leaf scald disease. The results presented here disclose that the disease is not associated with a single shared characteristic between the most pathogenic strains or a unique genomic pattern. Sequence data from four Brazilian strains are presented that differ in pathogenicity index: Xa04 and Xa11 are highly virulent, Xa26 is intermediate, and Xa21 is the least pathogenic strain, while, based on genome structure, Xa04 shares with Xa26, and Xa11 shares with X21 most of the genome features. Other than presenting more CRISPR clusters and prophages than the previously sequenced strains, the integration of aggressiveness and cell biology points out that disease establishment is not a result of a single determinant factor as in other xanthomonads.


Subject(s)
Genome, Bacterial , Plant Diseases , Saccharum , Xanthomonas , Brazil , Genomics , Xanthomonas/classification , Xanthomonas/genetics , Xanthomonas/pathogenicity , Saccharum/microbiology , Plant Diseases/microbiology , Genetic Variation , Phylogeny , Gene Expression Profiling , Transcriptome , Ribosomal Proteins/genetics , Ribosomal Proteins/metabolism , Multigene Family/genetics
14.
Biochimie ; 208: 86-92, 2023 May.
Article in English | MEDLINE | ID: mdl-36586564

ABSTRACT

Leishmania parasites have an oxidative and chemical defense mechanism called trypanothione system (T[SH]2), the most abundant thiol system in trypanosomatids. This system has a central role in processing pentavalent antimony and resistance has been related to a better capacity to metabolize it through the activation of T[SH]2 enzymatic cascade. A biochemical approach was applied to assess the effect of trivalent (SbIII) and pentavalent antimony (SbV) on Trypanothione Reductase (TR) activity of two Leishmania (Viannia) braziliensis clinical isolates, which were labeled as responder (R) and non-responder (NR) after patient treatment with Glucantime®. Both isolates were characterized based on in vitro susceptibility to SbIII and SbV and trypanothione reductase (TR) activity. SbIII and SbV discriminated susceptibility profiles in all parasite forms, since isolate NR had significantly higher EC50 values than isolate R. Differences were observed in TR activity between promastigotes, axenic amastigotes and intracellular amastigotes: R (0.439 ± 0.009, 0.103 ± 0.01 and 0.185 ± 0.01AU.min-1.µg of protein-1) and NR (1.083 ± 0.04, 0.914 ± 0.04 and 0.343 ± 0.04 AU. min-1.µg of protein-1), respectively. Incubation with SbIII and SbV using each form EC50 value caused a time-dependent differential effect on TR activity suggesting that oxidative defense is related to the antimony susceptibility phenotype. Data gathered here shows a biochemical approach able to discriminate two L. (V.) braziliensis clinical isolates measurements TR activity of promastigotes, axenic amastigotes and intracellular amastigotes.


Subject(s)
Leishmania braziliensis , Leishmania , Antimony/pharmacology , Meglumine Antimoniate
15.
Front Fungal Biol ; 4: 1276287, 2023.
Article in English | MEDLINE | ID: mdl-38186633

ABSTRACT

Brazil has a long history of using biological control and has the largest program in sugarcane agriculture to which a biocontrol program has been applied. This achievement is at least partly due to the utilization of the entomopathogenic fungus Metarhizium. This well-known fungal genus exhibits pathogenicity against a broad range of arthropod hosts and has been used globally as a biocontrol agent. This fungus is also a root symbiont, and in this capacity, it is a plant growth promoter. However, this feature (i.e., as a plant symbiont) has yet to be fully explored and implemented in Brazil, although the number of reports demonstrating Metarhizium's utility as a plant bioinoculant is increasing. The Brazilian bioproduct industry targets agricultural pests, and is limited to two Metarhizium species represented by four fungal isolates as active ingredients. Entomopathogenic fungi have also been successful in controlling arthropods of public health concern, as shown in their control of mosquitoes, which are vectors of diseases. The isolation of new indigenous Metarhizium isolates from a variety of substrates such as soil, insects, and plants shows the wide genetic diversity within this fungal genus. In this review, we emphasize the significance of Metarhizium spp. for the biological control of insects in Brazil. We also suggest that the experience and success of biological control with fungi in Brazil is an important resource for developing integrated pest management and sustainable strategies for pest control worldwide. Moreover, the future implementation prospects of species of Metarhizium being used as bioinoculants and possible new advances in the utility of this fungus are discussed.

16.
Biosci. j. (Online) ; 39: e39004, 2023. tab
Article in English | LILACS | ID: biblio-1415866

ABSTRACT

Lysine is an essential amino acid that is not biologically manufactured in the body. Different chemical methods for lysine production are expensive and give low yields. The present study was conducted with the purpose to evaluate the biochemical production of lysine by different carbon sources using bacterial isolates. Three carbon sources namely glucose, sucrose, and fructose were used to evaluate the biochemical production of lysine by Escherichia coli and Klebsiella spp. isolates. Optimum incubation periods were between 48-96 hours. An extensive amount of lysine was produced by all of these isolates in L6 fermentation medium. Maximum lysine was produced by Klebsiella isolate K1 6.48 g/L after 96 hours of incubation by using glucose as carbon source followed by 6.0 g/L by Klebsiella isolates K3 after 72 hours of incubation when sucrose was used as a carbon source at 37 °C. Highest amount of lysine was produced at 96 hours by Klebsiella isolates in addition to E. coli. From all three carbon sources using Klebsiella isolates and E. coli, glucose showed better lysine production.


Subject(s)
Biochemical Phenomena , Fermentation , Lysine
17.
Braz. dent. sci ; 26(3): 1-9, 2023. ilus, tab
Article in English | LILACS, BBO - Dentistry | ID: biblio-1509645

ABSTRACT

Objective:Streptococcus mutans is one of the etiological agents associated with caries due to its ability to metabolize carbohydrates and resist acidic environments. On the other hand, Streptococcus dentisani, shows characteristics associated with caries control due to its ability to inhibit growth of cariogenic bacteria. The aim of this work was to quantify the levels of Streptococcus mutans and Streptococcus dentisani from dental biofilm of children related to their caries situation. Material and Methods: After identification of morphologic characteristics of reference strains was performed, clinical isolates of biofilm compatible with these strains were selected and the Polymerase Chain Reaction technique was performed using species-specific primers. Biofilm samples from 25 children with caries and 21 without caries were collected to quantify the levels of S. mutans and S. dentisani. Results: There were statistically significant differences in the levels of S. mutans in the caries group and the levels increased as the severity of the carious lesion increased. By contrast, higher levels of S. dentisaniwere found in the caries-free group, although no statistically significant differences were found. In addition, the levels of S. dentisani decreased as the severity of the carious lesion increased. Conclusion: The increase in the frequency of S. dentisani in the caries-free group suggests the possibility of requiring minimum levels of this species in the dental biofilm to show an actual protective effect. It must also be considered that this effect might be related to intrinsic factors in children and the intraspecies genetic variability found in every individual. (AU)


Objetivo : Streptococcus mutans é um dos agentes etiológicos associados à cárie devido à sua habilidade de metabolizar carboidratos e resistir a ambientes ácidos. Já o Streptococcus dentisani , apresenta características associadas ao controle da cárie devido à sua capacidade de inibir o crescimento de bactérias cariogênicas. O objetivo deste trabalho foi quantificar os níveis de Streptococcus mutans e Streptococcus dentisani no biofilme dental de crianças em relação à situação de cárie destas. Material e Métodos: Após a identificação das características morfológicas das cepas de referência, foram selecionados do biofilme isolados clínicos compatíveis com essas cepas e realizada a técnica de Reação em Cadeia da Polimerase utilizando primers espécie-específicos. Amostras de biofilme de 25 crianças com cárie e 21 sem cárie foram coletadas para quantificar os níveis de S. mutans e S. dentisani. Resultados: Houve diferenças estatisticamente significativas nos níveis de S. mutans no grupo com cárie e os níveis aumentaram à medida que a gravidade da lesão cariosa aumentou. Por outro lado, foram encontrados níveis mais elevados de S. dentisani no grupo sem cáries, embora não tenham sido encontradas diferenças estatisticamente significativas. Além disso, os níveis de S. dentisani diminuíram à medida que a gravidade da lesão cariosa aumentava. Conclusão: O aumento da frequência de S. dentisani no grupo livre de cárie sugere a possibilidade de exigir níveis mínimos desta espécie no biofilme dental para mostrar um efeito protetor real. Deve-se considerar também que esse efeito pode estar relacionado a fatores intrínsecos nas crianças e à variabilidade genética intraespécie encontrada em cada indivíduo. (AU)


Subject(s)
Humans , Child , Streptococcus mutans , Biofilms , Dental Caries
18.
Dis Aquat Organ ; 152: 85-98, 2022 Dec 01.
Article in English | MEDLINE | ID: mdl-36453457

ABSTRACT

White spot syndrome virus (WSSV) infects several economically important aquaculture species, and has caused significant losses to the industry. This virus belongs to the Nimaviridae family and has a dsDNA genome ranging between 257 and 309 kb (more than 20 isolate genomes have been fully sequenced and published to date). Multiple routes of infection could be the cause of the high virulence and mortality rates detected in shrimp species. Particularly in Penaeus vannamei, differences in isolate virulence have been observed, along with controversy over whether deletions or insertions are associated with virulence gain or loss. The pathogenicity of 3 isolates from 3 localities in Mexico (2 from Sinaloa: 'CIAD' and 'Angostura'; and one from Sonora: 'Sonora') was evaluated in vivo in whiteleg shrimp P. vannamei infection assays. Differences were observed in shrimp mortality rates among the 3 isolates, of which Sonora was the most virulent. Subsequently, the complete genomes of the Sonora and Angostura isolates were sequenced in depth from infected shrimp tissues and assembled in reference to the genome of isolate strain CN01 (KT995472), comprising 289350 and 288995 bp, respectively. Three deletion zones were identified compared to CN01, comprising 15 genes, including 3 envelope proteins (VP41A, VP52A and VP41B), 1 non-structural protein (ICP35) and 11 other encoding proteins whose function is currently unknown. In addition, 5 genes (wsv129, wsv178, wsv204, wsv249 and wsv497) presented differences in their repetitive motifs, which could potentially be involved in the regulation of gene expression, causing virulence variations.


Subject(s)
Penaeidae , White spot syndrome virus 1 , Animals , White spot syndrome virus 1/genetics , Virulence/genetics , Aquaculture , Biological Assay/veterinary
19.
Chem Biol Interact ; 368: 110191, 2022 Dec 01.
Article in English | MEDLINE | ID: mdl-36181831

ABSTRACT

Praziquantel (PZQ) is the only drug available for community-based control programs which aim to reduce the prevalence and morbidity associated with schistosomiasis. Here, we synthesized and evaluated the schistosomicidal, biochemical and cytotoxic activities of EF24, a synthetic curcumin analog, against different isolates of Schistosoma mansoni. EF24 elicited marked phenotypic alterations at 10 µM against schistosomula and 42-day-old adult worms of the Naval Medical Research Institute (NMRI) isolate. EF24 had 50% effective concentration (EC50) values of <10 µM against the Luis Evangelista (LE), Sergipe (SE), Belo Horizonte (BH) and Belo Horizonte less sensitive to PZQ (BH < PZQ) isolates of adult S. mansoni; however, the respective sensitivities of these isolates differed. Changes in the parasite included, vacuolization of the tegument and focal lysis of the interstitial tissue and muscle layers. Against 28-day-old juvenile worms (LE isolate), EF24 was about three times more potent than PZQ. After 6 h at 12.5 µM, EF24 increased reactive oxygen species (ROS) and the activity of the antioxidant enzyme, glutathione-S-transferase (GST), by 32 and 19% in female and male adult worms, respectively. By contrast, after 6 h at 12.5 µM glutathione reductase (GR) activity decreased by 43 and 30%, and glutathione peroxidase (GPx) activity decreased by 67 and 44% in females and males, respectively. EF24 was less cytotoxic to mammalian host cells than to S. mansoni, with selectivity indexes (SIs) of 1.8-3.4 and 2.7-7.5 for juvenile and adult worms, respectively. Given the current evidence for the in vitro schistosomicidal effect of EF24, the structure-activity relationship of additional analogs to identify new candidates for schistosomiasis treatment is warranted.


Subject(s)
Curcumin , Schistosoma mansoni , Schistosomicides , Animals , Female , Male , Antioxidants/metabolism , Curcumin/analogs & derivatives , Curcumin/pharmacology , Mammals , Praziquantel/pharmacology , Schistosoma mansoni/drug effects , Schistosomiasis/drug therapy , Schistosomicides/pharmacology , Glutathione Peroxidase/metabolism , Glutathione Transferase/metabolism , Reactive Oxygen Species/metabolism , Glutathione Reductase/metabolism
20.
Rev. chil. infectol ; Rev. chil. infectol;39(5): 623-629, oct. 2022. ilus, tab, mapas
Article in Spanish | LILACS | ID: biblio-1431690

ABSTRACT

La disponibilidad de cepas bacteriana para el estudio de la resistencia bacteriana es clave para los avances en la investigación básica y clínica respecto del tema. Existen pocos biorrepositorios o bancos de bacterias con mecanismos de resistencia conocidos, aisladas de infecciones clínicamente significativas. Una revisión de la literatura revela que sólo en los Estados Unidos de América existe un biobanco de aislados resistentes disponibles para estudios. En esta publicación se cuenta cómo se creó el primer biorrepositorio de bacterias resistentes en Chile asociados a la Red de Laboratorios MICROB-R, con la participación de 11 centros distribuidos a lo largo del país, que a la fecha cuenta con más de 3.500 aislados bacterianos estudiados fenotípica y genotípicamente, disponibles para la comunidad científica chilena.


The availability of bacterial strains for the study of bacterial resistance is key to advances in basic and clinical research. There are few biobanks of bacteria with known resistance mechanisms, isolated from clinically significant infections. A review of the literature reveals that only in the United States of America is there a biobank of resistant isolates. This publication shows the creation of the first biorepository of resistant bacteria Chile associated with the MICROB-R Laboratory Network, with the participation of 11 centers distributed throughout the country, which to date has more than 3,500 bacterial isolates studied phenotypically and genotypically, available to the Chilean scientific community.


Subject(s)
Bacteria/isolation & purification , Biological Specimen Banks , Drug Resistance, Bacterial , Chile , Laboratories
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