scholarly journals Use of Metagenomics and Isolation of Actinobacteria in Brazil’s Atlantic Rainforest Soil for Antimicrobial Prospecting

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Danyelle Alves Martins Assis ◽  
Rachel Passos Rezende ◽  
João Carlos Teixeira Dias
Keyword(s):  
Promising Result ◽  
Multidrug Resistant ◽  
Fosmid Library ◽  
Atlantic Rainforest ◽  
Human Pathogen ◽  
Culture Methods ◽  
Antimicrobial Substances ◽  
Salmonella Choleraesuis ◽  
Acai Berry ◽  
Number Of Microorganisms

Modern techniques involving molecular biology, such as metagenomics, have the advantage of exploiting a higher number of microorganisms; however, classic isolation and culture methods used to obtain antimicrobials continue to be promising, especially in the isolation of Actinobacteria, which are responsible for the production of many of these compounds. In this work, two methodologies were used to search for antimicrobial substances—isolation of Actinobacteria and metagenomics of the Atlantic Rainforest soil and of the cultivation of cocoa intercropped with acai berry in the Atlantic Rainforest. The metagenomic libraries were constructed with the CopyControl Fosmid Library kit EPICENTRE, resulting in a total of 2688 clones, 1344 of each soil sample. None of the clones presented antimicrobial activity against the microorganisms tested: S. aureus, Bacillus subtilis, and Salmonella choleraesuis. A total of 46 isolates were obtained from the isolation of soil Actinobacteria: 24 isolates from Atlantic Rainforest soil and 22 isolates from the intercrop cultivation soil. Of these, two Atlantic Rainforest soil isolates inhibited the growth of S. aureus including a clinical isolate of S. aureus MRSA—a promising result, since it is an important multidrug-resistant human pathogen.

scholarly journals Occurrence, antimicrobial susceptibility patterns and genotypic relatedness of Salmonella spp. isolates from captive wildlife, their caretakers, feed and water in India

2018 ◽  
Vol 146 (12) ◽  
pp. 1543-1549
Author(s):  
Arockiasamy Arun Prince Milton ◽  
Rajesh Kumar Agarwal ◽  
Govindarajan Bhuvana Priya ◽  
Cheruplackal Karunakaran Athira ◽  
Mani Saminathan ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Animal Species ◽  
Multidrug Resistant ◽  
Wild Animal ◽  
Diffusion Method ◽  
Pcr Assay ◽  
Direct Pcr ◽  
Salmonella Spp ◽  
Culture Methods ◽  
Faecal Samples

AbstractOccurrence of Salmonella spp. in captive wild animal species in India is largely unknown. The purpose of this study was to determine the occurrence of different Salmonella serotypes, antimicrobial resistance patterns and genotypic relatedness of recovered isolates. A total of 370 samples including faecal (n = 314), feed and water (n = 26) and caretakers stool swabs (n = 30) were collected from 40 different wild animal species in captivity, their caretakers, feed and water in four zoological gardens and wildlife enclosures in India. Salmonellae were isolated using conventional culture methods and tested for antimicrobial susceptibility with the Kirby–Bauer disc diffusion method. Salmonella isolates were serotyped and genotyping was performed using enterobacterial repetitive intergenic consensus (ERIC) PCR and 16S rRNA sequencing. Animal faecal samples were also subjected to direct PCR assay. Salmonella was detected in 10 of 314 (3.1%) faecal samples by isolation and 18 of 314 (5.7%) samples by direct PCR assay; one of 26 (3.8%) feed and water samples and five of 30 (16.7%) caretakers stool swabs by isolation. Salmonella was more commonly isolated in faecal samples from golden pheasants (25%; 2/8) and leopard (10%; 2/20). Salmonella enterica serotypes of known public health significance including S. Typhimurium (37.5%; 6/14), S. Kentucky (28.5%; 4/14) and S. Enteritidis (14.3%; 2/14) were identified. While the majority of the Salmonella isolates were pan-susceptible to the commonly used antibiotics. Seven (43.7%; 7/16) of the isolates were resistant to at least one antibiotic and one isolate each among them exhibited penta and tetra multidrug-resistant types. Three S. Kentucky serotype were identified in a same golden pheasants cage, two from the birds and one from the feed. This serotype was also isolated from its caretaker. Similarly, one isolate each of S. Typhimurium were recovered from ostrich and its caretaker. These isolates were found to be clonally related suggesting that wildlife may serve as reservoir for infections to humans and vice versa. These results emphasise the transmission of Salmonella among hosts via environmental contamination of feces to workers, visitors and other wildlife.

scholarly journals Environmental Isolation of Candida auris from the Coastal Wetlands of Andaman Islands, India

mBio ◽  
2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Parth Arora ◽  
Prerna Singh ◽  
Yue Wang ◽  
Anamika Yadav ◽  
Kalpana Pawar ◽  
...  
Keyword(s):  
Salt Marsh ◽  
Coastal Wetlands ◽  
Ecological Niche ◽  
Multidrug Resistant ◽  
Sandy Beaches ◽  
Andaman Islands ◽  
Human Pathogen ◽  
Candida Auris ◽  
Content Type ◽  
The Indian Ocean

ABSTRACT Candida auris is a multidrug resistant pathogen that presents a serious global threat to human health. As C. auris is a newly emerged pathogen, several questions regarding its ecological niche remain unexplored. While species closely related to C. auris have been detected in different environmental habitats, little is known about the natural habitat(s) of C. auris. Here, we explored the virgin habitats around the very isolated Andaman Islands in the Indian Ocean for evidence of C. auris. We sampled coastal wetlands, including rocky shores, sandy beaches, tidal marshes, and mangrove swamps, around the Andaman group of the Andaman & Nicobar Islands, Union Territory, in India. Forty-eight samples of sediment soil and seawater were collected from eight sampling sites representing the heterogeneity of intertidal habitats across the east and west coast of South Andaman district. C. auris was isolated from two of the eight sampling sites, a salt marsh and a sandy beach. Interestingly, both multidrug-susceptible and multidrug-resistant C. auris isolates were found in the sample. Whole-genome sequencing analysis clustered the C. auris isolates into clade I, showing close similarity to other isolates from South Asia. Isolation of C. auris from the tropical coastal environment suggests its association with the marine ecosystem. The fact that viable C. auris was detected in the marine habitat confirms C. auris survival in harsh wetlands. However, the ecological significance of C. auris in salt marsh wetland and sandy beaches to human infections remains to be explored. IMPORTANCE Candida auris is a recently emerged multidrug-resistant fungal pathogen capable of causing severe infections in hospitalized patients. Despite its recognition as a human pathogen a decade ago, so far the natural ecological niche(s) of C. auris remains enigmatic. A previous hypothesis suggested that C. auris might be native to wetlands, that its emergence as a human pathogen might have been linked to global warming effects on wetlands, and that its enrichment in that ecological niche was favored by the ability of C. auris for thermal tolerance and salinity tolerance. To understand the mystery of environmental niches of C. auris, we explored the coastal wetland habitat around the very isolated Andaman Islands in the Indian Ocean. C. auris was isolated from the virgin habitats of salt marsh area with no human activity and from a sandy beach. C. auris isolation from the marine wetlands suggests that prior to its recognition as a human pathogen, it existed as an environmental fungus.

scholarly journals Convergent Evolution Driven by Rifampin Exacerbates the Global Burden of Drug-Resistant Staphylococcus aureus

mSphere ◽  
2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Romain Guérillot ◽  
Anders Gonçalves da Silva ◽  
Ian Monk ◽  
Stefano Giulieri ◽  
Takehiro Tomita ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Multidrug Resistant ◽  
Clinical Impact ◽  
Drug Resistant ◽  
Deleterious Mutations ◽  
Human Pathogen ◽  
Laboratory Studies ◽  
Increased Risk ◽  
Rifampin Resistance ◽  
Recent Laboratory

Increasing antibiotic resistance in the major human pathogen Staphylococcus aureus is threatening the ability to treat patients with these infections. Recent laboratory studies suggest that mutations in the gene commonly associated with rifampin resistance may also impact susceptibility to other last-line antibiotics in S. aureus; however, the overall frequency and clinical impact of these mutations are unknown. By mining a global collection of clinical S. aureus genomes and by mutagenesis experiments, this work reveals that common rifampin-induced rpoB mutations promote phenotypic plasticity that has led to the global emergence of stable, multidrug-resistant S. aureus lineages that are associated with increased risk of therapeutic failure through coresistance to other last-line antimicrobials. We recommend decreasing susceptibility breakpoints for rifampin to allow phenotypic detection of critical rpoB mutations conferring low resistance to rifampin and reconsidering the appropriate use of rifampin to reduce the fixation and spread of these deleterious mutations globally.

scholarly journals Complete Genome Sequence of Neisseria gonorrhoeae NCCP11945

2008 ◽  
Vol 190 (17) ◽  
pp. 6035-6036 ◽  
Author(s):  
Gyung Tae Chung ◽  
Jeong Sik Yoo ◽  
Hee Bok Oh ◽  
Yeong Seon Lee ◽  
Sun Ho Cha ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Neisseria Gonorrhoeae ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Multidrug Resistant ◽  
Human Pathogen ◽  
Korean Patient

ABSTRACT Neisseria gonorrhoeae is an obligate human pathogen that is the etiological agent of gonorrhea. We explored variations in the genes of a multidrug-resistant N. gonorrhoeae isolate from a Korean patient in an effort to understand the prevalence, antibiotic resistance, and importance of horizontal gene transfer within this important, naturally competent organism. Here, we report the complete annotated genome sequence of N. gonorrhoeae strain NCCP11945.

scholarly journals CHLOROFORM FRACTION OF PARKIA JAVANICA BARK POSSESSES ANTIBACTERIAL ACTIVITY AGAINST MULTIDRUG RESISTANT GRAM NEGATIVE BACTERIA PREDOMINANTLY FOUND IN SKIN WOUND

2018 ◽  
Vol 8 (5) ◽  
pp. 184-189
Author(s):  
S Saha ◽  
P Karmakar ◽  
Samir Kumar Sil
Keyword(s):  
Antibacterial Activity ◽  
Cell Damage ◽  
Multidrug Resistant ◽  
Skin Wound ◽  
Lag Phase ◽  
Klebsiella Pneumonia ◽  
Chloroform Fraction ◽  
Bacterial Strains ◽  
Gram Negative ◽  
Antimicrobial Substances

Aim: To evaluate the antibacterial activity of Parkia javanica against gram negative MDR bacterial strains which are predominantly found in skin wound. Methods: The 5 different solvent fractions of Parkia javanica were screened for antibacterial activity against gram negative multi drug resistant bacterial strains namely Enterobacter aerugenes, Pseudomonas aeruginosa and Klebsiella pneumonia by serial dilution technique. Growth kinetics study was performed and percentage of ROS production was measured by NBT reduction assay. Results: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were obtained with a range of IC100 0.08-0.31 mg/ml in case of MDR bacterial strains. The lag phase of all extract treated bacteria is extended compared to untreated cells. The normalized % of ROS is increased in presence of Parkia javanica extract. Conclusions: This study suggests that, chloroform fraction of Parkia javanica possesses promising antimicrobial substances which are having activity against MDR bacterial strains and ROS induced bacterial cell damage could be the possible mediator of its antimicrobial activity. Keywords:   Parkia javanica, antibacterial activity, MDR bacterial strains, growth curve, ROS.

scholarly journals Identification of a novel subfamily of bacterial AAT-fold basic amino acid decarboxylases and functional characterization of its first representative:Pseudomonas aeruginosaLdcA

2018 ◽  
Author(s):  
Diego Carriel-Lopez ◽  
Pierre Simon Garcia ◽  
Florence Castelli ◽  
Patricia Lamourette ◽  
François Fenaille ◽  
...  
Keyword(s):  
Amino Acid ◽  
Functional Characterization ◽  
Acid Stress ◽  
Basic Amino Acid ◽  
Multidrug Resistant ◽  
Human Pathogen ◽  
Short Forms ◽  
Charged Macromolecules ◽  
Functionally Diverse

SummaryPolyamines are small amino-acid derived polycations capable of binding negatively charged macromolecules. Bacterial polyamines are structurally and functionally diverse, and are mainly produced biosynthetically by PLP-dependent amino acid decarboxylases referred to as LAOdcs (Lysine-Arginine-Ornithine decarboxylases). In a phylogenetically limited group of bacteria, LAOdcs are also induced in response to acid stress. Here, we performed an exhaustive phylogenetic analysis of the AAT-fold LAOdcs which showcased the ancestral nature of their short forms inCyanobacteriaandFirmicutes,and emergence of distinct subfamilies of long LAOdcs inProteobacteria.We identified a novel subfamily of lysine decarboxylases, LdcA, ancestral inBetaproteobacteriaandPseudomortadaceae {Gammaproteobacteria).We analyzed the expression of LdcA fromPseudomonas aeruginosa,and uncovered its role, intimately linked to cadaverine production, in promoting growth and reducing persistence of this multidrug resistant human pathogen during carbenicillin treatment. Finally, we documented a certain redundancy in the function of the three main polyamines - cadaverine, putrescine and spermidine - inP. aeruginosaby demonstrating the link between their intracellular level, as well as the capacity of putrescine and spermidine to complement the growth phenotype of theIdcAmutant.

scholarly journals CsrA Coordinates Compatible Solute Synthesis in Acinetobacter baumannii and Facilitates Growth in Human Urine

2021 ◽  
Author(s):  
Josephine Joy Hubloher ◽  
Kim Schabacker ◽  
Volker Müller ◽  
Beate Averhoff
Keyword(s):  
Acinetobacter Baumannii ◽  
Multidrug Resistant ◽  
Compatible Solute ◽  
Human Pathogen ◽  
Content Type ◽  
Clinical Environments ◽  
Resistant Strains ◽  
Opportunistic Human Pathogen ◽  
Mrna Binding ◽  
Optimal Adaptation

The opportunistic human pathogen Acinetobacter baumannii has become one of the leading causes of nosocomial infections around the world due to the increasing prevalence of multidrug-resistant strains and their optimal adaptation to clinical environments and the human host. Recently, it was found that CsrA, a global mRNA binding posttranscriptional regulator, plays a role in osmotic stress adaptation, virulence, and growth on amino acids of A. baumannii AB09-003 and 17961.

scholarly journals Molecular Identification and Susceptibility Pattern of ClinicalNocardiaSpecies: Emergence ofNocardia crassostreaeas an Agent of Invasive Nocardiosis

2013 ◽  
Vol 24 (2) ◽  
pp. e33-e38 ◽  
Author(s):  
Saad J Taj-Aldeen ◽  
Anand Deshmukh ◽  
Sanjay Doiphode ◽  
Atqah Abdul Wahab ◽  
Mona Allangawi ◽  
...  
Keyword(s):  
Molecular Identification ◽  
Ribosomal Rna ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
16S Ribosomal Rna ◽  
Resistant Organism ◽  
Human Pathogen ◽  
Resistant Species ◽  
Nocardia Otitidiscaviarum

BACKGROUND:Nocardiaspecies are rare, opportunistic organisms that cause disease in both immunocompetent and immunocompromised individuals.OBJECTIVE: To investigate the clinical presentations of variousNocardiainfections based on the 16S ribosomal RNA gene of the isolate, as well as related risk factors and susceptibility patterns to antimicrobial agentsMETHODS: Thirteen patients with a diagnosis of nocardiosis were included in the present study. SevenNocardiaspecies were identified by 16S ribosomal RNA. Susceptibility testing was performed using six antimicrobial agents.RESULTS: Five patients were immunocompromised, and eight were immunocompetent with predisposing factors including cystic fibrosis, tuberculosis and ophthalmic infections.Nocardiacaused pulmonary infections in eight patients (61.5%), invasive systemic infections in three patients (23%) and local (ophthalmic) infections in two patients (15.4%). In the patients with pulmonary disease, nocardiosis was caused by six species (Nocardia cyriacigeorgica,Nocardia otitidiscaviarum,Nocardia farcinica,Nocardia carnea,Nocardia testaceaandNocardia asiatica). The seventh species identified in the present study wasNocardia crassostreae.DISCUSSION:N crassostreaeis a multidrug-resistant organism that was reported to be an emerging human pathogen causing invasive nocardiosis in a patient with non-Hodgkin’s lymphoma.N farcinicawas isolated from blood in a patient with breast cancer. None of theNocardiaisolates were resistant to linezolid. OneN otitidiscaviarumisolate was a multidrug-resistant organism. All patients in the present study were treated with the appropriate antibiotics and their condition resolved without further sequelae.CONCLUSIONS: The present study is the first report onN crassostreaeas a human pathogen. The detection of multidrug-resistant species necessitate molecular identification and susceptibility testing, and should be performed for allNocardiainfections. Nocardiosis manifests various clinical features depending on theNocardiaspecies and underlying conditions.

scholarly journals Three New Integration Vectors and Fluorescent Proteins for Use in the Opportunistic Human Pathogen Streptococcus pneumoniae

Genes ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 394 ◽  
Author(s):  
Lance E. Keller ◽  
Anne-Stéphanie Rueff ◽  
Jun Kurushima ◽  
Jan-Willem Veening
Keyword(s):  
Streptococcus Pneumoniae ◽  
Transformation Efficiency ◽  
Fluorescent Proteins ◽  
Inducible Promoter ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Large Set ◽  
Human Pathogen ◽  
Wide Range ◽  
Opportunistic Human Pathogen

Here, we describe the creation of three integration vectors, pPEPX, pPEPY and pPEPZ, for use with the opportunistic human pathogen Streptococcus pneumoniae. The constructed vectors, named PEP for Pneumococcal Engineering Platform (PEP), employ an IPTG-inducible promoter and BglBrick and BglFusion compatible multiple cloning sites allowing for fast and interchangeable cloning. PEP plasmids replicate in Escherichia coli and harbor integration sites that have homology in a large set of pneumococcal strains, including recent clinical isolates. In addition, several options of antibiotic resistance markers are available, even allowing for selection in multidrug resistant clinical isolates. The transformation efficiency of these PEP vectors as well as their ability to be expressed simultaneously was tested. Two of the three PEP vectors share homology of the integration regions with over half of the S. pneumoniae genomes examined. Transformation efficiency varied among PEP vectors based on the length of the homology regions, but all were highly transformable and can be integrated simultaneously in strain D39V. Vectors used for pneumococcal cloning are an important tool for researchers for a wide range of uses. The PEP vectors described are of particular use because they have been designed to allow for easy transfer of genes between vectors as well as integrating into transcriptionally silent areas of the chromosome. In addition, we demonstrate the successful production of several new spectrally distinct fluorescent proteins (mTurquoise2, mNeonGreen and mScarlet-I) from the PEP vectors. The PEP vectors and newly described fluorescent proteins will expand the genetic toolbox for pneumococcal researchers and aid future discoveries.

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