Detection of ESBLs and carbapenemases among Enterobacteriaceae isolated from diabetic foot infections in Ouargla, Algeria.

INTRODUCTION: The emergence and rapid spread of Enterobacteriaceae carrying extended spectrum beta-lactamases (ESBLs) and carbapenemases represent a great threat to clinical treatment due to their multi-drug resistance. This study investigated ESBLs and carbapenemases encoding genes in Enterobacteriaceae collected from diabetic foot infections (DFIs) in Ouargla, southern Algeria. METHODOLOGY: A total of 70 Enterobacteriaceae strains were recovered from 76 patients with DFI between February 2017 and April 2018. Antimicrobial susceptibility testing was performed using the disc diffusion method, and the presence of bla genes was detected using polymerase chain reaction (PCR) and DNA sequencing. The genetic transfer of the plasmids was carried out by conjugation using the broth mating method. RESULTS: The most common isolate was Proteus mirabilis, followed by Escherichia coli, Morganella morganii and Klebsiella pneumoniae. The prevalence of ESBL and carbapenemase-producing Enterobacteriaceae was 11.42% and 2.85 % respectively. Plasmid-mediated AmpC was detected in 5.71% isolates. Conjugation experiments showed the transferability of blaCTX-M-2. CONCLUSIONS: Our findings support the view that various pathogens found in DFIs differ from one part of the country to another. This study reports the first description of metallo-ß-lactamase NDM-5 producing Klebsiella pneumoniae clinical isolate in Algeria.

Pharmacological studies on aqueous extract of Launaea arborescens from southwest Algeria.

ETHNOPHARMACOLOGICAL RELEVANCE: Launaea arborescens, its vernacular name is Mol-albina belonging to asteracaea family origin of the southwest of Algeria. This plant is used in folk medicines to treat gastroenteritis, diabetes, child aliment and other diseases; it is taken macerated or boiled. AIM: This study aims to evaluate the anti-inflammation an analgesic activity of the aqueous extract of Launaea arborescens (AqELA) and its pathway of action. METHODS: the investigation of anti-inflammatory and analgesic effects were done using formalin test, acetic acid test. For mechanism investigation, it was used hot plate test to induce opioid receptors, a histamine and serotonin test to induce edema paw, finally, for the TRPV1 receptor, it was used the capsaicin test. RESULTS: The aqueous extract of Launaea arborescens showed a significant inhibition of abdominal writhing test 95% and 100% inhibition of licking paw using acid acetic test and formalin test respectively (EC: 47 mg/kg and 104 mg/kg). The analgesic effect of the aqueous extract of Launaea arborescens showed inhibition of sensation of pain after 120 min compared to morphine effect. The aqueous extract of Launaea arborescens reduced paw volume after 180 min and 120 min for histamine and serotonin respectively with dose-dependent. Concerning of TRPV1 receptors, the inhibition was showed at doses 100 mg and 300 mg. CONCLUSION: Our results contribute towards validation of the traditional use of Launaea arborescens for inflammation ailment.

Antimicrobial and Antioxidant Effects of a Forest Actinobacterium V002 as New Producer of Spectinabilin, Undecylprodigiosin and Metacycloprodigiosin.

The aim of the study is the research and identification of a Streptomyces strain as a new producer of spectinabilin, undecylprodigiosin and metacycloprodigiosin. Among 54 actinomycete isolates isolated from El-Ogbane forest soils in Algeria, only one isolate, designated V002, was selected for its ability to produce prodigiosins. The selected strain was analysed for its ability to produce three different secondary metabolites as well as their biological activities. V002 belongs to the Streptomyces genus and has significant antimicrobial and antioxidant activities. The taxonomic position of V002 by 16S rRNA sequence analysis showed a similarity of 99.93% with Streptomyces lasiicapitis DSM 103124T and 98.96% with Streptomyces spectabilis DSM 40512T. Fractionation of crude secondary metabolites produced by the strain using HPLC-MS revealed the presence of spectinabilin, undecylprodigiosin and metacycloprodigiosin, which demonstrated significant activity. Strain V002 is considered a new producer of spectinabilin, undecylprodigiosin and metacycloprodigiosin with significant antimicrobial and antioxidant activity.

Petroleum degradation by endophytic Streptomyces spp. isolated from plants grown in contaminated soil of southern Algeria.

Petroleum hydrocarbons are well known by their high toxicity and recalcitrant properties. Their increasing utilization around worldwide led to environmental contamination. Phytoremediation using plant-associated microbe is an interesting approach for petroleum degradation and actinobacteria have a great potential for that. For this purpose, our study aimed to isolate, characterize, and assess the ability of endophytic actinobacteria to degrade crude petroleum, as well as to produce plant growth promoting traits. Seventeen endophytic actinobacteria were isolated from roots of plants grown naturally in sandy contaminated soil. Among them, six isolates were selected on the basis of their tolerance to petroleum on solid minimal medium and characterized by 16S rDNA gene sequencing. All petroleum-tolerant isolates belonged to the Streptomyces genus. Determination by crude oil degradation by gas chromatorgraph-flame ionization detector revealed that five strains could use petroleum as sole carbon and energy source and the petroleum removal achieved up to 98% after 7 days of incubation. These isolates displayed an important role in the degradation of the n-alkanes (C6-C30), aromatic and polycyclic aromatic hydrocarbons. All strains showed a wide range of plant growth promoting features such as siderophores, phosphate solubilization, 1-aminocyclopropane-1-carboxylate deaminase, nitrogen fixation and indole-3-acetic acid production as well as biosurfactant production. This is the first study highlighting the petroleum degradation ability and plant growth promoting attributes of endophytic Streptomyces. The finding suggests that the endophytic actinobacteria isolated are promising candidates for improving phytoremediation efficiency of petroleum contaminated soil.

Genetic characterisation of carbapenem-resistant Gram-negative bacteria isolated from the University Hospital Mohamed Boudiaf in Ouargla, southern Algeria.

OBJECTIVES: The aim of this study was to investigate the molecular epidemiology and the genetic support of carbapenem-resistant Enterobacteriaceae and Acinetobacter spp. isolates collected in the University Hospital of Ouargla, southern Algeria. METHODS: A total of 99 Gram-negative bacteria (GNB) were collected from stool samples of colonised patients and from inanimate surfaces in the hospital environment between December 2014 and August 2015. Selected Enterobacteriaceae and Acinetobacter spp. isolates with reduced susceptibility to carbapenems were subjected to phenotypic study, including antibiotic susceptibility testing according to CA-SFM-EUCAST 2015 guidelines and modified Carba NP test. Genes encoding carbapenemases, extended-spectrum ß-lactamases (ESBLs) and AmpC ß-lactamases were screened by PCR and sequencing. Clonal relatedness was determined by multilocus sequencing typing (MLST). RESULTS: Of the 99 GNB isolates, 10 (10.1%) showed reduced susceptibility to carbapenems were studied further, including 7 Acinetobacter baumannii, 1 Acinetobacter nosocomialis, 1 Escherichia coli and 1 Klebsiella pneumoniae. PCR and sequencing showed that four A. baumannii isolates and the single A. nosocomialis isolate harboured blaNDM-1. In addition, blaOXA-23 was observed in three A. baumannii isolates, and blaOXA-48 was detected in the two Enterobacteriaceae isolates. MLST assigned the K. pneumoniae to ST999 and the E. coli to ST38. The seven A. baumannii isolates belonged to ST85 and ST2. CONCLUSIONS: This study describes the epidemiology of carbapenemases produced by Enterobacteriaceae and Acinetobacter spp. in southern Algeria and reports the first description of metallo-ß-lactamase NDM-1-producing A. nosocomialis in Algeria.