Lotfi Loucif

This study aimed to evaluate the antibacterial activity of three Algerian medicinal plants (Brocchia cinerea, Artemisia campestris and Origanum vulgare) against multi-drug-resistant Gram-negative bacteria isolates from different food samples. Essential oils of the mentioned plants were extracted by hydro-distillation of the aerial parts. The chemical composition of the obtained oils was then determined using Gas Chromatography/Mass Spectrometry analysis. Moreover, the antibacterial activity was performed against different antibiotic-resistant bacterial species using the broth micro-dilution method. Gas Chromatography/Mass Spectrometry analysis showed a dominance of beta-thujone for the Brocchia cinerea essential oil. Whereas, the major components of Origanum vulgare and Artemisia campestris essential oils were carvacrol and thymol as well as 5(6 H)-benzocyclooctenone − 7,8- dihydro-8,8-dimethyl, 1,2,3,4,4a,9,10,10a-(trans- 4a,10a)-octahydro-9- oxophenanthrene, Germacrene D, respectively. The antibacterial activity results revealed that all the tested essential oils were active against the tested bacteria, including those resistant to last-resort antibiotics with minimum inhibitory concentrations ranging between 1.333 µg/mL and 170.72 µg/mL. In addition, Brocchia cinerea essential oil showed the best activity with the lowest minimum inhibitory concentration followed by Origanum vulgare and Artemisia campestris essential oils, respectively. Our study confirmed that the tested essential oils had an interesting activity against the tested antibiotic-resistant bacteria and may act as a potential source of natural effective antibacterial molecules that could be further investigated to help prevent bacterial proliferation in food products.

The aim of this study was to investigate the distribution and genetic determinants of carbapenemase production and colistin resistance among Acinetobacter baumannii isolates recovered from three health care facilities in the city of Batna, Algeria. A prospective study was conducted between 2021 and 2022 on 46 Acinetobacter baumannii clinical isolates, which were collected and identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Antibiotic susceptibility testing was performed using the disk diffusion method and colistin minimum inhibitory concentrations (MICs) were determined by broth microdilution method. Carbapenemase and colistin resistance determinants were detected by qPCR. The 46 clinical isolates were mainly from the intensive care unit (52.17%) and the burns unit (17.39%). The strains were collected primarily from pus samples (34.78%) and blood samples (17.39%). Eleven strains were classified as colistin-resistant, with MICs ranging from 4 to 128 μg/mL. The blaOXA-24 gene was detected in 63.04% of the isolates, followed by the blaOXA-23 gene (43.47%). Nine strains were positive for both blaOXA-23-like and blaOXA-24-like genes. The blaNDM gene was detected in eight isolates (17.39%), including two which co-expressed a blaOXA-24 gene. In contrast, all strains were negative for the plasmid-mediated colistin resistance mcr-1 to mcr-5 and mcr-8. Here, we report a high prevalence of carbapenemases-producing A. baumannii isolates in Batna hospitals. Notably, this study is the first to identify A. baumannii isolates co-producing OXA-24 and NDM carbapenemases and to report the first detection of colistin-resistant A. baumannii co-producing OXA-24 and OXA-23 carbapenemases from a patient in Algeria.

Background: Lactic acid bacteria (LAB) are central to the food and health sectors due to their capacity to enhance the safety, shelf-life, and functional properties of fermented foods, while also exerting probiotic effects that support immune function and gut health. This study aimed to isolate and characterize LAB from dromedary camel milk (DCM) collected in semi-arid regions of Algeria, a traditional yet underexplored matrix, eval-uating their technological features and antimicrobial potential. Methods: Thirty-one LAB strains were isolated from raw DCM samples. Species identification was performed using MALDI-TOF MS. Isolates were assessed for acidification kinetics, proteolytic, lipolytic, and amylolytic activities, production of exopolysaccharides (EPS) and acetoin, as well as an-timicrobial activity against common foodborne pathogens. Results: Four LAB species were identified, with Enterococcus italicus reported for the first time in this niche. All functional traits showed significant inter-strain variability (p < 0.0001). Three strains (BLC9, BLC12, BLC14) acidified milk rapidly to pH 4.6 within 12 hours. Proteolytic ac-tivity was present in 87.10% of isolates, while EPS and acetoin were produced by 29.03% and 48.39%, respectively. Most strains (74.19%) exhibited antimicrobial activity against at least one pathogen, with variable inhibition zones. Conclusion: Algerian dromedary camel milk represents a valuable reservoir of LAB strains with functional, antimicrobial, and potential probiotic attributes. These native strains hold promise for the development of additive-free fermented foods and natural biopreservation strategies, ultimately supporting human health and contributing to sustainable, culturally rooted dietary practices.

The escalating prevalence of drug-resistant Gram-negative bacteria represents a widespread threat to global public health. This study aimed to investigate the occurrence and genetic determinants of extended-spectrum β-lactamases (ESBLs), carbapenemases, and mcr-encoding genes in GNB isolates from companion animals in the city of Batna, Algeria. In total, 50 faecal samples were collected from various veterinary clinics and pet shops, including direct collection of faecal droppings (n = 22) and rectal swabs (n = 28). After selective isolation, Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry was used to identify representative colonies. Susceptibility testing was carried out using the disk diffusion method. ESBLs, carbapenemases, and colistin resistance determinants were searched for by real-time PCR. Thirty-seven strains were isolated and were mostly identified as Escherichia coli (n = 21). Molecular analysis revealed that 23 isolates carried only the bla_ESBL genes. The genes which were most detected were bla_CTX-M-A (n = 14) and bla_CTX-M-A associated with bla_TEM (n = 8). Two Enterobacter cloacae isolates were positive for the bla_OXA-48 gene while one of them additionally carried the bla_CTX-M-A gene. The mcr-1 gene was detected in one Enterobacter kobei isolate. To the best of our knowledge, this is the first report of E. kobei carrying the mcr-1 gene from rectal swabs of cats.

As defined by the World Health Organization, stillbirth refers to the delivery of a baby without signs of life at or after 28 weeks of gestation. This study aims to predict stillbirth occurrences by identifying influencing factors. Interviews were conducted with 786 pregnant women from Eastern Algeria between 2011 and 2015, resulting in a 9.03% stillbirth rate. Utilizing statistical techniques, a predictive model with a 95% confidence interval was developed. Findings indicated a significantly higher likelihood of stillbirths among pregnant women aged over 34 (OR 8.77; 1.70-161.28, p=0.039) and those with a high BMI of (25-30) (OR 1.73; 0.83-3.78, p=0.153) and (30-35) (OR 4.16; 2.04-8.93, p<0.001). Additionally, associations were observed with stress (OR 2.19; 1.15-4.49, p=0.023) and diabetes (OR 2.68; 1.15-5.90, p=0.017). Moderate correlations were noted with meat consumption (OR 0.53; 0.21-1.19, p=0.144), high fish intake (OR 3.02; 1.04-8.62, p=0.039), dairy products (OR 1.81; 0.94-3.67, p=0.085), and sweets (OR 0.47; 0.24-0.88, p=0.021). This study underscores that identifying these risk factors aids in preventing stillbirths, thereby safeguarding the well-being of both mothers and infants. Future research should explore the broader implications of these findings and potential avenues for further investigation.

BackgroundCarbapenemase-producing Enterobacterales isolates are associated with significant mortality and have emerged as a major problem in healthcare settings worldwide.ObjectiveOur aim was to investigate the epidemiological and genotypic characteristics of carbapenemase-positive Enterobacterales isolates from patients hospitalised in three hospitals in the city of Batna, Algeria.MethodsBetween 2016 and 2019, a total of 5,316 clinical isolates were obtained. The collected isolates were identified using the VITEK-2 system. Demographic and microbiological data were collected as well as the antimicrobial susceptibility testing, phenotypic and molecular characterisation of carbapenemase and mcr-1 genes were performed.ResultsOut of the 5,316 isolates, 201 were carbapenem-resistant Enterobacterales isolates and 179 of them (89.05%) were positive for the production of carbapenemase, of which Klebsiella pneumoniae, Escherichia coli and Enterobacter cloacae were the most common. The blaOXA-48-like gene alone was detected in 147 isolates (82.12%) moreover, the blaNDM gene was detected in ten isolates (5.59%). Dual and triple combinations of carbapenemase genes were also observed here for the first time in Algeria: blaVIM and blaOXA-48-like; blaKPC, blaVIM and blaOXA-48-like; blaVIM and blaNDM; blaKPC, blaNDM and blaVIM; blaNDM and blaOXA-48-like genes. In addition, resistance to both colistin and carbapenem antibiotics was detected in eight isolates, however none of them was positive for the mcr-1 gene.ConclusionThis is the first study reporting the detection of carbapenemase genes in Klebsiella aerogenes, Enterobacter sakazakii, Raoultella ornithinolytica, Serratia ficaria, and Serratia marcescens and specific carbapenemase gene combinations in Algeria. The present study revealed that blaOXA-48-like were found to be the predominant carbapenemase genes in Batna hospitals.

Antibiotic-resistant-bacteria are being considered as emerging environmental contaminants where the importance of the surrounding environment in their emergence and dissemination has been emphasized. The aim of this study was to screen for the presence and diversity of carbapenem- and colistin-resistant Gram-negative bacteria (GNBs) in different aquatic environments. Water samples were collected in Batna, Algeria. Carbapenem- and colistin-resistant GNBs were selectively isolated and then identified using matrix-assisted laser desorption and ionization time-of-flight mass spectrometry. After phenotypic antibiotic susceptibility testing, the molecular mechanisms of β-lactams and colistin-resistance were investigated by PCR and sequencing. The clonality of mcr-1 positive Escherichia coli was determined by multi-locus sequence typing. We noticed a high level of resistance in both tap water and wastewater. The most commonly found carbapenem-resistance mechanism was the OXA-48 enzyme, but other carbapenemases were also detected. In addition, the mcr-1 gene was detected in 18 E. coli of different sequence types. Our findings highlight the role of aquatic environments in the dissemination of resistant-bacteria, especially considering that water is a connecting medium between different ecological systems and can easily transmit resistant-bacteria and promote horizontal gene transfer. Thus, the development of effective treatment strategies for eliminating antibiotic-resistance is seriously needed.

The aim of this study was to screen for extended spectrum cephalosporin-, carbapenem- and colistin-resistant Gram-negative bacteria in fresh vegetables in Batna, Algeria. A total of 400 samples of fresh vegetables were collected from different retail stores. Samples were immediately subjected to selective isolation, then the representative colonies were identified using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF–MS). Phenotypic and genotypic analyses were carried out in terms of species identification and relative antibiotic resistance. Transferability of the carbapenemase and mcr-bearing plasmids was verified by conjugation. The clonal relationships of carbapenemase and mcr-positive Escherichia coli isolates were studied by multi-locus sequence typing (MLST). Sixty-seven isolates were characterised and were mostly isolated from green leafy vegetables, where the dominant species identified included Citrobacter freundii, Klebsiella pneumoniae, Enterobacter cloacae, Stenotrophomona maltophilia, E. coli and Citrobacter braakii. PCR and sequencing results showed that E. coli was the bacterial species presenting the highest antibiotic resistance level in parallel to bla_TEM (n = 16) and bla_CTX-M-15 (n = 11), which were the most detected genes. Moreover, five isolates carried carbapenemase genes, including the bla_OXA-48 and/or bla_VIM-4 genes. The mcr-1 gene was detected in two E. coli isolates. MLST analysis revealed three different E. coli sequence types: ST101 (n = 1), ST216 (n = 1) and ST2298 (n = 1). Conjugation assays confirmed the transferability of the bla_OXA-48 and mcr-1 genes. In this study we report, for the first time, the detection of the bla_OXA-48 gene in E. coli and C. braakii isolates and the bla_VIM-4 gene in vegetables. To the best of our knowledge, this is the first report on the detection of mcr-1 genes from vegetables in Algeria.

Multidrug-resistant (MDR) Enterobacterales remain an increasing problem in Algeria, notably due to the emergence of carbapenemase producers. We investigated the molecular characteristics of carbapenem-resistant Enterobacterales isolates recovered from outpatients and inpatients in Eastern Algeria. Non-repetitive Enterobacterales with reduced susceptibility to carbapenems were consecutively collected from clinical specimens in Annaba University Hospital (Algeria) between April 2016 and December 2018. Isolates were characterized with regard to antibiotic resistance, resistome and virulome content, clonality, and plasmid support. Of the 168 isolates analyzed, 29 (17.3%) were carbapenemase producers and identified as K. pneumoniae (n = 23), E. coli (n = 5), and E. cloacae (n = 1). bla_OXA-48 was the most prevalent carbapenemase-encoding gene (n = 26/29), followed by bla_NDM-1 gene (n = 3/29). K. pneumoniae isolates harbored some virulence traits (entB, ugeF, ureA, mrkD, fimH), whereas E. coli had a commensal origin (E, A, and B1). Clonality analysis revealed clonal expansions of ST101 K. pneumoniae and ST758 E. coli. Plasmid analysis showed a large diversity of incompatibility groups, with a predominance of IncM (n = 26, 89.7%). A global dissemination of OXA-48-producing Enterobacterales in the Algerian hospital but also the detection of NDM-1-producing E. coli in community settings were observed. The importance of this diffusion must be absolutely investigated and controlled.