Prevalence and Genomic Investigation of Salmonella Isolates Associated with Watermelons and Their Environmental Reservoirs in Bejaia, Algeria.

This study was conducted in Bejaia, Algeria, to determine the presence of Salmonella in fresh watermelon (n = 105), soil (n = 23), and irrigation water samples (n = 17) collected from two different farms. After isolation, antimicrobial susceptibility testing, serotype determination, multilocus sequence typing, antimicrobial resistance genes detection, and whole genome sequencing were performed. Twenty watermelon samples (19%) were contaminated with Salmonella, but none were found in the soil or irrigation water. Among the 20 Salmonella isolates, 2 serovars were identified (Salmonella Liverpool and Salmonella Anatum), belonging to sequence types ST1959 and ST64, respectively. Ten Salmonella isolates showed significant resistance to nalidixic acid, ofloxacin, and ciprofloxacin but were susceptible to all other antibiotics. The coexistence of point mutations (parC:p.T57S) in Quinolone Resistance-Determining Regions and the qnrB19 gene may contribute to quinolone resistance. The study identified 164 virulence genes in the Salmonella isolates. Our study found Salmonella in fresh watermelon during the preharvest season in Bejaia, Algeria. Our study indicates a relatively high prevalence of Salmonella on watermelon samples before harvest. Although we cannot directly compare our results with previous studies, it is crucial to recognize that the absence of comprehensive comparative data underscores the need for further research and surveillance.

Nontyphoid Salmonella in farm animals and food products in the Middle East and North Africa: a systematic review.

Aim: This study aimed to document the prevalence, serotype distribution and antibiotic resistance of nontyphoidal Salmonella in animal food products from Middle East/North Africa (MENA) countries. Methods: Peer-reviewed articles published from 1 January 2011 to 7 March 2023 were included and the data were narratively synthesized and statistically analyzed to estimate and compare the overall prevalence. Results: The authors found a high prevalence of Salmonella in MENA countries (12.80%), with the highest prevalence in Lebanon (41.10%). Poultry had a higher prevalence of Salmonella (14.49%) than livestock (9.62%). Salmonella enteritidis was the most commonly identified serotype (21.99%), and sulfamethoxazole had the highest resistance rate (78.81%). Conclusion: The authors emphasize the importance of implementing control measures in MENA countries to limit the spread of the Salmonella pathogen.

Comprehensive Genomic Characterization of Antibiotic Resistance, Virulence, and Clonality in Salmonella Isolates from Wild Animals in Algeria.

This study investigated Salmonella spp. in wild animals in Algeria, focusing on their prevalence, serotypes, antibiotic resistance, and virulence profiles. From fecal samples collected between May 2021 and June 2022, 1.9% showed Salmonella shedding. The identified serotypes included S. Bredeney, S. Enteritidis, S. Altona, and S. Virchow. Except for S. Altona, all isolates were resistant to quinolones, with S. Bredeney strains, exhibiting multidrug resistance. Whole-genome sequencing revealed various resistance genes and mutations in gyrA or parC genes. Additionally, plasmids IncX1 and ColpVC were detected in several isolates. A comprehensive analysis identified 201 virulence genes. These findings contribute to understanding Salmonella in wild animal populations and their potential impact on public health.

Whole Genome Sequencing of Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Isolated from Neonatal Bloodstream Infections at a Neonatal Care Unit, Algeria.

Aims: Neonatal bloodstream infections (BSIs) are an important cause of mortality among neonates. Besides, extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-Kp) is one of the most frequent pathogens causing neonatal BSIs. This study aimed to characterize ESBL-Kp strains recovered from neonatal BSI and to investigate risk factors associated with ESBL-Kp BSI at the neonatal care unit of Elmeki Hospital, Bejaia, Algeria. Methodology: After isolation, identification, and antibiotic susceptibility testing, the ESBL-Kp strains were characterized by whole genome sequencing. The genomes were then analyzed using bioinformatic tools to determine the resistome, virulome, and phylogenetic relatedness. Results: From September 2019 to May 2020, 27 (8.2%) out of 328 neonates were infected by ESBL-Kp strains. These strains displayed a multidrug-resistant phenotype, and on further investigation, were found to carry an array of antibiotic resistance genes. All ESBL-Kp strains harbored the blaCTX-M-15 gene. Using in silico multilocus sequence typing analysis, six sequence types (STs) were detected with ST268 being the most frequent (56%, n = 15) indicating a local outbreak, confirmed by single nucleotide polymorphism analysis. The yersiniabactin and colibactin gene clusters were identified in six and two ESBL-Kp strains, respectively. Conclusion: This study showed a high prevalence of CTX-M-15-producing K. pneumoniae strains coharboring different antibiotic resistance mechanisms from neonatal BSIs in Algeria. Screening of health care personnel and mothers for ESBL carriage before delivery, isolation of carriers, barrier precautions, antimicrobial usage, and control of hygiene are needed to prevent the dissemination of these pathogens.

First Report of CC5-MRSA-IV-SCCfus "Maltese Clone" in Bat Guano.

Methicillin-resistant Staphylococcus aureus (MRSA) is a widespread pathogen that could cause different illnesses in both human and animals. Presence of MRSA in animals raises concerns of their capacity to act as reservoirs, particularly in wild animals. This study aimed to characterize the resistance and virulence patterns of S. aureus strains isolated from bat guano in Algeria. From March to May 2016, 98 bat guano samples from Aokas's cave (Bejaia, Algeria) were collected. Swabs were taken for microbiological studies. Isolates were identified by Vitek® MS system, and antibiotic susceptibility was determined by disk diffusion method. The clonal origin, virulence and antibiotic resistance profiles of S. aureus isolates were characterized by whole genome sequencing. Eleven S. aureus strains were obtained from the 98 guano samples. Seven isolates were sensitive to all antibiotics tested and four (36.3%) were resistant to penicillin G, cefoxitin and fusidic acid. The four MRSA isolates were assigned to the sequence type ST149 and related to spa type t010. These isolates harbored a SCCmecIV element and the fusidic acid resistance element Q6GD50 (fusC). They carried different virulence genes including several enterotoxins (sea, egc enterotoxin locus, sec, sel), and the toxic shock syndrome toxin (tst). Our results highlight that bat guano may constitute an important reservoir of MRSA strains.

Epidemiological and Genetic Features of Plasmids Carrying blaNDM Genes: An In Silico Analysis with Emphasis on Replicon Types, and Resistome.

Background: New Delhi metallo-ß-lactamase (NDM) is a metallo-ß-lactamase that has been disseminated worldwide. Plasmids harboring the blaNDM gene belonged to many incompatibility groups, of which IncX3, IncF, and IncA/C were the most represented. This in silico study aimed at analyzing a set of 649 plasmids carrying NDM-type carbapenemase (pNDMs) previously assigned in GenBank. Materials and Methods: The selected plasmids were analyzed by ResFinder (antibiotic resistome identification), BacMet (metal/biocides resistome identification), PlasmidFinder/PLSDB (replicon typing), TAfinder (toxin-antitoxin system [TAS] identification), and OriTfinder (prediction of the transferability). Results: We found that Escherichia coli and Klebsiella pneumoniae amounted to about 68.6% of all reported species. The distribution of these plasmids by samples showed a diversity of origins. Many plasmids carried different genes encoding resistance to antibiotics, heavy metals, and biocides with different frequencies. The TAfinder allowed the identification of a TAS in 292 plasmids (45%). Twenty-four different incompatibility groups were predicted, of which IncX3 (34.2%; n = 222), IncC (10.9%, n = 71), and IncFII (9.9%, n = 64) were the most often described. Besides, 23.6% (n = 151) of pNDMs were recognized as multireplicon plasmids. Conclusion: This study has shown the importance of plasmids in the dissemination of the NDM carbapenemase and raises the importance of monitoring these elements to better understand the evolution of the antibiotic resistance threat.

Fitness Cost of Antibiotic Resistance in Staphylococcus aureus: A Systematic Review.

Background: Recent reports have shown the potential of Staphylococcus aureus for acquiring resistance to last-resort antibiotics. However, most antibiotic resistance mechanisms were associated with a fitness cost that was typically observed as a reduced bacterial growth rate. This systematic review aimed to address the fitness cost of antibiotic resistance in S. aureus that emerged by mutations. Methods: A systematic review was conducted after searching in two databases (PubMed and Scopus) using specific keywords. We included peer-reviewed articles published only in English. All studies describing the fitness cost associated with antibiotic resistance in S. aureus were selected. For each article, the results of fitness testing, minimum inhibition concentrations of mutants, the position of mutation, and the appearance of compensatory mutations were recorded. Results: At all, 35 articles were recorded in the final analysis examining the fitness cost associated with antibiotic resistance in S. aureus that conferred by mutations. Analysis of the data showed that 26 studies reported that the emergence of antibiotic resistance was frequently associated with a fitness cost. Conclusion: This review summarized that the antibiotic resistance selection caused in the majority of cases a substantial fitness cost. Further in vivo experiments revealed that these mutations affected bacterial virulence and the ability to establish a successful infection.

Plasmid-Determined Colistin Resistance in the North African Countries: A Systematic Review.

We have conducted a systematic review to update available information on plasmid-mediated colistin resistance (mobilized colistin resistance [mcr]) genes in North African countries. We have searched the articles of PubMed, Scopus, and Web of Science databases reporting plasmid-mediated colistin resistance bacteria isolated in North African countries. After searching and selection, 30 studies that included 208 mcr-positive isolates were included. Different mcr-positive strains frequencies were recorded and ranged from 2% in clinical isolates to 12.3% in environmental samples. Escherichia coli was the predominant species recorded and these microorganisms showed high resistance to ciprofloxacin and cotrimoxazole. IncHI2 plasmids are probably the key vectors responsible for the dissemination of mcr genes in these countries. This review highlighted that the mcr-positive isolates are circulating in different ecological niches with different frequencies. Therefore, actions should be implemented to prevent the dissemination of the mcr genes within and outside of these countries, such as microbiological and molecular surveillance programs and restriction use of colistin in farming.

Genomic analysis of a multidrug-resistant Klebsiella pneumoniae ST11 strain recovered from Barbary deer (Cervus elaphus barbarus) in Akfadou Forest, Algeria.

OBJECTIVES: The emergence and worldwide spread of carbapenemase-producing Enterobacterales (CPE) is a great public-health concern. This study aimed to screen for the presence of CPE isolates from Barbary deer in Akfadou Forest, Béjaïa (Algeria). METHODS: Faecal samples (n=39) were obtained from Barbary deer in Akfadou Forest between March-June 2018. Whole-genome sequencing (WGS) was performed to characterise one representative strain of Klebsiella pneumoniae. Data analysis was performed using online tools. RESULTS: A total of 13 carbapenem-resistantK. pneumoniae isolates were obtained. The isolates showed an identical antimicrobial resistance pattern and were susceptible to colistin and fosfomycin. WGS analysis revealed the complete resistome of K. pneumoniae strain CF21, including blaNDM-1, blaCTX-M-15, blaSHV-182, blaDHA-1, blaOXA-1, aac(3)-IIa, aac(3)-IId, aac(6')-Ib-cr, rmtC, sul1, qnrB9, fosA, tetA, dfrA14, catA2, catB3 and mphA. Multilocus sequence typing (MLST) analysis assigned this strain to the international clone ST11. Plasmid analysis showed that this K. pneumoniae strain possesses five different plasmids including IncA/C2, IncFIA(HI1), IncFIB(K), IncFII(K) and ColRNAI. CONCLUSION: This study reports a multidrug-resistantK. pneumoniae strain recovered from Barbary deer in Algeria and confirms that wild animals could serve as a reservoir of antimicrobial resistance genes.

Fecal Carriage of Extended-Spectrum ß-Lactamase and Carbapenemase-Producing Enterobacterales Strains in Patients with Colorectal Cancer in the Oncology Unit of Amizour Hospital, Algeria: A Prospective Cohort Study.

Background: The prevalence of extended-spectrum ß-lactamase (ESBL)-producing Enterobacterales (ESBL-E) and carbapenemase-producing Enterobacterales (CPE) is now disseminated worldwide. This study aims to describe the prevalence of ESBL and CPE fecal carriage in colorectal cancer patients. Methods: All patients admitted to the oncology service of Amizour hospital (Algeria) for colorectal cancer chemotherapy from March to May 2019 were screened for ESBL-E or CPE fecal carriage. After culturing on chromogenic media, the presumptive colonies were identified by mass spectroscopy. Antibiotic susceptibility testing was performed according to the European Committee on Antimicrobial Susceptibility Testing. The ß-lactamases encoding genes and plasmid-mediated quinolone-resistant genes were screened by PCR and sequencing. Results: ESBL-E strains were recovered from rectal swabs in 6 patients (14.3%) and only 1 patient (2.4%) was found a carrier for OXA-48-producing Klebsiella pneumoniae. The most frequently encountered species among ESBL-E was Escherichia coli (n = 5), followed by K. pneumoniae (n = 1). PCR and sequencing showed that four isolates harbored the blaCTX-M-15 gene and two strains harbored the blaCTX-M-14 gene. Also, one strain of K. pneumoniae was found to harbor both qepA and qnrS genes. Conclusion: This study highlighted the fecal carriage of ESBL-E and OXA-48-producing Enterobacterales strains in colorectal cancer patients.

Methicillin-Resistant Staphylococcus aureus ST80 Clone: A Systematic Review.

This review assessed the molecular characterization of the methicillin-resistant Staphylococcus aureus (MRSA)-ST80 clone with an emphasis on its proportion of total MRSA strains isolated, PVL production, spa-typing, antibiotic resistance, and virulence. A systematic review of the literature was conducted on MRSA-ST80 clone published between 1 January 2000 and 31 August 2019. Citations were chosen for a review of the full text if we found evidence that MRSA-ST80 clone was reported in the study. For each isolate, the country of isolation, the sampling period, the source of isolation (the type of infection, nasal swabs, or extra-human), the total number of MRSA strains isolated, number of MRSA-ST80 strains, antibiotic resistance patterns, PVL production, virulence genes, and spa type were recorded. The data from 103 articles were abstracted into an Excel database. Analysis of the data showed that the overall proportion of MRSA-ST80 has been decreasing in many countries in recent years. The majority of MRSA-ST80 were PVL positive with spa-type t044. Only six reports of MRSA-ST80 in extra-human niches were found. This review summarizes the rise of MRSA-ST80 and the evidence that suggests that it could be in decline in many countries.

Epidemiology of carbapenemase-producing Enterobacterales in the Middle East: a systematic review.

Introduction: The Middle East is actually recognized as endemic for carbapenemases-producing Enterobacterales (CPE) including at least OXA-48-like and NDM-like.Areas covered: We performed a search of PubMed and Scopus using relevant keywords. We included peer-reviewed articles published only in English reporting any data on carbapenemase-producing bacteria from Middle East countries. The last literature search was performed on 26 October 2019. All studies describing carbapenemase-producing Enterobacterales isolated from humans, animals or environmental samples from the Middle East were included.Expert opinion: The Middle-East is considered an endemic region for CPE strains and the extensive international exchange could facilitate the spread of CPE from these countries to other parts of the Globe in which the prevalence of the CPE is low. The expansion of the Middle East conflict has been associated with the rapid collapse of the existing health care system of the concerned countries. Considering that Millions of refugees have fled their country, they could introduce these CPE strains in countries with low endemicity. In conclusion, the health care system actors should take in a count the endemicity of CPE in these countries and develop local surveillance programs to limit the spread of these MDR bacteria.

Carbapenemase-Producing Enterobacterales in Algeria: A Systematic Review.

The dramatic increase in prevalence and clinical impact of infections caused by carbapenemase-producing Enterobacterales (CPE) are major and ongoing public health problems globally. The carbapenemase groups most commonly encountered include Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-ß-lactamase (NDM), and OXA-48, with OXA-48 thought to be endemic to North Africa. These enzymes are frequently found on mobile genetic elements and have the potential to be widespread over the world, while the situation in Algeria is not well documented. In this article, we provide an in-depth review of CPE in Algeria, providing up-to-date information on the subject.

Distribution of Toxinogenic Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus from Different Ecological Niches in Algeria.

The diffusion of Panton-Valentine leukocidin (PVL)-positive methicillin-resistant S. aureus (MRSA) is a health problem in Algeria. The objectives of the study were to investigate the global distribution of methicillin-susceptible S. aureus (MSSA) and MRSA isolates in different ecological niches in this country. In total, 2246 samples were collected from humans, livestock, wild animals, pets, food products and the aquatic environment, from 12 Algerian provinces. A total of 312 S. aureus were detected from 2446 samples (12.7%) in the screened niches. We observed the emergence of toxinogenic S. aureus representing 41% of the isolates. Among them, we noted the diffusion of ST80-IV CA-MRSA PVL + strains isolated in human, animals, and food and genetic diversity of MSSA PVL + isolates. This study suggests an alarming dissemination of MRSA-ST80 PVL + in both human and extra-human sources in Algeria. Moreover, MSSA may become a permanent reservoir of the PVL genes necessary for human infections.

OXA-48-producing Enterobacterales in different ecological niches in Algeria: clonal expansion, plasmid characteristics and virulence traits.

OBJECTIVES: To investigate the prevalence and molecular characteristics of OXA-48-carbapenemase-producing Enterobacterales strains recovered from various ecological niches in Algeria. METHODS: In total, 3309 samples were collected from different ecological niches (human carriage, animal farms, wild animals, pets, food products, aquatic environment and wastewater treatment plants) distributed among six provinces in Algeria between December 2015 and April 2017. The potential presence of OXA-48-producing Enterobacterales isolates was screened on selective medium. Resistance and virulence profiles were characterized by PCR and sequencing. The clonal relatedness of the different isolates was studied using Rep-PCR and MLST. Conjugation was performed for all OXA-48-producing isolates. The plasmids were analysed by PCR-based replicon typing and WGS. RESULTS: A total of 78 OXA-48-producing Enterobacterales isolates were detected from 3309 samples (2.4%). OXA producers were observed in all the screened sources. The blaCTX-M-15 gene was only observed in two isolates. Clonality analysis revealed distinct lineages of the isolates and a clonal expansion of Klebsiella pneumoniae ST13. K. pneumoniae and Escherichia coli had few virulence factors. Plasmid analysis confirmed that all the isolates harboured a very similar transferable plasmid (belonging to IncL) with a similar structure to the pOXA-48a plasmid carried by K. pneumoniae strain Kp11978. CONCLUSIONS: This study suggests a global dissemination of OXA-48-producing Enterobacterales in different niches due mainly to the spread of an epidemic plasmid. Furthermore, it clearly shows that K. pneumoniae and commensal E. coli can be reservoirs of the blaOXA-48 gene, contributing to the dissemination and transfer of this gene to diverse bacteria among different sources.

Performance of a new in-house medium Carba MTL-broth for the rapid detection of carbapenemase-producing Enterobacteriaceae.

INTRODUCTION: The spread of carbapenemase-producing Enterobacteriaceae (CPE) represents a major public health issue. Methods allowing rapid detection of carbapenemases in developing countries are therefore urgently needed. In the current study, we developed a new in-house medium for the rapid detection of CPE isolates, especially OXA-48 producers. METHODOLOGY: A panel of 144 clinical strains previously characterized was tested on in-house Carba MTL-broth medium using four different concentrations of ertapenem (0.5 to 2 mg/L), and compared to chromID® OXA-48 and chromID® CARBA (BioMérieux) media. RESULTS: Comparative evaluation of the Carba MTL-broth with chromID® OXA-48 and chromID® CARBA showed that chromID® OXA-48 and Carba MTL-broth had the highest sensitivity for detection of OXA-48 producers (93.9% and 100%, respectively) comparatively to chromID® CARBA (21.2%). The chromID® OXA-48 had the highest specificity (100%), as compared to the Carba MTL-broth (65.5%) and chromID® CARBA (84.4%) for the detection of OXA-48 producers. CONCLUSIONS: The in-house Carba MTL-broth developed in this study is sensitive, inexpensive, an easy-to-use phenotypic method for the detection of OXA-48-producing enterobacteria. Given the burden of pan-drug resistance, its implementation in the microbiology laboratory of developing countries could be a useful tool for rapid detection of these bacteria.

Carbapenemase-producing Enterobacteriaceae among pregnant women and newborns in Algeria: Prevalence, molecular characterization, maternal-neonatal transmission, and risk factors for carriage.

The diffusion of carbapenemase-producing Enterobacteriaceae (CPE) represents a worldwide public health problem. This study revealed that the prevalence of OXA-48-producing enterobacteria was 4.6% (19/414) and 1.6% (7/422) in mothers and newborns, respectively, from 2 maternity units in Algeria. Previous hospital admission was an independent factor associated with an increased risk of CPE carriage in the mothers (P = .021). The low birth weight was significantly associated with this carriage in the newborns (P = .008). The screening of these bacteria is essential to prevent the dissemination of CPE.

OXA-48-like carbapenemases producing Enterobacteriaceae in different niches.

The emergence of carbapenem-resistant enterobacterial species poses a serious threat to public health worldwide. OXA-48-type carbapenem-hydrolyzing class D ß-lactamases are widely distributed among Enterobacteriaceae, with significant geographical differences. To date, 11 OXA-48-like variants have been identified, with classical OXA-48 being the most widespread. These enzymes show high-level hydrolytic activity against penicillins and low-level hydrolysis towards carbapenems. Since the first description of the OXA-48 carbapenemase in Turkey, bacterial strains producing the enzyme have been extensively reported in nosocomial and community outbreaks in many parts of the word, particularly in the Mediterranean area and European countries. The rapid spread of Enterobacteriaceae producing OXA-48-like enzymes in different ecosystems has become a serious issue recently. The number of reservoirs for such organisms is increasing, not only in hospitals, but also in the community, among animals (e.g., livestock, companion animals, and wildlife) and in the environment. This review aims to summarize the main characteristics of the OXA-48-type carbapenemases, covering genetic and enzymatic traits, their epidemiology, clonality and associated genes, correlation with extended-spectrum ß-lactamases (ESBLs) or plasmidic AmpC (pAmpC) in different bacterial species worldwide.

Extended spectrum ß-lactamase and plasmid mediated quinolone resistance in Escherichia coli fecal isolates from healthy companion animals in Algeria.

The aim of this study was to evaluate the rate of fecal carriage of Escherichia coli strains producing Extended-spectrum ß-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR) isolated from healthy pets (dogs and cats) in Algeria. Fecal samples from 171 healthy pets (102 dogs and 69 cats) in one veterinary practice and private owners were included. After isolates identification, antibiotic susceptibility was determined by disk diffusion procedure. ESBL were detected by combination disk tests. PCR and sequencing were used to characterize genes encoding ESBLs and PMQR. Transfer of ESBL and PMQR genes was assessed by conjugation experiments. Phylogenetic groups of E. coli were determined by PCR. Of the 171 animals, 20 carried an ESBL producing E. coli giving a prevalence of ESBL fecal carriage of 11.7%. All isolates were susceptible to carbapenems, cefoxitin, piperacillin-tazobactam, amikacin and fosfomycine. For the rest of the tested ß-lactams, susceptibility rates ranged from 35% to 70% for cefepime and amoxicillin-clavulanic acid respectively. Concerning the non-beta-lactams antibiotics, the rates of susceptibility ranged between 5% to trimethoprim and 95% for chloramphenicol. The beta-lactamase genes identified in E. coli isolates were blaCTX-M-15, blaCTX-M-1, blaSHV-12 and blaTEM-1. The PMQR determinants aac(6')-Ib-cr, qnrS1 and qnrB5 genes were identified in 15 isolates. Transconjugants were obtained for two isolates. Phylogenetic analysis showed that E. coli isolates belong to commensal phylogroups of A and B1. We reported here for the first time in Algeria ESBL and PMQR-producing E. coli in healthy cats and dogs.