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Objectives To determine the genetic context of genes conferring antibiotic resistance on the carbapenem-resistant Acinetobacter baumannii Cl415, recovered in 2017 at El Youssef Hospital Centre in Akkar Governorate, North Lebanon. Methods Antibiotic resistance phenotype for 22 antibiotics was determined using disc diffusion or MIC determination. The whole-genome sequence of Cl415 was determined using a combination of the Illumina MiSeq and Oxford Nanopore (MinION) platforms. Complete genome was assembled using Unicycler and antibiotic resistance determinants and ISs were identified using ResFinder and ISFinder, respectively. Results Cl415 is a global clone 2 (GC2) strain and belongs to the most common STs of this clone, ST2IP and ST218OX. Cl415 is resistant to several antibiotics, including aminoglycosides and carbapenems to a high level. Genomic analysis of Cl415 revealed that it carries four chromosomal AbaR4 copies. One copy was found in the comM gene replacing the AbGRI1 island. Cl415 also contains a novel variant of AbGRI2, herein called AbGRI2-15, carrying only the blaTEM and aphA1 resistance genes. Cl415 belongs to a subclade of GC2 strains that appear to have diverged recently with a wide geographical distribution. Conclusions The resistance gene complement of Cl415 was found in the chromosome with four oxa23 located in AbaR4 copies and the remaining genes in a novel variant of the AbGRI2 resistance island. Cl415 was isolated in Lebanon, but phylogenetic analysis suggests that Cl415 represents a new lineage with global distribution within GC2.

Globally, more than billion people suffer from fungal infections each year. The early diagnosis of aspergillosis is mandatory for successful treatment outcome. As careful testing takes time, epidemiological surveillance is crucial to guide individual patient therapy and to promote a high standard of health care. In this paper, we first present current trends in the epidemiology and antifungal susceptibility patterns of Aspergillus spp. in Middle Eastern and North African (MENA) countries in order to support infectious disease specialists and health workforces in this geographic area to treat adequately patients with aspergillosis. Then we discuss the existing literature data regarding the available diagnostic tools and antifungal resistance mechanisms of Aspergillus spp. Although a limited number of studies were reviewed here, the currently available data show that Aspergillus infections are not negligible in the MENA region, and that the emergence of antifungal resistance is a growing health issue, especially among immunocompromised patients. Keywords: Aspergillus spp., Epidemiology, Antifungal resistance, Molecular mechanisms, MENA region

While cigarette smoke compounds are known to have immunosuppressive effects on the oral mucosa, the relationship between in vivo immune dysfunction caused by smoking and the development of oral Candida infections remains largely unexplored. In a recent issue of The Journal of Cellular and Molecular Medicine, Ye and colleagues provide evidence that smoking increases oral mucosa susceptibility to Candida albicans infection via the activation of the Nrf2 pathway, which in turn negatively regulates the NLRP3 inflammasome. This opens new perspective in considering Nrf2 as a relevant target for smoking‐induced C. albicans‐related oral diseases.

The outstanding ability of Acinetobacter baumannii to cause outbreaks and acquire multidrug resistance motivated the development of a plethora of typing techniques, which can help infection preventionists and hospital epidemiologists to more efficiently implement intervention controls. Nowadays, the world is witnessing a gradual transition from traditional typing methodology to whole genome sequencing-based approaches. Such approaches are opening new prospects and applications never achieved by existing typing methods. Herein, we provide the reader with an updated review on A. baumannii typing methods recapping the added value of well-established techniques previously applied for A. baumannii and detailing new ones (as clustered regularly interspaced short palindromic repeats-based typing) with a special focus on whole genome sequencing.

1. The real burden of Campylobacter spp. in Lebanon is still unknown. The aims of this study were to unravel the epidemiology of Campylobacter spp. in broilers at slaughterhouses in Tripoli, North of Lebanon and to characterise their antibiotic resistance profiles.2. From May to November 2015, sampling was performed through five repeated surveys from 15 slaughterhouses that sold chicken directly to Lebanese customers. Isolates were subjected to pulsed field gel electrophoresis (PFGE) and flaA-restriction fragment length polymorphism (flaA-RFLP).3. All investigated slaughterhouses were found to be positive for Campylobacter spp. Campylobacter coli was the predominant species (38 isolates) followed by C. jejuni (eight isolates). A noticeable level of resistance was detected among isolates against ciprofloxacin (97% of C. coli and 87.5% of C. jejuni), amoxicillin (89% of C. coli and 75% of C. jejuni), gentamicin (79% of C. coli and 50% of C. jejuni), and co-amoxiclav (24% of C. coli and 25% of C. jejuni). Erythromycin and ertapenem resistance were observed only in C. coli with the following percentages 74% and 13% respectively, but not in C. jejuni. PFGE and flaA-RFLP using DdeI as restriction enzyme divided the strains into 27 and 25 types respectively.4. The high observed genetic diversity of Campylobacter spp. revealed the complexity of the spread of this genus in broilers. This study highlighted the pressing need to monitor antibiotic resistance and to ensure food safety from 'farm to fork' in Lebanon.

Mastitis is an infectious disease encountered in dairy animals worldwide that is currently a growing concern in Lebanon. This study aimed at investigating the etiology of the main mastitis-causing pathogens in Northern Lebanon, determining their antimicrobial susceptibility profiles, and identifying their antimicrobial resistance (AMR) genes. A total of 101 quarter milk samples were collected from 77 cows and 11 goats presenting symptoms of mastitis on 45 dairy farms. Bacterial identification was carried out through matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibility was tested by disc diffusion and broth microdilution methods. Molecular characterization included polymerase chain reaction (PCR) screening for genes encoding extended-spectrum beta-lactamases (ESBLs) and plasmid-mediated AmpC among Enterobacterales isolates, and virulence factors among Staphylococcus isolates. Escherichia coli isolates were subjected to phylogenetic typing by a quadruplex PCR method. The most frequently identified species were Streptococcus uberis (19.2%), Streptococcus agalactiae (15.1%), E. coli (12.3%), and Staphylococcus aureus (10.96%). Gram-positive bacteria were resistant to macrolides and tetracycline, whereas gram-negative bacteria displayed resistance to ampicillin and tetracycline. Two ESBL genes, blaTEM (83.3%) and blaOXA (16.7%), and one AmpC beta-lactamase gene, blaCMY-II (16.7%), were detected among six E. coli isolates, which mainly belonged to phylogenetic group B1. Among Staphylococcus spp., the mecA gene was present in three isolates. Furthermore, four isolates contained at least one toxin gene, and all S. aureus isolates carried the ica operon. These findings revealed the alarming risk of AMR in the Lebanese dairy chain and the importance of monitoring antimicrobial usage.

Objectives Knowledge on the dynamic of MDR Escherichia coli in the human community is still limited, especially in low- and middle-income countries. Our goal was to decipher the dynamics of E. coli lineages and plasmids resistant to ESC, carbapenem and colistin within and between food workers in Lebanon using genomic-based approaches. Methods Eighty-four healthy adults working in three bakeries were sampled twice at a 6 monthly interval. E. coli resistant to ESC (ESC-E), carbapenem (CP-E) and colistin (CO-E) were collected on selective plates. Non-duplicate isolates were whole-genome sequenced using the Illumina technology and plasmid transmission was assessed by long-read sequencing. Data were analysed using bioinformatics tools and SNP-based phylogeny. Results ESC-E carriage rate reached 34.5% (t0) and 52.9% (t6), and 15 workers were positive at both t0 and t6. Carbapenem resistance (blaOXA-181, blaOXA-204, blaNDM-5) was found in five workers at t0 and two at t6, while colistin resistance (mcr-1.1) was found in five workers at t0 and one at t6. Forty-seven different STs were identified, of which three STs were predominant (ST131, n = 9; ST10, n = 5; ST69, n = 5). One worker presented the same ESC-E clone at t0 and t6. Twelve different events of clonal transmission among individuals were exemplified while plasmid transmission was only shown once. Conclusions Our study revealed a high carriage rate of MDR E. coli (60.7%) and the emergence of CP and colistin resistance in the Lebanese community. Incidental and long-term ESC-E carriage was observed in 41.7% and 17.9% of the workers, respectively. The high clonal diversity suggests an important dynamic of acquisition and loss of MDR E. coli and limited plasmid spread.

Aim: We sought to provide first insights into the epidemiology and antifungal susceptibility patterns of the aspergilli in Lebanon. Materials & methods: After species identification, antifungal susceptibility was investigated according to EUCAST recommendations. CYP51A gene was sequenced in resistant isolates and its expression level was evaluated by Reverse transcription-quantitative PCR. Results: Among the 73 Aspergillus isolates studied (mostly from ears), the predominant species was Aspergillus niger (54.8%). The overall drug resistance was highest for amphotericin B (38.4%), followed by itraconazole (31.5%), posaconazole (30.1%) and voriconazole (23.3%). In addition, CYP51A gene mutations were not the major cause of azole resistance among these isolates. Conclusion: Our findings indicate the paramount need for an integral One Health strategy and a national reference center for invasive mycoses and antifungals.

The Shigella genus includes serious foodborne disease etiologic agents, with 4 species and 54 serotypes. Identification at species and serotype levels is a crucial task in microbiological laboratories. Nevertheless, the genetic similarity between Shigella spp. and Escherichia coli challenges the correct identification and serotyping of Shigella spp., with subsequent negative repercussions on surveillance, epidemiological investigations, and selection of appropriate treatments. For this purpose, multiple techniques have been developed historically ranging from phenotype-based methods and single or multilocus molecular techniques to whole-genome sequencing (WGS). To facilitate the selection of the most relevant method, we herein provide a global overview of historical and emerging identification and serotyping techniques with a particular focus on the WGS-based approaches. This review highlights the excellent discriminatory power of WGS to more accurately elucidate the epidemiology of Shigella spp., disclose novel promising genomic targets for surveillance methods, and validate previous well-established methods.

Endocannabinoids are host-derived lipid hormones that fundamentally impact gastrointestinal (GI) biology. The use of cannabis and other exocannabinoids as anecdotal treatments for various GI disorders inspired the search for mechanisms by which these compounds mediate their effects, which led to the discovery of the mammalian endocannabinoid system. Dysregulated endocannabinoid signaling was linked to inflammation and the gut microbiota. However, the effects of endocannabinoids on host susceptibility to infection has not been explored. Here, we show that mice with elevated levels of the endocannabinoid 2-arachidonoyl glycerol (2-AG) are protected from enteric infection by Enterobacteriaceae pathogens. 2-AG directly modulates pathogen function by inhibiting virulence programs essential for successful infection. Furthermore, 2-AG antagonizes the bacterial receptor QseC, a histidine kinase encoded within the core Enterobacteriaceae genome that promotes the activation of pathogen-associated type three secretion systems. Taken together, our findings establish that endocannabinoids are directly sensed by bacteria and can modulate bacterial function.

Aim: To characterize the epidemiology of group A Streptococcus (GAS) involved in nonpharyngeal infections sparingly addressed in Lebanon. Materials & methods: A collection of 63 nonpharyngeal GAS isolates recovered between 2010 and 2019 from northern Lebanon were analyzed through emm typing, virulence gene profiling, FCT typing and antibiotic susceptibility analysis. Results & conclusion: A total of 29 emm subtypes was detected, with emm1 being the most dominant. A great intraclonal divergence driven by the loss and gain of superantigens or by the structural variability within the FCT regions was unraveled. The resistance rates for erythromycin and tetracycline were 8 and 20.6%, respectively. The 30-valent vaccine coverage was 76%. This study evidences the complexity of the neglected GAS pathogen in Lebanon.

In recent years, the increased use of macrolides was linked with the emergence of resistance Streptococcus pneumoniae worldwide. The main aim of this study was to determine the prevalence of S. pneumoniae resistant to macrolides and to identify the macrolide resistance genotypes among clinical isolates collected in North Lebanon. Disk diffusion susceptibility method was performed for 132 strains of S. pneumoniae isolated over a period of 5 years in North Lebanon. Polymerase Chain Reaction followed by pyrosequencing was carried out for confirmation of phenotypic diagnosis. The macrolide resistance genotypes were also identified by using PCR amplification of genes implicated in this resistance: erm(A), erm(B), erm(C), msr(A), lin(A) and mef(A/E). Macrolide resistance was found in 34.1% of S. pneumoniae isolates. We observed that the cMLSB phenotype (31/45, 68.9%) was the most common in these pneumococci and erm(B) was the most common resistance gene (32/45, 71.1%). This study shows that macrolide resistance in S. pneumoniae in North Lebanon is mainly related to target site modification with predominance of cMLSb phenotype but is also mediated by efflux pumps. lin(A) gene was reported for the first time in one S. pneumoniae strain in combination with erm(B) and mef(A/E) genes. Keywords: Streptococcus pneumoniae, Macrolide, erm(B), mef(A/E), lin(A), Lebanon

Background. Moraxella catarrhalis is an important bacterial pathogen. Although national data have shown an increase in the levels of antimicrobial resistance in clinical settings in Lebanon, there is a lack of data regarding this human pathogen. This study aimed to determine for the first time the antimicrobial susceptibility profiles of M. catarrhalis isolates in Lebanon. Methods. A total of 34 M. catarrhalis strains were isolated from clinical specimens during the period from November 2010 to March 2019. Bacterial identification was carried out using matrix assisted laser desorption ionization–time of flight mass spectrometry. Antibiotic susceptibility of all isolates was performed according the recommendations of the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Results. A total of 34 non-duplicated M. catarrhalis strains were isolated from nose (n=19), ear (n=7), sputum (n=5), blood (n=1), eye (n=1), and throat (n=1) of patients referred to Nini Hospital in Tripoli, North governorate of Lebanon. Regarding antibiotic susceptibility rates, the percent susceptibility is 100% to the majority of antibiotics, except ampicillin (7.4%), trimethoprim-sulfamethoxazole (85.3%), nalidixic acid (85.3%), and ciprofloxacin (97.1%). Conclusion. To our knowledge, this study is the first investigation regarding the antimicrobial susceptibility patterns of M. catarrhalis isolates in Lebanon. In addition to the high level of resistance to ampicillin, our findings showed the emergence of resistance to trimethoprim-sulfamethoxazole, nalidixic acid and ciprofloxacin. Even if this study provides useful information to develop effective empirical treatment, we recommend the implementation of reliable diagnostic tools to guide appropriate treatment.

International audience; Blastocystis sp. is frequently identified in a wide range of animal hosts, including bovids. Because of its burden and zoonotic potential, this parasite has been sought in domestic cattle from various countries, since this livestock may also represent a possible reservoir of human infection. However, epidemiological data regarding the prevalence and ST distribution of Blastocystis sp. in this animal group is lacking in Lebanon. Therefore, faecal samples were collected from a total of 254 dairy cattle raised on 55 farms located in the North Lebanon region and screened for the presence of the parasite by quantitative real-time PCR. The overall prevalence of Blastocystis sp. was shown to reach 63.4% in cattle livestock. Sequence analysis of positive samples indicated the presence of seven STs, with predominance of ST10 (44.0%) and ST14 (36.8%) and lower proportions of ST2 (8.0%), ST1 (7.2%), ST5 (2.4%), ST3 and ST7 (0.8% each). This survey was the first conducted worldwide reporting ST2 and ST7 in domestic cattle and confirmed that ST10 and ST14 represent cattle-adapted STs in view of their high prevalence. Faecal samples from in-contact dairy farmers and patients hospitalised in the same Lebanese gov-ernorate who reported no contact with cattle livestock were also analysed for the presence of Blastocystis sp. The same three STs were identified in both human cohorts, with predominance of ST3, followed either by ST1 or ST2 depending of the group. No other STs, including ST10 or ST14, have been reported. Moreover, even though ST1, ST2 and ST3 were found to be common to dairy cattle and farmers cohorts, only one ST3 isolate showed 100% sequence identity between both hosts. Consequently, the presence and low prevalence of ST1, ST2, ST3, ST5 and ST7 identified herein in domestic cattle, most of which exhibit low host specificity, could be derived from occasional direct exposure to faecal material from human and non-human hosts or by ingestion of contaminated drinking water or food in the enclosure of the farms. Together with the absence of ST10 and ST14 in the human population, these data suggest that cattle play a negligible role as zoonotic reservoirs of Blastocystis sp.

Quinolones are an important antimicrobial class used widely in the treatment of enterobacterial infections. Although there are multiple mechanisms of quinolone resistance, attention should be paid to plasmid-mediated genes due to their ability to facilitate the spread of quinolone resistance, the selection of mutants with a higher-level of quinolone resistance, and the promotion of treatment failure. Since their discovery in 1998, plasmid-mediated quinolone resistance (PMQR) mechanisms have been reported more frequently worldwide especially with the extensive use of quinolones in humans and animals. Nevertheless, data from the Arab countries are rare and often scattered. Understanding the prevalence and distribution of PMQR is essential to stop the irrational use of quinolone in these countries. This manuscript describes the quinolone resistance mechanisms and particularly PMQR among Enterobacteriaceae as well as their methods of detection. Then the available data on the epidemiology of PMQR in clinical and environmental isolates from the Arab countries are extensively reviewed along with the other associated resistance genes. These data shows a wide dissemination of PMQR genes among Enterobacteriaceae isolates from humans, animals, and environments in these countries with increasing rates over the years and a common association with other antibiotic resistance genes as blaCTX-M-15. The incontrovertible emergence of PMQR in the Arab countries highlights the pressing need for effective stewardship efforts to prevent the selection of a higher rate of quinolone resistance and to preserve these crucial antibiotics.

International audience; Campylobacter jejuni is recognized as the most common foodborne pathogen associated with human gastroenteritis worldwide. Broilers are frequently infected by the bacteria and are considered the main source of exposure to humans. However, despite its public health impact, no recent data are currently available in Lebanon about Campylobacter spp. in poultry and human population. Therefore, this study aimed to determine the prevalence and genetic diversity of Campylobacter spp. in 227 ceca and on 227 carcasses of broiler chickens collected in Lebanese slaughterhouses. Overall, the prevalence of Campylobacter was shown to reach 67.0% in ceca and 17.2% on carcasses of Lebanese poultry. The only 2 Campylobacter species identified were C. jejuni and C. coli, with a slightly higher prevalence of C. coli in ceca and of C. jejuni on carcasses. A high level of genetic diversity was reported among the 51 C. jejuni isolates selected, since 25 distinct profiles were identified according to the comparative genomic fingerprinting typing method based on a subset of 40 genes using the 90% similarity threshold. Predominant clusters observed in Lebanese poultry isolates were also frequently found among French human clinical cases, highlighting that broiler chickens represent a potential reservoir for human campylobacteriosis. In addition, a significantly higher prevalence of Campylobacter spp. was found in slaughterhouse workers than in a cohort of hospitalized patients with no contact with poultry, confirming that contaminated broiler chickens in slaughterhouse appeared to be a non-negligible source of Campy-lobacter spp. transmission. Interestingly, a significant association between Campylobacter spp. and Blastocys-tis sp. has been observed. This correlation suggested that the presence of Campylobacter spp. would be favored when Blastocystis sp. is present and, similarly, the absence of one would favor the absence of the other. This is the first large-scale investigation focusing on the impact of Campylobacter spp. in broiler chickens in Lebanon and confirmed the need to implement prevention and control measures in the poultry production to reduce the burden of campylobacteriosis in the human population.

Purpose. The aim was to evaluate several microbiological tools for the identification of non-gonococcal Neisseria spp. isolated from semen samples from Lebanese men and to determine the putative link between the presence of Neisseria commensal species and infertility. Methodology. Within a cross-sectional retrospective study design, the whole population included in this investigation was divided in 2 categories: 173 patients with symptoms of infertility and 139 patients with normal seminograms. Epidemiological and microbiological investigations were performed for 59 strains of Neisseria through several phenotypic and genotypic tools, including seminograms, an analytical profile index of Neisseria and Haemophilus (API-NH), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), porA PCR, 16S rRNA and rplF gene sequencing, and antimicrobial susceptibility testing. Results. The risk of Neisseria infection was twice as high in infertile patients compared to the control group [odds ratio (OR): 1.95, confidence interval (CI): 1.05–3.65, P =0.03]. Unreliable diagnosis of Neisseria urogenital infection has serious health and social consequences. Our findings showed that API-NH and 16S rRNA sequencing are poor tools to identify Neisseria at the species level. Therefore, reliable diagnosis of cases using MALDI-TOF MS and/or rplF sequencing is needed to provide critical treatment decisions and prevent antimicrobial resistance spreading in the community. Conclusion. This work predicted a strong and significant association between the presence of Neisseria spp. in semen and male infertility among the Lebanese population. For a better understanding of this association, it is recommended that more genomic and large-scale epidemiological investigations are undertaken to reach definitive conclusions.

Background: Mycobacterium spp. are responsible for several diseases, particularly in immunocompromised populations. The spreading of the resistance to antimycobacterial drugs is a significant problem to the public health and requires to find out a new and innovative alternative for the treatment of drug resistant mycobacterial strains. In this study, the antimycobacterial activity of Micromeria barbata, Eucalyptus globulus and Juniperus excelsa essential oils extracted from Lebanese plants was investigated against selected Mycobacterium spp. strains. Methods: Several dilutions of the three aforementioned essential oils were studied for antimycobacterial activity against four Mycobacterium spp. strains: Mycobacterium tuberculosis subsp. tuberculosis (ATCC® 27294™), multidrug-resistant M. tuberculosis (CMUL 157), Mycobacterium kansasii Hauduroy (ATCC® 12478™) and Mycobacterium gordonae Bojalil et al. (ATCC® 14470™). Results: Even with high dilutions, all tested essential oils showed a high antimycobacterial activity against targeted strains. Our data showed that M. barbata, E. globulus and J. excelsa essential oils totally inhibit the mycobacterial growth whatever the tested strains for the dilution 1/250, 1/100 and 1/250, respectively. Conclusion: To our knowledge, this is the first study regarding the antimycobacterial activity of essential oils in Lebanon. Our data show promising results, and encourage to investigate more on these medicinal plants, especially M. barbata. Keywords: Essential oils, Antimycobacterial activity, Micromeria barbata, Eucalyptus globulus, Juniperus excelsa

Several parasites including the protozoa Blastocystis sp. and Cryptosporidium spp. may be causative agents of gastrointestinal symptoms in domestic dogs, and there may be a potential risk of transmission to owners. While France is one of the largest European countries in terms of its canine population, little data is available about the molecular epidemiology of these two parasites. The purpose of this study was to determine the prevalence of intestinal parasites in household dogs in France, and to evaluate the zoonotic risk of Blastocystis sp. and Cryptosporidium spp. by genotyping the corresponding isolates. To this end, 116 faecal samples were collected from household dogs regardless of breed, age or gender, living in the Lyons area, France. Various intestinal protozoa and helminths were identified by light microscopy. Screening for Blastocystis sp. and Cryptosporidium spp. were subsequently performed by PCR targeting the small subunit (SSU) rDNA coding region, followed by direct sequencing of the PCR products and analysis of the sequences obtained for genotyping. The overall prevalence of dogs infected with at least one gastrointestinal parasite was 42.2% (49/116). After light microscopy examination of faecal samples, the most common parasites found were the protozoa Giardia sp. (25.0%) and Cystoisospora sp. (19.8%). Using molecular methods, four dogs (3.4%) were shown to be infected by Blastocystis sp. and carried either subtype (ST) 2, commonly identified in various animal groups, or ST10, frequently found in bovids. Three dogs (2.6%) were positive for C. canis, infecting humans episodically. The low prevalence of both parasites, combined with the identification of C. canis and Blastocystis sp. ST2 and ST10 in the canine population, strongly suggests that dogs play a negligible role as zoonotic reservoirs for both parasites and do not seem to be natural hosts of Blastocystis sp.

Despite increasing reports that Blastocystis infection is associated with digestive symptoms, its pathogenicity remains controversial. We report appendicular peritonitis in a 9-year-old girl returning to France from Morocco. Only Blastocystis parasites were detected in stools, appendix, peritoneal liquid, and recto-uterine pouch. Simultaneous gastroenteritis in 26 members of the child's family suggested an outbreak.

Acinetobacter baumannii is an opportunistic bacterium involved in several types of infection with high mortality and morbidity, especially in intensive care units. Treatment of these infections remains a challenge due to the worldwide emergence of broad-spectrum resistance to many antibiotics. Following the implementation of molecular techniques to study A. baumannii outbreaks, it has been shown that they are mainly caused by specific clones such as international clones I, II and III. The present work aims to review the available data on the mechanisms underlying antimicrobial resistance in A. baumannii, with a special focus on the molecular epidemiology of this species in Lebanon.

International audience; Cryptosporidium represents a major cause of gastrointestinal illness in humans and animals including domestic, wild, and in captivity animals, and more than 30 validated species of Cryptosporidium are recognized as infectious to different hosts such as mammals, birds, reptiles, amphibians, and fish. Therefore, numerous investigations have been conducted worldwide in order to shed light on the epidemiology of this parasite and to explore its potential reservoirs. Few surveys, targeting humans and animals have been carried out regarding the epidemiology of Cryptosporidium spp. in France and no data are available about the circulation of this parasite in French zoological gardens. Herein, we determined the prevalence of Cryptosporidium in animals housed in two French zoos. A total of 307 fecal samples belonging to 161 species were screened by nested PCR. Overall, Cryptosporidium DNA was detected in 1.9% of the 161 species and 1% of the total number of fecal samples tested. Additionally, three Cryptosporidium species were identified: C. galli, C. andersoni, and C. tyzzeri. To our knowledge, this is the first molecular study focused on Cryptosporidium infection in captivity animals in France. This study is of interest considering the exposure of a large number of humans and animals to this waterborne protozoan, found ubiquitously in the environment.

Brucella melitensis is the main causative agent of the zoonotic disease brucellosis. This study aimed at typing and characterizing genetic variation in 33 Brucella isolates recovered from patients in Lebanon. Bruce-ladder multiplex PCR and PCR-RFLP of omp31, omp2a and omp2b were performed. Sixteen representative isolates were chosen for draft-genome sequencing and analyzed to determine variations in virulence, resistance, genomic islands, prophages and insertion sequences. Comparative whole-genome single nucleotide polymorphism analysis was also performed. The isolates were confirmed to be B. melitensis. Genome analysis revealed multiple virulence determinants and efflux pumps. Genome comparisons and single nucleotide polymorphisms divided the isolates based on geographical distribution but revealed high levels of similarity between the strains. Sequence divergence in B. melitensis was mainly due to lateral gene transfer of mobile elements. This is the first report of an in-depth genomic characterization of B. melitensis in Lebanon.

The emergence and spread of carbapenem-resistant bacteria are a significant clinical and public health concern. The aim of the study is to determine the prevalence of intestinal carriage of carbapenem-resistant bacteria and yeasts in cancer patients under chemotherapy. 41 stool samples collected from cancer patients in Nini hospital in Tripoli, North Lebanon have been analyzed. After isolating yeasts and carbapenem-resistant bacteria, a biochemical identification and antimicrobial susceptibility profile were determined. The mechanism of enzymatic carbapenem-resistance was detected by searching for carbapenemases by both Hodge test and PCR assays. The association of several mechanisms of resistance was also searched. 46.3% (19/41) of patients were colonized by yeast. Candida glabrata (6/19) was the major species. The prevalence of carbapenem-resistant bacteria was 24.4% (10/41) including Escherichia coli (5/10), Enterobacter cloacae (1/10), Enterobacter aerogenes (1/10) Edwardsiella hoshinae (1/10) Pantoea agglomerans (1/10) and Pseudomonas stutzeri (1/10). PCR and sequencing of the amplified fragments revealed that Pseudomonas stutzeri (1/1) carried VIM gene and Enterobacter aerogenes (1/1) and E. coli (1/5) carried OXA-48 gene. The other Enterobacteriaceae were resistant to carbapenems by mechanisms other than a carbapenemase including hyperproduction of cephalosporinase (4/10), extended spectrum beta-lactamases (1/10) and both cephalosporinase and extended spectrum beta-lactamases (2/10). High prevalence of intestinal carriage of carbapenem-resistant bacteria and yeasts were detected in cancer patients under chemotherapy. In order to prevent the development of endogenous infection and the dissemination of antimicrobial resistance, an implementation of antibiotic stewardship programs and infection control measures is required in hospitals particularly in the department of chemotherapy. Keywords: Cancer, Yeasts, Carbapenemases, Multidrug resistant bacteria, Colonization

Background. Urinary tract infection (UTI) is common infection feature worldwide. Infected patients are usually treated empirically treated without culture or antibiotics susceptibility testing, and which may lead to increase antibiotic resistance level. This study aims to determine the prevalence and antibiotic susceptibility patterns of common uropathogenic bacteria isolated at Youssef Hospital Center, Akkar governorate, North of Lebanon. Methods. Spot midstream of urine samples from 9662 patients with UTI symptoms who came for medical investigation to Youssef Hospital Center located in Akkar governorate. Urine specimens were collected in sterile plastic bottles. Culture, identification and antibiotic susceptibility testing were performed using conventional tools according to the recommendations of the European Committee on Antimicrobial Susceptibility Testing. Results. A total of 1009 bacterial uropathogens were isolated. Escherichia coli was most presented (72.5%) of all isolates, followed by Klebsiella pneumoniae (8.2%), Enterococcus spp. (5.5%), Pseudomonas aeruginosa (4.5%), Proteus spp. (3%), Enterobacter spp. (2%), Staphylococcus aureus (2%), Streptococcus agalactiae (1.6%), Staphylococcus saprophyticus (0.4%), Acinetobacter baumannii(0.2%) and Providencia rettgeri (0.1%). Moreover, the mean antibiotic resistance rates of isolates was relatively high, but comparable to previously published data in Lebanon. Conclusion. To the best of our knowledge, this is the first investigation reporting epidemiological data regarding the prevalence and antibiotic susceptibility patterns of bacterial uropathogens isolated from patients in the Akkar governorate. Our data indicated the urgent need of a strategic plan to tackle antibiotic resistance, particularly in deprived regions with poor healthcare centers. Keywords: Uropathogens, Epidemiology, Antibiotic susceptibility, Risk factors, Akkargovernorate, North Lebanon.

Background. Urinary tract infection (UTI) is a severe public health problem. However, infected patients are usually treated empirically without preceding culture or antibiotics susceptibility testing, which may increase the antibiotic resistance level. The aim of this study is to determine the prevalence and antibiotic susceptibility patterns of common bacterial uropathogens isolated in Akkar governorate, North Lebanon. Methods. Spot midstream from urine samples from 9662 patients presenting UTI symptoms who came to Youssef Hospital Center located in Akkar governorate, were collected in sterile plastic cups. Culture, identification and antibiotic susceptibility testing were performed through conventional tools according to the manufacturer’s recommended procedures and the recommendations of the European Committee on Antimicrobial Susceptibility Testing. Results. Overall, a total of 1009 bacterial uropathogens were isolated. Escherichia coli was predominant and represented 72.5% of all isolates , followed by Klebsiella pneumoniae (8.2%), Enterococcus spp. (5.5%), Pseudomonas aeruginosa (4.5%), Proteus spp. (3%), Enterobacter spp. (2%), Staphylococcus aureus (2%), Streptococcus agalactiae (1.6%), Staphylococcus saprophyticus (0.4%), Acinetobacter baumannii (0.2%) and Providencia rettgeri (0.1%). Moreover, the mean antibiotic resistance rates of isolates was relatively high, but similar to previous investigations reported in our country. Conclusion. To our knowledge, this is the first investigation reporting epidemiological data regarding the prevalence and antibiotic susceptibility patterns of uropathogens isolated from patients in Akkar governorate. Our data indicated the urgent need of a strategic plan to tackle antibiotic resistance, particularly in deprived regions with poor healthcare structures such as Akkar governorate. DOI: http://dx.doi.org/10.3823/801

International audience; Blastocystis sp. is a common intestinal parasite infecting humans and a wide range of animals worldwide. It exhibits an extensive genetic diversity and 17 subtypes (STs) have thus far been identified in mammalian and avian hosts. Since several STs are common to humans and animals, it was proposed that a proportion of human infections may result from zoonotic transmission. However, the contribution of each animal source to human infection remains to be clarified. Therefore, the aim of this study was to expand our knowledge of the epidemiology and host specificity of this parasite by performing the largest epidemiological survey ever conducted in animal groups in terms of numbers of species screened. A total of 307 stool samples from 161 mammalian and non-mammalian species in two French zoos were screened by real-time PCR for the presence of Blastocystis sp. Overall, 32. 2% of the animal samples and 37. 9% of the species tested were shown to be infected with the parasite. A total of 111 animal Blastocystis sp. isolates were subtyped, and 11 of the 17 mammalian and avian STs as well as additional STs previously identified in reptiles and insects were found with a varying prevalence according to animal groups. These data were combined with those obtained from previous surveys to evaluate the potential risk of zoonotic transmission of Blastocystis sp. through the comparison of ST distribution between human and animal hosts. This suggests that non-human primates, artiodactyls and birds may serve as reservoirs for human infection, especially in animal handlers. In contrast, other mammals such as carnivores, and non-mammalian groups including reptiles and insects, do not seem to represent significant sources of Blastocystis sp. infection in humans. In further studies, more intensive sampling and screening of potential new animal hosts will reinforce these statements and expand our understanding of the circulation of Blastocystis sp. in animal and human populations.

Background. – Enteric infections are a leading cause of diarrheal disease and death in infants and young children, particularly in the developing regions. The aim of this study is to determine the antibiotic susceptibility of three Enterobacteriaceae enteropathogens (Salmonella spp., Shigella spp. and enteropathogenic Escherichia coli (EPEC)) isolated from hospitalized diarrheic children in North Lebanon during the period of 7 years.Methods and Findings. – 172 Enterobacteriaceae enteropathogens strains were isolated from fecal or blood specimens of 158 hospitalized diarrheic children collected at Nini hospital in Tripoli, North Lebanon. Culture, identification and antibiotic susceptibility of strains were carried out through conventional tools according to standard protocols proposed by the REMIC group and the recommendations of the European Committee on Antimicrobial Susceptibility Testing. Out of 172, 158 (91.9%) Enterobacteriaceae enteropathogens strains were isolated from stool samples and only 14 (8.1%) were isolated from blood cultures. EPEC were predominant (78/172; 45.3%), followed by Salmonella spp. (64/172; 37.2%) and Shigella spp. (30/172; 17.5%). According to API identification and immunological serotyping reports of Salmonella isolates, Salmonella Typhi was the predominant serotype (32/64; 50%). During the study period, antibiotics susceptibility testing showed that around 50% and 25% of the isolates were resistant to ampicillin and extended-spectrum cephalosporins, respectively. Overall, Salmonella Typhi strains were the most susceptible to antibiotics.Conclusion. – Our study revealed a relatively high antibiotic resistance rate of EPEC, Shigella spp. and non-Typhi Salmonella strains among studied population. The findings of this study provide useful information to survey, prevent and control diarrheal disease and antimicrobial resistance in Lebanon.

Introduction: Bacteria of Aeromonas genus are ubiquitous organisms in aquatic environments. This work aims to explore the presence of four clinically relevant species in Lebanese waters and to study their susceptibility to antibiotics. Methods: One hundred water samples are collected in northern Lebanon from different sources (river, springs, wells, ponds, sea, chlorinated water and sewage) and analyzed by culture methods. The isolates were identified at genus level by a PCR assay targeting gcat gene. The presence of four species (A. hydrophila, A. caviae, A. media and A. veronii) was investigated by a multiplex PCR. The antibiotic susceptibility was also studied. Results: Aeromonas spp. was detected in 28% of investigated samples. In total, 38 isolates were identified as Aeromonas spp (10 out of 7 river water samples, 9 out of 35 spring, 9 among 26 wells, 5 out of 4 sewage samples, 2 out of 5 ponds, 2 out of 8 seawater samples and 1 out of 15 chlorinated water samples). Aeromonas hydrophila was the predominant species (19 isolates; 50%). The 3 other species were found to a lesser extent: A. caviae (3 isolates; 7.9%), A. veronii (2 isolates; 5.3%) and A. media (2 isolates; 5.3%). 12 other isolates (31.5%) remained unidentified by the used multiplex PCR technique. All isolates were resistant to more than one antibiotic. The most common resistance concerned beta-lactams. Conclusion: This study has highlighted the interesting distribution of these species in aquatic biotopes in Lebanon and the threat of potential transmission of these resistant strains to humans.

Cryptosporidium spp. are enteroparasites with worldwide distribution that infect the gastrointestinal tract of several vertebrates including humans. Human to human, zoonotic, foodborne and waterborne are reported as the main transmission routes of this parasite. Cryptosporidium spp. have been recognized as the predominant cause of waterborne and foodborne outbreaks. However, the epidemiological situation of cryptosporidiosis is not well known in Lebanon, a developing country with a population often affected by intestinal parasitic infections. This study was devoted to determine the prevalence and the genetic diversity of Cryptosporidium spp. in symptomatic hospitalized patients and in two children populations with different socio-economic level in North-Lebanon, as well as the risk factors associated with cryptosporidiosis. Fecal samples obtained from these populations were examined microscopically by modified Ziehl-Neelsen staining as well as nested PCR were done for the detection of Cryptosporidium oocysts. Out of 163 symptomatic hospitalized patients and 249 children, Cryptosporidium was present in 11% and 10.4% respectively according to microscopy examination and/or molecular tests. The genotyping showed the predominance of Cryptosporidium hominis in both populations. Subgenotype analysis of the isolates at the gp60 locus identified three subtypes IdA19, IbA10G2 and IaA18R3 for C. hominis and two subtypes IIaA15G1R1 and IIaA15G2R1 for C. parvum. Moreover, cryptosporidiosis was correlated with having meals outside home and presence of gastrointestinal symptoms especially diarrhea (p

Background. – Staphylococcus aureus is one of the most significant pathogens causing significant morbidity and mortality. Moreover, the incidence of MLS-resistant Staphylococcus aureus infections continues to grow globally.Objective. – The aim of this study is to examine the expression of resistance of Staphylococcus aureus (S. aureus) isolates to MLS and the prevalence of genes involved in this resistance by PCR.Methods. – 38 strains of S. aureus MLS-resistant (resistant at least for one macrolide) were isolated in the sector of microbiology at Nini Hospital in North Lebanon. The disk diffusion method was used to determine the phenotype of the MLS resistance. The resistance genes involved were detected by PCR using specific gene primers for ermA, ermB, ermC, msrA, linA, mefA, vat and vgb genes.Results. – A total of 55.3% of the isolates were positive for inducible phenotype (iMLSB), 15.8% for the constitutive phenotype (cMLSB), 23.7% for MSB phenotype and 5.2% for L phenotype. The ermC gene was the most prevalent (52.6%), while ermA, ermB, msrA and linA genes were observed with lower prevalence. However, a combination of several of these genes was detected. The vgb, vat and mefA genes were not detected in any of the clinical isolates.Conclusion. – To our knowledge, this study is the first investigation regarding characterization of MLS resistance genes in clinical isolates of S. aureus in Lebanon. The study revealed a high prevalence of the inducible resistance to lincosamides (iMLSB phenotype) and the most prevalent resistance determinants was ermC.

International audience; BackgroundThe association between Cryptosporidium and human colon cancer has been reported in different populations. However, this association has not been well studied. In order to add new strong arguments for a probable link between cryptosporidiosis and colon human cancer, the aim of this study was to determine prevalence and to identify species of Cryptosporidium among Lebanese patients.Methodology and principal findingsOverall, 218 digestive biopsies were collected in Tripoli, Lebanon, from three groups of patients: (i) patients with recently diagnosed colon intraepithelial neoplasia/adenocarcinoma before any treatment (n = 72); (ii) patients with recently diagnosed stomach intraepithelial neoplasia/adenocarcinoma before any treatment (n = 21); and (iii) patients without digestive intraepithelial neoplasia/adenocarcinoma but with persistent digestive symptoms (n = 125). DNA extraction was performed from paraffin-embedded tissue. The presence of the parasite in tissues was confirmed by PCR, microscopic observation and immunofluorescence analysis. We identified a high rate (21%) of Cryptosporidium presence in biopsies from Lebanese patients with recently diagnosed colonic neoplasia/adenocarcinoma before any treatment. This prevalence was significantly higher compared to 7% of Cryptosporidium prevalence among patients without colon neoplasia but with persistent gastrointestinal symptoms (OR: 4, CI: 1.65–9.6, P = 0.001). When the comparison was done against normal biopsies, the risk of infection increased 11-fold in the group of patients with colon adenocarcinoma (OR: 11.315, CI: 1.44–89.02, P = 0.003).ConclusionsThis is the first study performed in Lebanon reporting the prevalence of Cryptosporidium among patients with digestive cancer. These results show that Cryptosporidium is strongly associated with human colon cancer being maybe a potential etiological agent of this disease.

Cryptosporidium spp. represent a major public health problem worldwide and infect the gastrointestinal tract of both immunocompetent and immunocompromised persons. The prevalence of these parasites varies by geographic region, and no data are currently available in Lebanon. To promote an understanding of the epidemiology of cryptosporidiosisin this country, the main aim of this study was to determine the prevalence Cryptosporidium in symptomatic hospitalized patients, and to analyze the genetic diversity of the corresponding isolates. Fecal specimens were collected in four hospitals in North Lebanon from 163 patients (77 males and 86 females, ranging in age from 1 to 88 years, with a mean age of 22 years) presenting gastrointestinal disorders during the period July to December 2013. The overall prevalence of Cryptosporidium spp. infection obtained by modified Ziehl-Neelsen staining and/or nested PCR was 11%, and children

Cryptosporidium species are worldwide spread apicomplexan protozoan. These parasites constitute a significant risk to humans and animals. They cause self-limited diarrhea in immunocompetent hosts and a life threatening disease in immunocompromised hosts. Interestingly, Cryptosporidium parvum has been related to digestive carcinogenesis in humans. Consistently with a potential tumorigenic role of this parasite, in an original reproducible animal model of chronic cryptosporidiosis based on dexamethasone-treated or untreated adult SCID mice, we formerly reported that C. parvum (strains of animal and human origin) is able to induce digestive adenocarcinoma even in infections induced with very low inoculum. The aim of this study was to further characterize this animal model and to explore metabolic pathways potentially involved in the development of C. parvum-induced ileo-caecal oncogenesis. We searched for alterations in genes or proteins commonly involved in cell cycle, differentiation or cell migration, such as β-catenin, Apc, E-cadherin, Kras and p53. After infection of animals with C. parvum we demonstrated immunohistochemical abnormal localization of Wnt signaling pathway components and p53. Mutations in the selected loci of studied genes were not found after high-throughput sequencing. Furthermore, alterations in the ultrastructure of adherens junctions of the ileo-caecal neoplastic epithelia of C. parvum infected mice were recorded using transmission electron microscopy. In conclusion, we found for the first time that the Wnt signaling pathway, and particularly the cytoskeleton network seems to be pivotal for the development of C. parvum-induced neoplastic process and cell migration of transformed cells. Furthermore, this model is a valuable tool to contribute to the comprehension of the host-pathogen interactions associated to the intricate infection process due to this parasite, which is able to modulate host cytoskeleton activities and several host-cell biological processes and that remains a significant cause of infection worldwide.

International audience; The intestinal protistan parasite Blastocystis is characterized by an extensive genetic variability with 17 subtypes (ST1–ST17) described to date. Only the whole genome of a human ST7 isolate was previously sequenced. Here we report the draft genome sequence of Blastocystis ST4-WR1 isolated from a laboratory rodent at Singapore.

International audience; Blastocystis is the most common eukaryotic parasite in the intestinal tract of humans. Because of its potential impact in public health, we acquired the first data concerning the prevalence of this parasite and the frequency of the Blastocystis subtypes (STs) in the Lebanese population. In this study, fecal samples from 220 Lebanese symptomatic and asymptomatic patients were collected and a total of 42 patients (19%) were identified as positive for this parasite by direct-light microscopy of smears. Among these, 36 Blastocystis isolates were genotyped using partial small subunit ribosomal RNA gene sequencing. The ST distribution in the present Lebanese population was as follows: ST3 (33.3%), ST2 (33.3%), ST1 (30.6%), and ST4 (2.8%). These data were compared with those available in other Middle Eastern and neighboring countries. Finally, ST1 was significantly more prevalent among symptomatic patients of this Lebanese population.