Search

Objective: To study the prevalence and incidence of extended-spectrum beta-lactamase -producing microorganisms, microbiological characteristics and antimicrobial-susceptibility patterns. Methods: A prospective study involving all cases of extended-spectrum beta-lactamase -producing microorganisms among all bacteriological samples collected over a 1-year period from 11 November 2015 to 10 November 2016, in the Clinical and Molecular Microbiology Laboratory at King Abdulaziz University Hospital, Jeddah. Detection of extended-spectrum beta-lactamase-producing microorganisms and antimicrobial-susceptibility profiles were done using automated Vitek 2 system. Clinical data such as recent use of antibiotics or invasive devices were investigated as risk factors for multidrug resistance. Results: The prevalence of extended-spectrum beta-lactamase-producing microorganisms was 5.4% (95% CI = 4.7% – 6.2%); for an incidence = 54 per 1,000 isolates-years. Distribution by species showed 70.0% Escherichia coli, 28.5% Klebsiella pneumoniae and 1.5% Proteus mirabilis. Vitek 2 system showed 6% of false positive ESBL detections by reference to confirmatory E-test. Antimicrobial- susceptibility tests showed that 86.5% of beta-lactamase-producing strains were resistant to ≥ 1 other antimicrobial class and 20% were multidrug resistant. Univariate logistic regression showed that the presence of multidrug resistance was significantly predicted by age (OR = 1.02; P = 0.026), use of urinary catheter (OR = 2.05; P = 0.046) and number of devices used (OR = 1.60; P = 0.046); only age (OR = 1.02; P = 0.022) was significant in the multivariate model. Conclusion: Clinicians and microbiologists should maintain a high level of alertness and contribute for effective screening and adequate treatment of infections caused by beta-lactamase-producing organisms according to international guidelines and the local epidemiological picture.

Acinetobacter baumannii has emerged as a problematic multidrug-resistant (MDR) pathogen worldwide. The aim of this study was to characterize clinical isolates of A. baumannii from Jeddah, a coastal city in the western region of Saudi Arabia that has an ∼50% expatriate population. In this study, 135 nonrepetitive clinical isolates of A. baumannii were identified through MALDI-TOF and susceptibility was determined with VITEK-2 systems and broth microdilution method. Genotypic characterization of the isolates was performed by using M13 primer typing and polymerase chain reaction screening was performed for carbapenemase genes, insertion sequences, metallo-β-lactamases, and cephalosporinase genes. The isolates were recovered from heterogeneous clinical specimens, and the majority of the cases of A. baumannii infection were acquired in the hospital and predominantly involved patients who were older than 50 years. Total, 58.5% of the isolates were MDR, and 55.6% isolates were resistant to carbapenem antibiotics. Approximately half of the isolates were resistant to ceftazidime, and cefepime among the β-lactam antibiotics and ciprofloxacin from the quinolone group. The blaOXA-23-like gene and ISAba1 upstream of blaOXA-23-like were detected in 92% of the carbapenem-resistant isolates, while all carbapenem-resistant isolates were found to carry blaOXA-51-like, and blaADC-type cephalosporinase gene. The blaIMP gene was detected in 84% isolates, and two isolates carried the blaNDM-1 gene. Data demonstrate the coexistence of multiple carbapenem resistance determinants in A. baumannii from the western region of Saudi Arabia.

Objectives: A rapid molecular typing system was used to determine the impact of mass migration on the clonal variation of Staphylococcus aureus isolates recovered from King Abdulaziz University Hospital (KAUH) Jeddah, in the western region of Saudi Arabia. This region experiences an annual influx of millions of pilgrims. Methods: SmaI-multiplex PCR typing (SMT) was used for the initial analysis of strains and the resulting data subsequently supported by Multi-Locus Sequence Typing (MLST). Results: A total of 89 S. aureus isolates were SMT typed and revealed a high degree of genetic variation, with 40 SMT profiles detected among the isolates. Representatives of all forty SMT types were subsequently analysed by MLST, identifying 26 sequence types. A novel sequence type (ST), named ST3303, was identified in two methicillin-sensitive S. aureus (MSSA) isolates. MSSA strains exhibited more diversity than methicillin-resistant S. aureus (MRSA) strains, with community acquired MSSA and MRSA strains reaching alarmingly high levels. Conclusion: The relatively high degree of genetic diversity found among S. aureus isolates of single hospital was attributed to the fact that Jeddah is the principal gateway to Mecca, visited each year by millions of pilgrims from many countries. The observed diversity clearly reflects the impact of such mass migrations in the rapid dissemination of strains world-wide. Our findings suggest the importance of surveillance programmes in locations affected by mass migrations, both to monitor their impact on endemic strains and for the detection of pandemic strains. SMT provides a cost-effective and sensitive typing method for achieving this objective. Keywords: SmaI-multiplex-PCR, Molecular typing, Epidemiology, Genotyping, S. aureus

Background: Tuberculosis (TB) is one of the deadliest health problems. Accurate rapid diagnosis and prompt treatment are crucial for cure. The aim of this study was to investigate the role of GeneXpert PCR in the rapid detection of MTBC directly from clinical samples and to determine the anti-TB drug resistance patterns in pulmonary tuberculosis. Methods: This is a cross-section analysis, involving a total of 92 patients with pulmonary TB (primary TB resistance); 37 Saudis and 55 non-Saudis. Respiratory specimens were processed, and examined by flurochrome stain. Culture and susceptibility testing was performed by VersaTREK , a liquid culture system. Adding, Mycobacterium tuberculosis complex (MTBC) was directly detected from the sample by GeneXpert PCR. Results: GeneXpert assay achieved 96 % sensitivity and 100 % specificity for the detection of MTBC in sputum. Resistance to pyrazinamide was (6.5 %), followed by rifampicin (2.2 %), streptomycin and isoniazid (1.1 %). Mono-resistance to pyrazinamide and streptomycin (5.4%) and (1.1%) respectively. MDR-TB was (1.1%) and there was no substantial difference in anti-TB resistance between Saudi and non-Saudi TB patients. Conclusions: The GeneXpert PCR is a very helpful tool for the detection of MTBC in respiratory specimens with a high sensitivity, specificity and accuracy within a shorter time as compared to conventional methods.Recommendations: Costs-benefit analysis of GeneXpert versus other recently deployed TB diagnostic systems may be required on a wider scale.

The goal of this study was to genotypically characterize extended-spectrum β-lactamase-producing Escherichia coli isolates from the western region of Saudi Arabia and to identify active antibiotics against these isolates using phenotypic and molecular modeling. In total, 211 ESBL-producing E. coli isolates recovered from heterogeneous clinical specimens were identified by MALDI-TOF. Thirty-two sequence types (STs) were identified from a multilocus sequence typing (MLST) analysis of ESBL-producing E. coli, including a novel ST (ST8162). The most common ST in the Saudi and expatriate population was ST131, followed by ST38. All the isolates were multidrug resistant (MDR), and >95% of the isolates were resistant to third-generation (ceftriaxone and ceftazidime) and fourth-generation (cefepime) cephalosporins. The ESBL-positive E. coli isolates primarily harbored the blaCTX-M and blaTEM genes. No resistance was observed against the carbapenem antibiotic group. All the ESBL-producing E. coli isolates were observed to be susceptible to a ceftazidime/avibactam combination. Molecular interaction analyses of the docked complexes revealed the amino acid residues crucial for the binding of antibiotics and inhibitors to the modeled CTX-M-15 enzyme. Importantly, avibactam displayed the most robust interaction with CTX-M-15 among the tested inhibitors in the docked state (∆G = −6.6 kcal/mol). The binding free energy values for clavulanate, tazobactam and sulbactam were determined to be −5.7, −5.9 and −5.2 kcal/mol, respectively. Overall, the study concludes that ‘ceftazidime along with avibactam’ should be carefully used as a treatment option against only carbapenem-resistant MDR ESBL-producing E. coli in this region.

We report here the main characteristics of 'Beduinibacterium massiliense' strain Marseille-P3337T gen. nov., sp. nov., 'Massilimaliae massiliensis' Marseille-P2963T gen. nov., sp. nov., 'Provencibacterium massiliense' Marseille-P2780T gen. nov., sp. nov. and 'Oscilibacter massiliensis' Marseille-P2778T sp. nov., all isolated from the stool of a Bedouin from Saudi Arabia. We used a bacterial culturomics approach combined with taxonogenomics.

International audience; We report here the main characteristics of 'Arabia massiliensis' strain Marseille-P3078(T) gen. nov., sp. nov., 'Gordonibacter massiliensis' Marseille-P2775(T) sp. nov. and 'Bacilliculturomica massiliensis' strain Marseille-P3303 gen. nov., sp. nov. The culturomics approach combined with taxonogenomics was used to characterize these strains, which were all isolated from a faecal specimen of a 50-year-old Saudi Bedouin woman.

Background Honey has been increasingly recognized as a potential therapeutic agent for treatment of wound infections. There is an urgent need for assessment and evaluation of the antibacterial properties against wound pathogens of honeys that have not yet been tested. Methods Ten Saudi honeys collected from different geographical locations were screened initially for their antibacterial potential against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA) by the agar well diffusion method. Manuka honey (UMF-12) was used for comparison. Of the tested honeys, the honey that exhibited the greatest antibacterial activity in the agar well diffusion assay was further evaluated for its minimum inhibitory concentration (MIC) against ten MRSA clinical isolates and three American Type Culture Collection (ATCC) reference strains by the microbroth dilution method. Results Locally produced honeys exhibited variable antibacterial activity against the tested isolates in the agar well diffusion assay. They were unable to exhibit antibacterial activity against MSSA and MRSA at 25% dilutions (w/v) in catalase solution. However, Sumra and Talha honeys showed a zone of inhibition at 50% dilutions (w/v) in catalase solution. This finding means that both honeys possess weak non-peroxide-based antibacterial activity. Moreover, Sumra honey showed a larger inhibition zone at 50 and 25% dilutions (w/v) in distilled water than Manuka honey against both MSSA and MRSA. This result demonstrates that Sumra honey has more hydrogen peroxide-related antibacterial activity or total antibacterial activity than Manuka honey. In addition, MIC results obtained through a microbroth dilution assay showed that Sumra honey inhibited the growth of all MRSA clinical isolates (n = 10) and reference strains [MRSA (ATCC 43300) and MSSA (ATCC 29213)] at lower concentrations (12.0% v/v) than those required for Manuka honey-mediated inhibition (14.0% v/v). This result means that Sumra honey has more peroxide or synergistic antibacterial activity than Manuka honey. An equivalent MIC (15.0% v/v) was observed for E. coli (ATCC 25922) between Manuka honey and Sumra honey. Conclusions Sumra honey may be used as an alternative therapeutic agent for infected wounds and burns, where additional hydrogen peroxide-related antibacterial activity is needed. In the future, the physiochemical characteristics of Sumra honey may be evaluated and standardized.

Objectives Acinetobacter baumannii has emerged as a prevalent multidrug-resistant nosocomial pathogen worldwide. Here we report the draft genome sequence of A. baumannii strain Ab174 isolated from a neonatal patient diagnosed with acute peritonitis. Methods The draft genome sequence of A. baumannii Ab174 was determined using a MiSeq platform (Illumina Inc., San Diego, CA) using v.3, 2 × 30-bp chemistry. Genomic assembly was performed using SPAdes 3.9 algorithm. Results The draft genome of A. baumannii Ab174 is 3 747 065 bp in length and was classified as a new sequence type (ST1688). The genome of A. baumannii Ab174 has a G + C content of 39% and harbours two plasmids. The antimicrobial resistance gene blaADC-25 and the virulence factor gene for penicillin-binding protein G (pbpG) as well as 17 genomic islands and 14 insertion sequences were identified in the genome of A. baumannii Ab174. Conclusion The genome sequence of A. baumannii strain Ab174 can be used as a reference sequence for the new ST1688. These data will facilitate further understanding of genomic variation in isolates from different geographical regions.

International audience; Background/objectives High salt intake has been linked to several diseases including obesity and an increased risk of death; however, fecal salinity and the ability of salt to alter the gut microbiota, which was recently identified as an instrumental factor for health and disease, remains poorly explored. Methods/subjects We analyzed the fecal samples of 1326 human individuals for salinity by refractometry, 572 for gut microbiota by culturomics, and 164 by 16S rRNA-targeted metagenomics. Geographical origin, age, gender, and obesity were tested as predictors of fecal salinity and halophilic diversity. All halophilic isolates were characterized by tax-onogenomics and their genome sequenced. Results Fecal salinity was associated with obesity independently of geographical origin, gender, and age. The first 2 human-associated halophilic archaeal members were isolated along with 64 distinct halophilic species, including 21 new species and 41 known in the environment but not in humans. No halophiles grow in less than 1.5% salinity. Above this threshold, the richness of the halophilic microbiota was correlated with fecal salinity (r = 0.58, p < 0.0001). 16S metagenomics linked high fecal salinity to decreased diversity (linear regression, p < .035) and a depletion in anti-obesity Akkermansia muciniphila and Bifidobacterium, specifically B. longum and B. adolescentis. Genomics analysis suggested that halophilic microbes are not only transient passengers but may be residents of the human gut. Conclusions High salt levels are associated with alteration of the gut microbial ecosystem and halophilic microbiota, as discovered during this study. Further studies should clarify if the gut microbiota alterations associated with high salt levels and the human halophilic microbiota could be causally related to human disease, such as obesity.

There is an evident difference in the intensity of morbidity caused by Schistosoma haematobium in North-African zones compared to Sub-Saharan ones. Clinical outcome dichotomy corresponds to two geographically distinct intermediate host snail species that are only infected by the related strain of the parasite. In concert, there is a manifest hybridization of the parasite with other Schistosoma species confined to certain regions of Africa. This raises a reasonable suggestion that S. haematobium has no less than two phylogenetic clusters that have different virulence. The aim of the study was to examine the possible diversity among S. haematobium using simultaneous amplification of genomic DNA of selected isolates. Random amplified polymorphic DNA-polymerase chain reaction markers were used to study the genetic diversity among S. haematobium natural isolates from selected regions of Africa (Egypt, Zimbabwe, and South Africa) that represent different ecological conditions, different species of intermediate host, and different possibilities of field hybridization with other schistosomes. A moderate to high level of genetic diversity was evident among the three isolates. More bands were shared by the isolates from Zimbabwe and South Africa (similarity index = 0.721) than those shared by each with the Egyptian isolate (similarity index = 0.551 and 0.566, respectively), suggesting that at least two phylogenetic groups of S. haematobium do exist in distinct geographic regions of Africa. The elucidation of the possible genetic diversity among S. haematobium parasites may explain many ambiguous aspects of the biology of the parasite-like virulence, immune evasion and drug resistance.

Microbial culturomics is a new field of omics sciences that examines the bacterial diversity of human gut coupled with a taxono-genomic strategy. Using microbial culturomics, we report here for the first time a novel Gram negative, catalase- and oxidase-negative, strict anaerobic bacilli named Beduini massiliensis gen. nov., sp nov. strain GM1 (= CSUR P1440 = DSM 100188), isolated from the stools of a female nomadic Bedouin from Saudi Arabia. With a length of 2,850,586 bp, the Beduini massiliensis genome exhibits a G + C content of 35.9%, and contains 2819 genes (2744 protein-coding and 75 RNA genes including 57 tRNA and 18 rRNA genes). It is composed of 6 scaffolds (composed of 6 contigs). A total of 1859 genes (67.75%) were assigned a putative function (by COGs or by NR blast). At least 1457 (53%) orthologous proteins were not shared with the closest phylogenetic species. 274 genes (10.0%) were identified as ORFans. These results show that microbial culturomics can dramatically improve the characterization of the human microbiota repertoire, deciphering new bacterial species and new genes. Further studies will clarify the geographic specificity and the putative role of these new microbes and their related functional genetic content in health and disease. Microbial culturomics is an emerging frontier of omics systems sciences and integrative biology and thus, warrants further consideration as part of the postgenomics methodology toolbox.

International audience; We report here the main characteristics of 'Blautia phocaeensis' strain Marseille-P3441 sp. nov. and 'Lachnoclostridium edouardi' strain Marseille-P3397 sp. nov., that were isolated from a faecal specimen of a 42-year-old female Saudi Bedouin. We used a bacterial culturomics approach combined with taxono-genomics.

We report here the main characteristics of a new bacterium named ‘Beduinella massiliensis’ strain Marseille-P2846T (CSURP2846P) that was isolated from a faecal specimen of a 50-year-old Saudi Bedouin female and propose the creation of a new family ‘Beduinellaceae’.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Tuberculosis (TB) represents a significant challenge to public health authorities, especially with the emergence of drug-resistant (DR) and multidrug-resistant (MDR) isolates of Mycobacterium tuberculosis. We sought to examine the genomic variations among recently isolated strains of M. tuberculosis in two closely related countries with different population demography in the Middle East. Clinical isolates of M. tuberculosis from both Egypt and Saudi Arabia were subjected to phenotypic and genotypic analysis on gene and genome-wide levels. Isolates with MDR phenotypes were highly prevalent in Egypt (up to 35%) despite its relatively stable population structure (sympatric pattern). MDR-TB isolates were not identified in the isolates from Saudi Arabia despite its active guest worker program (allopatric pattern). However, tuberculosis isolates from Saudi Arabia, where lineage 4 was more prevalent (>65%), showed more diversity than isolates from Egypt, where lineage 3 was the most prevalent (>75%). Phylogenetic and molecular dating analyses indicated that lineages from Egypt were recently diverged (~78 years), whereas those from Saudi Arabia were diverged by over 200 years. Interestingly, DR isolates did not appear to cluster together or spread more widely than drug-sensitive isolates, suggesting poor treatment as the main cause for emergence of drug resistance rather than more virulence or more capacity to persist.

Background The present study was conducted to discover genetic imbalances such as DNA copy number variations (CNVs) associated with gastric cancer (GC) and to examine their association with different genes involved in the process of gastric carcinogenesis in Saudi population. Methods Formalin-fixed paraffin-embedded (FFPE) tissues samples from 33 gastric cancer patients and 15 normal gastric samples were collected. Early and late stages GC samples were genotyped and CNVs were assessed by using Illumina HumanOmni1-Quad v.1.0 BeadChip. Results Copy number gains were more frequent than losses throughout all GC samples compared to normal tissue samples. The mean number of the altered chromosome per case was 64 for gains and 40 for losses, and the median aberration length was 679115bp for gains and 375889bp for losses. We identified 7 high copy gain, 52 gains, 14 losses, 32 homozygous losses, and 10 copy neutral LOHs (loss of heterozygosities). Copy number gains were frequently detected at 1p36.32, 1q12, 1q22, 2p11.1, 4q23-q25, 5p12-p11, 6p21.33, 9q12-q21.11, 12q11-q12, 14q32.33, 16p13.3, 17p13.1, 17q25.3, 19q13.32, and losses at 1p36.23, 1p36.32, 1p32.1, 1q44, 3q25.2, 6p22.1, 6p21.33, 8p11.22, 10q22.1, 12p11.22, 14q32.12 and 16q24.2. We also identified 2 monosomy at chromosome 14 and 22, 52 partially trisomy and 22 whole chromosome 4 neutral loss of heterozygosities at 13q14.2-q21.33, 5p15.2-p15.1, 5q11.2-q13.2, 5q33.1-q34 and 3p14.2-q13.12. Furthermore, 11 gains and 2 losses at 1p36.32 were detected for 11 different GC samples and this region has not been reported before in other populations. Statistical analysis confirms significant association of H. pylori infection with T4 stage of GC as compare to control and other stages. Conclusions We found that high frequency of copy number gains and losses at 1p36.23, 1p32.1, 1p36.32, 3q25.2, 6p21.33 and 16q24.2 may be common events in gastric cancer. While novel CNVs at 1p36.32 harbouring PRDM16, TP73 and TP73-AS1 genes showed 11 gains and 2 losses for 11 different GC cases and this region is not reported yet in Database of Genomic Variants may be specific to Saudi population.

A Streptomyces strain MS-6-6 with promising anti-tuberculous activity was isolated from soil samples in Saudi Arabia. The nucleotide sequence of its 16S rRNA gene (1426 bp) evidenced a 100% similarity to Streptomyces mutabilis. Through an anti-tuberculous activity-guided approach, a polyketide macrolide was isolated and identified as treponemycin (TP). The structure of the isolated compound was determined by comprehensive analyses of its 1D and 2D NMR as well as HRESI-MS. In addition to the promising anti-tuberculous activity (MIC = 13.3 µg/mL), TP showed broad spectrum of activity against the Gram positive, Gram negative strains, and Candida albicans. Improvement of TP productivity (150%) was achieved through modification in liquid starch nitrate medium by replacing KNO3 with corn steep liquor and yeast extract or tryptone, and removing CaCO3 and K2HPO4. The follow up of TP percentage as well as its metabolites profile for each media was assessed by LC/DAD/MS.

This work is dedicated to the memory of Marcel Sy (PhD student died on 15 September 2016); Marcel Sy has made a substantial contribution to this work, especially in fungal culturomics.; International audience; Herein, the mycobiota was characterized in fecal samples from sick patients and healthy subjects, collected from different geographical locations and using both culturomics and amplicon-based metagenomics approaches. Using the culturomics approach, a total of 17,800 fungal colonies were isolated from 14 fecal samples, and resulted in the isolation of 41 fungal species, of which 10 species had not been previously reported in the human gut. Deep sequencing of fungal-directed ITS1 and ITS2 amplicons led to the detection of a total of 142 OTUs and 173 OTUs from the ITS1 and ITS2 regions, respectively. Ascomycota composed the largest fraction of the total OTUs analyzed (78.9% and 68.2% of the OTUs from the ITS1 and ITS2 regions, respectively), followed by Basidiomycota (16.9% and 30.1% of the OTUs from the ITS1 and ITS2 regions, respectively). Interestingly, the results demonstrate that the ITS1/ITS2 amplicon sequencing provides different information about gut fungal communities compared to culturomics, though both approaches complete each other in assessing fungal diversity in fecal samples. We also report higher fungal diversity and abundance in patients compared to healthy subjects. In conclusion, combining both culturomic and amplicon-based metagenomic approaches may be a novel strategy towards analyzing fungal compositions in the human gut.

Background and Objectives: Helicobacter pylori (H. pylori) infection is cause of several gastrointestinal diseases in humans. Virulence genes of H. pylori are associated with severity of disease and vary geographically. The aim of present study was to detect H. pylori in formalin-fixed paraffin-embedded (FFPE) tissues and further investigate prevalence of babA2, cagA, iceA1, iceA2, vacA s1/s2 and vacA m1/m2 genotypes in H. pylori from gastric cancer (GC) and gastric ulcer (GU) patients’ biopsy samples. Methods: We used FFPE tissues of 35 GC and 10 GU patients’ biopsy samples. Using Polymerase Chain Reaction (PCR), detection of H. pylori strain was performed by using specific primers targeting 16S rRNA and ureC encodes for phosphoglucosamine mutase genes. We have identified different virulence genes of H. pylori by PCR. Results: Of all the 45 samples tested, 20 GC and all 10 GU samples were positive for identification of H. pylori using specific genes (16S rRNA and ureC). The prevalence of babA2(100%) was significantly higher in GC as compared to GU (40%) samples. The rate of virulence genes vacAs1 was higher in both GU 8 (80%) and GC (100%). Conclusions: Our study finds that vacAs1am1 and babA2 are most prominent genotypes and may play role in development of Gastric cancer.

Background and objectives Helicobacter pylori (H. pylori) infection is cause of several gastrointestinal diseases in humans. Virulence genes of H. pylori are associated with severity of disease and vary geographically. The aim of present study was to detect H. pylori in formalin-fixed paraffin-embedded (FFPE) tissues and further investigate prevalence of babA2, cagA, iceA1, iceA2, vacA s1/s2 and vacA m1/m2 genotypes in H. pylori from gastric cancer (GC) and gastric ulcer (GU) patients' biopsy samples. Methods We used FFPE tissues of 35 GC and 10 GU patients' biopsy samples. Using Polymerase Chain Reaction (PCR), detection of H. pylori strain was performed by using specific primers targeting 16S rRNA and ureC encodes for phosphoglucosamine mutase genes. We have identified different virulence genes of H. pylori by PCR. Results Of all the 45 samples tested, 20 GC and all 10 GU samples were positive for identification of H. pylori using specific genes (16S rRNA and ureC). The prevalence of babA2 (100%) was significantly higher in GC as compared to GU (40%) samples. The rate of virulence genes vacAs1 was higher in both GU 8 (80%) and GC (100%). Conclusions Our study finds that vacAs1am1 and babA2 are most prominent genotypes and may play role in development of Gastric cancer.

Background Progression to a chronic state after an initial, mostly asymptomatic, acute phase is the normal fate of Toxoplasma infection in immunocompetent hosts. The inter-conversion between tachyzoite and bradyzoite life stages is a key to Toxoplasma survival and pathogenesis. Chronic toxoplasmosis is resistant to all commercially available medications. Reactivation of chronic stage has serious complications on immunocompromised hosts including encephalitis and death. Cyclic AMP, TNF-α and Th1 immune response have pivotal roles in establishing chronic disease. Rolipram, a PDE4 inhibitor, has regulatory roles on these factors. This is the first study to investigate a possible modulatory effect of rolipram on Toxoplasma infection in an experimental murine model. The ability, of this novel regimen to prevent or reverse Toxoplasma-induced pathology was evaluated. Materials and methods Toxoplasma-infected mice received rolipram (10 mg/kg) daily for three weeks. Life expectancy, serum ALT and histopathology of liver and brain were the parameters used to evaluate therapeutic and/or toxic impact of rolipram in comparison with infection and healthy control groups of mice. Results Toxoplasma-induced pathology was never completely cleared by rolipram. However, a significant impact in aborting chronic state and in mitigating pathology was detected. The drug almost normalized Toxoplasma-induced high ALT levels reversing the hepatotoxic effect of the parasite. Conclusion This study is the first to demonstrate relative merits of rolipram therapy to toxoplasmosis with illustration of its efficacy and toxicity. It reports a novel approach with potential future benefits using PDE inhibitors to abort resistant latent phases of biologically related infectious agents.

Corynebacterium jeddahense sp. nov., strain JCB(T), is the type strain of Corynebacterium jeddahense sp. nov., a new species within the genus Corynebacterium. This strain, whose genome is described here, was isolated from fecal flora of a 24-year-old Saudi male suffering from morbid obesity. Corynebacterium jeddahense is a Gram-positive, facultative anaerobic, nonsporulating bacillus. Here, we describe the features of this bacterium, together with the complete genome sequencing and annotation, and compare it to other member of the genus Corynebacterium. The 2,472,125 bp-long genome (1 chromosome but not plasmid) contains 2,359 protein-coding and 53 RNA genes, including 1 rRNA operon.

International audience; We report here the main characteristics of 'Jeotgalicoccus saudimassiliensis' strain 13MG44_air(T) (CSUR P1221), a new species of the Jeotgalicoccus genus that was isolated from air samples in the city environment of Makkah, Saudi Arabia, during the pilgrim period of Hajj 2012.

International audience; As part of the culturomics project aiming at describing the human microbiota, we report in this study the description of the new bacterial genus Raoultibacter gen. nov. that includes two new species, that is, R. massiliensis sp. nov. and R. timonensis sp. nov. The R. massiliensis type strain Marseille‐P2849T was isolated from the fecal specimen of a healthy 19‐year‐old Saudi Bedouin, while R. timonensis type strain Marseille‐P3277T was isolated from the feces of an 11‐year‐old pygmy female living in Congo. Strain Marseille‐P2849T exhibited 91.4% 16S rRNA sequence similarity with Gordonibacter urolithinfaciens, its phylogenetic closest neighbor with standing in nomenclature. As well, strain Marseille‐P3277T exhibited 97.96% 16S rRNA similarity with strain Marseille‐P2849T. Both strains were Gram‐positive, motile, nonspore‐forming rod and form transparent microcolonies on blood agar in both anaerobic and microaerophilic atmospheres. The genome sizes of strain Marseille‐P2849T and strain Marseille‐P3277T were 3,657,161 bp and 4,000,215 bp, respectively. Using a taxono‐genomic approach combining the phenotypic, biochemical, and genomic characteristics, we propose the genus Raoultibacter gen. nov., which contains strains Marseille‐P2849T (= CSUR P2849T, = DSM 103407T) and Marseille‐P3277T (=CCUG 70680T, =CSUR P3277T) as type strains of the species R. massiliensis sp. nov., and R. timonensis sp. nov., respectively.

Clostridium dakarense strain FF1(T), is the type strain of Clostridium dakarense sp. nov., a new species within the genus Clostridium. This strain, whose genome is described here, was isolated from the fecal flora of a 4-month-old Senegalese child suffering from gastroenteritis. C. dakarense sp. nov. strain FF1(T) is an obligate anaerobic Gram-positive bacillus. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 3,735,762 bp long genome (1 chromosome but no plasmid) exhibits a G+C content of 27.98% and contains 3,843 protein-coding and 73 RNA genes, including 8 rRNA genes.

AimThe aim of this study was to evaluate the diagnostic accuracy of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) in comparison to procalcitonin (PCT) and C-reactive protein (CRP) in early diagnosis of septic meningitis and its usefulness to distinguish between septic and aseptic meningitis.Patients and methodsThis study included 26 patients with septic meningitis, 32 patients with aseptic meningitis and 15 controls. Besides Gram staining, cultures of blood and CSF and latex agglutination test of CSF, CRP, serum PCT and sTREM-1 measurement were done on admission and after 48–72h of treatment.ResultsSeptic meningitis was diagnosed in 26 (44.82%) of the studied cases. Patients with septic meningitis had a significant increase in serum sTREM-1 and PCT levels at the time of admission (mean 32.99±19.7ng/ml and 97.9±86.35ng/ml, respectively), while patients with aseptic meningitis showed (6.8±5.67ng/ml and 1.88±1.6ng/ml, respectively). Control group showed sTREM-1 and PCT levels (6.6±4.6ng/ml and 0.58±0.36ng/ml, respectively) (P

Host genetics, environment, lifestyle and proximity between hosts strongly influence the composition of the gut microbiome. To investigate the association of dietary variables with the gut microbiota, we used 16S rDNA sequencing to test the fecal microbiome of Bedouins and urban Saudis and we compared it to the gut microbiome of baboons living in close contact with Bedouins and eating their leftovers. We also analyzed fermented dairy products commonly consumed by Bedouins in order to investigate their impact on the gut microbiome of this population. We found that the gut microbiomes of westernized urban Saudis had significantly lower richness and biodiversity than the traditional Bedouin population. The gut microbiomes of baboons were more similar to that of Bedouins compared to urban Saudis, probably due the dietary overlap between baboons and Bedouins. Moreover, we found clusters that were compositionally similar to clusters identified in humans and baboons, characterized by differences in Acinetobacter, Turicibacter and Collinsella. The fermented food presented significantly more bacteria genera common to the gut microbiome of Bedouins compared to urban Saudis. These results support the hypothesis that dietary habits influence the composition of the gut microbiome.

Recent studies have shown that bacterial and viral infections are risk factors for various neurodegenerative diseases such as Amyotrophic lateral sclerosis (ALS), Multiple Sclerosis (MS), Alzheimer's disease (AD), and Lyme disease (LD). However, it is still controversial how the infections play a role in neurological diseases progression. Infections in central nervous system may lead multiple damages in infected and neighboring cells. The infection leads to the activation of inflammatory processes and host immune responses, which acts as defense mechanism and also causes damage to the host neuronal functions and viability. Several bacterial and viral pathogens have been reported for neurodegeneration, such as the production and deposit of misfolded protein aggregates, oxidative stress, deficient autophagic processes, synaptopathies and neuronal death. These effects may act in combination with other factors, like aging, metabolic diseases and the genetic makeup of the host. We will focus in this review on the possible link between neurodegeneration and infections particularly Chlamydophila pneumoniae, Borrelia burgdorferi, Mycoplasma etc.

International audience; Culturomics is a new omics subspecialty to map the microbial diversity of human gut, coupled with a taxono-genomic strategy. We report here the description of a new bacterial species using microbial culturomics: strain S5(T), (=CSUR P1091=DSM 28586) isolated from a stool specimen of a 25-year-old obese patient from Saudi Arabia. The strain S5(T) was a Gram-positive, strictly aerobic rod, which was motile by a polar flagellum, spore-forming, and exhibited catalase and oxidase activities. It grows optimally at 37 degrees C, with a pH of 7.5 and 10% of NaCl. 16S rRNA gene-based identification revealed that strain S5(T) has 98.6% 16S rRNA sequence similarity with the reference O. oncorhynchi, phylogenetically the closest validated Oceanobacillus species. Here, we further describe the phenotypic characteristics of this organism and its complete genome sequence and annotation. The 5,388,285bp long genome exhibits a G+C content of 37.24% and contains 5109 protein-coding genes and 198 RNA genes. Based on the characteristics reported here, we propose classifying this novel bacterium as representative of a new species belonging to the genus Oceanobacillus, Oceanobacillus jeddahense sp. nov. In a broader context, it is noteworthy that halophilic bacteria have long been overlooked in the human gut, and their role in human health and disease has not yet been investigated. This study thus further underscores the usefulness of the culturomics approach exploring the bacterial diversity of the gut.

We propose the description of a new bacterial genus and new bacterial species, “Raoultibacter massiliensis,” isolated from a faecal specimen of a 19-year-old healthy Saudi Bedouin.

This study aims at investigating the status of bacterial contamination of four daily used objects, computer keyboards, computer mice, elevator buttons and shopping carts handles. A total of 400 samples were collected from 4 different objects; 100 from each. Samples were collected from different places (offices, internet cafes, homes, buildings and supermarkets) in the city of Jeddah, Saudi Arabia. 95.5% of the total samples collected were contaminated with mixed bacterial growth. Coagulase-negative staphylococci dominated the isolates. The second most common bacterial growth in all specimens was Gram-positive bacilli. Potential pathogens isolated from all specimens were: Staphylococcus aureus, Pseudomonas spp. and Gram negative bacilli. Results indicate that internet cafe computer keyboards and mice showed 100% contamination in comparison with other objects. The presence of pathogenic and commensal bacteria on the four objects indicates that they might act as environmental vehicles for the transmission of potentially pathogenic bacteria. Key words: Bacterial contamination, computers, public surfaces.

Introduction: Toxoplasma gondii, a common parasitic infection, has a special affinity to the brain. It has a lifelong existence without an apparent clinical disease. While the etiology of bipolar disorder (BD) remains unclear, epidemiological studies suggest a role for infections. Central nervous system is particularly susceptible to oxidative stress (OS) because of its high metabolic rate and its low levels of antioxidant defenses. OS is a contributor to the initiation and progression of many neurological illnesses. OS injury is a constantly and compelling finding associated with BD and toxoplasmosis. Aim: This cross-sectional study has investigated a possible role of toxoplasma-induced OS in the development of BD. Methods: Healthy controls and BD patients were examined for anti-Toxoplasma immunoglobulin-G (IgG) and two protein (3-nitrotyrosine) and DNA (8-hydroxy-2’ deoxyguanosine [8-OHdG]) OS markers. Results: Toxoplasma positivity was higher (40%) among BD patients compared to controls (12%). Significantly higher levels of anti-Toxoplasma IgG were detected in BD patients compared to controls. Nitrotyrosine (796.7 ± 106.28) and especially 8-OHdG (20.31 ± 8.38) were significantly higher among toxo-positive BD compared to toxo-negative BD (675.97 ± 144.19 and 7.44 ± 2.86) and healthy controls (464.02 ± 134.6 and 4.17 ± 1.43). Conclusion: These findings might indicate a role for Toxoplasma infection in the development of BD, possibly through creating a highly oxidative brain environment.

Oceanobacillus picturae is a strain of a moderately halophilic bacterium, first isolated from a mural painting. We demonstrate, for the first time, the culture of human Oceanobacillus picturae, strain S1T, whose genome is described here, from a stool sample collected from a 25-year-old Saoudian healthy individual. We used a slightly modified standard culture medium adding 100 g/L of NaCl. We provide a short description of this strain including its MALDI-TOF spectrum, the main identification tool currently used in clinical microbiology. The 3,675,175 bp long genome exhibits a G + C content of 39.15 % and contains 3666 protein-coding and 157 RNA genes. The draft genome sequence of Oceanobacillus picturae has a similar size to the Oceanobacillus kimchii (respectively 3.67 Mb versus 3.83 Mb). The G + C content was higher compared with Oceanobacillus kimchii (respectively 39.15 % and 35.2 %). Oceanobacillus picturae shared almost identical number of genes (3823 genes versus 3879 genes), with a similar ratio of genes per Mb (1041 genes/Mb versus 1012 genes/Mb). The genome sequencing of Oceanobacillus picturae strain S1 isolated for the first time in a human, will be added to the 778 genome projects from the gastrointestinal tract listed by the international consortium Human Microbiome Project. Electronic supplementary material The online version of this article (doi:10.1186/s40793-015-0081-2) contains supplementary material, which is available to authorized users.

Cell phones are commonly used in healthcare settings for rapid communication within hospitals. Concerns have been increased about the use of these devices in hospitals, as they can be used everywhere, even in toilets. Therefore, they can be vehicles for transmitting pathogens to patients. This study aimed to examine the presence of pathogenic bacteria on the surfaces of cell phones that are used frequently by preclinical medical students. This cross-sectional study identified both pathogenic and nonpathogenic bacteria on cell phones of 105 medical students at King Abdulaziz University, Jeddah, Saudi Arabia, using standard microbiological methods. Out of 105 cell phones screened, 101 (96.2%) were contaminated with bacteria. Coagulase-negative staphylococci were the most abundant isolates (68%). Seventeen (16.2%) cell phones were found to harbor Staphylococcus aureus. Gram-positive bacilli were isolated from 20 (19%) samples. Viridans streptococci and Pantoea species were also isolated but at lower levels. Our findings indicate that cell phones can act as reservoirs of both pathogenic and nonpathogenic organisms. Therefore, full guidelines about restricting the use of cell phones in clinical environments, hand hygiene, and frequent decontamination of mobile devices are recommended at an early stage in medical schools, to limit the risk of cross-contamination and healthcare-associated infections caused by cell phones.

Background Early diagnosis of hepatocellular carcinoma (HCC) is the most important step in successful treatment. However, it is usually rare due to the lack of a highly sensitive specific biomarker so that the HCC is usually fatal within few months after diagnosis. The aim of this work was to study the role of plasma nuclear factor kappa B (NF-?B) and serum peroxiredoxin 3 (PRDX3) as diagnostic biomarkers for early detection of HCC in a high-risk population. Materials and methods Plasma nuclear factor kappa B level (NF-?B) and serum peroxiredoxin 3 (PRDX3) levels were measured using enzyme linked immunosorbent assay (ELISA), in addition to alpha-fetoprotein (AFP) in 72 cirrhotic patients, 64 patients with HCC and 29 healthy controls. Results NF-?B and PRDX3 were significantly elevated in the HCC group in relation to the others. Higher area under curve (AUC) of 0.854 (for PRDX3) and 0.825 (for NF-?B) with sensitivity of 86.3% and 84.4% and specificity of 75.8% and 75.4% respectively, were found compared to AUC of alpha-fetoprotein (AFP) (0.65) with sensitivity of 72.4% and specificity of 64.3%. Conclusions NF-?B and PRDX3 may serve as early and sensitive biomarkers for early detection of HCC facilitating improved management. The role of nuclear factor kappa B (NF-?B) as a target for treatment of liver fibrosis and HCC must be widely evaluated.

Background: Candidemia is a major cause of morbidity and mortality in hospitalized patients. The spectrum of candidemia has been changed especially among critically ill patients due to emergence of non-albicans Candida (NAC) species. The increasing use of azole agents is suggested to be responsible for this epidemiological shift. NAC species are of special concern because of their high drug-resistance and increasing prevalence.The aim of this study was to detect antifungal-susceptibility patterns, treatment outcomes and associated risk factors in patients with candidemia who were admitted to King Abdulaziz University Hospital (KAUH), Jeddah, Kingdom of Saudi Arabia (KSA) .Methods: This work represents a cross sectional study done in the Clinical and Microbiology Laboratory at KAUH, during the periodfrom March 2012 till February 2014 on a total of 141 patients with candidemia. They were 31(22%) Saudi and 110(87%) non-Saudi patients with age ranged from 1 day - 102 years. Blood cultures were collected for suspected cases of candidemia, followed by subculture on SDA. Identifiation was done by VITEK MS (MALDI-TOF MS), and confimation of Candida isolates and antifungal-susceptibility testing were performed by using VITEK ®2 system.Results: C.albicans isolates accounted for 39.7%, followed by C. tropicalis (21.3%), C. galabrata (18.4%) and C. parapsiliosis (14.9%). Additionally, C. dublinsis, C. krusei and C. famata were representing 2.1%, 2.1% and 1.4%, respectively. All Candida isolates were 100% susceptible to amphotericin B. The best susceptibility to flconazole was detected among each C. dubliensis and C. famata (100%). All C. krusei isolates were resistant to flconazole, while they were susceptible  to other antifungal agents. All isolates were susceptible to flcytosine,except C. albicans and C. dubliensis which were susceptible 92.9%and 66.7%, respectively. All isolates were susceptible to itraconazol,except C. albicans and C. tropicalis which were susceptible 94.6% and 96.7%, respectively. The percentage of deceased patients with candidemia was signifiantly higher than the survivors among age group >64 years, particularly those who were mechanically ventilated and those understeroid therapy. The percentage of deceased patients was signifiantly higher than survivors among those admitted to adult ICUs (73.78%vs 26.23%) .Conclusion: This study shows an epidemiological shift to higher NAC species isolation rates, with 100% susceptibility to amphotericin B in all isolates either C. albicans or NAC species, and 100% susceptibility to flconazole among C. dubliensis and C. famata. Patients aged > 64 years, admission to adult ICUs, mechanical ventilation and steroid therapy were signifiant risk factors for increased mortality due to candidemia.

Background Rheumatoid arthritis (RA) is the most common chronic inflammatory joint disease, with a worldwide prevalence of 0.5% to 1%. Anti-cyclic citrullinated peptide antibody (anti-CCP-2 Ab) is a marker of choice for diagnosing early and late RA. Anti-oxidant enzymes activity decreases in RA patients. Till now, the relationship between the rheumatoid factor (RF) and anti-CCP-2 Ab, anti-oxidant activity and polymorphism of paraoxenase-1 (PON-1) 192 Q/R in patients with RA has not been investigated. In this study, we aimed to determine the serum level of RF and anti-CCP-2 Ab, PON-1 activity and 192 Q/R polymorphism and arylesterase (ARE) activity in patients with RA. Also, we studied RA markers in different genotypes of PON-1 of RA patients. Methods A total of 120 RA patients and 90 healthy persons were subjected to full clinical examinations and routine laboratory tests. PON-1 and ARE activities were determined using an enzymatic spectrophotometric method. PON-1 192 gene polymorphism was determined using polymerase chain reaction based restriction fragment analysis. RF was measured by immunoturbidimetry method and anti-CCP-2 Ab was assayed by enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed using SPSS for windows 20.0. Results The sensitivity and specificity of anti-CCP-2 Ab for the diagnosis of RA were 76.2% and 100% respectively. PON-1 and ARE activities were statistically lower (P

The incidence of Candida infections have increased substantially in recent years due to aggressive use of immunosuppressants among patients. Use of broad-spectrum antibiotics and intravascular catheters in the intensive care unit have also attributed with high risks of candidiasis among immunocompromised patients. Among Candida species, C. albicans accounts for the majority of superficial and systemic infections, usually associated with high morbidity and mortality often caused due to increase in antimicrobial resistance and restricted number of antifungal drugs. Therefore, early detection of candidemia and correct identification of Candida species are indispensable pre-requisites for appropriate therapeutic intervention. Since blood culture based methods lack sensitivity, and species-specific identification by conventional method is time-consuming and often leads to misdiagnosis within closely related species, hence, molecular methods may provide alternative for accurate and rapid identification of Candida species. Although, several molecular approaches have been developed for accurate identification of Candida species but the internal transcribed spacer 1 and 2 (ITS1 and ITS2) regions of the rRNA gene are being used extensively in a variety of formats. Of note, ITS sequencing and PCR-RFLP analysis of ITS region seems to be promising as a rapid, easy, and cost-effective method for identification of Candida species. Here, we review a number of existing techniques ranging from conventional to molecular approaches currently in use for the identification of Candida species. Further, advantages and limitations of these methods are also discussed with respect to their discriminatory power, reproducibility, and ease of performance.

Coronary artery disease (CAD) is the leading cause of death in many countries. The underlying mechanism of the chronic inflammatory process in atherosclerosis is still unknown. As a possible trigger, different viruses and bacteria may be associated with atherosclerotic diseases. The aim of this work was to investigate the association of chronic infection with C pneumoniae, H pylori and cytomegalovirus (CMV) infections and CAD. Fifty patients [20 with acute coronary artery disease (ACAD) and 30 with chronic coronary artery disease (CCAD)] in addition to 15 healthy individuals as a control group were involved in this study. The studied individuals were subjected to complete history taking, thorough physical examination, electrocardiography, echocardiography and coronary angiography (for patients). Assessment of blood glucose level, lipid profile and creatine kinase (CK) was performed. Determination of hsCRP was done by nephlemetry, while C pneumoniae-, H pylori- and CMV-specific IgG antibodies was done by enzyme immunoassay. Results showed that the levels of cholesterol, triglycerides, LDL-c and hsCRP were significantly higher, while HDL-c was significantly lower among patients compared to that of the controls. A significantly (P0.05) higher perecentage of patients had C pneumoniae and H pylori-specific IgG antibodies as compared to that of the controls. Higher percentage of patients had CMV-specific IgG antibody, however, there was no significant difference between the 2 groups. The levels of C pneumoniae and H pylori-specific IgG antibodies were significantly (P0.001) higher among patients with CAD when compared to that of the controls. CMV-specific IgG level in patients was higher compared to that of the controls, however, the difference was not statistically significant. Among acute CAD patients, C pneumoniae-specific IgG was positively correlated with hsCRP (P0.05), cholesterol (p0.01) and HDL-c (P0.05), while H pylori-specific IgG was positively correlated with triglyceride level (P0.05). Among patients with CCAD, hsCRP was negatively correlated with HDL-c (P0.05). There was no significant correlation between the levels of CMV-specific IgG and lipid profile or hsCRP. In conclusion, the level of C pneumoniae and H pylori-specific IgG antibodies are elevated among CAD patients and their presence was associated with development of the disease. They were significantly correlated to cholesterol level. Moreover, C pneumoniae-specific IgG was significantly correlated with hsCRP among ACAD patients, suggesting an important role of these organisms in the development of CAD by altering lipid profile and induction of inflammation.