CHROMagar Orientation Research Articles

Comparison of CHROMagar Orientation Medium with Traditional Methods in Urinary Tract Infection Cultures.

Urinary tract infections (UTIs) are the most common infections in adults, and urine culture is the parameter that uses the most time and cost in microbiology laboratories. For this reason, the selection of fast and cost-effective methods in the evaluation of urine samples is one of the priority issues of microbiology laboratories. The aim of this study was to investigate the compatibility and cost-effectiveness of routinely used Blood Agar (BA), Eosin Methylene Blue (EMB) medium, and CHROMagar Orientation Medium (CO Medium) in the identification of microorganisms in urine samples. Consecutive urine samples (n: 700) sent to our laboratory were simultaneously inoculated onto BA/EMB media and CO medium. Urine samples were evaluated after 18 - 24 hours of incubation at 37℃ and the compatibility of the two methods was compared. The use of 104 Gram stains, 198 biochemical tests, and 9 identification kits was required with BA/EMB agar. When 104 colonies with single growth were evaluated, presumptive identification with CO medium was found to be 100% compatible with the VITEK 2 system. The most isolated 62 Escherichia coli (E. coli) colonies gave dark pink-red color and were found to be fully compatible with the VITEK 2 system. Compatibility of BA and EMB medium evaluations with VITEK 2 system; E. coli (n: 62), KES group (n: 26), Pseudomonas spp. (n: 6) and Proteus spp. For (n: 2), it was determined as 69.3%, 57.69%, 100%, and 100%, respectively. According to the results of our study, when BA/EMB and CO Medium methods were compared, 182 Euro (€) savings were achieved in 700 urine cultures with CO Medium. It was estimated that the amount of savings could be 15,600 € per year. CHROMagar Orientation Medium method can be used routinely with its advantages such as being cost-effective, reducing the workload, and not requiring additional operations. CHROMagar Orientation Medium can also be considered as an easily accessible method and opportunity that does not require infrastructure and trained personnel, especially for laboratories with low test capacity and having problems with the supply of com-mercial kits and automated systems.

Characterizing the impact of Enterococcus cecorum infection during late embryogenesis on disease progression, cecal microbiome composition, and early performance in broiler chickens

Enterococcus cecorum (EC) has been associated with septicemia and early mortality in broiler chickens. There is limited research investigating the pathogenicity of EC field strains obtained from affected birds. The purpose of this study was to evaluate the effect of in-ovo administration into the amnion with different EC field isolates at d 18 of embryogenesis (DOE18). In Exp 1, 7 EC field isolates alone or in combination (EC1-EC3, EC4-EC5, EC6, and EC7) were selected based on phenotypic characteristics and evaluated at different concentrations (1×102, 1×104, and 1×106 CFU/200 µL/embryo) to assess the impact on early performance and macroscopic lesions. Three isolates (n=3; EC2, EC5, EC7) were selected for additional evaluation based on the significant (P < 0.05) BWG reduction (d 0-21) compared to the negative control (NC) and the presence of macroscopic lesions observed during posting sessions at d 14 and d 21. An additional isolate associated with enterococcal spondylitis was included in Exp 2 (EC11B). Treatment groups for Exp 2 include: 1) NC, 2) EC2, 3) EC5, 4) EC7, and 5) EC11B (n=90-120/embryos/group). Groups 2 to 5 were challenged at 1×102 CFU/200 µL/embryo by in-ovo injection into the amnion at DOE18. Chicks were placed in battery cages for the duration of the study (21 d), and pen weights were recorded at d 0, d 7, d 14, and d 21 to calculate average BW and BWG. At d 14 and d 21 posthatch, liver, spleen, free thoracic vertebrae (FTV), and femoral head (FH) were aseptically collected to enumerate Enterococcus spp. using Chromagar Orientation as the selective media. Cecal contents were collected at d 21 to evaluate the effect of EC challenge on the cecal microbiome composition. There was a significant (P < 0.05) reduction in BW at d 21, and BWG from d 14 to 21 and d 0 to 21, for EC7 and EC11B. Enterococcus cecorum was recovered from the FTV of all challenged groups at d 14 and d 21. The most representative lesions were pericarditis, hydropericardium, focal heart necrosis, and FH osteomyelitis. However, lesions were not uniform across challenged groups or ages (d 14 and d 21). Alpha diversity of the cecal contents was markedly lower in EC5 and EC11B compared to all treatment groups suggesting that EC exposure during late embryogenesis affect the cecal microbiome up to 21 d posthatch. Additionally, these results highlight the differences in pathogenicity of EC strains isolated from field cases and suggest that hatchery exposure to EC during late embryogenesis is a potential route of introduction into a flock.

Diagnostic Accuracy of CHROMagar Orientation for Identification of Uropathogens

Objective: To determine the diagnostic accuracy of CHROMagar Orientation for its ability to support the growth and identification of uropathogens by keeping API 20E/20NE as a gold standard.&#x0D; Study Design: Cross-sectional study.&#x0D; Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi Pakistan, from Jun to Nov 2019.&#x0D; Methodology: A total of 470 midstream specimen of urine from patients suspected of UTIs were analyzed. Urine specimens were inoculated by 1µL calibrated sterilized loop each on CLED Agar, Blood agar and CHROMagar Orientation media plates.Culture plates were incubated at 35.00±2.00ºC for 18 to 24 hours. As per standard protocols, isolates were identified by colony morphology, Gram stain, catalase, oxidase, DNAase, coagulase, and API 20E/20NE. In addition, the appearance of microorganism on CHROMagar Orientation media were identified according to the manufacturer’s instructions.&#x0D; Results: Out of 470 samples, 211 showed no growth, 90 were classified as mix growth (polymicrobial), and 169 samples were positive (unimicrobial) for the growth of uropathogens. All the samples positive for bacterial growth on conventional media were also positive on CHROMagar orientation and vice versa. The sensitivity and specificity of CHROMagar Orientation for differentiation of uropathogens were 99% and 100%, respectively.&#x0D; Conclusion: The overall findings of our study suggested that CHROMagar supports the growth of all uropathogens. It provided the rapid identification of organisms based on their colour morphology, even in a mixed bacterial culture, growth within 24 hours easily.

KPC-Producing Enterobacterales from Douro River, Portugal-Persistent Environmental Contamination by Putative Healthcare Settings.

Carbapenemase-producing Enterobacterales (CPE) are a growing concern, representing a major public health threat to humans, especially in healthcare settings. In the present study, we evaluated the persistent contamination by carbapenem-resistant Enterobacterales in water from Douro River, Portugal. KPC-producing Enterobacterales were detected in five water samples separated chronologically by 15 days each. Susceptibility testing was performed by disk-diffusion-method according to Clinical and Laboratory Standards Institute (CLSI), phenotypic carbapenemase activity was evaluated by carbapenem inactivation method, presumptive identification of the isolates was performed by CHROMagar orientation and confirmed by API-20E. Carbapenemase genes were screened by PCR and the clonality of all isolates was assessed by XbaI-Pulsed Field Gel Electrophoresis (PFGE). Fifteen KPC-producing Enterobacterales isolates were selected, identified as multidrug-resistant and showed a resistance profile to non-beta-lactam antibiotics: sulfamethoxazole + trimethoprim (7/15), ciprofloxacin (3/15), fosfomycin (3/15) and chloramphenicol (2/15). Isolates were identified as (6) Escherichia coli and (9) Klebsiella pneumoniae. Our results suggest a punctual contamination with KPC-producing Enterobacterales continued through the time. The absence of clonality between the isolates suggests a circulation of mobile genetic element harbouring KPC gene in the origin of contamination. This work provides a better understanding on the impacts of water pollution resulting from human activities on aquatic environments.

S2.5a Magnusiomyces spp.

S2.5 Rare yeasts, September 21, 2022, 3:00 PM - 4:30 PM Magnusiomyces clavatus and Magnusiomyces capitatus are emerging yeasts with intrinsic resistance to many commonly used antifungal agents. Identification is difficult, and the determination of susceptibility patterns with commercial and reference methods is equally challenging. For this reason, few data on invasive infections by Magnusiomyces spp. are available. We, therefore, determined the epidemiology and susceptibility of Magnusiomyces isolates from bloodstream infections (BSI) isolated in Germany and Austria from 2001 to 2020. A total of 34 Magnusiomyces BSI were analyzed; isolates were identified by internal transcribed spacer (ITS) sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Antifungal susceptibility was determined by EUCAST broth microdilution and gradient tests. Of the 34 isolates, M. clavatus was more common (n = 24) than M. capitatus (n = 10). BSI by Magnusiomyces spp. were more common in men (62%) and mostly occurred in patients with hemato-oncological malignancies (79%). The highest in vitro antifungal activity against M. clavatus/M. capitatus was observed for voriconazole (MIC50, 0.03/0.125 mg/l), followed by posaconazole (MIC50, 0.125/0.25 mg/l). M. clavatus isolates showed overall lower MICs than M. capitatus. With the exception of amphotericin B, a low essential agreement between gradient test and microdilution was recorded for all antifungals (0%-70%). Both species showed distinct morphologic traits on ChromAgar Orientation medium and Columbia blood agar, which can be used for differentiation if no MALDI-TOF MS or molecular identification is available. In conclusion, most BSI were caused by M. clavatus. The lowest MICs were recorded for voriconazole. Gradient tests demonstrated unacceptably low agreement and should preferably not be used for susceptibility testing of Magnusiomyces spp.

P15 What is the value of direct-from-blood culture ASTs in the study of bacterial susceptibility to antibiotics during bacteraemia?

ObjectivesTo evaluate the results obtained with direct-from-blood culture antimicrobial susceptibility tests (ASTs) for the study of bacterial susceptibility to antibiotics during bacteraemia.Material and methodsThis was a prospective study in which 124 direct ASTs were performed directly from positive blood cultures according to the CLSI technique using Mueller–Hinton CHROMagar Orientation medium. Twenty-one antibiotics were tested. The resulting diameters were read after 8 h and 18 h of incubation and interpreted according to the CLSI breakpoints. Thus, a standard AST was performed and interpreted for all isolated strains according to CLSI recommendations. The results were analysed by calculating the rates of categorical agreements (CA) and disagreements represented by minor errors (mE), major errors (ME) and very major errors (VME).ResultsOne hundred and twenty-four strains were isolated, including Escherichia coli (26; 20.97%), Klebsiella pneumoniae (25; 20.16%) and SCN (21; 16.94%). More than half of the isolates (69; 55.65%) were MDR bacteria. It was found that the overall percentage CA (%CA) obtained at 18 h (94.43%) was higher than the one obtained at 8 h (87.32%). Disagreement rates obtained at 8 h and 18 h remain acceptable. The best %CA was obtained with non-fermenting GNB (98.74%) and the lowest with Staphylococcus species (90.70%). For Enterobacteriaceae, an excellent overall %CA (94.32%) was found at 18 h with 96.3% for CTX, 95.38% for ETP, 95.31% for CRO and 92.19% for MEM. Lower rates were noted with IPM (88.71%), CAZ (87.5%) and CIP (85.71%); due to mE for CAZ (12.5%) and mE and ME for IPM (9.68% mE; 1.85% ME) and CIP (12.7% mE; 3.23% ME). Particular attention should be given to clindamycin and teicoplanin tested with Staphylococcus species as they record low categorical agreements at 8 h and 18 h.ConclusionsThe encouraging results obtained at 18 h suggest a possible future implementation of the direct-from-blood culture AST as a routine technique for studying susceptibility to antibiotics during bacteraemia. However, the standard AST remains the reference technique.

Remarkable genomic diversity among Escherichia isolates recovered from healthy chickens.

The genus Escherichia has been extensively studied and it is known to encompass a range of commensal and pathogenic bacteria that primarily inhabit the gastrointestinal tracts of warm-blooded vertebrates. However, the presence of E. coli as a model organism and potential pathogen has diverted attention away from commensal strains and other species in the genus. To investigate the diversity of Escherichia in healthy chickens, we collected fecal samples from antibiotic-free Lohmann Brown layer hens and determined the genome sequences of 100 isolates, 81 of which were indistinguishable at the HC0 level of the Hierarchical Clustering of Core Genome Multi-Locus Sequence Typing scheme. Despite initial selection on CHROMagar Orientation medium, which is considered selective for E. coli, in silico phylotyping and core genome single nucleotide polymorphism analysis revealed the presence of at least one representative of all major clades of Escherichia, except for E. albertii, Shigella, and E. coli phylogroup B2 and cryptic clade I. The most frequent phylogenomic groups were E. coli phylogroups A and B1 and E. ruysiae (clades III and IV). We compiled a collection of reference strains isolated from avian sources (predominantly chicken), representing every Escherichia phylogroup and species, and used it to confirm the phylogeny and diversity of our isolates. Overall, the isolates carried low numbers of the virulence and antibiotic resistance genes typically seen in avian pathogenic E. coli. Notably, the clades not recovered are ones that have been most strongly associated with virulence by other studies.

Culturomics Approach to Identify Diabetic Foot Infection Bacteria.

The main goal of the study was to evaluate the usefulness of the culturomics approach in the reflection of diabetic foot infections (DFIs) microbial compositions in Poland. Superficial swab samples of 16 diabetic foot infection patients (Provincial Polyclinical Hospital in Toruń, Poland) were subjected to culturing using 10 different types of media followed by the identification via the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and Biotyper platform. Identified 204 bacterial isolates representing 18 different species—mostly Enterococcus faecalis (63%) and Staphylococcus aureus (44%). Most of the infections (81%) demonstrated a polymicrobial character. Great differences in the species coverage, the number of isolated Gram-positive and Gram-negative bacteria, and the efficiency of the microbial composition reflection between the investigated media were revealed. The use of commonly recommended blood agar allowed to reveal only 53% of the entire microbial composition of the diabetic foot infection samples, which considerably improved when the chromagar orientation and vancomycin-resistant enterococi agar were applied. In general, efficiency increased in the following order: selective < universal < enriched < differential media. Performed analysis also revealed the impact of the culture media composition on the molecular profiles of some bacterial species, such as Corynebacterium striatum, Proteus mirabilis or Morganella morganii that contributed to the differences in the identification quality. Our results indicated that the culturomics approach can significantly improve the accuracy of the reflection of the diabetic foot infections microbial compositions as long as an appropriate media set is selected. The chromagar orientation and vancomycin-resistant enterococi agar media which were used for the first time to study diabetic foot infection microbial profiles demonstrate the highest utility in the culturomics approach and should be included in further studies directed to find a faster and more reliable diabetic foot infection diagnostic tool.

PCR detection of quorum sensing autoinducer (AI) type one among gram negative bacteria

Any bacterial species is found to be regulating their activities like pathogenicity, antibiotic resistant, biofilm production, and others by phenomenon system called quorum sensing. This system is dependent on production signaling molecules called autoinducers (AI); consequently, we tested the presence of QS genes by polymerase chain reaction (PCR) among different gram negative bacterial pathogens.Material and Methods: Fifty isolates of each of Citrobacter spp., E. coli, Salmonella typhimurium, and Pseudomonas auroginosa were obtained from different private clinical laboratories in Baghdad and Al-Najaf cites in the period of June 2020- February 2021. Bacteria were identified using CHROM agar Orientation followed by Vitek2 system. Quorum sensing genes (abaI 382bp and AHL gene 498bp) were screened by PCR.

Highland Influence on Chronic-bacterial-prostatitis and Developing-reason of Prostate-cancer, KSA

This work was for "Highland (HL) Influence on Chronic-bacterial-prostatitis (CBP) and Developing-reason of Prostate-cancer (PCa), KSA"; the purpose was to determine CBP percentage at HL "Taif"; and influence of location factors on CBP percentage and indication of its developing-reason to causing PCa. It may negatively affected other body organs and may reach death of males' society, affected marital and community life. Used medical methods for cases taster (CsT) from patients were diagnosed as suffered from CBP. That were used direct and indirect identification by "CHROMagar Orientation Medium", and "VITEK 2 System (BioMerieux, France)". The pathogenic bacteria were found in more than a quarter of the CsT as (26 and 29%). That were Staphylococcous aureus (Staph. aureus), Escherichia coli (E. coli), Staphylococcous epidermidis (Staph. epidermidis) and Streptococcus species (Strept. Spp) in (36.2%, 9.5%, 5.7% and 4.3%) respectively. Pathogenic bacteria Staph. aureus represented more than third of CsT. E. coli represented tenth and it was dangerous for its transmission from UT and GT easily. Staph. epidermidis was present on the skin and represented normal flora can easily contaminated. Strept. Spp represented less than tenth, can transferred to prostate tissue and cause CBP leading to PCa. Concluded the CBP infection was present in HL and had developing-reason for PCa, the conditions must followed to reduce it to prevent condition of turning into PCa. Recommended best periodic follow-up to detect CBP infection presence to protect against infection and its PCa developing-reason. Cases CBP infection must treated and PCa cases should not neglected or tolerated.

Main bacterial species causing clinical disease in ornamental freshwater fish in Brazil.

Bacterial diseases are common in ornamental fish, more frequently associated with ubiquitous bacteria from the aquarium environment. The disease can lead to fish mortality and cause high economic losses if not rapidly controlled. The aim of this study was to identify the main causative bacterial agents of infection in ornamental fish with different clinical signs. A total of 126 freshwater fish, from 12 families and 38 species, with clinical signs were collected in a wholesaler in São Paulo, SP, Brazil. Samples were taken from the eye, skin ulcers, kidneys, and gills, plated on MacConkey, CHROMagar Orientation, and blood agar and incubated under aerobic and anaerobic conditions. Bacterial identification was performed by MALDI-TOF mass spectrometry. From the 126 studied animals, 112 were positive for bacterial isolation. Among the positive animals, 32.1% presented infection caused by a single bacterial species, while in the remaining 67.9%, two to six different bacterial species were identified. A total of 259 bacterial strains were obtained and classified among 46 bacterial species. The species of higher frequency were Aeromonas veronii (26.3%), Aeromonas hydrophilla (16.2%), Shewanella putrefaciens (7.3%), Citrobacter freundii (8.1%), Vibrio albensis (5.8%), and Klebsiella pneumoniae (4.2%). MALDI-TOF MS showed to be a rapid method for diagnosis of bacterial disease outbreaks in ornamental fish establishments.

Evaluation of the NG-Test CARBA 5 Kit for Rapid Detection of Carbapenemase Resistant Enterobacteriaceae.

We evaluated NG-Test CARBA 5, a new phenotypic carbapenemase detection assay, and compared it to the routine Xpert CARBA-R polymerase chain reaction assay. Furthermore, we tested the kit's performance after bacterial growth on 4 different solid media. Seventy carbapenem resistant Enterobacteriaceae (CRE) isolates (60 were carbapenemase producers) were collected at the Poriya Baruch Padeh Medical Center. All isolates were grown on 4 types of agar media-BD BBL CHROMagar carbapenem resistant Enterobacteriaceae, BD CHROMagar Orientation, BD MacConkey II agar, and BD Trypticase Soy Agar II with 5% sheep blood-and were then subjected to NG-Test CARBA 5 kit analysis. The NG-Test CARBA 5 specificity was 100% for all 4 media. However, the sensitivity was higher when bacteria were grown on TSA with 5% sheep blood (98.3%) as compared with the Orientation medium (88.3%), the CPE medium (84.7%), and the MacConkey medium (83.6%). In addition, some of the carbapenemase mechanisms such as Verona Integron-Mediated Metallo-β-lactamase were detected with low agreement levels in specific media but higher agreement levels in the other media. NG-Test CARBA 5 may enable faster detection of carbapenemase producing CRE, which will be of value for treatment adjustment and prevention control. However, the medium type on which the bacteria are grown affects kit sensitivity.

Effectiveness of two bed bath methods in removing microorganisms from hospitalized patients: A prospective randomized crossover study

Few studies have compared the effectiveness of washing with either soap and water or disposable wet wipes. The objective of this study was to compare the effectiveness of washing with either soap and water or disposable wet wipes in reducing microorganisms in the groin and perineum of hospitalized patients, which could potentially reduce the risk of hospital-acquired urinary tract infections. In this crossover, block-randomized trial, skin swabs from the groin and perineum areas of patients were obtained before and after these areas were washed with either soap and water or disposable wet wipes. Columbia agar plates and CHROMagar Orientation Medium (Becton Dickinson; Franklin Lakes, NJ) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry procedures were used to identify species of microorganisms. Fifty-eight paired skin swabs were obtained. Both washing methods resulted in a statistically significant reduction in the amount of all microorganisms, including microorganisms with the potential to cause urinary tract infections. New species were observed after using both washing methods. No statistically significant difference in the removal of microorganisms was observed between the two washing methods. The two washing methods appear to be equally efficient in removal of microorganisms in the groin and perineum areas, including microorganisms that potentially could cause hospital-acquired urinary tract infections.

Integrated aquaculture contributes to the transfer of mcr-1 between animals and humans via the aquaculture supply chain

BackgroundSince its discovery in 2015, the mobile colistin resistance gene mcr-1 has been reported in bacteria from >50 countries. Although aquaculture-associated bacteria may act as a significant reservoir for colistin resistance, systematic investigations of mcr-1 in the aquaculture supply chain are scarce. ObjectivesWe investigated the presence of colistin resistance determinants in the aquaculture supply chain in south China and determined their characteristics and relationships. MethodsA total of 250 samples were collected from a duck-fish integrated fishery, slaughter house, and market in Guangdong Province, China, in July 2017. Colistin-resistant bacteria were isolated on colistin-supplemented CHROMagar Orientation plates, and the species were identified by matrix-assisted laser desorption/ionization time-of-flight assay. The presence of mcr genes was confirmed by polymerase chain reaction analysis. We examined the minimum inhibitory concentrations (MICs) of 16 antimicrobial agents against the isolates using agar diffusion and broth microdilution methods. Whole-genome sequencing (WGS) was used to explore the molecular characteristics and relationships of mcr-1-positive Escherichia coli (MCRPEC). ResultsOverall, 143 (57.2%) colistin-resistant bacteria were isolated, of which, 56 (22.4%, including 54 Escherichia coli and two Klebsiella pneumoniae) and four Aeromonas species were positive for mcr-1 and mcr-3, respectively. The animal-derived MCRPEC were significantly more prevalent in integrated fishery samples (40.0%) than those in market (4.8%, P<0.01) samples but not in slaughter house (28.0%, P=0.164). All MCRPEC were highly resistant to ampicillin, tetracycline, and compound sulfamethoxazole (>90%) but were susceptible to carbapenems and tigecycline. WGS analysis suggested that mcr-1 was mainly contained on plasmids, including IncHI2 (29.6%), IncI2 (27.8%), IncX4 (14.8%), and IncP (11.1%). Genomic analysis suggested mcr-1 transmission via the aquatic food chain. ConclusionsMCRPEC were highly prevalent in the aquaculture supply chain, with the isolates showing resistance to most antibiotics. The data suggested mcr-1 could be transferred to humans via the aquatic food chain. Taking the “One Health” perspective, aquaculture should be incorporated into systematic surveillance programs with animal, human, and environmental monitoring.

Detection of lasB Gene of Pseudomonasaeruginosa Causing Different Infection

The result obtained 75 isolate of Pseudomonasaeruginosa included: 28 isolates from otitis media, 23 isolates from burn infections, 10 isolates from wound infections, 8 isolates from urinary tract infections and 6 isolates from blood. The isolates were identified by culturing on MacConkey agar, Blood agar, Cetrimide agar, Pseudomonas agar and CHROMagar Orientation then identified by performing biochemical tests including (oxidase test and catalase test) and further identification by using a API20E system. The result obtained Pseudomonasaeruginosa that have Las B included : 23 isolates from otitis media 82.14%, 19 isolates from burn infections 82.60%,10 isolates from wound infections 100%, 5 isolates from urinary tract infections 62.5 % and 6 isolates from blood 100%.The result revealed that the lasB gene was present in 63 isolates 84% of Pseudomonasaeruginosa. The gel electrophoresis showed that the molecular weight of alg D gene was 300bp.

The use of light to potentiate antidepressant medication: Preclinical evidence

Experimental animal models are indispensable components of preclinical sepsis research. Reproducible results highly rely on defined and invariant baseline conditions. Our hypothesis was that the murine gut microbiota varies among different distributors of laboratory animals and that these variations influence the phenotype of abdominal sepsis derived from a bacterial inoculum model (intraperitoneal stool injection).Male C57BL/6 mice (8-wk old) purchased from Charles River (CR), Janvier (J), and Harlan (H) were sacrificed, and the bacterial composition of feces was analyzed using CHROMagar orientation medium. Stool was injected intraperitoneally into CR mice, followed by clinical observation and gene expression analysis. Experiments were repeated 16 mo later under the same conditions.Stool analysis revealed profound intervendor differences in bacterial composition, mainly regarding Staphylococcus aureus and Bacillus licheniformis. Mice challenged with CR as well as H feces developed significantly higher severity of disease and died within the observation period, whereas stool from J mice did not induce any of these symptoms. Real-time polymerase chain reaction revealed corresponding results with significant upregulation of proinflammatory cytokines and vascular leakage-related mediators in CR and H injected animals. Sixteen months later, the bacterial fecal composition had significantly shifted. The differences in clinical phenotype of sepsis after intraperitoneal stool injection had vanished.We are the first to demonstrate vendor and time effects on the murine fecal microbiota influencing sepsis models of intraabdominal stool contamination. The intestinal microbiota must be defined and standardized when designing and interpreting past and future studies using murine abdominal sepsis models.

ЭФФЕКТИВНОСТЬ КОМПЛЕКСА ЛЕЧЕБНЫХ МЕРОПРИЯТИЙ ХРОНИЧЕСКОГО ГИНГИВИТА У ПАЦИЕНТОВ С ХРОНИЧЕСКИМ ГАСТРОДУОДЕНИТОМ

Subject. The article presents clinical, biochemical, immunological and microbiological results of a complex of therapeutic measures of chronic gingivitis in patients with chronic gastroduodenitis.&#x0D; The aim of the study was to study the effectiveness of treatment of chronic gingivitis in patients with chronic gastroduodenitis with the use of the proposed method.&#x0D; Methodology. The subject of the study was 155 persons aged 19 to 25 years with diagnosed chronic gingivitis and chronic gastroduodenitis, which were divided by stratified randomization into three groups: 1 group (primary) - 52 patients; 2 group (positive control) - 52 patients; Group 3 (negative control) - 51 patients. All groups were comparable in age (F = 2,355, p = 0,092) and sex (χ2 = 2,133, p = 0,334).&#x0D; To study the state of free radical oxidation, the method of chemiluminescence (CL) of the oral liquid was applied on the device HL-003. The quantitative and qualitative content of oral microflora was determined using nutrient media: CHROMagar Orientation, CHROMagar Candida (France). The concentration in the oral fluid of the intercellular adhesion molecules sICAM-1 was carried out on the automatic enzyme immunoassay analyzer Lazurit and test systems manufactured by Vector-Best CJSC (Russia) and eBioscience (Austria).&#x0D; Results. The use of the complex proposed by us significantly improved the parameters of the clinical examination (dental indices). In addition, the state of free-radical oxidation has normalized: at previously increased values of the light-emission sum (S) and its maximum value (I max), during the time of the study, their decrease occurred, while the lowered values - the increase to the level of normal values. The level of microorganisms of the orange and red complex decreased to normal levels, and the level of sICAM-1 also significantly decreased.&#x0D; Conclusions. The scheme of treatment of chronic gingivitis with the use of NUTRIEL with laminaria and tooth elixir with chlorophyll in patients with chronic gastroduodenitis led to an increase in the duration of remission compared with the traditional treatment regimen.

Impact of selective media for detecting fluoroquinolone-insusceptible/extended-spectrum beta-lactamase-producing Escherichia coli before transrectal prostate biopsy.

To investigate the prevalence of fluoroquinolone-insusceptible and/or extended-spectrum beta-lactamase-producing Escherichia coli colonizing in the male rectum before transrectal prostate biopsy. We carried out a prospective cohort study of men undergoing transrectal prostate biopsy. CHROMagar Orientation originally supplemented with levofloxacin and CHROMagar Orientation/extended-spectrum beta-lactamase were used for detecting fluoroquinolone-insusceptible and extended-spectrum beta-lactamase-producing Escherichia coli. Rectal specimens were collected before prostate biopsy, and the results of cultures in the selective medium were compared with drug susceptibility measured by standard methods. Targeted prophylactic antimicrobials were administered to patients with drug-resistant Escherichia coli and the incidence of postoperative prostatitis was investigated. In the case of prostatitis, pathogens preoperatively isolated from the rectum and those from urine were compared using pulsed-field gel electrophoresis. Rectal colonization of fluoroquinolone-insusceptible or extended-spectrum beta-lactamase-producing Escherichia coli was detected in 217 of 694 (31.3%) and 85 of 640 (13.3%) participants, respectively. The sensitivity and specificity of fluoroquinolone-insusceptible selective media were 96.8% and 88.2%, respectively. A total of 618 participants underwent transrectal prostate biopsy, and postoperative acute prostatitis was observed in four of 618 (0.6%) participants. Escherichia coli strains isolated preoperatively from the rectum and postoperatively from urine were found to be identical. The present findings showed accuracy and performance of the selective media. Screening cultures before transrectal prostate biopsy using selective media seems to be helpful for guiding antibiotic prophylaxis and thus decreasing the rate of post-biopsy acute prostatitis.

Vendor effects on murine gut microbiota influence experimental abdominal sepsis

BackgroundExperimental animal models are indispensable components of preclinical sepsis research. Reproducible results highly rely on defined and invariant baseline conditions. Our hypothesis was that the murine gut microbiota varies among different distributors of laboratory animals and that these variations influence the phenotype of abdominal sepsis derived from a bacterial inoculum model (intraperitoneal stool injection). Materials and methodsMale C57BL/6 mice (8-wk old) purchased from Charles River (CR), Janvier (J), and Harlan (H) were sacrificed, and the bacterial composition of feces was analyzed using CHROMagar orientation medium. Stool was injected intraperitoneally into CR mice, followed by clinical observation and gene expression analysis. Experiments were repeated 16 mo later under the same conditions. ResultsStool analysis revealed profound intervendor differences in bacterial composition, mainly regarding Staphylococcus aureus and Bacillus licheniformis. Mice challenged with CR as well as H feces developed significantly higher severity of disease and died within the observation period, whereas stool from J mice did not induce any of these symptoms. Real-time polymerase chain reaction revealed corresponding results with significant upregulation of proinflammatory cytokines and vascular leakage-related mediators in CR and H injected animals. Sixteen months later, the bacterial fecal composition had significantly shifted. The differences in clinical phenotype of sepsis after intraperitoneal stool injection had vanished. ConclusionsWe are the first to demonstrate vendor and time effects on the murine fecal microbiota influencing sepsis models of intraabdominal stool contamination. The intestinal microbiota must be defined and standardized when designing and interpreting past and future studies using murine abdominal sepsis models.

Travelers Can Import Colistin-Resistant Enterobacteriaceae, Including Those Possessing the Plasmid-Mediated mcr-1 Gene.

Stool samples from 38 travelers returning from India were screened for extended-spectrum cephalosporin- and carbapenem-resistant Enterobacteriaceae implementing standard selective plates. Twenty-six (76.3%) people were colonized with CTX-M or DHA producers, but none of the strains was colistin resistant and/or mcr-1 positive. Nevertheless, using overnight enrichment and CHROMagar Orientation plates supplemented with colistin, four people (10.5%) were found to be colonized with colistin-resistant Escherichia coli One cephalosporin-susceptible sequence type 10 (ST10) strain carried a 4,211-bp ISApl1-mcr-1-ISApl1 element in an IncHI2 plasmid backbone.