High MIC Values Research Articles

Surveillance for azoles resistance in Aspergillus spp. highlights a high number of amphotericin B-resistant isolates.

Aspergillus spp. are the most common invasive mould infection and are responsible for high mortality. Aspergillus fumigatus is currently of interest because resistance to azole antifungals has emerged. The Campinas University Hospital (HC-UNICAMP) receives high-risk patients susceptible to opportunistic infections but there have been no reports of resistant A.fumigatus. This study aimed to assess the susceptibility profile of Aspergillus isolates, specifically looking for azole resistance. ITS and β-tubulin DNA sequencing was performed on 228 sequential clinical isolates. Broth microdilution susceptibility testing was performed for all isolates. A.fumigatus represented 74% of the isolates followed by Aspergillus flavus (12%). Nine A.fumigatus isolates from 9 different patients showed high MIC values to at least 1 azole, but cyp51A polymorphisms were detected in only 6 isolates and none correlated with known resistance mutations. The most troubling observation was that the minimum inhibitory concentration for amphotericin B was elevated (≥2mgL-1 ) in 87% of patients with A.flavus isolates and 43% with Aspergillusfumigatus isolates. Given that amphotericin B is used to treat azole-resistant infections, these data highlight the need for continuous surveillance in Aspergillus for all antifungal resistance to implement correct treatment strategies for the management of these pathogens.

Growth media and assay plate material can impact on the effectiveness of cationic biocides and antibiotics against different bacterial species

There are an increasing number of reports of bacterial strains that are multi-drug resistant. The use of biocides as part of infection control is crucial in helping to combat the spread of these particular strains. Unlike for antibiotics, there are few standardized measuring techniques to understand if an isolate has become more resistant to biocides. This study demonstrates the importance of media composition and plate material on variation and reporting of susceptibility of several bacterial species to specific cationic biocides. It is a useful comparison study to highlight the need to standardize biocide susceptibility testing.

Prevalence of carbapenem resistance genes and corresponding MIC90 in Enterobacteriaceae at a tertiary care center in Lebanon.

The aim of this study was to correlate genes involved in carbapenem resistance to MIC levels among clinical ESBL and non-ESBL producing carbapenem resistant Enterobacteriaceae (CRE) isolates of Escherichia coli and Klebsiella pneumoniae. E. coli (n = 76) and K. pneumoniae (n = 54), collected between July 2008 and July 2014, were analyzed. The MICs were determined against ertapenem (ERT), imipenem (IMP) and meropenem (MER). PCR was performed on all 130 isolates to amplify the resistance and outer membrane proteins (OMPs) encoding genes: blaOXA-48, blaNDM-1, blaTEM-1, blaCTX-M-15, ompC and ompF. Sequencing was performed on selected isolates. The prevalence of blaOXA-48, blaNDM-1, blaTEM-1, and/or blaCTX-M-15 among E. coli isolates were 36%, 12%, 20% and 80%, respectively, while among K. pneumoniae they were 37%, 28%, 28% and 72%, respectively. K. pneumoniae isolates positive for any of these genes had an MIC90 > 32μg/mL against ERT, IMP and MER, while in E. coli isolates there was a variation in the MIC90 values. Porin impermeabilities were due to mutations in ompC and ompF genes in E. coli, and loss of ompC and ompF genes in K. pneumoniae, and increased MIC90 values. The presence of more than one carbapenem resistance encoding gene and/or ESBL encoding gene did not have an effect on the MIC90 value in K. pneumoniae isolates, while in E. coli it showed higher MIC90 values.

Efficacy of different fungicides against the leaf spot of lettuce caused by Allophoma tropica

The efficacy of different fungicides, belonging to different chemical groups, was in vitro and in vivo tested in glasshouse trials against Allophoma tropica, the causal agent of a new leaf spot on lettuce. The in vitro trials showed that only a few fungicides were effective in inhibiting the mycelial growth of the four tested strains of A. tropica: the most effective fungicide, with the lowest ED50 and MIC values, was penconazole. Prochloraz, propiconazole, boscalid and fludioxonil showed low ED50 values, but high MIC values. The results from the in vivo trials showed that all the tested fungicides significantly reduced Phoma leaf spot incidence and severity, although differences were observed in the achieved disease pressure in the four trials. Generally, in the presence of an average disease severity in the untreated control, the best disease severity reduction (up to 90%) was provided 9 days after the application of fludioxonil, mancozeb, pyraclostrobin + boscalid, propiconazole and iprodione. Fludioxonil and iprodione showed a persistent effect (up to 90%) still being evident 12 days after the application. Generally, in the presence of a high disease severity in the untreated control, 9 days after the application of fludioxonil, mancozeb, iprodione and pyraclostrobin + boscalid provided a disease severity reduction up to 80%. Moreover, copper provided a disease severity reduction from 84 to 89%, applied as preventative treatment, resulting of special interest, particularly for crops grown under organic farming rules.

Candida infanticola and Candida spencermartinsiae yeasts: Possible emerging species in cancer patients

Opportunistic infections due to Candida species occur frequently in intensive care settings. We investigated the prevalence of Candida species among 65 clinical specimens obtained from 200 cancer patients by phenotypic and molecular (ITS sequencing and AFLP) methods. Among the 65 yeast isolates, Candida albicans was the most commonly isolated species (n = 34, 52.3%), whereas other Candida species comprised 47.7% (n = 31) and consisted of Candida glabrata (n = 14, 21.5%), Candida tropicalis (n = 5, 7.7%) and uncommon Candida species (n = 12, 18.5%) such as Candida pelliculosa (n = 3, 4.6%), Pichia kudriavzevii (= Candida krusei, n = 2, 3.1%), Candida orthopsilosis (n = 2, 3.1%), Candida parapsilosis (n = 1, 1.5%), Candida infanticola (n = 2, 3.1%), Candida spencermartinsiae (n = 1, 1.5%), and Kluyveromyces marxianus (=Candida kefyr, n = 1, 1.5%). Candida infanticola and Candida spencermartinsiae were recovered from oral lesions of cancer patients. Matrix assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) easily confirmed these isolates as less common Candida isolates (4.6%). The in vitro antifungal susceptibilities of C. spencermartinsiae and the two strains of C. infanticola were determined according to CLSI guidelines (M27-A3). MIC results among these isolates showed they were susceptible to isavuconazole, posaconazole and voriconazole, however, fluconazole and caspofungin had high MIC values. These Candida species that may occur more commonly in infections remain unnoticed using commonly used phenotypical methods in routine microbiology laboratories. MALDI-TOF MS proved to be a more fast and robust diagnostic technique for identification of the yeasts isolated from different clinical specimens of cancer patients.

Antibiotic susceptibility profiles of Mycoplasma synoviae strains originating from Central and Eastern Europe

BackgroundMycoplasma synoviae causes infectious synovitis and respiratory diseases in chickens and turkeys and may lead to egg shell apex abnormalities in chickens; hence possesses high economic impact on the poultry industry. Control of the disease consists of eradication, vaccination or medication. The aim of the present study was to determine the in vitro susceptibility to 14 different antibiotics and an antibiotic combination of M. synoviae strains originating from Hungary and other countries of Central and Eastern Europe.ResultsMinimal inhibitory concentration (MIC) values of a total of 41 M. synoviae strains were determined by the microbroth dilution method. The strains were collected between 2002 and 2016 and originated from Hungary (n = 26), Austria (n = 3), the Czech Republic (n = 3), Slovenia (n = 3), Ukraine (n = 3), Russia (n = 2) and Serbia (n = 1). Tetracyclines (with MIC50 values of 0.078 μg/ml, ≤0.25 μg/ml and 0.5 μg/ml for doxycycline, oxytetracycline and chlortetracycline, respectively), macrolides (with MIC50 values of ≤0.25 μg/ml for tylvalosin, tylosin and tilmicosin), pleuromutilins (with MIC50 values of 0.078 μg/ml and ≤0.039 μg/ml for tiamulin and valnemulin) and the combination of lincomycin and spectinomycin (MIC50 1 μg/ml (0.333/0.667 μg/ml)) were found to be the most effective antibiotic agents against M. synoviae in vitro. High MIC values were detected in numerous strains for fluoroquinolones (with MIC50 values of 1.25 μg/ml and 2.5 μg/ml for enrofloxacin and difloxacin), neomycin (MIC50 32 μg/ml), spectinomycin (MIC50 2 μg/ml), lincomycin (MIC50 0.5 μg/ml) and florfenicol (MIC50 4 μg/ml). Nevertheless, strains with elevated MIC values were detected for most of the applied antibiotics.ConclusionsIn the medical control of M. synoviae infections the preliminary in vitro antibiotic susceptibility testing and the careful evaluation of the data are crucial. Based on the in vitro examinations doxycycline, oxytetracycline, tylvalosin, tylosin and pleuromutilins could be recommended for the therapy of M. synoviae infections in the region.

Predominance of Serogroup 19 CC320/271 among Penicillin-Nonsusceptible Streptococcus pneumoniae Isolates after Introduction of the PCV7 Vaccine in Several Regions of Japan.

Multidrug-resistant Streptococcus pneumoniae serogroup 19, including serotypes 19A and 19F, associated with clonal complex 320/271 (CC320/271), has been previously shown to be predominant in many countries after introduction of a 7-valent pneumococcal conjugate vaccine (PCV7). However, in Japan there has been no epidemiological research focused on penicillin-nonsusceptible isolates after this event. Therefore, we aimed to characterize penicillin-nonsusceptible S. pneumoniae (PNSSP; penicillin minimum inhibitory concentration [MIC] ≥ 4.0 μg/ml) after the introduction of PCV7 in Japan. Throughout Japan, we collected 1,057 pneumococcal isolates from 2010 to 2014. We then evaluated MICs and performed serotyping, multilocus sequence typing, and sequencing of penicillin-binding protein genes in 51 isolates (penicillin MIC ≥ 2.0 μg/ml). Twenty-three isolates (2.2%) showed penicillin nonsusceptibility (penicillin MIC ≥ 4.0 μg/ml). Serotypes 19F (14 isolates, 60.9%) and 23F (4 isolates, 17.4%), which are covered by the vaccine, were predominant among PNSSP strains. Only 3 isolates belonged to nonvaccine serotype 19A. Among the PNSSP isolates, CC320/271 (16/23 strains, 69.6%) was the most prevalent clone. Moreover, CC320/271 clones showed high MIC values of a third-generation cephalosporin. Thus, we demonstrated clonal predominance of serogroup 19 CC320/271 with strong resistance to β-lactams including a third-generation cephalosporin among PNSSP isolates.

Antibacterial Activity of Nanoparticles Produced by Ethanol Extract of Boesenbergia rotunda Rhizome Loaded with Chitosan and Alginic Acid Against Streptococcus mutans by In Vitro

The antibacterial activity of three Cupressaceae plants (Thujaoccidentalis,ThujaorientalisandChamaecyparisobtusa) was tested against three bacteria using the agar diffusion method. The ether and ethylacetate fraction of crude methanol extract from the three plants showed potent antibacterial activity against the tested microorganisms. The result showed that Staphylococcus aureus revealed the most sensitivity among the tested bacteria. Thujaoccidentalisether fraction and Thujaorientalis hexane fraction exhibited the highest antibacterial activity against Staphylococcus aureus. E. coli was shown the highest MIC values compared to the other two tested bacteria, which indicates the lowest antibacterial activity against the bacterium. This study promises an interesting future for designing a potentially active antibacterial agent from the three Cupressaceae plants.

Kinetics and mechanisms of mercury biosorption by an exopolysaccharide producing marine isolate Bacillus licheniformis.

Eight exopolysaccharide (EPS) producing metal-removing marine bacteria were screened for mercury (Hg) sorption. Bacillus licheniformis with the highest MIC values and Hg sorption ability was selected for further study. Biosorption of Hg from aqueous solution by Bacillus licheniformis was studied with respect to the metal concentration, adsorbent concentration, pH, different contact times, and in the presence of other metal ions. Under optimum conditions, more than 70% mercury was removed by 25mg dried biomass of Bacillus licheniformis at pH 7.0 after 1h of contact time. Freundlich adsorption isotherm was acceptable at studied Hg concentrations as compared to Langmuir isotherm model. Pseudo-second-order kinetic model was found to be more suitable for data presentation in contrast to pseudo-first-order kinetic model. Involvement of external mass transfer was prominent as compared to intraparticle diffusion model. Desorption of Hg was more effective with acids from all the studied eluents, showing 49.36 and 33.8% eluting capacity for 0.1N HCL and 0.1N HNO3, respectively. Scanning electron microscopy exhibited altered cell surface morphology of the cells under the influence of mercury. The spectral images of energy dispersive spectroscopy showed the presence of metal ions on the surface of cells.

Anticandidal activity of ethanolic root extract of Juglans regia (L.): Effect on growth, cell morphology, and key virulence factors

The emergence of drug-resistant strains has encouraged several studies on natural products with antifungal activity and low toxicity.In this study, the antifungal effect of methanolic root extract of Juglans regia (JRE) was investigated against 9 strains of Candida (one reference and 8 clinical strains) through MIC90 and spot assays. To gain insight into the mechanism of antifungal action, we carried out confocal scanning laser microscopy (CLSM), transmission electron microscopy (TEM), and then examined the effect of JRE on hydrolytic enzyme secretion. Additionally, JRE was subjected to various phytochemical tests, chemically characterized by gas chromatography-mass spectrometry analysis (GC-MS) and its toxicity was tested against H9c2 rat cardiac myoblasts. The phytochemical tests showed the presence of phenols, alkaloids, steroids, saponins, and tannins in JRE. In the GC-MS analysis, a total of 40 compounds were identified. JRE was found to be effective in liquid media with MICs ranging from 300 to 700μg/mL. Spot assay results revealed that Candida cells show increased sensitivity to JRE. CSLM experiments showed that cells exposed to JRE (MIC) exhibited cell membrane disruption. TEM micrograph of treated cells showed extensive breakage in the cell wall and cell membrane. Average inhibition of proteinase and phospholipase secretion (of five C.albicans strains) at MIC/2 values of JRE was 45.17%, and 34.29%, respectively. Cellular toxicity of JRE against H9c2 rat cardiac myoblasts was less than 10% at the highest MIC value. These findings encourage further development of JRE.

Characterization of Corynebacterium diphtheriae, C. confusum and C. amycolatum isolated from sows with genitourinary infection

Porcine Corynebacterium infection is still poorly studied, even though the pig has been described as an asymptomatic carrier of Corynebacterium species, including the zoonotic species C. ulcerans, C. confusum and C. amycolatum. Here we present the identification, molecular and antimicrobial susceptibility characterization of coryneform bacteria isolated from sows with urinary tract infection. C. diphtheriae, C. confusum and C. amycolatum were isolated from sows with urinary infection and metritis. Corynebacterium species were identified by MALDI-TOF MS and confirmed by 16S rRNA and rpoB sequencing. All porcine C. diphtheriae strains were further characterized as non-toxigenic (tox−). SE-AFLP genotyping was also performed and enabled not only Corynebacterium species differentiation but also the assessment of C. amycolatum genetic heterogeneity. All studied porcine Corynebacterium strains presented alarming resistance profiles with high MIC values for macrolides/lincosamide, tetracyclines and quinolones, which can be related with high usage in both veterinary and human medicine. Isolation of zoonotic Corynebacterium species from commercial swine is important for assessing the potential zoonotic risk for farmers and further implication for both human and animal treatment.

Antimicrobial Activities of Daucus Carota Seeds on Selected Pathogenic Micro-organisms

Background: The antibacterial/antifungal toxicity of Daucus carota (carrot) seeds was evaluated using selected multi-drug resistant bacteria and yeast of clinical origin. Methods: The active constituents of the Daucus carota seeds were extracted using conventional Plant Tissue Homogenization method using cold distilled water, Ethanol and Methanol as solvents. Varying concentrations (5-250 mg/ml) of the three extracts were assayed for antimicrobial activity against the selected isolates- Salmonella typhi, Staphylococcus aureus, Escherichia coli, Klebseilla pneumoniae and Candida albicans; the agar well diffusion method was used. The antibiogram profile of the organisms was also obtained through disc diffusion method. Results: Similar activity was observed in the methanolic and ethanolic extracts while cold distilled water showed no activity on any of the isolates. The antibiotic susceptibility results showed that the isolates used are highly multi-drug resistant. Ofloxacin exhibited the most pronounced activity against all the isolates. Gentamicin and erythromycin both showed activity on Escherichia coli and Salmonella typhi. Lower concentrations of both extracts presented no inhibitory effects on the test organisms, thus resulting in high MIC values recorded for both extracts. Also, the extracts showed no bactericidal action against the isolates. Conclusions: Observations from this research therefore affirm that Daucus carota seeds possess antimicrobial properties that may be explored as a source of future antimicrobial compounds.

Chemical Composition, Antimicrobial and Antioxidant Activities of Essential Oils from Two Avicennia schaueriana Stapf & Leechm. Ex Moldenke (Acanthaceae) Populations.

Background: Avicennia schaueriana Stapf & Leechm. ex Moldenke (Acanthaceae) is a native species from the Brazilian mangroves presenting ecological and economic significance. This study compared the composition and the biological activities from the essential oils obtained from two A. schaueriana populations collected at Jureia-Itatins and Ilha do Cardoso. Methods: Essential oils were obtained by conventional means, and their compositions were analyzed by GC-MS. Screening assays for antimicrobial activity were carried out by the microdilution method and the antioxidant potential was assessed by the DPPH scavenging method. Results: The GC-MS analysis indicated that the Jureia oil (1) was composed mostly of the fatty acids palmitic (46.5%) and myristic (11.6%) acids, while the main components for the Ilha do Cardoso oil (2) were eugenol (19.7%), eugenol acetate (12.9%) and palmitic acid (15.1%). The oils showed an IC50 of 0.9 ± 0.011 mg/mL for 1 and 1.13 ± 0.028 mg/mL for 2 in the DPPH assay. The antimicrobial assay indicated MIC > 217 µg/mL for all tested microorganisms. Conclusions: The different essential oil composition may indicate the presence of chemotypes for A. schaueriana. The antioxidant activity of the oils was weak if compared with flavonoids. Despite the high MIC values, these oils presented some antibacterial potential against Pseudomonas aeruginosa.

In vitro susceptibility of methicillin-resistant Staphylococcus aureus isolates from skin and soft tissue infections to vancomycin, daptomycin, linezolid and tedizolid

IntroductionTreatment of multidrug-resistant Gram-positive infections caused by Staphylococcus aureus remains as a clinical challenge due to emergence of new resistance mechanisms. Tedizolid is a next-generation oxazolidinone, recently approved for skin and soft tissues infections. We conducted a study to determine in vitro susceptibility to vancomycin, daptomycin, linezolid and tedizolid in MRSA clinical isolates from adult patients with skin and soft tissue infections. Material and methodsMethicillin-resistant S. aureus isolates were collected in three tertiary-care hospitals of Medellin, Colombia, from February 2008 to June 2010 as part of a previous study. Clinical characteristics were assessed by medical records and MIC values were determined by Epsilometer test. Genotypic analysis included spa typing, MLST, and SCCmec typing. ResultsA total of 150 MRSA isolates were evaluated and tedizolid MIC values obtained showed higher in vitro activity than other antimicrobials, with MIC values ranging from 0.13μg/mL to 0.75μg/mL and lower values of MIC50 and MIC90 (0.38μg/mL and 0.5μg/mL). In contrast, vancomycin and linezolid had higher MIC values, which ranged from 0.5μg/mL to 2.0μg/mL and from 0.38μg/mL to 4.0μg/mL, respectively. Tedizolid MICs were 2- to 5-fold lower than those of linezolid. Clinical characteristics showed high previous antimicrobial use and hospitalization history. The majority of the strains belong to the CC8 harboring the SCCmec IVc and were associated with the spa t1610 (29.33%, n=44). ConclusionIn vitro effectiveness of tedizolid was superior for isolates from skin and soft tissue infections in comparison with the other antibiotics evaluated. The above added to its less toxicity, good bioavailability, daily dose and unnecessity of dosage adjustment, make tedizolid in a promising alternative for the treatment of infections caused by MRSA.

A Probable Case of Scrub Typhus Meningoencephalitis – In the Era of Dual Seropositivity and Possible Emerging Doxycycline Resistance - Are Dictums Changing?

Scrub typhus is an acute febrile illness which has varied manifestation from febrile illness to a more severe multi-organ dysfunction including CNS involvement in the form of meningitis and meningoencephalitis and in extremes cases, septic shock and death. Scrub typhus is prevalent in India and there have been reported case series from north-eastern regions of India and also from various institutes of south India. In sharp contrast, our case is a single incidence and classically showed features of lung involvement which predisposes to CNS involvement (in the form of meningitis or meningoencephalitis) later. All the published case reports/series from the subcontinent, it is unanimously stated that response is dramatic upon starting of oral doxycycline and the clinical scenario changes within 48- 72 hours. However in this anecdotal case of ours, intravenous doxycycline was started and still patient went downhill before he stabilised after 10 days of the therapy. Although doxycycline failures have been reported from Korea and the medical scientists have put forward their explanations about the questionable emerging resistance to doxycycline, still this case remains important because it is one of the first reported cases from the Indian subcontinent where the clinical features deteriorated even after starting Doxycycline (The erstwhile gold standard antibiotic against Scrub Typhus) and that too via intravenous route. The case presents a dilemma in the minds of the clinician as to what strategy may be taken to achieve higher MIC values of Doxycycline (to overcome the emerging resistance) and also triggers the search for new antibiotic regimen which can help us to control scrub typhus early and prevent the more severe and life threatening complications of the disease.

High vancomycin MICs predict the development of infective endocarditis in patients with catheter-related bacteraemia due to methicillin-resistant Staphylococcus aureus.

It has been suggested that there is an increased risk of treatment failure in episodes of MRSA bloodstream infection (BSI) caused by strains with high vancomycin MICs. However, it is unknown if this phenomenon may also act as a risk factor for the development of infective endocarditis (IE). We analysed 207 episodes of catheter-related (CR)-BSI recruited from June 2008 to December 2009 within a prospective study on MRSA BSI in 21 Spanish hospitals. Vancomycin susceptibility was centrally tested. The impact of high vancomycin MIC values (≥1.5 mg/L by Etest) on the subsequent development of IE was investigated by Cox regression. High vancomycin MIC values were observed in 46.9% of the isolates. Initial therapy consisted of vancomycin [99 episodes (44.7%)], daptomycin [25 (12.1%)], linezolid [18 (8.7%)] and other antistaphylococcal agents [16 (7.7%)]. Haematogenous complications occurred in 41 patients (19.8%), including 10 episodes complicated by IE. Early (48 h) and late (30 day) all-cause mortality were 3.4% and 25.1%, respectively. High vancomycin MIC isolates were more common among patients that developed IE compared with those free from this complication [90.9% (9/10) versus 44.7% (88/197); P = 0.007]. This association remained significant after adjusting for multiple confounders (including initial antibiotic therapy and catheter removal) in different models (minimum hazard ratio: 9.18; 95% CI: 1.16-72.78; P = 0.036). There were no differences in mortality according to vancomycin MIC values. Decreased susceptibility to vancomycin acted as a predictor of the development of IE complicating MRSA CR-BSI.

Molecular Characterization of Macrolide Resistance Genes among the Clinical Isolates of Enterococci.

Enterococci have emerged as an important pathogen due to their intrinsic and acquired resistance to several antibiotics. Macrolides were used to treat most of the gram positive infections. However, recently there is an increase in acquired resistance towards these antibiotics which leads to therapeutic challenge. This study was carried out to screen clinical isolates of enterococci for erythromycin resistance phenotypes and molecular characterization of macrolide resistance genes. A total of 97 enterococcal isolates were included, among which 60.8% of isolates were identified as E. faecium followed by 30.9% of E .faecalis. 75.2% (70/97) were identified as MDR isolates. 3.09% were iMLSB, 55.6% were cMLSB and 38.8% isolates neither belong to iMLSB/cMSLB phenotypes when tested by double disk diffusion method. The MIC50 and MIC90 are > 256 µg/ml concentration. Molecular analysis of ermgenes revealed that 48.45% isolates were positive for erm B gene while other tested genes such aserm A, erm C and mef A genes were not amplified by PCR. In our study, the predominant species identified was E. faecium. Constitutive erythromycin resistance was higher and only erm B gene was amplified among our isolates. Relatively, higher percentage of erythromycin resistance with higher MIC value was observed among our isolates.

Molecular and antimicrobial susceptibility characterization of Globicatella sulfidifaciens isolated from sow's urinary tract infection

ABSTRACTBackground: The Globicatella genus comprises Gram-positive, facultative anaerobic, α-hemolytic and catalase negative cocci morphologically and phenotypically very similar to Streptococcus and Aerococcus genus which can lead to misidentification and underestimation of this pathogen. Globicatella species have already been isolated from human and animals with heart and brain disorders. Their clinical relevance in animals, and its zoonotic potential, remains unknown due to the difficulty in their identification.Objective: To present the isolation, phenotypic and molecular characterization of G. sulfidifaciens from urinary tract infection in sows.Materials and Methods: Urine samples from 140 sows of two swine herds located in São Paulo State (Brazil) yielded the isolation of three presumptive G. sulfidifaciens strains. Identification and species confirmation were done by MALDI-TOF MS and 16S rRNA sequencing. Strains were further characterized by single enzyme amplified fragments length polymorphism (SE-AFLP) and broth microdilution techniques.Results: All three isolates were confirmed as G. sulfidifaciens. The SE-AFLP genotyping resulted in distinct fingerprint patterns for each strain. All isolates presented high MIC values to tetracycline, sulphonamides, aminoglycosides and tylosin tartrate, which present high usage in human and animal medicine.Conclusions: Globicatella sulfidifaciens could be related to sporadic urinary tract infections in swine and appear to present alarming antimicrobial susceptibility profile. It is necessary to differentiate Streptococcus-like microorganisms in routine laboratory diagnostics for the correct identification of underestimated species potentially pathogenic to animals.

Correlation Between qacE and qacE∆1 Efflux Pump Genes, Antibiotic and Disinfectant Resistant Among Clinical Isolates of E.coli.

Antiseptics and disinfectants have been used widely in hospitals and other health care settings to control the growth of microorganisms. However, some disinfectant resistant strains were reported. The objectives of our study were to evaluate correlation between the efflux pump genes, drugs and disinfectant resistant among clinical isolates of E.coli. A total of 102 of E. coli strains were isolated from urine sample of hospitalized patients. The antibiotic susceptibility was carried out by disc diffusion method. Didecyl di-methyl ammonium chloride (DDDMAC) was used as Quaternary ammonium compound (QAC) disinfectant which was used in Heart Center Hospital. PCR reaction was carried out for detection of qacE and qac∆E efflux pump genes. Almost all the strains had higher resistance to ampicillin, ciproflaxacin, cotrimaxazole and cephalothin. Totally 49% (n: 50) of strains were produced ESBL. Almost all the strains have MIC value between 0.00195 to 0.0078 mg/l for DDDMAC. Correlation between presence of qacE and qac∆E genes and antibiotic resistance was perceived. Presence of qacE and qac∆E genes among strains that have high disinfectant MIC value were 96.9% and 93.7% respectively. In addition, 98% of ESBL producing strains harbored qacE gene and 94% of ESBL producing strains harbored qac∆E gene. Our study indicated that there was a strong correlation between presence of qacE and qac∆E genes with resistance to some antibiotics and growth in media which contain high concentration of disinfectant. In conclusion, other mechanisms also play important role in resistant to antimicrobial agents but the role of efflux pumps in resistant to antimicrobial agents should not be neglected.

Surveillance of Helicobacter pylori Antibiotic Susceptibility in Indonesia: Different Resistance Types among Regions and with Novel Genetic Mutations.

Information regarding Helicobacter pylori antibiotic resistance in Indonesia was previously inadequate. We assessed antibiotic susceptibility for H. pylori in Indonesia, and determined the association between virulence genes or genetic mutations and antibiotic resistance. We recruited 849 dyspeptic patients who underwent endoscopy in 11 cities in Indonesia. E-test was used to determine the minimum inhibitory concentration of five antibiotics. PCR-based sequencing assessed mutations in 23S rRNA, rdxA, gyrA, gyrB, and virulence genes. Next generation sequencing was used to obtain full-length sequences of 23S rRNA, infB, and rpl22. We cultured 77 strains and identified 9.1% with clarithromycin resistance. Low prevalence was also found for amoxicillin and tetracycline resistance (5.2% and 2.6%, respectively). In contrast, high resistance rates to metronidazole (46.7%) and levofloxacin (31.2%) were demonstrated. Strains isolated from Sumatera Island had significantly higher metronidazole resistance than those from other locations. Metronidazole resistant strains had highly distributed rdxA amino acid substitutions and the 23S rRNA A2143G mutation was associated with clarithromycin resistance (42.9%). However, one strain with the highest MIC value had a novel mutation in rpl22 without an A2143G mutation. Mutation at Asn-87 and/or Asp-91 of gyrA was associated with levofloxacin-resistance and was related to gyrB mutations. In conclusions, although this is a pilot study for a larger survey, our current data show that Indonesian strains had the high prevalence of metronidazole and levofloxacin resistance with low prevalence of clarithromycin, amoxicillin, and tetracycline resistance. Nevertheless, clarithromycin- or metronidazole-based triple therapy should be administered with caution in some regions of Indonesia.