Threefold Increase in the Number of Drug Resistant TB Cases after Introduction of Universal Drug Susceptibility Testing: Experiences from Two South India Districts

Background: In India, tuberculosis (TB) is a major public health problem, and the advent of drug resistance TB (DR-TB) has worsened the situation. The Revised National TB Control Programme (RNTCP) has introduced universal drug susceptibility testing (UDST) for all diagnosed TB cases in 2018. We conducted this study to know the advantage of implementing UDST when compared to selective testing existent in 2017 on key diagnostic cascade parameters and to identify the challenges in the implementation of UDST. Methods: The study was conducted in two districts of Karnataka, India during January 2017-December 2018. The quantitative part consisted of before-and-after design and the qualitative part consisted of descriptive design. Results: In 2017 (during selective testing/“before” period) out of the 2440 TB patients, 80 (3%) were diagnosed with Isoniazid and Rifampicin resistance patients;in contrast in 2018 (during UDST/“after” period) of the 5129 TB patients 258 (5%) were diagnosed with Isoniazid and Rifampicin resistance. However, the proportion of eligible patients tested for rifampicin resistance during the “after” period was 60% when compared to 100% during the “before” period and median turnaround time for testing was also longer during the “after” period when compared to the “before” period (32.5 days vs 27.5 days). Major reasons for these two gaps were found to be difficulties in collecting sputum specimens and transportation. Conclusion: The rollout of UDST has led to a three-fold increase in a number of DR-TB cases detected in the region. There is a need for the programme to increase the proportion tested for DST by increasing the laboratory capacity and address the challenges in sputum collection and transportation.

Importance of antimicrobial susceptibility testing for the management of eradication in Helicobacter pylori infection

The management of Helicobacter pylori(H. pylori) infection treatment differs from the common treatment protocol for other infectious diseases. Because culture-or molecular-guided approaches face several practical issues, such as the invasive procedures required to obtain gastric biopsy specimens and the lack of availability of routine laboratory testing in some places, H. pylori treatment includes the administration of two or three empirically selected antibiotics combined with a proton pump inhibitor rather than evidence-based eradication treatment. The efficacy of empirical therapy is decreasing, mostly due to increasing multiple resistance. Multiresistance to levofloxacin, clarithromycin, and metronidazole, which are commonly used in empirical treatments, appears to have increased in many countries. Mutations play a primary role in the antimicrobial resistance of H. pylori, but many different mechanisms can be involved in the development of antibiotic resistance. Determining and understanding these possible mechanisms might allow the development of new methods for the detection of H. pylori and the determination of antimicrobial resistance. A treatment based on the detection of antimicrobial resistance is usually more effective than empirical treatment. Nevertheless, such an approach before treatment is still not recommended in the Maastricht guidelines due to the difficulty associated with the routine application of available cultureor molecular-based susceptibility tests, which are usually administered in cases of treatment failure. The management of first and rescue treatments requires further research due to the steadily increase in antimicrobial resistance.

Expert Consensus on Polymyxin Antimicrobial Susceptibility Testing and Clinical Interpretation

The polymyxins are important antimicrobial agents against antibiotic-resistant gram-negative bacilli.In 2020,the Clinical and Laboratory Standards Institute modified the clinical breakpoints for polymyxin susceptibility test by eliminating the"susceptible"interpretive category,only reporting intermediate(≤2 mg/L)and resistant(≥4 mg/L).However,the European Committee on Antimicrobial Susceptibility Testing recommended the use of clinical breakpoints of W2 mg/L as susceptible and>2 mg/L as resistant.The first-line laboratorians and clinicians in China have been perplexed by the inconsistence of international polymyxin clinical breakpoints and discouraged by the difficulty of conducting polymyxin susceptibility testing.Therefore,it is urgently needed to make it clear for the laboratorians in China to know how to accurately carry out polymyxin susceptibility testing and standardize the interpretation of susceptibility testing results.To this end,the experts from relevant fields were convened to formulate this consensus statement on the testing and clinical interpretation of polymyxin susceptibility.Relevant recommendations are proposed accordingly for laboratorians and clinicians to streamline their daily work.

Detection of Beta-Lactamase and Extended-Spectrum Beta-Lactamase of Pathogens Isolated from Pig and Chicken and Their Antibiotic Susceptibility Test

The antibacterial activity of beta-lactam antibiotics or their combinations with inhibitor sulbactum against non-lactamase- producing strains, lactamase-producing and ESBLs-producing isolates was evaluated with twofold dilution method after pathogens isolated from pigs and chickens were detected, respectively, for beta-lactamase and extended-spectrum beta- lactamases （ESBLs）, The results revealed that most of 43 clinically isolated strains could produce beta-lactamase and 3 strains of shigella isolated from chicken samples produced ESBLs. All of 30 lactamase-producing strains isolated and only one of 16 non-lactamase-producing strains were resistant to amoxicillin and ampicillin. MICs of ampicillin against lactamaseproducing isolates decreased 10-40 and 10-20 times respectively, when it was conbined with sulbactam at ration of 1：2 and 1：4. All clinical isolates were susceptible to third-generation cephalosporins. The MICs of third-generation cephalosporins against lactamase-producing isolates did not change when they were conbined with sulbactam. MICs of ceftiofur and ceftriaxone against ESBLs-producing isolates decreased 2-4 times when they were conbined with sulbactam.

Isolation,Identification and Antibiotics Susceptibility Test of Citrobacter freundii from Procambarus clarkia

This experiment was conducted to clarify species and drug resistance of pathogen from the diseased Procambarus clarkia. Pathogenic bacteria from hepatopancreas of the diseased P. clarkia were examined using conventional methods,and then were isolated. The further tests and analysis of the isolated strain were developed,including the regression experiment to P. clarkia,the morphology,physiological and biochemical characteristics,sequence analysis of their 16 S rRNA and gyr B genes,and the susceptibility test to antibiotics. Large colonies with similar morphology and color were obtained. Strain X120523 was identified as Citrobacter freundii,proved to have strong pathogenicity,and was susceptible to quinolones and aminoglycosides.

Drug Susceptibility Test of Volatile Oil of Artemisiaargyi to Avian Pathogenic Escherichia coli

The volatile oil of Artemisia argyi was extracted by ultrasonic assisted extraction, and the extraction rate of volatile oil was 0.68%. Thevolatile oil of A. argyi was emulsified with 1% Tween-80, and drug susceptibility test was conducted with avian Escherichia coli. The results showedthat the volatile oil of A. argyi had antibacterial effect against avian E. coli, and the minimal inhibitory concentration was 50 mg/mL. Taking sixcommon antibiotics as the control, drug susceptibility test was conducted with volatile oil of A. argyi. The results showed that 10 strains of E. coliwere sensitive to the volatile oil of A. argyi, three of which had different degrees of resistance and one had the tendency of resistance.

Assessment of Clinical Presentation, Performance of Diagnostic Methods and Antibiotic Susceptibility Testing for Salmonella among Patients Attending Kangema Sub-County Hospital, Kenya

Background: Typhoid disease remains a major public health problem globally, especially in developing countries in sub-Saharan Africa. Symptoms associated with typhoid disease mimic those of other febrile illnesses and are thus difficult to make an accurate diagnosis. A confirmed diagnosis requires the determination or isolation of the bacteria in well-equipped laboratories. Developing countries are faced with a huge limitation of the laboratory infrastructure to diagnose typhoid disease, which would otherwise guide in treating, managing, controlling, and halting the spread of drug resistant mutants. Objective: This study, therefore, was aimed at determining the clinical presentation, performance of diagnostic tests and antibiotic susceptibility testing of Salmonella among adults attending Kangema Sub-County Hospital. Study Population: The study population was residents of Kangema Sub-County in Murang’a County, Kenya while the target population was adults. Methods: The study adopted a cross-sectional study design that employed a systematic random sampling procedure. The study took place between April and June 2021. The sample size was 97 respondents who all consented and were enrolled in the study. Interviewing the respondents was carried out by administering structured questionnaires to collect quantitative data. Stool samples were obtained and cultured in Cary Blair transport media and then cultured in appropriate media at the Murang’a County Referral Hospital Laboratory. A rapid Salmonella Antigen (SAT) test was also performed on all the stool samples. Data Analyses: Word Statistics and Data (STATA) v 13 was used for statistical analysis. Results: The prevalence of Typhoid Fever was at 6.2% (95% CI) which included S. Typhi (n = 1;16.7%) and S. Paratyphi B (n = 5;83.3%). No isolate showed resistance to Ciprofloxacin. The sensitivity of SAT is 100% and a specificity of 98.9% with a kappa statistic of almost perfect agreement (0.9641) with culture. Patients who had fever p = 0.001, abdominal distention p = 0.028, diarrhoea p = 0.038, loose or watery stool p = 0.021 and mild general condition p = 0.02 remained independently associated with Salmonella infection. Conclusion: Typhoid Fever being endemic, laboratory diagnosis was a key for confirmation after clinical diagnosis. SAT can accurately be used to detect the disease where culture is unavailable. However, antibiotic sensitivity tests were crucial when determining the drug of choice as Salmonella isolates were multi-drug resistant. Establishment of prescribing antimicrobial policies and guidelines can periodically monitor the antibiogram patterns.

Performance of the microscopic observation drug susceptibility assay in pyrazinamide susceptibility testing for Mycobacterium tuberculosis

Background Drug susceptibility assay is very important in tuberculosis therapy. Pyrazinamide is a first line antituberculosis drug and diagnosis of its resistance in Mycobacterium tuberculosis （M. tuberculosis） is difficult and time consuming by conventional methods. In this study, we aimed to evaluate the performance of the microscopic observation drug susceptibility （MODS） assay in the detection of pyrazinamide resistance in M. tuberculosis relative to the conventional Wayne assay and Lowenstein-Jensen （L J） proportion method. Methods M. tuberculosis clinical isolates （n=132） were tested by the MODS and the Wayne assay： the results were compared with those obtained by the LJ proportion method. Mutations in the gene were identified by direct sequencing of the pncA genes of all isolates in which pyrazinamide resistance was detected by any of the three methods. Results Compared to the LJ results, the sensitivity and specificity of the MODS assay were 97.8% and 96.5% respectively; the sensitivity and specificity of the Wayne assay were 87.0% and 97.7% respectively. Mutations in the pncA gene were found in 41 of 46 strains that were pyrazinamide resistant （3 tests）, in 1 of the 4 strains （LJ only）, in 42 of 48 strains （at least I test）, but no mutations in 1 strain sensitive according to the MODS assay only. The MODS assay, Wayne assay and LJ proportion method provided results in a median time of 6, 7 and 26 days respectively. Conclusions MODS assay offers a rapid, simple and reliable method for the detection of pyrazinamide resistance in M. tuberculosis and is an optimal alternative method in resource limited countries.

Rapid, automated, and reliable antimicrobial susceptibility test from positive blood culture by CAST‐R

Antimicrobial susceptibility tests(ASTs)are pivotal in combating multidrug resistant pathogens,yet they can be time‐consuming,labor‐intensive,and unstable.Using the AST of tigecycline for sepsis as the main model,here we establish an automated system of Clinical Antimicrobials Susceptibility Test Ramanometry(CAST‐R),based on D2O‐probed Raman microspectroscopy.Featuring a liquid robot for sample pretreatment and a machine learning‐based control scheme for data acquisition and quality control,the 3‐h,automated CAST‐R process accelerates AST by>10‐fold,processes 96 paralleled antibiotic‐exposure reactions,and produces high‐quality Raman spectra.The Expedited Minimal Inhibitory Concentration via Metabolic Activity is proposed as a quantitative and broadly applicable parameter for metabolism‐based AST,which shows 99%essential agreement and 93%categorical agreement with the broth microdilution method(BMD)when tested on 100 Acinetobacter baumannii isolates.Further tests on 26 clinically positive blood samples for eight antimicrobials,including tigecycline,meropenem,ceftazidime,ampicillin/sulbactam,oxacillin,clindamycin,vancomycin,and levofloxacin reveal 93%categorical agreement with BMD‐based results.The automation,speed,reliability,and general applicability of CAST‐R suggest its potential utility for guiding the clinical administration of antimicrobials.

Intraprocedural gastric juice analysis as compared to rapid urease test for real-time detection of Helicobacter pylori

BACKGROUND Endofaster is an innovative technology that can be combined with upper gastrointestinal endoscopy(UGE)to perform gastric juice analysis and real-time detection of Helicobacter pylori(H.pylori).AIM To assess the diagnostic performance of this technology and its impact on the management of H.pylori in the real-life clinical setting.METHODS Patients undergoing routine UGE were prospectively recruited.Biopsies were taken to assess gastric histology according to the updated Sydney system and for rapid urease test(RUT).Gastric juice sampling and analysis was performed using the Endofaster,and the diagnosis of H.pylori was based on real-time ammonium measurements.Histological detection of H.pylori served as the diagnostic gold standard for comparing Endofaster-based H.pylori diagnosis with RUT-based H.pylori detection.RESULTS A total of 198 patients were prospectively enrolled in an H.pylori diagnostic study by Endofasterbased gastric juice analysis(EGJA)during the UGE.Biopsies for RUT and histological assessment were performed on 161 patients(82 men and 79 women,mean age 54.8±19.2 years).H.pylori infection was detected by histology in 47(29.2%)patients.Overall,the sensitivity,specificity,accuracy,positive predictive value,and negative predictive value(NPV)for H.pylori diagnosis by EGJA were 91.5%,93.0%,92.6%,84.3%,and 96.4%,respectively.In patients on treatment with proton pump inhibitors,diagnostic sensitivity was reduced by 27.3%,while specificity and NPV were unaffected.EGJA and RUT were comparable in diagnostic performance and highly concordant in H.pylori detection(κ-value=0.85).CONCLUSION Endofaster allows for rapid and highly accurate detection of H.pylori during gastroscopy.This may guide taking additional biopsies for antibiotic susceptibility testing during the same procedure and then selecting an individually tailored eradication regimen.

Rapid antimicrobial susceptibility testing for mixed bacterial infection in urine by AI-stimulated Raman scattering metabolic imaging

Urinary tract infection with mixed microorganisms may lead to false-positive resistance detection.Current antimicrobial susceptibility testing(AST)performed in clinical laboratories is based on bacterial culture and takes a long time for mixed bacterial infections.Here,we propose a machine learning-based single-cell metabolism inactivation concentration(ML-MIC)model to achieve rapid AST for mixed bacterial infections.Using E.coli and S.aureus as a demonstration of mixed bacteria,we performed feature extraction and multi-feature analysis on stimulated Raman scattering(SRS)images of bacteria with the ML-MIC model to determine the subtypes and AST of the mixed bacteria.Furthermore,we assessed the AST of mixed bacteria in urine and obtained single-cell metabolism inactivation concentration in only 3 h.Collectively,we demonstrated that SRS imaging of bacterial metabolism can be extended to mixed bacterial infection cases for rapid AST.

Isolation,identification,and virulence gene analysis of pathogenic Aeromonas dhakensis in Macrobrachium rosenbergii and histopathological observation

To identify the cause of mass mortality of adult Macrobrachium rosenbergii in a farm in Gaoyou City,Jiangsu Province,China,a dominant strain named DKQ-1 was isolated from the hepatopancreas of dying M.rosenbergii and identified as Aeromonas dhakensis by purification culture,biochemical characterization,and 16S rRNA and gyrB gene sequence analysis.The results of the challenge test revealed that the strain was highly pathogenic and the 50%lethal dose(LD_(50))in 72 h to M.rosenbergii was 1.54×10^(5)CFU/mL.The amplification results of virulence genes show that strain DKQ-1 carried 9 virulence genes,including ascV,aexT,aer,act,lip,ompAI,gcaT,acg,and exu,supporting the strong virulence of strain DKQ-1 to M.rosenbergii.Histopathological observation of the hepatopancreas,gills,and intestines indicated that DKQ-1 injection into M.rosenbergii could cause serious tissue damage,which further supported the strong virulence of this strain.In addition,a drug susceptibility test revealed that strain DKQ-1 was sensitive to 16 kinds of antibiotics,resistant to 9 kinds of antibiotics,and had intermediate resistance to spectinomycin and kanamycin.This study is the first report of A.dhakensis isolated from M.rosenbergii and provided a reference for the pathogen identification of bacterial diseases in M.rosenbergii,and for the prevention and treatment caused by A.dhakensis.

EXTREMELY LOW FREQUENCY MAGNETIC FIELD SUSCEPTIBILITY OF VISUAL DISPLAY UNITS

Extremely low frequency (ELF) magnetic field susceptibility is an index of visual display unit (VDU) quality and performance. This paper provided field measured data on the susceptibility for a large variety of VDUs. A test rig was built to study the susceptibility of VDUs to magnetic fields at fundamental and third harmonic frequencies. It was found that the susceptibility level is largely dependent on refresh rate of the VDU and the orientation of the external ELF field. It was also found that the VDU susceptibility is significantly increased in the presence of harmonic frequency magnetic fields. About 30% of the tested samples have susceptibility levels higher than that stated in IEC 1000-4-8 standard.

A Retrospective Study on the Pyogenic Pathogens and Their Antibiotic Susceptibility Patterns along with the ES<i>β</i>L Production

Pyogenic infections are caused by various pathogens leading to pus formation and that can be attributed due to a wound either through accident or during surgery leading to infection spread. There are pathogenic strains that are not uncommon in hospital settings like Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter etc., that are multidrug resistant (MDR) and are a cause of concern. The bacteriological profile in the pyogenic infections tends to be same but there is a vast difference with the antibiotic resistant patterns in different hospital settings. Hence, the aim was to study the antibiotic susceptibility profiles and Extended spectrum βeta Lactamases (ESβL) production in these pathogens. A prospective study was carried out in Silchar Medical College and Hospital Assam, India, over a four-month period from February to May 2021. The samples were processed using Blood and MacConkey’s agar. Further, these isolated pathogens were identified by standard morphological, cultural and biochemical tests. The antibiotic susceptibility test was conducted by Kirby Bauer disc diffusion method and ESβL production was detected by using combined disk diffusion test. It was observed that the identified pathogens had an incidence rate of 84.2% and further revealed that Gram negative had a higher incidence rate compared to Gram positive with 59.8%. The pathogens isolated from pus samples had a maximum of Klebsiella sps (19.64%) and the lowest was E. coli with 5.36%. Antibiotic susceptibility test (AST) of Gram-negative bacterial isolates showed the highest incidence with aztreonam (40.6%) and the lowest was observed in Piperacillin/Tazobactam with 7.5%. The only Gram positive was observed in our study, Staphylococcus aureus had the highest resistance in amikacin with 80% and interestingly, all the isolates were sensitive to Linezolid with 100%. There is a high rise and spreading with the multi-drug resistance (MDR) strains along with ESβL production and it was observed in our studies that these pathogens had an incidence rate of 18.5%. The highest was 58.1% in Pseudomonas sps. None of Proteus sps were found to be ESβL producers. To combat resistance, the irrational use of antibiotics should be avoided and surveillance of the rising multidrug species regularly helps in implementing better therapeutic options to reduce the morbidity and mortality.

Causative Microorganisms Isolated from Patients with Intra-Abdominal Infections and Their Drug Resistance Profiles:An 11-Year(2011–2021)Single-Center Retrospective Study

Objective To investigate the distribution and antimicrobial susceptibility of causative microorganisms recovered from patients with intra-abdominal infections(IAIs).Methods A total of 2,926 bacterial and fungal strains were identified in samples collected from 1,679 patients with IAIs at the Peking Union Medical College Hospital between 2011 and 2021.Pathogenic bacteria and fungi were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.Antimicrobial susceptibility testing(AST)was performed using the VITEK 2 compact system and the Kirby–Bauer method.AST results were interpreted based on the M100-Ed31 clinical breakpoints of the Clinical and Laboratory Standards Institute.Results Of the 2,926 strains identified,49.2%,40.8%,and 9.5%were gram-negative bacteria,gram-positive bacteria,and fungi,respectively.Escherichia coli was the most prevalent pathogen in intensive care unit(ICU)and non-ICU patients;however,a significant decrease was observed in the isolation of E.coli between 2011 and 2021.Specifically,significant decreases were observed between 2011 and 2021 in the levels of extended-spectrumβ-lactamase(ESBL)-producing E.coli(from 76.9%to 14.3%)and Klebsiella pneumoniae(from 45.8%to 4.8%).Polymicrobial infections,particularly those involving co-infection with gram-positive and gram-negative bacteria,were commonly observed in IAI patients.Moreover,Candida albicans was more commonly isolated from hospital-associated IAI samples,while Staphylococcus epidermidis had a higher ratio in community-associated IAIs.Additionally,AST results revealed that most antimicrobial agents performed better in non-ESBL-producers than in ESBL-producers,while the overall resistance rates(56.9%–76.8%)of Acinetobacter baumanmii were higher against all antimicrobial agents than those of other common gram-negative bacteria.Indeed,Enterococcus faecium,Enterococcus faecalis,S.epidermidis,and S.aureus were consistently found to be susceptible to vancomycin,teicoplanin,and linezolid.Similarly,C.albicans exhibited high susceptibility to all the tested antifungal drugs.Conclusion The distribution and antimicrobial susceptibility of the causative microorganisms from patients with IAls were altered between 2011 and 2021.This finding is valuable for the implementation of evidence-based antimicrobial therapy and provides guidance for the control of hospital infections.

Analysis on Antimicrobial Resistance of Clinical Bacteria Isolated from County Hospitals and a Teaching Hospital

The distinction of antimicrobial resistance of clinical bacteria isolated from county hospitals and a teaching hospital was investigated. Disc diffusion test was used to study the antimicrobial resistance of isolates collected from county hospitals and a teaching hospital. The data was analyzed by WHONET5 and SPSS statistic software. A total of 655 strains and 1682 strains were collected from county hospitals and a teaching hospital, respectively, in the year of 2003. The top ten pathogens were Coagulase negative staphylococci （CNS）, E. coil, Klebsiella spp. , S. areus, P. aeruginosa, Enterococcus spp. , Enterobacter spp. , otherwise Salmonella spp. , Proteus spp. , Shigella spp. in county hospitals and Streptococcus spp. , Acinetobacter spp. , X. maltophilia in the teaching hospital. The prevalence of multi-drug resistant bacteria was 5 % （4/86） of methicillin-resistant S. areus （MRSA）, 12 % （16/133） and 15.8 % （9/57） of extended-spectrum β-lactamases producing strains of E. coil and Klebsiella spp. , respectively, in county hospitals. All of the three rates were lower than that in the teaching hospital and the difference was statistically significant （P〈0. 01）. However, the incidence of methicillin-resistant CNS （MRCNS） reached to 70 % （109/156） in the two classes of hospitals. Generally, the antimicrobial resistant rates in the county hospitals were lower than those in the teaching hospital, except the resistant rates of ciprofloxacin, erythromycin, clindamycin, SMZco which were similar in the two classes of hospitals. There were differences between county hospitals and the teaching hospital in the distribution of clinical isolates and prevalence of antimicrobial resistance. It was the basis of rational use of antimicrobial agents to monitor antimicrobial resistance by each hospital.

Comparison of a glucose consumption based method with the CLSI M38-A method for testing antifungal susceptibility of Trichophyton rubrum and Trichophyton mentagrophytes

Background The prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document （M38-A） in 2002 by the Clinical and Laboratory Standards Institute （CLSI）, dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay --glucose consumption method （GCM） for dermatophytes. Methods In this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole （ITC）, voriconazole （VOC）, econazole nitrate （ECN） and terbinafine （TBF） by glucose consumption method （GCM）, in comparison to the Clinical and Laboratory Standards Institute （CLSI） M38-A method. Twenty-eight dermatophyte isolates, including Trichophyton rubrum （T. rubrum） （n=-14） and Trichophyton mentagrophytes （T. mentagrophytes） （n=-14）, were tested. In the GCM, the minimum inhibitory concentrations （MICs） were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually. Results Comparison revealed best agreement for TBF against T. mentagrophytes and T. rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. rubrum, high MICs were tested by GCM （0.125-16 pg/ml） but not M38-A method （0.5-1 IJg/ml）. The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively. Conclusion Measurement of glucose uptake can predict the susceptibility of T. rubrum and T. mentagrophytes to ECN and TBF.

Isolation and Identification of Photobacterium damselae subsp.damselae(PDD) from Tongue Sole

In May 2016,an epizootic occured among cultured tongue soles caused mass deaths in a fish farm in Qinhuangdao,China.In order to find out the etiological agent,a bacterial strain was isolated from ascites and other tissues of sick tongue sole aseptically collected.The isolate was identified as Photobacterium damselae subsp.damsela（PDD） by isolation culture,Gram staining,physiological identification,morohological observation,biochemical identification and 16S rDNA sequence analysis.The results showed that the isolate shared 99.6% homology with the reference strain in GenBank.The animal regression test displayed that the isolate had very strong pathogenicity to tongue sole.The LD（50） was 3.1 × 10~4 CFU/mL,and it showed pathogenicity to mammals.The antimicrobial susceptibility test showed the isolate was highly sensitive to nrofloxacin,Norfloxacin,Ciprofloxacin,Mequindox;moderately sensitive to Cefradine,Doxycycline;and insensitive to Gentamicin,Ceftriaxone,Tilmicosin,etc..

Analysis of Etiology and Drug Resistance of Biliary Infections

Summary: The bile was collected from fro patients with biliary infections, with the bacterium isolated to study the sensitivity of each kind of the bacterium to several antibiotics in common use. Except G-bacterium, we also found some kinds of G+ bacterium in infection bile. G-bacterium were not sensitive to Clindamycin, G+ bacterium were sensitive to Ciprofloxacin. Escherichia coli, Xanthomonas maltophilia, Enterobacter cloacae, Pseudomonas aeruginosa were sensitive to Ampicillin. G+ bacterium were not sensitive to Azactam. Enterococcus faecalis, Enterococcus faecium, Enterobacter cloacae were not sensitive to Ceftazidime. Enterococcus faecalis, Staphylococcus coagulase negative, Staphylococcus epidermidis, Pseudomonas aeruginosa were not sensitive to Ceftriaxone Sodium. We didn't found any bacterium resistance Imipenem. The possibility of the existence of G+ bacterium as well as drug resistance should be considered n patients with biliary infections. The value of susceptibility test should be respected to avoid drug abuse of antibiotics.

Isolation of Pathogenic Gram-Negative Bacteria from Urinary Tract Infected Patients

This study investigated the susceptibility pattern of different bacteria isolated from urinary tract infection to different antibiotics. 83 uropathogen bacteria were isolated from 300 urine samples taken from patients attended to Tikrit Teaching Hospital from March, 2011 through February, 2012. The patients were males and females aged between 4 days to 95 years. Bacteria obtained from urine samples were cultured and tested for antimicrobial susceptibility to 16 kinds of antibiotics. Urine samples were cultured on different media and incubated, thereafter bacteria were isolated and purified by streaking four times on the same media;isolates were identified depending on morphological, microscopic, and biochemical characteristics. The isolated strains of bacteria were tested for their susceptibility to some antibiotics using disk diffusion method. The antagonistic activity was evaluated by observing a clear zone of inhibition growth. The results showed that the bacterial species of Eschericia coli, Proteus mirabilis, Klebsiella pneumonia, Citrobacter diversus, Citrobacter freundii, Enterobacter aerogenes, Yersinia pestis, Pseudomonas aeruginosa, Klebsiella oxytoca and Hafnia alvei were identified in 44 (53%), 18 (21.7%), 4 (4.8%), 4 (4.8%), 3 (3.6%), 3 (3.6%), 3 (3.6%), 2 (2.4%), 1 (1.2%) and 1 (1.2%), respectively, of the isolates. The results of antimicrobial susceptibility test showed that 83 (100%) isolates were resistant to Ampicillin, Rifampicin and Erythromycin. 75 (90.3%) isolates were resistant to Cefotaxime, 67 (80.7%) isolates were resistant to Tobramyci. 66 (79.5%), 65 (78.3%), 56 (67.4%) and 48 (57.8%) isolates showed susceptibility to Nalidixic acid, Tetracycline, Nitrofurantoin, Chloramphenicol, respectively. 45 (54.2%) isolates were resistant to Azithromycin, Norfloxacin and Ciprofloxacin. Meropenem, Gentamicin, Amikacin, and Imipenem show significant effect on 35 (42.1%), 32 (38.5%), 27 (32.5%) and 1 (1.2%) isolates, respectively. In conclusion, significant bacteria count isolated from urine samples is pathogenic. The most effective antibiotic in inhibiting the bacterial growth was Imipenem while Ampicillin, Rifampicin and Erythromycin showed no effect on all 83 isolates (100%).