Rapid bacterial identification and antimicrobial susceptibility testing assay in positive blood cultures / 预防医学

Objective@#To establish a rapid bacterial identification and antimicrobial susceptibility testing assay in positive blood cultures, so as to provide insights into timely diagnosis and treatment of bloodstream infections.@*Methods@#A total of 1 154 blood culture samples were collected from inpatients in Zhejiang Hospital from February to May, 2022. The bacterial isolates were enriched and purified using improved separation gel method, and bacterial identification and antimicrobial susceptibility tests were performed using VITEK2 mass spectrometry system and VITEK2 Compact automated microbiology system. The accuracy of the new assay for bacterial identification and antimicrobial susceptibility tests was evaluated with the conventional VITEK 2 compact system as the standard. @*Results@#Of 1 154 blood culture specimens, the conventional VITEK 2 compact system detected 174 positives and 980 negatives. The new assay and the conventional VITEK 2 compact system identified consistent bacterial isolates in 165 out of 174 positive blood culture samples, and the accuracy of bacterial identification was 94.83% for the new assay, with a 99.21% accuracy for identifying Gram-negative bacteria and 82.22% for Gram-positive bacteria. Antimicrobial susceptibility tests were performed in 158 bacterial isolates, and the new assay presented a 90.17% accuracy, with a 90.27% accuracy for Gram-negative bacteria and 89.74% for Gram-positive bacteria. The conventional VITEK 2 compact system required 30 hours and longer to complete bacterial identification and antimicrobial susceptibility tests, and the new assay required 9 to 18 hours.@*Conclusions@#The new rapid bacterial identification and antimicrobial susceptibility testing assay shortens the time of bacterial culture, achieves rapid bacterial identification and antimicrobial susceptibility testing in blood culture specimens and has a high accuracy that meets clinical needs, which facilitates rapid diagnosis and treatment of bloodstream infections.

Detection of pathogenic Vibrio species and antibiogram activity in Asian Seabass (Lates calcarifer) in Tumpat, Kelantan

@#Some of Vibrio species is well known as pathogenic bacteria in aquaculture and the marine industry. Its infection is able to generate a massive outbreak and affect the fish population, especially for net caged fish such as seabass. This study was conducted to investigate the prevalence of Vibrio spp. isolated from seabass (Lates calcarifer) in Sri Tujuh Lagoon, Tumpat, Kelantan. Then, to determine the antibiotic resistance in Vibrio isolates. Polymerase chain reaction (PCR) was used to detect Vibrio species using specific primer VR169 and VR744 with estimation base pair size band, 597 bp and further identified by sequencing. On the other hand, antibiotic susceptibility tests were continued by using 13 types of antibiotics; kanamycin (K30), chloramphenicol (C30), neomycin (N10), ampicillin (AMP10), nitrofurantoin (F300), tetracycline (TE30), streptomycin (S10), norfloxacin (NOR10), ciprofloxacin (CIP5), nalidixic acid (NA30), gentamicin (CN10), doxycycline (DO30) and sulfamethoxazole (SXT100). As a result, 14 Vibrio isolates were identified, including Vibrio fluvialis (n=6), Vibrio parahaemolyticus (n=3), Vibrio harveyi (n=2) and each isolate for Vibrio vulnificus, Vibrio alginolyticus and Vibrio spp. The results showed that all isolates were sensitive to most antibiotics except ampicillin, neomycin and streptomycin. The MAR index value was ranging from 0 to 0.31. This study demonstrates the prevalence of Vibrio spp. in seabass and the report on multidrug resistance strains that could be of concern to the fish farmers. In addition, data from this study can be further used in fish disease management plans.

Disseminated cryptococcosis caused by Cryptococcus neoformans a case report and review / 中国热带医学

@#Abstract: To analyze the clinical, therapeutic and laboratory characteristics of disseminated cryptococcosis caused by Cryptococcus neoformans invading the blood stream in patient with liver cirrhosis and splenectomy. A 30-year-old male underwent splenectomy plus pericardial devascularization due to "splenomegaly and hypersplenism" in March in 2016. The patient had intermittent fever after operation for many times, and successively accompanied with back pain, left lower limb abscess and right hip pain. The highest body temperature was 39 ℃. CT and MRI revealed the lung lesion and multiple bone destruction. During that period, the effect of antibiotics was not good. On April 19th, 2017, Gram's stain, India ink stain, API 32C, Vitek 2 Compact, ribosomal ITS and IGS sequence analysis were performed to identify the strain isolated from the pus and blood stream. The serum of the patient was detected for cryptococcal antigen. Antifungal susceptibility test was used to determine drug sensitivity and minimum inhibitory concentration (MIC). The Cryptococcus neoformans isolated from fresh pus specimen showed a prominent, thick capsule after India ink stain. The colonies isolated from pus and blood stream were identified Cryptococcus neoformans using API 32C, Vitek 2 Compact, and sequence analysis of rDNA ITS and IGS. Cryptococcal capsule antigen was positive. The minimal inhibitory concentrations of 5-Flucytosine, amphotericin B, fluconazole, itriconazole, voriconazole against the isolate were <4 μg/mL, <0.5 μg/mL, 4 μg/mL, ≤0.25 μg/mL, 0.125 μg/mL respectively. The patient was initially treated with intravenous amphotericin B and flucytosine. After anti-Cryptococcus treatment for two months, the patient clinically improved, and the lesions were reduced on a follow-up CT scan. The patient made a full functional recovery after treatment for six months. Cryptococcosis has hidden onset, atypical clinical symptoms and lack of specificity. Blood stream is the main channel for Cryptococcus to spread and involve many organs of the whole body, including skin, bone and so on. Therefore, early use of blood culture to monitor blood flow dissemination, actively removing the primary focus and cutting off the infection route in time and carrying out effective anti-Cryptococcus treatment are conducive to the patient's early recovery.

Distribution and drug resistance of nontuberculous mycobacteria / 公共卫生与预防医学

Objective To analyze the distribution characteristics and drug resistance of nontuberculous mycobacteria(NTM),and to provide guidance for the selection of targeted agents in clinical treatment. Methods The clinical data of inpatients in our hospital from April 2019 to February 2021 were collected,the culture and strain identification of non tuberculosis mycobacteria were carried out,the drug sensitivity test of anti tuberculosis drugs was carried out,and the drug resistance of non tuberculosis mycobacteria to first-line anti tuberculosis drugs was analyzed. Results A total of 1 326 strains of mycobacterium were isolated,including 1 154(87.03%)strains of mycobacterium tuberculosis and 172(12.97%)strains of non-mycobacterium tuberculosis.Nine species of nontuberculous mycobacteria were detected,including slow-growing mycobacteria such as Mycobacterium kansasii and Mycobacterium avium-intracellulare complex,belonging to Groups I-III,and fast-growing mycobacteria such as Mycobacterium chelonae and Mycobacterium smegmatis,belonging to Group IV. Among them , Mycobacterium avium-intracellulare complex and Mycobacterium chelonae were dominant,accounting for 26.16%and 36.63%,respectively.Drug susceptibility tests showed that the resistance rate of Mycobacterium avium-intracellulare complex to streptomycin was 100.00%,the drug resistance rate of Mycobacterium chelonae to isoniazid,rifampicin and streptomycin was 100.00%,and the drug resistance rate of Mycobacterium smegmatis and Mycobacterium abscessus to most antibacterial drugs was 100.00%.The resistance rate of major NTM bacteria to clarithromycin was relatively low.There was no statistically significant difference in the susceptibility rates of slow and fast-growing mycobacteria to isoniazid and clarithromycin(P>0.05) ; The susceptibility rates of slow-growing mycobacteria to amikacin,clarithromycin and rifambutin were 62.86%,92.86%and 72.86%,all above 50.00%.The susceptibility rate of the fast-growing mycobacteria to clarithromycin was also more than 50.00%,being 87.25%.The susceptibility rate of slow-growing mycobacteria to other antibiotics was higher than that of fast-growing mycobacteria(P<0.05).The drug resistance of Mycobacterium tuberculosis to first-line anti tuberculosis drugs was significantly lower than that of non Mycobacterium tuberculosis(P<0.05). Conclusion Non-tuberculous mycobacteria have high drug resistance,especially fast-growing mycobacteria,so drug susceptibility tests are of great value in clinical treatment.

BRCAR: herramienta de soporte en la evaluación del riesgo para cáncer de mama / BRCAR: Support Tools for Risk Evaluation of Breast Cancer

Introducción: El cáncer de mama es el tumor maligno más frecuente en el mundo, en Cuba, es la segunda causa de muerte en mujeres. La insuficiencia en el diagnóstico precoz y la existencia de novedosas estrategias de tratamiento plantean la necesidad de establecer formas eficaces para identificar el riesgo en personas sanas, sin embargo en nuestro país no se cuenta con un método eficaz para predecir el riesgo y direccionar las acciones preventivas y terapéuticas. Objetivo: Crear un estándar nacional orientado a la identificación del cáncer de mama como soporte a la práctica médica y como herramienta de apoyo en la evaluación del riesgo. Método: Se combinaron 28 variables (determinadas por los factores de riesgo de cáncer de mama) a las cuales se les atribuyeron parámetros de ponderación asociados al nivel de incidencia registrado en la literatura médica, utilizando un algoritmo de votación como elemento matemático central. Resultados: Se desarrolló un sistema computarizado para la evaluación del riesgo de cáncer de mama en personas sanas. Conclusiones: BRCAR es una herramienta de soporte para objetivar el riesgo del cáncer de mama, al determinar el impacto de determinados factores de riesgo, con el fin de direccionar los métodos de estudio para la detección precoz(AU)

Introduction: Breast cancer is the most frequent malignant tumour in the world; it is the second cause of women death in Cuba. The insufficiency in early diagnosis and the existence of novel treatment strategies raise the need to establish effective ways to identify risk in healthy people, however in our country there is no effective method to predict risk and direct preventive actions and therapeutic. Objective: To create a national standard aimed at identifying breast cancer as a support to medical practice and support tool in risk assessment. Method: 28 variables (determined by risk factors for breast cancer) were combined; assigning to those variables weighting parameters associated with the level of incidence recorded in the medical literature, using a voting algorithm as the central mathematical element. Results: A computerized system was developed to assess the risk of breast cancer in healthy people. Conclusions: BRCAR is a support tool to objectify the risk of breast cancer, by determining the impact of certain risk factors, in order to direct study methods for early detection(AU)

Evaluation of rapid identification methods for foodborne diarrhea Campylobacter and detection of drug resistance / 中华检验医学杂志

Objective:To explore and evaluate a appropriate suitable method for detection of Campylobacter and antibiotic sensitivity test for foodborne diarrhea in clinical laboratories. Methods:Pre-experiment:a total number of 400 fecal samples of patients with foodborne diarrhea were prospectively collected from the intestinal disease clinic of Beijing Tongren Hospital from September 2017 to January 2018. Double-hole filtration culture method and modified cefoperazone charcoal deoxycholate (CCD) agar culture method were used for fecal culture in micro-aerobic environment for 48 hours, and then suspicious colonies were identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Meanwhile, C. jejuni and C. coli were detected by real-time quantitative polymerase chain reaction(qPCR). Large sample verification: 2 062 fecal samples of patients with foodborne diarrhea in three hospitals of different levels in different areas of Beijing were collected for qPCR detection and culture from April 2018 to March 2019. The antimicrobial sensitivity test (AST) of C. jejuni and C. coli was performed according to the disk diffusion method and agar dilution method recommended by Clinical and Laboratory Standards Institute and National Antimicrobial Resistance Monitoring System for Enteric Bacteria. The results of the three detection methods and the consistency of the two antibiotic sensitivity tests were compared. Results:In the pre-experiment, the positive rates of Campylobacter ( jejuni/coli) detected of qPCR, double-hole filtration culture and modified CCD agar culture were 9.0% (36/400), 5.0% (20/400)and 3.5% (14/400), and the difference was statistically significant ( P<0.01). The samples with negative result of qPCR were negative by both culture methods. The total positive rates of Campylobacter detected by qPCR was 8.1% (168/ 2 062)including 7.0% (144/2 062) for C. jejuni and 1.2% (24/2 062) for C. coli. The samples with positive qPCR results were cultured by double-hole filtration culture method and the positive rate was 61.9%(104/168), among which, the positive rate of C. jejuni and C. coli were 58.3%(84/144) and 83.3%(20/24) respectively, which was not significantly different from the detection rate and culture positive rate in the pre-test ( P>0.1). The resistance rates of C. jejuni and C. coli to ciprofloxacin were 94.0%(94/100) and 100.0%(24/24) and to erythromycin were 6.0%(6/100) and 33.3%(8/24). The results from two antibiotic sensitivity test methods were consistent (Kappa>0.75). Conclusions:qPCR is rapid, sensitive and easy to operate, so it is suitable for routine development in clinical laboratories. The double-hole filtration culture method is beneficial to the acquisition of strains and is essential for the further study of Campylobacter. There was no significant difference between agar dilution method and disk diffusion method in antibiotic sensitivity test. Campylobacter showed a very high resistance rate to quinolones, which was no longer suitable for the treatment of Campylobacter foodborne diarrhea in Beijing area. Macrocyclic lipid antibiotics should be preferred.

Efficacy of precision treatment in multidrug-resistant pulmonary tuberculosis based on proportion method for drug susceptibility test / 中华传染病杂志

Objective:To explore the precision treatment effect of multidrug-resistant pulmonary tuberculosis (MDR-PTB) based on the proportion method for drug susceptibility test, and to provide a scientific basis for formulating MDR-PTB treatment plan.Methods:One hundred and eighty patients with MDR-PTB treated in Shenzhen Center for Chronic Disease Control from January 5, 2016 to April 30, 2018 were enrolled. The initial treatment plan after diagnosis was six months of amikacin (AM), pyrazinamide (Z), levofloxacin (LFX), ethambutol (E), prothionamide (PTO) and 18 months of Z, LFX, E, PTO. According to whether proportion method for drug susceptibility test for 10 commonly used drugs was implemented, patients were divided into precision treatment group and empirical treatment group. In the precision treatment group, the treatment plans were adjusted according to the results of the drug susceptibility test. The treatment plans and disease outcomes of the two groups of patients were retrospectively analyzed. Chi-square test was used for statistical analysis.Results:Among the 180 patients, there were 113 patients in the precision treatment group and 67 patients in the empirical treatment group. The drug resistance rates of the precision treatment group from low to high were: capromycin (CM) (0, 0/113), AM (2.65%, 3/113) and kanamycin (KM) (2.65%, 3/113), para-aminosalicylic acid (PAS) (7.96%, 9/113), PTO (11.50%, 13/113), ofloxacin (OFX)(38.05%, 43/113), E (39.82%, 45/113), and streptomycin(S) (76.99%, 87/113). In the precision treatment group, the drugs were adjusted for 104 person-times according to the proportion method for drug susceptibility test during the treatment, from low to high: AM (3 person-times), PTO (13 person-times), LFX (43 person-times) and E (45 person-times). The treatment success rate of the precision treatment group was 78.8%(89/113), which was higher than that of the experience treatment group (52.2%(35/67)), the difference was statistically significant ( χ2=13.805, P=0.000 2). In the precision treatment group and empirical treatment group, there were no statistically significant differences of alanine aminotransferase elevated (32.3%(31/96) vs 34.0%(18/53)), serum creatinine elevated (4.2%(4/96) vs 5.7%(3/53)), and white blood cell count decreased (24.0%(23/96) vs 22.6%(12/53)) (all P>0.05). Conclusion:The traditional treatment plan based on the proportion method for drug susceptibility test has a high success rate in the treatment of MDR-PTB, which is still a worthy choice.

Evaluation of in vitro activity of ceftolozane-tazobactam against recent clinical bacterial isolates from Brazil - the EM200 study

ABSTRACT Background: The emergence of antibiotic resistance is increasing and there are few effective antibiotics to treat infections caused by resistant and multidrug resistant bacterial pathogens. This study aimed to evaluate the in vitro activity of ceftolozane-tazobactam against clinical bacterial isolates from Brazil. Methods: A total of 673 Gram-negative bacterial isolates including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and other Enterobacterales collected from 2016 to 2017 were tested, most of them isolated from patients in intensive care units. Minimum inhibitory concentrations (MIC50/90) were determined by broth microdilution for amikacin, aztreonam, cefepime, cefotaxime, cefoxitin, ceftolozane-tazobactam, ceftazidime, ceftriaxone, ciprofloxacin, colistin, ertapenem, imipenem, levofloxacin, meropenem, and piperacillin-tazobactam using dried panels. Antimicrobial susceptibility results were interpreted according to Clinical and Laboratory Standards Institute criteria. Results: Susceptibility rates to ceftolozane-tazobactam ranged from 40.4% to 94.9%. P. aeruginosa susceptibility rate to ceftolozane-tazobactam was 84.9% (MIC50/90, 1/16 µg/mL) and 99.2% to colistin. For E. coli, ceftolozane-tazobactam inhibited 94.9% (MIC50/90, 0.25/1 µg/mL) of the microorganisms. The susceptibility rate of K. pneumoniae to ceftolozane-tazobactam was 40.4% (MIC50/90, 16/>32 µg/mL). Other Enterobacterales have shown susceptibility rates of 81.1% (MIC50/90, 0.5/16 µg/mL) to ceftolozane-tazobactam, 93.9% to meropenem, 90.9% to amikacin (90.9%), and 88.6% to ertapenem. In non-carbapenemase producing isolates, AmpC mutations were found three isolates. Conclusions: Ceftolozane-tazobactam has shown relevant activity against a large variety of the analyzed microorganisms collected from multiple centers in Brazil, showing promising results even in multidrug resistant strains.

Isolation, identification and antibiogram profiling of bacteria isolated from water, seafood, and macroplastic samples from Baseco Beach, Manila Bay

Background and Objective@#Manila Bay plays an important role both in economics and ecology because it serves as the major economic center of the Philippines and as it harbors different habitats and biodiversity. Unfortunately, it is threatened by various pollutions including the unregulated discharge of wastewater from industrial, agricultural, and household sectors and improper disposal of trash such as macroplastics among others. All these contributes to the current state of Manila Bay. This study identified bacteria isolated from water, seafood and floating macroplastic samples from Baseco Beach, Manila Bay and determined their antibiogram profiles. @*Methodology@#Bacterial isolates were obtained from water, seafoods and macroplastic samples from Baseco Beach, Manila Bay using conventional culture techniques. Identification of the isolates was done using Vitek-2 Automated System and antibiogram profiling was done using Kirby-Bauer Disk Diffusion Susceptibility Test. @*Results and Conclusions@#A total of 30 bacterial isolates were obtained from different samples from water, seafood and macroplastic samples from Baseco Beach, Manila Bay. These isolates were identified and found to belong to 13 different bacterial species with Bacillus spp. comprising 33.33% of the isolates (10 out of 30), and Vibrio alginolyticus comprising 23.33% of the isolates (7 out of 30) and the other species comprise the remaining 43.34% (Pseudomonas spp., Vibrio fluvialis, Klebsiella pneumoniae, Shewanella alga, Sphingomonas paucimobilis, Staphylococcus haemolyticus, Chryseobacterium indologenes, Myroides sp. and Aeromonas salmonicida). Of these, six out of 30 isolates (20%) showed susceptibility to all six representative antibiotics used (Cefazolin 30μg, Gentamicin 10 μg, Chloramphenicol 30 μg, ampicillin 10 μg, Cefuroxime 30 μg, Ceftazidime 30 μg) while 7 isolates (23.33%) were resistant to only one class of antibiotic. Moreover, 17 out of 30 isolates (56.66%) were resistant to two or more classes of antibiotic while only one isolate (3.33%) was found to be resistant to gentamicin. All 30 isolates (100%) were susceptible to chloramphenicol. Interestingly, three antibiotic resistant (AMR) bacteria were isolated from macroplastics namely Pseudomonas oleovorans (S2), Vibrio alginolyticus (S5), and Pseudomonas alcaligenes (S29) which were all resistant to ampicillin and cefazolin. This is the first study in the Philippines to isolate AMR bacteria from macroplastics from Manila Bay. The presence of AMR bacteria in macroplastics shows that these materials can be a reservoir for its dynamics and distribution. Lastly, with the emergence of antimicrobial resistant bacteria, the elucidation of the antibiogram profile of bacteria is necessary to determine its implication sand threats to public health. This study served as a baseline study of presence of AMR bacteria in macroplastic samples from Manila Bay.

A Study on Burden of Carbapenem-Resistant Pseudomonas aeruginosa and Acinetobacter baumannii Infections in a Tertiary Care Hospital of Eastern India

Introduction: Healthcare associated infections(HAI) bymulti-drug resistant organisms(MDRO) are major cause ofmortality and morbidity having significant impact on qualityof life and economic burden. HAI by carbapenem-resistantPseudomonas aeruginosa (CRPsA) and Acinetobacterbaumannii (CRAB) are emerging threat for their highantibiotic resistance and spread via mobile genetic elements.Objectives of this study were to detect prevalence of CRPsAand CRAB infections in a tertiary care hospital of EasternIndia and to determine their antimicrobial resistance profile.Material and methods: This observational study was done inDepartment of Microbiology from January 2018-June 2019.From HAI patients, different clinical samples were collected.Culture and identification by standard conventional methodsand antimicrobial susceptibility tests by modified KirbyBauer disc-diffusion method following CLSI guidelines wereperformed. CRPsA and CRAB cases were identified whenisolates were resistant to ≥1 carbapenem, 10µg imipenemdisc(zone diameter ≤15mm for P. aeruginosa or ≤18mm forA. baumanii) or meropenem disc (≤15mm for P. aeruginosa or≤14mm for A. baumanii).Result: From 27,043 clinical samples, 1785(6.6%)Acinetobacter baumannii and 777(2.87%) Pseudomonasaeruginosa were isolated. CRAB and CRPsA prevalencewere 74.17% and 62.29% respectively. Carbapenemresistance were further categorised into imipenem-resistantmeropenem-resistant (IRMR) (A. baumanii-63.19%, P.aeruginosa-51.61%), imipenem-resistant-meropenemsensitive (IRMS) (A. baumanii-10.48%, P. aeruginosa-9.13%), meropenem-resistant-imipenem-sensitive (MRIS)(A. baumanii -0.51%, P. aeruginosa -1.54%) phenotypes.Fourth category was imipenem-sensitive-meropenemsensitive (ISMS) (A. baumanii -25.82%, P. aeruginosa-37.71%). Carbapenem-resistant groups showed significantlyhigh resistance for all antibiotics excepting colistin.Conclusion: Carbapenems are often used for treatingMDRO. But high carbapenem-resistance in HAI is alarming,warranting judicious use of antibiotics.

Salmonella spp. prevalence, antimicrobial resistance and risk factor determination in Colombian swine farms / Salmonella spp. prevalência, resistência antimicrobiana e determinação de fatores de risco em granjas suínas colombianas

To determine Salmonella spp. prevalence/seroprevalence, antimicrobial resistance patterns and risk factor identification associated with its presence in Colombian swine farms. 504 samples (Faeces, swabs and environment samples) were obtained from 21 farms distributed in four geographical regions in Colombia. Salmonella spp. microbiological and molecular detection were determined by two Salmonella spp. MDS3M™ and MALDI-TOF MS assays, respectively. In addition, for serological evaluation 231 serum samples were analyzed employing ELISA Salmonella Pigtype®-Salmonella Ab (QUIAGEN®). Additionally, 41 isolates were tested for antimicrobial susceptibility using broth microdilution technique (Panel B1016-180 Beckman Coulter NC72®) and verified with WHONET 2016 software. Risk factors were assessed from a survey and analyzed for statistical significance by U Mann-Whitney test. An 8.9% prevalence (n=45) and 38.1% (n=88) seroprevalence were determined. All isolates presented 100% antimicrobial susceptibility against amikacin. However, resistance against penicillin, tetracycline, cefuroxime and trimethoprim/sulfamethoxazole was present in more than 50% of evaluated strains. Risk factors associated with Salmonella spp. presence were surface water use, rough-surfaced on floors, presence of hoppers as feeders and worker's boots. Bacteria were present in animals and environmental samples from evaluated farms. Animal contact and/or exposure with the microorganism were also evident in obtained serological response. Bacteria presence depended on management practices and infrastructure, likewise antibiotic use, supplemented in the diet may have induced an increase in Salmonella spp. antimicrobial resistance.(AU)

Para determinar Salmonellaspp. prevalência/soroprevalência, padrões de resistência antimicrobiana e identificação de fatores de risco associados à sua presença em granjas suínas colombianas. Foram obtidas 504 amostras (fezes, zaragatoas e amostras do ambiente) de 21 fazendas distribuídas em quatro regiões geográficas da Colômbia. Salmonella spp., a detecção microbiológica e molecular foi determinada por 2 Salmonella spp. Ensaios MDS3M™ e MALDI-TOF MS, respectivamente. Além disso, para avaliação sorológica, foram analisadas 231 amostras de soro empregando ELISA Salmonella Pigtype® - Salmonella Ab (QUIAGEN®). Além disso, 41 isolados foram testados quanto à suscetibilidade antimicrobiana usando a técnica de microdiluição em caldo (Painel B1016-180 Beckman Coulter NC72®) e verificados com o software WHONET 2016. Os fatores de risco foram avaliados em uma pesquisa e analisados quanto à significância estatística pelo teste U Mann-Whitney. Foram determinadas prevalências de 8,9% (n=45) e 38,1% (n=88). Todos os isolados apresentaram 100% de suscetibilidade antimicrobiana à amicacina. No entanto, resistência à penicilina, tetraciclina, cefuroxima e trimetoprim/sulfametoxazol estava presente em mais de 50% das cepas avaliadas. Fatores de risco associados à Salmonella spp., presença de uso de água de superfície, superfície áspera no chão, presença de tremonhas como alimentadores e botas de trabalho. Bactérias estavam presentes em animais e amostras ambientais de fazendas avaliadas. O contato animal e/ou a exposição ao microrganismo também foram evidentes na resposta sorológica obtida. A presença de bactérias dependia de práticas de manejo e infraestrutura, assim como o uso de antibióticos suplementados na dieta pode ter induzido um aumento de Salmonella spp. resistência antimicrobiana.(AU)

Helicobacter pylori: resistencia múltiple en pacientes de Bogotá, Colombia / Helicobacter pylori: Multiple resistance in patients from Bogotá, Colombia

Resumen Introducción. La resistencia a los antibióticos es la principal causa del fracaso del tratamiento contra Helicobacter pylori; la claritromicina y el metronidazol son los antibióticos que generan mayor resistencia. En Colombia, la resistencia primaria a estos dos antibióticos y el uso excesivo de levofloxacina han alcanzado los límites aceptados (13,6, 83 y 16 %, respectivamente). A pesar de ello, se usa el tratamiento empírico combinando estos antibióticos en pacientes en los que ha fallado anteriormente. Objetivo. Determinar la resistencia a los antibióticos en pacientes previamente tratados para H. pylori en Bogotá, Colombia. Materiales y métodos. Se llevó a cabo un estudio descriptivo en el que se evaluó mediante dilución en agar la resistencia a la amoxicilina, la claritromicina, la levofloxacina y el metronidazol en 10 aislamientos provenientes de 5 pacientes con tres o cuatro tratamientos fallidos para H. pylori. La resistencia a los antibióticos se confirmó mediante secuenciación de ADN (Magrogen, Korea). Resultados. Ocho de los aislamientos presentaron resistencia a dos o más antibióticos y todos fueron resistentes a la levofloxacina. Los patrones de sensibilidad de los aislamientos provenientes del antro pilórico y del cuerpo del estómago, fueron diferentes en tres de los pacientes. Conclusión. Hasta donde se sabe, esta es la primera evidencia de resistencia múltiple de H. pylori en Colombia en pacientes previamente tratados. Los resultados evidenciaron las consecuencias del uso de un esquema ineficaz de tratamiento antibiótico y la necesidad de evaluar la sensibilidad a los antibióticos en diferentes sitios anatómicos del estómago. La resistencia múltiple limita el número de antibióticos útiles para erradicar H. pylori.

Abstract Introduction: The main cause for Helicobacter pylori infection treatment failure is antibiotic resistance, where clarithromycin and metronidazole play the main role. In Colombia, primary resistance as a consequence of the use of these two antibiotics and excessive levofloxacin use is above the accepted limit (13.6%, 83%, and 16%, respectively). Despite this fact, empirical therapies that include the combination of these antibiotics are used in patients with previous therapeutic failure. Objective: To determine antibiotic resistance in patients previously treated for H. pylori in Bogotá, Colombia. Materials and methods: We conducted a descriptive study that included ten isolates obtained from five patients with three or four previous failed treatments for H. pylori. Antibiotic resistance to amoxicillin, clarithromycin, levofloxacin, and metronidazole was investigated by agar dilution and confirmed by DNA sequencing (Magrogen, Korea). Results: Eight isolates were resistant to two or more antibiotics. All isolates were resistant to levofloxacin. Susceptibility patterns in isolates from the gastric antrum and the body of the stomach were different in three patients. Conclusion: As far as we know, this is the first evidence of multiple H. pylori resistance in Colombia in previously treated patients. Results demonstrated the consequences of using an ineffective antibiotic scheme and the need to assess antibiotic susceptibility in different anatomical sites of the stomach. The consequences of multiple resistance decrease possible antibiotic effectiveness to eradicate H. pylori in the future.

A Prospective Study on Prevalence of Micro- Organisms Isolated from Ear Swab in a tertiary Care Teaching Hospital in Mathura

Background: Ear infection is an inflammation of the ear. Ear discharge is one of the most common symptoms of ear infection. Approximately, 65-330 million people suffer from ear infection worldwide. Around 60% of them had significant hearing impairment Methods: The duration of study was over a period of one year.400 cases were included with ear infection. This study was conducted in the Department of ENT, K M Medical College & Hospital, Mathura. Result: This study suggested that 40% bacteria were P. aeruginosa followed by Proteus mirabilis, Klebsiella pneumoniae, E. coli & Citrobacter spp.in gram negative bacteria. Conclusion: This study suggested that Ciprofloxacin and gentamycin were the most sensitive against pseudomonas aeruginosa and Staphylococcus aureus. Use of antibiotics for treating Pseudomonas aeruginosa and Staphylococcus aureus infections in otitis media, alert us against indiscriminate usage of antibiotics to prevent development of resistance.

Drug sensitivity test of human derived Brucella / 中华地方病学杂志

Objective To screen the most suitable medium for Brucella drug susceptibility test, and observe the resistance of human derived Brucella to different antibiotics. Methods Totally 180 strains of Brucella isolated from 25 provinces (municipalities, autonomous regions) in recent years were taken as observation objects. Mueller-Hinton ( MH ) agar , MH blood agar and Brinell agar were used to carried out the drug susceptibility test in vitro, and to compare the results of drug susceptibility test of different medium; the most suitable Brucella drug susceptibility test medium was used to detect the resistance of human derived Brucella to Doxycycline, Rifampicin, Streptomycin, Levofloxacin, Moxifloxacin, Ceftriaxone sodium, Co-trimoxazole and Amoxicillin/Clavulanic acid by K-B drug sensitive paper, and to observe the formation of antibacterial ring around the drug sensitive paper. Results The growth of Brucella on the MH agar and MH blood agar were slower than that on the Brinell agar, and the antibacterial rings were not obvious. All the 180 strains of Brucella were sensitive to seven antibiotics such as Doxycycline, Rifampicin, Streptomycin, Levofloxacin, Moxifloxacin, Ceftriaxone sodium, and Amoxicillin/Clavulanic acid; and 70 strains of Brucella were resistant to Co-trimoxazole, accounting for 39％ (70/180); Brucella strains resistant to Co-trimoxazole were found in 21 provinces. Conclusions Brinell agar is the most suitable medium for Brucella susceptibility test. The human derived Brucella is resistant to Co-trimoxazole; the resistant strains are distributed in 21 provinces ( municipalities , autonomous regions ) . It is recommended that relevant departm ents of the province ( municipalities , autonomous regions ) carry out epidemiological investigations on the resistance of Brucella, and strengthen the monitoring of drug resistance in clinical drugs of brucellosis patients.

Investigation and analysis of infection and antibiotic resistance of Helicobacter pylori in armed policemen in Shaanxi province / 解放军医学杂志

Objective To investigate and analyze the situation of Helicobacter pylori (Hp) infection and antibiotic resistance in armed police officers and soldiers (APOSs) in Shaanxi province, and provide a theoretical basis for the rational selection of drugs for the eradication of Hp in the armed police forces in this region. Methods Two hundred and three APOS patients from the Shaanxi Armed Police Corps Hospital who underwent gastroscopy for gastrointestinal symptoms from June 2016 to December 2017 were enrolled in this study. Biopsy samples were extracted by gastroscope and Hp was isolated, identified and cultured. Four types of antibiotics were used to analyze the sensitivity of resistance to drugs: amoxicillin, clarithromycin, levofloxacin and metronidazole. Results A total of 119 Hp strains were isolated from 203 endoscopic biopsies. The positivity rate was 58.6%. The positivity rate of Hp isolation and culture in patients with peptic ulcer was 58.1% (36/62), which was significantly higher than that in patients with chronic gastritis (35.2%, 32/91, χ2=7.832, P=0.005); There was no statistical difference compared with other pathological types of diseases (P>0.05). The resistance rates of Hp to amoxicillin, clarithromycin, levofloxacin, and metronidazole were 1.7% (2/119), 10.1% (12/119), 14.3% (17/119) and 28.6% (34/119), respectively. There were 12 of 119 strains were detected to be resistant to two drugs, of them 7 were resistant to clarithromycin and metronidazole, and 5 were resistant to levofloxacin and clarithromycin. We did not find the strains which are antibiotic resistant to three or more drugs. There was no significant difference in Hp resistance between different pathological disease types (P>0.05). Conclusions The role of Hp in the development of peptic ulcers among local armed police officers is very important. The current first-line therapy should be selected for eradication treatment of APOS patients in the region.

Distribution of bacteria of wound secretion and drug susceptibility analysis in infected patients from Department of Hand Surgery / 吉林大学学报(医学版)

Objective: To analyze the results of bacteria distribution of wound secretion and the drug susceptibility test of the infected patients from Department of Hand Surgery, and to provide the evidence for antibiotic selection in the infected patients in early stage of treatment. Methods: The clincal data of hospitalized patients from Department of Hand Surgery were collected. The patients were divided into three groups according to the wound conditions. The patients in Group 1 who had no wound while being in hospital were infected after operation; the patients in Group 2 had the contaminated wound but no infection while being in hospital; the patients in Group 3 had definite infection while being in hospital. The results of bacterium culture of wound secretion and drug susceptibility test of the patients in three groups were analyzed. Results: The clinical data of 297 patients were obtained, including 17 patients from Group 1, 201 patients from Group 2, and 79 patients from Group 3. A total of 406 strains of 54 species of bacteria were isolated from 297 patients, including 178 strains (43. 84%, 178/406) gram-positive bacteria and 228 strains (56. 2%, 228/406) gram-negative bacteria. The four most common strains were Staphylococcus aureus (15.8%, 64/406), Staphylococcus epidermidis (14.0%, 57/406), Enterobacter cloacae (10.3%, 42/406), and Serratia marcescens (9.9%, 40/406). The results of drug susceptibility test showed that the detection rates of methicillin-resistant staphylococcus aureus (MRSA) and staphylococcus epidermidis were 9. 4% (6/64) and 72. 0% (46/64), respectively. There were no vancomycin-resistant, linezolidresistant, and tigecycline-resistant positive strains among Staphylococcus susceptibility; and there were no the carbapenem-resistant positive strains among Enterobacter cloacae and Serratia marcescens. The susceptibility rate of Enterobacter cloacae to ceftriaxone was 83. 3% (35/42), and its susceptibility to ceftazidime was 90. 5% (38/42). The susceptibility rate of Serratia marcescens to ceftriaxone was 100.0% (40/40), and its susceptibility to ceftazidime was 97.5% (39/40). Conclusion: The first generation of cephalosporin and penicillinase-stabilized penicillin can be used as the preferred empirical antibiotics for the infected patients from Deparment of Hand Surgery in our hospital. The third and fourth generation cephalosporins and quinolones antibiotics can be used as the preferred antibiotics for the infected patients with open trauma as well as the possibility of G- bacillus infection.

Molecular epidemiological characteristics of Streptococcus pyogenes causing scarlet fever and angina in children / 中华微生物学和免疫学杂志

Objective@#To investigate the antimicrobial resistance, macrolide-resistance genes, virulence genes and emm types in Streptococcus pyogenes isolates.@*Methods@#A total of 247 oropharyngeal swab specimens were collected from pediatric outpatients (aged 2-11 years) who were clinically diagnosed as scarlet fever in Children′s Hospital of Fudan University from January to December, 2018. These specimens were timely sent to the Microbiology Laboratory for isolation and identification of Streptococcus pyogenes strains were isolate after culturing and identified with bacitracin susceptibility test. Moreover, the diameter of bacitracin inhibition zone was measured by vernier caliper. Their susceptibility to seven antibiotics, including erythromycin, clarithromycin, clindamycin, ampicillin, ceftriaxone, norfloxacin and chloramphenicol, were measured using KB method. Macrolide-resistance genes (mefA, ermA, ermB and Tn916 transposon) and virulence genes (speA, speB, speC, speG, speH, speI, speJ and speK) were detected by PCR. Amplification and sequencing of emm gene were conducted according to the protocol in the website of Centre for Disease Control and Prevention (CDC).@*Results@#A total of 86 strains of Streptococcus pyogenes were isolated from the 247 specimens. Their resistance rates to erythromycin, clarithromycin and clindamycin were 89.5%, 95.3% and 96.5%, respectively. However, these isolates showed high susceptibility to ampicillin (100.0%), ceftriaxone (100.0%), norfloxacin (90.7%) and chloramphenicol (95.3%). The positive rates of mefA, ermA, ermB and Tn916 genes were 20.9%, 24.4%, 98.8% and 97.7%. There was significant difference in the mefA-carrying rates between patients with scarlet fever and angina. The positive rates of virulence genes of speA, speB, speC, speG, speH, speI, speJ, speK, speL and speM were 8.1%, 100.0%, 95.3%, 100.0%, 80.2%, 90.7%, 10.5%, 100.0%, 5.8% and 5.8%. Seven emm types were identified and the predominant types were emm12.0 (75.6%), emm12.19 (9.3%) and emm1.0 (8.1%). The diameter of bacitracin inhibition zone was smaller in isolates of emm1.0 type than in emm12.0 type strains. The profile of virulence genes varied in the strains of different emm types. All types of strains carried speB, speG and speK genes. The isolates of emm1.0 type carried speA virulence gene, while speB, speC, speG, speH, speI and speK genes were more often identified in emm12.0 type isolates.@*Conclusions@#This study showed that emm types were associated with the profile of virulence genes and the diameter of bacitracin inhibition zone. It was recommended that the diameter of bacitracin inhibition zone should be measured in bacitracin inhibition susceptibility test apart from only observing the formation of inhibition zone.

Epidemiological and pathogenic features of a bacillary dysentery outbreak in a boarding school caused by Shigella sonneri / 中华流行病学杂志

Objective@#To investigate the etiologic and epidemiologic features of an infectious diarrhea outbreak in a boarding school in Fuyang city, Anhui province.@*Methods@#Traceability hypothesis of this study was tested according to the epidemiological characteristics of the cases. Feces, anal swabs, water samples and food residues related to the patients and chefs were collected for pathogen isolation and detection. Biochemical identification, virulence gene detection, drug susceptibility test, PFGE and multilocus sequence typing were performed.@*Results@#The incidence rate (3.41%) of different dormitory buildings within the water supply area by shallow wells was higher than that (0.98%) of the deep wells, with statistical significance (χ2=17.215, P<0.001). Sixteen strains belonged to the Shigella Sonneri family were isolated from the patient’s samples, and all carrying the ipaH gene. Seven strains belonged to sen and ial genes. Set1 gene that did not appear in all the 16 strains were highly resistant to ampicillin, tetracycline, compound xinnomine, cefazoline, cefotaxime, gentamicin, naphthidinic acid and streptomycin, including 9 strains to doxycycline. The pulse field pattern of the 16 strains of Shigella sonneri appeared the same, with the ST type as ST152.@*Conclusion@#When combined data from the etiological and epidemiological investigation, it was confirmed that Shigella sonneri was the pathogen of this outbreak, and water from the shallow wells might be responsible for the source of infection.

Trends in Bloodstream Infections and Antimicrobial Susceptibilities at a University Hospital in Korea Between 2007 and 2016

BACKGROUND: Blood culture is an important method for identifying infectious microorganisms and confirming that a selected antimicrobial treatment is appropriate. In this study, we investigated the annual changes in the frequencies of blood isolates and antibiotic susceptibility test (AST) results. METHODS: We created a large database comprising data on all patient-unique blood cultures obtained from January 2007 through December 2016. Blood specimens were cultured using the BD BACTEC FX system, and species identification and AST were performed using the VITEK 2 system. RESULTS: During the 10-year study period, a total of 203,651 blood culture results were collected. Of these, gram-positive cocci, gram-negative rods, and fungi were isolated in 2.15%, 0.55%, and 0.12% of the blood cultures, respectively. Escherichia coli was the most commonly isolated species (22.8%), followed by Staphylococcus epidermidis (16.8%), Klebsiella pneumoniae (8.1%), and Staphylococcus aureus (8.0%). Fungal species were isolated in 3.0% of all positive blood cultures. Candida albicans was the most commonly isolated species (1.1%), followed by Candida parapsilosis (0.6%). Methicillin resistance was seen in 55.2% of S. aureus isolates. The frequencies of vancomycin-resistant Enterococcus (VRE) and carbapenem-resistant Pseudomonas aeruginosa (CRPA) were 13.1% and 10.9%, respectively. The isolation rates of MRSA, VRE, and CRPA showed different patterns each year. CONCLUSIONS: Among the isolates, E. coli was the most common, followed by S. epidermidis and K. pneumoniae. This study represents a long-term analysis of bloodstream infections, and the results can be used to identify trends in the microorganisms isolated and their drug resistance.

In vitro antifungal susceptibility of Candida speciesisolated from diabetic patients

Abstract INTRODUCTION This study aims to evaluate the antifungal susceptibility of different species of Candida isolated from diabetic patients against eight antifungal agents. METHODS Susceptibility testing of 111 clinical isolates of Candida species was performed against 8 antifungals using the M27-A3 protocol of the Clinical and Laboratory Standards Institute (CLSI). RESULTS Voriconazole, lanoconazole, and caspofungin showed the highest in vitro activity against all the isolates of C. albicans. Resistance against the tested antifungals was only observed in the C. albicans isolates. CONCLUSIONS Our finding revealed that resistance against amphotericin B, itraconazole, ketoconazole, posaconazole, and fluconazole can be observed in C. albicans.