Microbiological Profile and Antibiogram of Uropathogens Isolate in Tertiary Care Hospital

Introduction: Urinary tract infections (UTIs) are one of the most common bacterial infections in humans, both in the community and the hospital settings. The distribution of antimicrobial susceptibility data of UTI-causing microorganisms changes from time to time and from place to place. The susceptibility data provided by regional microbiology laboratories helps to choose the empirical antimicrobials to treat UTI. Materials and methods: Total 1381 urine were received in Bacteriology section of microbiology department from January 2022 to August 2022. Urine was processed for culture according to standard operating procedures at Microbiology laboratory, tertiary care hospital. Identification and antibiotic sensitivity testing was performed by automated VITEK-2 compact system. Results: In total, 1381 urine were received during the study period, 433(31.35%) cultures yielded significant pathogens and no organisms were isolated in 941 cultures and some sample was contaminate. The most commonly isolated organism were Escherichia coli (38.8%), followed by Candida spp. 88(20.32%), Klebsiellaspp (13.40%), Pseudomonas aeruginosa (12.24%), Gram positive organisms 12(2%) isolated. E. coli were most susceptible to Amikacin (87%), Fosfomycin (83%), colistin (82%), and tigecycline (78.39%). Klebsiella spp were most susceptible to colistin (72%). Nearly, 75% strains of Klebsiella spp were showing resistance to Carbapenems due to Carbapenemase production. Conclusion: Over enthusiastic use of the antibiotic has resulted in the emergence of drug-resistant bacterial strains in patients. The study of antimicrobial susceptibility patterns of uropathogens in a particular area can guide the clinicians in the rational choice of empirical treatment to prevent the misuse of antibiotics.

Bacteriuria asintomática en el embarazo. Serie de casos en una clínica privada de Uruapan, Michoacán / Asymptomatic bacteriuria in pregnancy. Cases series in a private clinic in Uruapan Michoacán

Resumen OBJETIVO: Determinar la prevalencia de bacteriuria asintomática, su adecuado tratamiento con base en el reporte del antibiograma de pacientes embarazadas que acudieron a control prenatal en una clínica privada. MATERIALES Y MÉTODOS: Estudio retrospectivo de una serie de casos, descriptivo y transversal efectuado en mujeres embarazadas que acudieron a control prenatal en la consulta externa del Hospital Fray Juan de San Miguel de Uruapan, Michoacán, entre el 1 de enero de 2019 y el 31 de diciembre de 2021. Variables de estudio: edad de las madres, semanas de embarazo, embarazos, bacteriuria, agente causal, antibiótico indicado y respuesta al tratamiento. Para interpretar los resultados obtenidos se aplicaron estadística descriptiva, medidas de tendencia central y de frecuencias simples. RESULTADOS: Se estudiaron 227 embarazadas; de ellas, 49 tuvieron un urocultivo positivo. La prevalencia de bacteriuria asintomática fue de 21.59. Escherichia coli fue el agente causal más reportado. De los urocultivos de control 36 de 37 resultaron negativos y solo 1 de 37 fue positivo. CONCLUSIÓN: La prevalencia de bacteriuria asintomática de este estudio fue de 21.59% y el principal agente etiológico asilado E. coli en la mayoría de los casos, con sensibilidad a nitrofurantoína en la mayoría de los casos.

Abstract OBJECTIVE: To determine the prevalence of asymptomatic bacteriuria, its adequate treatment based on the antibiogram report of pregnant patients who attended prenatal control in a private clinic. MATERIALS AND METHODS: Retrospective study of a case series, descriptive and cross-sectional carried out in pregnant women who attended prenatal control in the outpatient clinic of the Hospital Fray Juan de San Miguel de Uruapan, Michoacán, between January 1, 2019 and December 31, 2021. Study variables: mothers' age, weeks of pregnancy, pregnancies, bacteriuria, causative agent, indicated antibiotic and response to treatment. Descriptive statistics, measures of central tendency and simple frequencies were used to interpret the results obtained. RESULTS: A total of 227 pregnant women were studied; 49 of them had a positive urine culture. The prevalence of asymptomatic bacteriuria was 21.59. Escherichia coli was the most reported causative agent. Of the control urine cultures 36 of 37 were negative and only 1 of 37 was positive. CONCLUSION: The prevalence of asymptomatic bacteriuria in this study was 21.59% and the main etiologic agent was E. coli in most cases, with sensitivity to nitrofurantoin in most cases.

Antimicrobial Resistance Patterns of Bacterial Isolates from Tertiary Care Hospital of North India

Antimicrobial resistance is one of the serious health threats in the world and death due to it is projected to reach up to 10 million every year by 2050. In India, the government implemented a National Action Plan on Antimicrobial Resistance (NAP-AMR) intending to develop a laboratory-based AMR surveillance system, guidelines and practices to promote the awareness of the rational use of antibiotics in healthcare communities. In this plane, only five major hospitals are included which is not sufficient to provide the actual status of AMR. The present study focused on the AMR patterns analysis for bacteria isolated from the different health units in SSB heart and multispecialty hospital, Faridabad, Haryana. Clinically relevant samples such as pus & Body fluid, respiratory, urine and blood samples from the suspected patients were screened over a period of 5 months (February to June 2021) and AMR pattern analysis was carried out by using the disc diffusion method (Kirby–Bauer test) and interpretation was made as per the guidelines of recent Clinical and Laboratory Standards Institute. The study revealed that E. coli was the most dominated resistant bacteria. Moreover, aminoglycosides showed 55% resistance against maximum bacterial isolates from respiratory samples while fosfomycin had higher sensitivity i.e. 85% against pathogens from urine samples. The present study concluded that regular susceptibility testing should be conducted with a defined interval to detect the current trend of resistance which helps clinicians in the effective infectious disease management and leads to reducing the burden of AMR.

Antibiogram Profile of Enteric Pathogens Isolated from Fomites in Cross River University of Technology Medical Centre, Calabar, Nigeria

Background: Healthcare environments are considered as potential reservoirs for pathogenic microorganisms especially those responsible for nosocomial infections. Such microorganisms often present with varying degrees of drugs resistance. This study was aimed at evaluating the Cross River University of Technology (CRUTECH) Medical Center Environment for the presence of pathogenic bacterial contaminants and antibiotics susceptibility profile of such isolates. Materials and Methodology: A total of 72 swab samples were collected from nineteen frequently touched hospital surfaces and processed using the standard bacteriological procedures. The emergent bacterial colonies were identified using phenotypic and biochemical tests. Antibiotic susceptibility testing of the presumptively identified isolates was carried out using Kirby-Bauer抯 method. Results: Out of 72 swab samples collected, 44 (61.1%) were positive for Enteric bacterial pathogens. The mean viable count ranged from 1.8 x 106 Cfu/cm2 (weighing scales) to 2.41 x 107 Cfu/cm2 (wash sinks). The most prevalent isolate was E. coli (48 of 127, 37.8%) followed by Klebseilla sp (27 of 127, 21.3%), Salmonella sp (19 of 127, 14.9%), Proteus sp (12 of 127, 9.4%), Citrobacter sp (11 of 127, 8.7%), Enterobacter sp (7 of 127, 5.5%) while Shigella sp (3 of 127, 2.4%) was least isolated. Isolates demonstrated high level of susceptibility to Norfloxacin 124 (97.6%), Imipenem 116 (91.3%) and Chloramphenicol 105 (82.7%). Resistance to Erythromycin was 95 (74.8%), Gentamycin 84 (66.1%) and Amikacin 82 (64.6%). Conclusion: Contamination of healthcare surfaces by multi-drugs resistant pathogens is a potential risk, especially to hospitalized patients and health care workers. Thus, it is therefore imperative that appropriate hygienic measures be implemented to suppress any potential microbial cross-contamination.

Survey on the Inpatient’s Antibiogram Affected by Sampling Day Difference / 医薬品情報学

Objective: Antibiogram preparation is important for the proper selection of antimicrobial agent in empiric therapy. It is recommended to prepare the antibiogram separately for inpatients and outpatients. However, the antimicrobial agent susceptibility of bacteria detected from an inpatient is thought to be different when detected at an early date after admission and after a certain period after admission.Methods: In this study, we defined the bacteria detected from an inpatient within 3 days after admission as “brought bacteria” and those over 3 days after admission as “bacteria detected after admission.” Antimicrobial agent susceptibilities of brought bacteria and bacteria detected after admission in our hospital between April 2018 and March 2019 were compared for Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa.Results: The occurrence rate of Escherichia coli producing an extended β-lactamase of bacteria detected after admission was found to be significantly higher compared with that of brought bacteria. The sulbactam/ampicillin susceptibility rate of bacteria detected after admission was significantly lower than that of brought bacteria. The occurrence rate of the two drug-resistant Pseudomonas aeruginosa detected after admission tends to be higher compared with that of brought bacteria.Conclusion: Our findings showed that the susceptibility rate of some drugs differed substantially between brought bacteria and bacteria detected after admission. Therefore, the inpatient’s antibiogram is thought to be prepared by separating bacteria detected at an early date after admission and after a certain period after admission, leading to the proper selection of antimicrobial agent in empiric therapy tailored to a patient’s number of days in the hospital.

Efecto antimicrobiano in vitro de extractos de Caesalpinia Spinosa (algarrobo) sobre patógenos orales / In vitro antimicrobial effect of Caesalpinia Spinosa (carob tree) extracts on oral pathogens

La resistencia antimicrobiana es un problema de sa-lud pública mundial. Las infecciones por microorga-nismos resistentes pueden ser altamente transmisi-bles e incluso causar la muerte. Este hecho genera grandes costos para los pacientes y para los servi-cios de salud. El objetivo del presente trabajo fue de-terminar el efecto antimicrobiano in vitro de extractos etanólicos de Caesalpinia spinosa sobre el crecimien-to de Enterococcus faecalis, Staphylococcus aureus y Candida albicans. Se recolectaron y certificaron muestras de C. spinosa. Se obtuvieron extractos de hojas, vainas y semillas en concentraciones de 100%, 75%, 50% y 25%. Mediante Kirby - Bauer, se cargaron los discos con los extractos y se depositaron en el medio inoculado con cepas de E. faecalis, S. aureus y C. albicans; junto a un CP (antimicrobiano), y un CN (etanol). Las placas se incubaron a 370°C durante 24 horas, y posteriormente se midieron los halos de inhi-bición con un vernier digital. Destaca el valor del halo de extracto de vainas; superó al de Ampicilina 10mg, sobre el E. faecalis. El extracto de vainas presentó ma-yor diámetro de inhibición (19mm), el de semillas pre-sentó el más bajo (1mm). ANOVA arrojó diferencia es-tadísticamente significativa entre los datos obtenidos para todos los extractos. En conclusión, los extractos etanólicos de Caesalpinia spinosa tienen efecto anti-microbiano in vitro sobre Enterococcus faecalis, Sta-phylococcus aureus y Candida albicans. La actividad antimicrobiana del extracto es directamente propor-cional a su concentración. Los extractos de C. spinosa podrían ser utilizados como coadyuvantes en el trata-miento contra Enterococcus faecalis, Staphylococcus aureus, Candida albicans, que están relacionados con patologías orales (AU)

Antimicrobial resistance is a global public health problem. Infections with resistant microorganisms can be highly transmissible and even cause death. This fact generates great costs for patients and for health services. The objective of this work was to determine the in vitro antimicrobial effect of ethanolic extracts of Caesalpinia spinosa on the growth of Enterococcus faecalis, Staphylococcus aureus and Candida albicans. Samples of C. spinosa were collected and certified. Leaf, pod and seed extracts were obtained at concentrations of 100%, 75%, 50% and 25%. Using Kirby-Bauer, the disks were loaded with the extracts and deposited in the medium inoculated with strains of E. faecalis, S. aureus and C. albicans; together with a CP (antimicrobial), and a CN (ethanol). The plates were incubated at 370°C for 24 hours, then the inhibition halos were measured with a digital vernier. The value of the pod extract halo stands out, surpassing that of Ampicillin 10mg, over E. faecalis. The pod extract presented the greatest diameter of inhibition (19mm), the seed extract presented the lowest (1mm). ANOVA showed a statistically significant difference between the data obtained for all the extracts. In conclusion, the ethanolic extracts of Caesalpinia spinosa have an in vitro antimicrobial effect on Enterococcus faecalis, Staphylococcus aureus and Candida albicans. The antimicrobial activity of the extract is directly proportional to its concentration. C. spinosa extracts could be used as adjuvants in the treatment against Enterococcus faecalis, Staphylococcus aureus, Candida albicans, which are related to oral pathologies (AU)

Perfil de sensibilidade a antibióticos de um isolado de Providencia rettgeri proveniente de equino / Antibiotic sensitivity profile in isolate of Providencia rettgeri from a horse

Introdução: a meningite bacteriana em equinos é uma enfermidade frequente em animais jovens. Streptococcus spp., Staphylococcus aureus e Escherichia coli são as bactérias mais comumente isoladas nesses casos. Apesar da bactéria Providencia rettgeri já ter sido isolada em casos de meningite humana e de crocodilo, não há relatos de seu isolamento em equinos. Objetivo: relatar o isolamento e a identificação da bactéria P. rettgeri de um potro com sintomas neurológicos e avaliar o perfil de sensibilidade a antibióticos deste isolado. Metodologia: o isolamento foi realizado a partir do líquido cefalorraquidiano do potro, por meio de cultivo em meio ágar chocolate. Após isolamento, as colônias formadas foram identificadas por equipamento Biotyper, baseado em espectrometria de massa. O perfil de sensibilidade foi definido por teste de difusão em discos, seguindo metodologia relatada pelo CLSI M2-A8 em 2003, sendo a bactéria classificada como resistente, padrão indeterminado ou sensível aos antibióticos, de acordo com o descrito pelo EUCAST em 2021. Resultados: este é o primeiro relato do isolamento de P. rettgeri como agente etiológico de meningite em potro. Dos 15 antibióticos testados, a bactéria foi resistente a 9, sensível a 5 e com padrão indeterminado a 1 antibiótico. Conclusão: nossos resultados indicam que P. rettgeri deve ser considerada entre possíveis agentes etiológicos de quadros neurológicos em equinos e que testes de sensibilidade a antibiótico são fundamentais, uma vez que essa bactéria já apresenta resistência a diversos antibióticos disponíveis comercialmente.

Introduction: Bacterial meningitis in horses is a frequent disease in young animals. Streptococcus spp., Staphylococcus aureus and Escherichia coli are the most commonly isolated bacteria in these cases. Although Providencia rettgeri bacterium has already been isolated in cases of human and crocodile meningitis, there are no reports of its isolation in cases of meningitis in horses. Objective: to report isolation and identification of the P. rettgeri bacteria from a foal with neurological symptoms and to assess antibiotic sensitivity profile in isolate of it. Methods: isolation was performed from the foal's cerebrospinal fluid, through cultivation in chocolate agar medium. After isolation, formed colonies were identified by Biotyper equipment, based on mass spectrometry. Sensitivity profile was verified by disk diffusion test, according to methodology that was reported by CLSI M2-A8 in 2003, which classified bacteria as resistant, indeterminate pattern or sensitive to antibiotics, as described by EUCAST in 2021. Results: this is the first report on isolation of P. rettgeri as an etiologic agent of meningitis in foals. Among 15 antibiotics that were tested, results showed bacteria resistence to 9 antibiotics, bacteria sensitivity to 5, but undetermined pattern to 1 antibiotic. Conclusion: results indicate that P. rettgeri shall be considered among potential etiologic agents of neurological conditions in horses and that antibiotic sensitivity tests are essential, since this type of bacterium is already resistant to several commercially available antibiotics.

Efecto antibacteriano in vitro de extractos de Caesalpinia spinosa sobre cepas de Staphylococcus aureus resistentes a antibióticos betalactámicos / In vitro antimicrobial effect of Caesalpinia spinosa extracts against betalactam antibiotics resistant Staphylococcus aureus strains

Objetivo. Determinar el efecto antimicrobiano de extractos etanólicos de Caesalpinia spinosa sobre cepas de Staphylococcus aureus penicilino y meticilino resistentes. Métodos. Se utilizaron las cepas de S. aureus ATCC 11632 y 33592, los extractos etanólicos de hojas, vainas y semillas de C. spinosa, se obtuvieron por maceración en concentraciones de 25%, 50%, 75% y 100%. Se utilizó el método de Kirby-Bauer, los discos de papel filtro se cargaron con los extractos y se depositaron sobre el medio, inoculado con una suspensión de S. aureus a 0,5 McFarland. El control positivo fueron discos de ampicilina y el control negativo discos impregnados con etanol. Después de 24 horas se midieron los diámetros de los halos con un calibrador Vernier. Resultados. Se registraron halos de hasta 18 mm de diámetro con los extractos de hojas al 100%, 17 mm con extractos de vainas y 14 mm con extractos de semillas sobre S. aureus ATCC 33592. En el caso de la cepa ATCC 11632, se registraron halos de hasta 14 mm con extractos de hojas y vainas al 100%, y de hasta 8 mm con extractos de semillas. La prueba de ANOVA indicó que existieron diferencias significativas entre los halos obtenidos con los diferentes tipos de extractos, a diferentes concentraciones. Conclusión. Se determinó que todos los extrac- tos de C. spinosa poseen actividad antimicrobiana sobre las dos cepas estudiadas, con un patrón directamente proporcional entre el efecto y la concentración.

Objective. To determine the antimicrobial effect of ethanolic extracts from Caesalpinia spinosa against penicillin and methicillin resistant strains of Staphylococcus aureus penicillin and methicillin. Methods. S. aureus ATCC 11632 and 33592 strains were used, ethanolic extracts from C. spinosa leaves and pods were obtained by maceration at concentrations of 25%, 50%, 75% and 100%. The Kirby-Bauer method was used, where filter paper discs were loaded with leaf, pod and seed extracts and deposited on the medium, inoculated with a 0.5 McFarland suspension of S. aureus. Ampicillin discs were used as positive con- trol and ethanol-impregnated discs as negative control. After 24 hours, the diameters of the halos were measured with a Vernier caliper. Results. Haloes up to 18 mm in diameter with 100% leaf extracts, 17 mm with pod extracts and 14 mm with seed extracts were recorded against S. aureus ATCC 33592; for ATCC strain 11632, haloes up to 14 mm with 100% leaf and pod extracts and up to 8 mm with seed extracts were recorded. The ANOVA test indicated significant differences between the inhibition halos obtained with the different types of extracts, at different concentrations. Conclusion. It was determined that all C. spinosa extracts possess antimicrobial activity against the two strains studied, with a pattern directly proportional between the effect and concentration.

What Are the Roles of Carbapenems in an Institution-specific Epidemiological Antibiogram in East Trinidad?

ABSTRACT Objective: To provide an overview of the development of an institution-specific epidemiological antibiogram. Emphasis was on last-line antibiotics, such as carbapenems. Methods: In 2013, the antibiograms of various organisms were retrieved from the computerized database of the Microscan (Siemens Healthcare) at the Microbiology Laboratory of the Sangre Grande Hospital in East Trinidad, West Indies. These were divided into blood and urine specimen antibiograms. All the wards and hospital clinics were included. A 20% cut-off was used to determine that a particular antibiotic or antibiotic class could be used for empiric therapy. All the organisms were not chosen. Only the most common and clinically relevant organisms were chosen. Results: Blood: Escherichia coli, Klebsiella pneumonia, Proteus mirabilis: Imipenem, mero- penem, ertapenem showed greater than 80% sensitivity, respectively. Pseudomonas aeruginosa: ceftazidime, ciprofloxacin, gentamicin, levofloxacin and tazobactam/piperacillin showed 100%, 80%, 80%, 100% and 100% sensitivity, respectively. Urines: E. coli, Klebsiella pneumonia, Proteus mirabilis: Imipenem, meropenem, ertapenem, were greater than 80% sensitive. Enterobacter cloacae: Imipenem, meropenem were 92%, 100% sensitive. Pseudomonas aeruginosa: tazobactam-piperacillin and amikacin were both 85% susceptible. Acinetobacter baumanii/haemolyticus: All the antibiotics were above the 20% resistance threshold. Conclusion: Patient-specific antibiograms and unit-specific trends (eg, ICU, surgical wards and outpatient clinic) can be used as a guide in patients with less severe infections. Carbapenems can still be used empirically, in East Trinidad, for sepsis.

Evaluation of the antimicrobial susceptibility testing process in clinical microbiology laboratories at Niamey, Niger

Background: Risk assessment is the means of identifying and evaluating potential errors or problems that may occur in testing process. The aim of this study was to perform risk assessment of antimicrobial susceptibility testing (AST) process in clinical microbiology laboratories of Niamey, Niger Republic. Methodology: We conducted a descriptive cross-sectional study from October 1 to December 31, 2019, to evaluate AST performance in seven clinical microbiology laboratories at Niamey, the capital city of Niger republic. The evaluation focused on the determination of the criticality index (CI) of each critical point (frequency of occurrence of anomalies, severity of the process anomaly, and detectability of the anomaly during the process) in the AST process and the performance of the AST through an observation sheet using two reference strains; Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29213. Results: The criticality index (CI) was greater than 6 for most of the critical points related to material, medium, equipment, method and labour for the AST process in all the laboratories. A range of 18-100% errors on the inhibition zone diameters of the reference strains were observed. Major and/or minor categorization (Sensitive S, Intermediate I and Resistance R) discrepancies were found at all the laboratories for either one or both reference strains. The antibiotics most affected by the S/I/R discrepancies were trimethoprim (100%), vancomycin (100%), amoxicillin (80%) and amoxicillin + clavulanic acid (70%). Conclusion: This study showed a deficiency in the control of critical control points that impacts the performance of the AST reported by the laboratories in Niger. Corrective actions are needed to improve the performance of AST in clinical microbiology laboratories in Niger

Prevalence of symptomatic urinary tract infection and bacterial spectrum of diabetic and non-diabetic patients at the two teaching hospitals in Enugu, Nigeria

Background: Diabetes mellitus is a group of metabolic disorder characterized by relative or absolute lack of insulin. When this condition is not properly managed, it can lead to complications that make diabetic patients vulnerable to urinary tract infections (UTI). The objectives of this study are to determine the prevalence of microbiologically confirmed UTI and the spectrum of uropathogens in diabetic and non-diabetic patients with clinical features of UTI attending the two tertiary hospitals in Enugu State, Nigeria. Methodology: Clean catch specimen of single mid-stream urine sample was collected from each of 60 (22 males, 38 females) diabetic and 60 (22 males, 38 females) non-diabetic patients enrolled using stratified random sampling method. The samples were cultured on standard microbiological culture media (MacConkey and Blood agar plates) and incubated aerobically at 37◦C for 24 hours. Plates with significant bacteria growth (>105 CFU/ml) were processed further for bacterial identification using conventional biochemical test scheme. Antibiotic susceptibility test (AST) of each isolate to 17 selected antibiotics was performed by the modified disc diffusion method. Results: Of the total 120 patients enrolled, 101 had bacterial pathogens isolated from their voided urine samples; 51 of 60 (85.0%) diabetics and 50 of 60 (83.3%) non-diabetics (p=0.802). Bacteria were isolated in 59.1% (13/22) of diabetic and 54.5% (12/22) of non-diabetic male patients compared to 100% (38/38) isolation rate in diabetic and non-diabetic female patients. The most frequently isolated bacteria in the diabetic patients were Proteus spp (18.6%), Klebsiella spp (16.9%) and Escherichia coli (15.5%) while the most frequently isolated bacteria among the non-diabetic patients were E. coli (30.0%), Proteus spp (26.3%) and Enterobacter spp (14.0%). Apart from Klebsiella spp which was more frequently isolated from the diabetic (16.9%) than non-diabetic patients (6%) (p=0.039), the frequency other bacterial pathogen isolation such as Proteus spp, E. coli, Enterobacter spp, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus spp was not significantly different between the two population groups (p>0.05). The Gram-positive and Gram-negative bacteria were highly sensitive to imipenem in both diabetic and non-diabetic patients, but the isolates from both study groups exhibited low susceptibility to amoxicillin, nitrofuran- toin, cefixime and cefuroxime. Conclusion: Although the overall frequency of bacterial pathogen isolation in the diabetic and non-diabetic patients was not significantly different, females had a higher pathogen isolation rate than the males, and diabetic females had a higher frequency of polymicrobial infections compared to non-diabetic females and the male population. The high antimicrobial resistance of the isolated bacteria pathogens underscores the need for clinical microbiology laboratory testings to optimize the management of UTI in diabetic patients.

Investigation of the effects of some phytochemicals on Yersinia ruckeri and antimicrobial resistance / Investigação dos efeitos de alguns fitoquímicos em Yersinia ruckeri e resistência antimicrobiana

Abstract In this study, it is aimed to investigate the effects of Moringa oleifera and Sorbus domestica plant extracts on bacterial disease agents Yersinia ruckeri in aquaculture. Morphological and biochemical properties of 2 different Y. ruckeri isolates were determined. Then, Real-Time PCR analysis and gene sequencing of the isolates were identified. Phytochemicals (M. oleifera and S. domestica) and antibiotics (Oxytetracycline (OX) and Enrofloxacin (ENR)) were used together in the antibiogram test of antibiotics compared to the effect status of antibiotics. Also, the effects of phytochemicals on Y. ruckeri growth was examined comparatively by spectrophotometrically measuring at 600 nm wavelength every 2 hours according to bacterial growth densities with 10 different groups formed on TSB medium. As a result of the study, it was observed that the isolates formed Gram negative, catalase positive, oxidase negative, mobile and typical Y. ruckeri colonies. After the biochemical tests performed with Microgen ID panel, 99.85% similarity was determined. The isolates overlap with the 16S rRNA gene region after sequence analysis, and 99% of the isolates were similar in phylogenetic analysis. After the antibiogram test, Oxytetracycline and Enrofloxacin antibiotics were resistant to Y. ruckeri but the effects of phytochemicals were less on solid medium (MHA). As a result of the measurements carried out in liquid medium (TSB), it was observed that phytochemicals such as M. oliefera and S. domestica inhibit the growth of bacteria by 40-50%. As the importance of antibiotic resistance is increasing day by day, we believe that these phytochemicals will give positive results in treatment instead of using antibiotics.

Resumo Neste estudo, objetiva-se investigar os efeitos dos extratos de plantas de Moringa oleifera e Sorbus domestica sobre agentes bacterianos Yersinia ruckeri na aquicultura. Foram determinadas as propriedades morfológicas e bioquímicas de 2 isolados diferentes de Y. ruckeri. Em seguida, a análise de PCR em tempo real e o seqüenciamento genético dos isolados foram identificados. Fitoquímicos (M. oleifera e S. domestica) e antibióticos (Oxitetraciclina e Enrofloxacina) foram usados ​​juntos no teste de antibiograma dos antibióticos em comparação com o status de efeito dos antibióticos. Além disso, os efeitos dos fitoquímicos no crescimento de Y. ruckeri foram examinados comparativamente por medição espectrofotométrica no comprimento de onda de 600 nm a cada 2 horas de acordo com as densidades de crescimento bacteriano com 10 grupos diferentes formados no meio TSB. Como resultado do estudo, observou-se que os isolados formaram colônias Gram-negativas, catalase-positivas, oxidase-negativas, móveis e típicas de Y. ruckeri. Após os testes bioquímicos realizados com o painel Microgen ID, foi determinada uma similaridade de 99,85%. Os isolados se sobrepõem à região do gene 16S rRNA após a análise da sequência e 99% dos isolados foram semelhantes na análise filogenética. Após o teste do antibiograma, os antibióticos Oxitetraciclina e Enrofloxacina foram resistentes a Y. ruckeri, mas os efeitos dos fitoquímicos foram menores no meio sólido (MHA). Como resultado das medições realizadas em meio líquido (TSB), observou-se que os fitoquímicos inibem o crescimento de bactérias em 40-50%. Como a importância da resistência aos antibióticos está aumentando dia a dia, acreditamos que as plantas que são mais alternativas e mais adequadas para o uso de antibióticos hoje em dia darão resultados positivos no tratamento.

Validación de un dispositivo point-of-care para la detección rápida de infección urinaria y susceptibilidad antimicrobiana / Validation of point-of-care device for rapid detection of urinary tract infection and antibiotic susceptibility

Resumen Introducción: Las infecciones del tracto urinario (ITU) presentan una elevada prevalencia en el ámbito comunitario. Un rápido diagnóstico microbiológico es esencial para asegurar una terapia adecuada y efectiva. Objetivo: Evaluar un kit de antibiograma rápido (KAR®) en formato point-of-care para la detección rápida de ITU y sensibilidad antimicrobiana. Material y Métodos: El dispositivo KAR® se diseñó y desarrolló en colaboración con ingenieros técnicos y microbiólogos clínicos. Su evaluación se realizó a través de un estudio multicéntrico en el que participaron tres hospitales españoles. Para ello, se realizaron distintos ensayos in vivo con el fin de determinar la correlación del dispositivo con las técnicas microbiológicas de referencia. Resultados: Se ensayó un total de 400 muestras de orinas procedentes de pacientes con sospecha de ITU. El dispositivo KAR® proporcionó rápidos resultados (tiempo medio de positividad de 7,8 ± 1,5 h) con 97% de sensibilidad, 89% de especificidad y 87% de concordancia para la detección de bacteriuria significativa. Los porcentajes de especificidad para los antimicrobianos testados fueron: ciprofloxacina (97%), fosfomicina (94%), cotrimoxazol (84%), ampicilina (80%) y amoxicilina/ácido clavulánico (55%). Conclusión: El dispositivo KAR® puede ser una herramienta útil para el diagnóstico de ITU en pacientes ambulatorios, especialmente en áreas de bajo nivel socio-económico.

Abstract Background: Urinary tract infections (UTI) presents a high prevalence in the community setting. Rapid and accurate microbiological diagnosis is essential to ensure adequate and effective therapy. Aim: To evaluate a rapid antibiogram kit (KAR®) in point-of-care format for rapid detection of UTI and antibiotic susceptibility. Methods: The KAR® device has been designed and developed in collaboration with technical engineers and clinical microbiologists. Its evaluation has been carried out through a multicenter study in which three Spanish hospitals have participated. Thus, different in vivo tests have been implemented in order to determine device correlation with the reference microbiological techniques. Results: During the study period, a total of 400 urine samples from patients with suspected ITU were tested. The KAR® device provided fast results (mean positivity time of 7,8 ± 1,5 hours) with 97% sensitivity, 89% specificity and 87% agreement for the detection of significant bacteriuria. The percentages of specificity for the antibiotics tested were: ciprofloxacin (97%), fosfomycin (94%),cotrimoxazole (84%), ampicillin (80%) and amoxicillin/clavulanic acid (55%). Conclusion: The KAR® device could be a useful tool for diagnosing UTI in outpatients, especially in areas of low socio-economic level.

Study of the utility of blood culture in febrile children aged between 3 months to 12 years admitted in a tertiary centre

Background: Fever, the most common complaint that led patients to seek healthcare, indicates an underlying infection which could either be simple self-limiting viral infections or life threatening bacterial infections. It's greatest challenge is the risk of occult bacteraemia, for which blood culture is the gold standard for the diagnosis. Objectives was to determine the proportion of blood culture positives among febrile children and to describe the bacteriological profile and antibiogram of blood culture isolates.Methods: A cross sectional study was done in the Department of Paediatrics and Microbiology, RIMS Hospital, Imphal. After obtaining consent (verbal assent in >7 years), blood culture samples were drawn from 200 children aged between 3 months to 12 years. The data was analysed using descriptive statistics. Chi square test was used and p-value of less than 0.05 taken as statistically significant.Results: Culture positivity was seen in 17 cases (8.5%) of which, participants who were less than 1 year of age and without proper immunisation record showed the highest positivity rate. It was higher in fever with localizing signs (9.2%) than those with fever without focus (7.8%). Gram positives constituted 11 (64.7%) of the isolates while 35.3% were Gram negatives. Staphylococcus aureus was the only Gram positive isolate. Of them, 4 were MRSA but all the strains were sensitive to Vancomycin and Linezolid. The most common Gram negative isolate was Acinetobacter spp and 80% of them were sensitive to Aminoglycosides while most of the Gram negatives were resistant to Ampicillin and 3rd generation Cephalosporins. All Acinetobacter spp were sensitive to Carbapenems but the only Pseudomonas spp isolated was sensitive only to Colistin.Conclusions: Blood culture positivity rate is relatively low in this study. However, studies with larger sample sizes are recommended to validate the findings. We emphasise the need for antibiotic stewardship

Microbial Profile and Antibiogram of Pus Isolate in a Tertiary Care Hospital of Western Odisha.

Pus sample is one of the major samples received in Microbiology laboratory. Overall incidence of wound sepsis in India is 10-33%. A predictable microbial profile in wound infection is very much important for clinician to start empirical therapy and also to combat drug resistance. We wanted to assess the microbial profile and antibiogram in pus isolates causing wound infection in a tertiary care centre of Western Odisha.METHODSThis is a cross-sectional descriptive study. 287 pus samples in total received from different departments of Hitech Medical College, Rourkela, from June 2019 to Dec. 2019 were processed using standard microbial culture methods and antibiotic sensitivity was done as per CLSI guideline.RESULTSAmong 287 samples, 187 (65%) showed positive growth. Among 187 positive samples, 64% were from male population. Middle age group (21-40 and 41-60) was most affected age group (36% and 37.4% respectively). Most commonly isolated bacteria were Staphylococcus aureus (38%) followed by Pseudomonas species (17.10%). Least common bacteria were Streptococcus pyogenes (0.5%). Most of the gram-negative bacilli showed high sensitivity towards imipenem (53-93%), meropenem (80-100%), piperacillin tazobactam (67-100%) and amikacin (67-88%) but lower sensitivity to 3rd generation cephalosporins (36-73%), cotrimoxazole (60-80%), amoxiclav (20-75%) and fluoroquinolones (42-73%). Staphylococcus aureus showed high sensitivity to linezolid (92%), aminoglycoside (84%), clindamycin (88%) and cotrimoxazole (72%) but lower sensitivity to erythromycin (25%), amoxiclav (40%) and fluoroquinolones (44%).CONCLUSIONSStaphylococcus aureus and Pseudomonas species were the most common bacteria isolated in our study with variable antibiogram. Pus aspirate was better sample than pus swab. A continuous inspection should be carried out to monitor the antibiogram of wound isolate to formulate and implement antibiotic policy in our hospital.

Isolation of Pseudomonas aeruginosa from Bovine Mastitic Milk Sample Along with Antibiogram Study

A total of 4378 cattle and buffalo were screened during period of study (July 2018-June 19). Out of which 27 Pseudomonas aeruginosa were isolated. The isolates were confirmed phenotypically based on pigment production on nutrient agar. These were then confirmed by PCR amplification of species specific oligonucleotide sequences. All the 27 isolates amplified 956bp amplicon 16srRNA Pseudomonas aeruginosa species specific nucleotide sequence. The isolates were also checked for exo and aglD virulence associated genes. All of them amplified 540bp and 313bp amplicon of exo gene and aglD gene. All the isolates were subjected to antibiotic sensitivity testing. Most of the isolates showed highest sensitivity for levofloxacin, streptomycin and enrofloxacin followed by gentamicin, moxifloxacin and amikacin. Neomycin, cefoperazone and ceftriaxone were intermediate in action

Characterisation of Escherichia coli Recovered from Wild Animals Kept at Bikaner Zoo for Their Antibiotic Resistance

In the present study, a total of 41 E. coliisolates obtained from Boselaphus tragocamelus (5), Antelope cervicapra(18) and Gazella gazelle (18) maintained at Bikaner (Rajasthan) zoo were subjected to antibiogram determination against 15 antibiotics belonging to four different classes and were also genotyped for detecting presence of blaTEM,sul-2, strA and aadAgenes. Antibiogram study revealed highest efficacy of ciprofloxacin (90.2%) followed by nalidixic acid (75.6%) and chloramphenicol (68.4%) and high resistance to β lactam, Sulfamethoxazole and Aminoglycoside antibiotics. The overall presence of blaTEM, sul-2, strA and aadA genes was detected in 95.12%, 80.48%, 60.97% and 87.8% in isolates.

Hypoalbuminemia and Central Venous Catheter as Risk Factors for Multidrug-resistant Healthcare-associated Pneumonia in an Intensive Care Setting

The rapid emergence of antibiotic­resistant bacteria is a threat to global health particularly in the area of healthcare­associate pneumonia (HCAP) where there is high rate of mortality. In general, guidelines should serve as a framework that needs to be complemented by local antibiogram data due to multiple factors influencing the development of multidrug­resistant (MDR) HCAP. Failure to administer prompt and appropriate empirical therapy would often result in a high mortality rate. Based on these concerns, the aim of the study was to evaluate the appropriate empirical use of antibiotic and risk factors of MDR HCAP based on local pathogen resistant pattern. This was a retrospective analysis on HCAP in critical care of a tertiary­care hospital with data fromJanuary 2016 to December 2018. Patients diagnosed with HCAP: hospital­associated pneumonia (HAP) and ventilator­associated pneumonia (VAP), with positive bacterial cultures were included into the study. Of the 269 patients and isolates included, 160 (59.5%) had MDR strains. The top causative pathogens isolated were Acinetobacter baumannii(n=104, 38.7%), Pseudomonas aeruginosa(n=66, 24.5%), Klebsiella spp(n==55, 20.4%), and Staphylococcus aureus(n=16, 5.9%). The incidence of inappropriate empirical antibiotic was significantly higher in patients with MDR HCAP (n=135, 84.4%) compared to those with non­MDR HCAP (n=34, 31.2%) (p < 0.001). Mortality was significantly higher in patients receiving inappropriate empirical therapy (n = 118, 72.4%) compared to those receiving appropriate empirical antibiotic (n = 36, 54.5%) (P = 0.009). The independent risk factors for MDR HCAP identified in this study were hypoalbuminemia (odds ratio [OR] 3.43, 95% confidence interval [CI] 1.08 –10.87, p = 0.036) and indwelling central venous catheter (OR 5.65, 95% CI 1.13 –28.18, p = 0.035). This work serves as a basis for a center­specific guideline to improve antibiotic use among HCAP patients in intensive care setting.

Aetiological Profile and Antibiogram of Urinary Isolates Causing UTI in Patients Attending a Tertiary Care Hospital of Western Odisha

Although UTI is a female disease, males are also susceptible during the neonatal period and old age. Asymptomatic bacteriuria, cystitis and acute urethral syndrome are the most common clinical type. Etiological profile is variable in different geographical areas, but E. coli is the most common agent worldwide. Klebsiella, Proteus, Pseudomonas are important causes of hospital acquired UTI. Culture sensitivity of early morning mid-stream urine collected by clean catch technique is the gold standard method of diagnosis of UTI. Sensitivity to 3rd generation cephalosporins and cotrimoxazole is variable in different areas but aminoglycoside, nitrofurantoin and carbapenem are almost sensitive worldwide. Resistance to nitrofurantoin and carbapenem has been reported in many areas of world. Before starting empirical therapy, physician should know the local etiological profile and antibiotic sensitivity pattern of uropathogens. We wanted to study the etiological profile and antibiotic sensitivity pattern of urinary isolates in a tertiary care hospital of Western Odisha.METHODSEarly morning mid-stream urine samples of 730 clinically suspected UTI patients were collected by clean catch technique and sent to microbiology department. Cysteine lactose electrolyte deficient (CLED) agar media was seeded with urine with the help of 0.01 ml (4 mm) loop. After incubation for 24 hrs at 370C growth was observed and identified by Gram stain and biochemical tests. Antibiotic sensitivity was performed by disc diffusion method as per CLSI guidelines. Antibiotic sensitivity was performed for all Gram-negative bacteria, Enterococci and Staphylococcus.RESULTSAmong 730 samples, 238 (33%) showed significant bacteriuria and 63 % of significant bacteriuria samples were from female. Middle age females (36-50 yrs.) were more affected (38%) followed by old age (>50 yrs.) male (19%) and old age (>50 yrs.) female (18%). E. coli was the most common bacteria (31%) followed by Enterococci (18%). Fluoroquinolones like nalidixic acid and norfloxacin showed high resistance rate (31%, 42% in case of Gram-negative bacteria and 12%, 25% in case of Staphylococcus species respectively). Nitrofurantoin showed excellent sensitivity to both Gram-positive cocci and Gram-negative bacilli. (80% for gram-negative bacilli and 87 % for Staphylococcus species and 78% for enterococci species). Aminoglycoside and carbapenem showed excellent sensitivity to Gram-negative bacteria (81% and 92% respectively). Third generation cephalosporins showed poor sensitivity (48% to 53%).CONCLUSIONSEnterococci rather than Klebsiella species was the 2nd most common uropathogen in our study. Aminoglycoside was still useful for UTI. Nitrofurantoin was the best option for empirical therapy.

Hospital Antibiogram- A Tool for Safe Antimicrobial Therapy.

Monitoring emerging trends in antimicrobial resistance at local levels is a very important aspect for clinical decision making and infection control interventions in this era of rising superbugs. The hospital antibiogram constructed by standardized methods is a summary of antimicrobial susceptibilities of local bacterial isolates, with periodic review.METHODSThis antibiogram was prepared based on the Clinical and Laboratory Standards Institute (CLSI) M39 – A4 guidelines. The data was collected for a period of three years from May 2016 to April 2019. All the samples which were received in the microbiology laboratory for aerobic bacterial culture and sensitivity testing were considered. Antibiotic susceptibility data of the bacterial isolates processed by VITEK2 system was considered. A hospital antibiotic policy was formulated and implemented.RESULTSA total of 15,135 samples were analysed from different clinical departments for a period of three years, of which 5,638 were culture positive. A comparative analysis of the three years’ data during the introduction of antibiotic protocol showed a gradual decrease in the rate of infection with multidrug resistant organisms. The prevalence of Methicillin Resistant Staphylococcus aureus (MRSA) showed a decrease from 15% to 7%. Extended spectrum beta-lactamase (ESBL) producing Escherichia coli in urine decreased from 42.4% to 27%.CONCLUSIONSPrompt antimicrobial therapy in case of an infection makes a lot of difference between recovery and death and most of the time prevents long term disability. Hence, antibiotic policy is one of the mandatory requirements, and making an antibiogram is the first step before framing antibiotic policy.