Multidrug Resistant Gram Research Articles

Effectiveness of Minocycline in Multidrug Resistance Gram Negative Bacilli: A Cross-sectional Study

Introduction: Antibiotic resistance is a global public health threat and remains a challenge for the physicians. Due to increased incidence of resistance to the commonly prescribed antibiotics, a newer drug or a re-emerge of an older class of antibiotic will be a choice of treatment of the Multidrug Resistant (MDR) organisms. Aim: To determine the effectiveness of Minocycline in MDR gram negative bacterial isolates by determining its Minimum Inhibitory Concentration (MIC) and to compare its effectiveness with Imipenem and Meropenem. Materials and Methods: A cross-sectional periodical study was conducted during May 2016 to May 2017 using 150 non repetitive MDR gram negative bacterial isolates recovered from various clinical specimens sent to Central Laboratory, Department of Microbiology, Sri Ramachandra Medical College & Research Institute, SRIHER, Porur, Chennai, India. All the isolates were subjected for antibiotic susceptibility testing by disc diffusion method for the routine antibiotics and MIC determination by Epsilometry test (E-Strip) for Minocycline and Meropenem and interpreted as per Clinical and Laboratory Standards Institute (CLSI) guidelines 2016. Results: Out of the 150 study isolates minocycline was was susceptible in 105 (70%; n=150) followed by imipenem and meropenem both susceptible for 94 isolates (62.7%; n=150). Minocycline was also susceptible among the 84% (79; n=94) of meropenem susceptible strains with a statistically significant p-value of <0.05. Similarly 46.4% (26; n=56) of meropenem resistant strains were susceptible for minocycline which was also statistically significant with a p-value <0.05. Conclusion: Considering the cost of treatment with colistin which is the choice of treatment for carbapenemase producing gram negative bacteria’s, minocycline can be considered as it is cheaper and less toxic. The possibility to switch from injectable to oral formulation is also possible with minocycline and so can also be considered as an alternative for colistin in such conditions.

Microbiological Profile of Osteomyelitis and Antibiotic Resistance Pattern of Bacterial Isolates with Special Reference to MDR Strains at a Tertiary Care Hospital, Kanpur, Uttar Pradesh, India

Introduction: Osteomyelitis is an inflammatory process that affects bone due to the contiguous infection, direct inoculation, or haematogenous spread of microorganisms. It is an infectious disease that is difficult to diagnose and treatment is complex because of its heterogeneity, pathophysiology, clinical presentation and management. Aim: To determine microbiological profile osteomyelitis and antibiotic resistance pattern of bacterial isolates with special reference to Multidrug Resistance (MDR) strains. Materials and Methods: A cross-sectional study was conducted in the Department of Microbiology and Department of Orthopedics Rama Medical College Hospital and Research Centre Kanpur, Uttar Pradesh, India. A total of 100 samples from osteomyelitis cases were aerobically cultured and isolates from culture positives were identified by standard procedures. Antimicrobial Susceptibility Testing (AST) was done following Clinical and Laboratory Standards Institute (CLSI) guidelines. Staphylococcal isolates were screened for methicillin resistance and Gram negative bacilli were screened for MDR production. Results: Out of 100 samples, 76% were culture positive and 24% were culture negative. Males were more affected than females. Staphylococcal spp. (47.3%) was predominant, E. coli (14.4%) and Klebsiella spp. (11.8%), Pseudomonas spp. (9.2%), Proteus spp. (5.2%), Coagulase-Negative Staphylococci (CoNS) (3.9%). Among the MDR strains, Methicillin Resistant Staphylococcus aureus (MRSA) was 44.4%. All the MDR Staphylococcal isolates were 100% sensitive for linezolid. Among the MDR Gram negative bacilli were Extended Spectrum Beta Lactamases (ESBL) (50%), AmpC (17.6%) and Metallo Beta Lactamase (MBL) (14.7%) and they were 100% sensitive for polymixin B and colistin. Conclusion: The microbiological profile of osteomyelitis in the present study showed high prevalence of MRSA44% as the commonest agent, sensitive only to linezolid. E. coli ESBL (50%) and MBL-14.7% were sensitive only to colistin and polymixin B, therefore proper infection control practices and antibiotic policy has to be followed to reduce the incidence of MDR strains.

Prevalence of Multidrug Resistance Gram Negative Bacteria among Patients with Bacteremia in Erbil, Iraq

Background: The presence of microorganisms in a patient's blood is a critical determinant of the severity of the patient's illness and its complication is one of the main infective causes of mortality and morbidity. Early and prompt antibiotic therapy based on the knowledge of the prevalent microorganisms can help reduce this rate.
 Objectives: Our study aimed to carry out prevalence study on bacteremia isolated from blood samples among patients in Erbil city and analyze its antibiotics susceptibility pattern test and multi-drug resistance.
 Materials and Methods: A total of 1023 from 2015 until 2021 bacteremia were isolated from patients blood samples attending (Nanakali, Raparin, Maryamana) hospitals and BIO lab from both male and female. Only 57 cases had been identified as bacteremia isolates which were identified by using microscopical, macroscopical identification, cultural, BacT/ALERT and vitek 2 compact system. Also antibiotic susceptibility test was performed by vitek 2 compact on 13 antibiotics.
 Results: Only 57 bacteremia isolates were isolated from 1023 samples from (2015-2021), in our study the percentage of females infected with bacteremia were more than the males, females being 29/1023 (2.84%) and males being 28/1023 (2.74%), about the age groups in (2015-2021) infections by bacteremia were increased in (21-30) age people being 13/57 ( 22.81%) in total, in (2015-2021) the most common bacteria that cause bacteremia were Escherichia coli being 28/57(49.12%) followed by Klebsiella pneumonia being 11/57 (19.3%) , Enterobacter spp. being 7/57 (12.28%) , Salmonella typhi and Serratia marcescens both had 1/57 (1.75%) , Acinetobacter baumanni had 7/57 (12.3%) and finally Pseudomonas aeruginosa had 2/57 (3.5%) . Escherichia coli this bacteria mainly found in females 16/28 (57.14%), in ( 2015-2021), for cancer patients with bacteremia Escherichia coli was the most common isolated bacteria being 24/732 (3.30%), bacteremia mostly infected patients with acute myeloid leukemia were 12/38(31.58%) in Nanakali hospital, the bacteria that isolated from patients with bacteremia had resistance to more than three classes of antibiotics they were highly resistance to vancomycin 15 (100%), tetracycline12 (80%) followed by sulfamethoxazole 11 (73.33%) then to erythromycin 9 (60%) and both amoxicillin and clindamycin had 8 (53.33%) antibiotic resistance.
 Conclusions: The study showed that the rate of bacteremia increased in last few years in Erbil city especially in cancer patient and those having weak immunity, bacteria acquired resistance to antibiotics and this due to frequent use of antibiotics, morbidity attribute to antibiotic resistant is significant, if prevailing resistance trends continue, high societal and economic costs can be expected. Better management of antibiotic use, and infection control is needed to avoid infections that caused by drug resistant pathogens bacteremia.

Multidrug resistant and extended spectrum β-lactamase producing gram negative bacterial uropathogens among females in a tertiary hospital, Pokhara

Introduction: Extended spectrum β-lactamase producing gram negative bacilli has emerged and spread worldwide as an important cause of urinary tract infections and indicates a major threat. The study aimed to determine the prevalence of multidrug resistant and extended spectrum beta-lactamase producing gram negative bacterial uropathogens among females in a tertiary level hospital. Methods: A hospital-based cross-sectional study was conducted in Pokhara Academy of Health Sciences, Pokhara from February to November, 2019. A total number of 301 midstream urine samples were collected and examined using MacConkey agar and blood agar medium. Antimicrobial susceptibility testing was done by Kirby Bauer disk diffusion method on Mueller Hinton agar using Clinical and Laboratory Standards Institute guidelines. Results: Out of 301 mid-stream urine samples, 99(33%) sample showed significant bacterial growth. Among them, 78(79%) were gram negative bacteria. Escherichia coli were the predominant organism. Multidrug resistant gram negative isolates were 65.4%. Among 78 Gram negative isolates, 31(39.7%) were extended spectrum β-lactamase producers. Among extended spectrum β-lactamase producers, 27(87.1%) were MDR. Highest frequency of extended spectrum β-lactamases production was seen in E. coli, 23(74.2%). Majority of gram negative bacteria showed susceptibility toward colistin and nitrofurantoin. Ampicillin was found to be highly resistant towards gram negative uropathogen. Conclusions: This study found that higher proportion of multi-drug resistants were among gram negative isolates and further more among extended spectrum β-lactamase producing gram negative isolates. Thus, there is urgent need to address the issue of antimicrobial resistant and promote rational use of the antibiotics in our region.

Differential antimicrobial extract activity of Graviola (Annona muricata) on gram positive and gram- negative antibiotic-resistant bacteria

New agents are needed with the increasing prevalence of multidrug-resistant (MDR) bacteria. Identification of plants with activity against MDR bacteria is of increasing importance. The antimicrobial activity of commercially available Graviola preparations against 38 clinical and laboratory isolates, representing MDR and drug-sensitive Gram positive and Gram negative bacteria, was measured by broth microdilution. Commercially Graviola fruit, as well as leaves/stems preparations, exhibited both inhibitory and cidal activity. However, the fruit preparation showed significantly less activity than the leaves/stems preparation. The Graviola leaves/stems preparation activity titer ranged from 1:4 to 1:16 against MDR Gram positive and Gram negative bacteria; including methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum β-lactamase (ESBL) Escherichia coli, Acinetobacter baumannii, and Klebsiella pneumoniae carbapenemase (KPC). However, the preparation had no effect on Lactobacillus. In addition, further extractions of the leaves/stems were tested against MRSA and MSSA in an in-vitro biofilm wound infection model. The polar extracts of the stems/leaves exhibited the highest bactericidal activity in preventing and inhibiting biofilm formation. Thus, Graviola leaves/stems extracts showed differential activity for phytochemical utility against multidrug-resistant bacteria.

High prevalence of Panton-Valentine Leucocidin (PVL) toxin carrying MRSA and multidrug resistant gram negative bacteria in late onset neonatal sepsis indicate nosocomial spread in a Pakistani tertiary care hospital

BackgroundNeonatal sepsis has high incidence with significant mortality and morbidity rates in Pakistan. We investigated common etiological patterns of neonatal sepsis at a tertiary care setup. Methods90 pus and blood, gram negative and gram positive bacterial isolates were analyzed for virulence and antibiotic resistance gene profiling using PCR and disc diffusion methods. ResultsStaphylococcus aureus showed strong association with neonatal sepsis (43 %) followed by Citrobacter freundii (21 %), Pseudomonas aeruginosa (13 %), Escherichia coli (15 %) and Salmonella enterica (8 %). Molecular typing of E. coli isolates depicted high prevalence of the virulent F and B2 phylogroups, with 4 hypervirulent phylogroup G isolates. 76.9 % S. aureus isolates showed presence of Luk-PV, encoding for Panton-valentine leucocidin (PVL) toxin with majority also carrying MecA gene and classified as methicillin resistant S. aureus (MRSA). ecpA, papC, fimH and traT virulence genes were detected in E. coli and Salmonella isolates. 47 % Citrobacter freundii isolates carried the shiga like toxin SltII B. Antimicrobial resistance profiling depicted common resistance to cephalosporins, beta lactams and fluoroquinolones. ConclusionPresence of PVL carrying MRSA and multidrug resistant gram negative bacteria, all isolated from late onset sepsis neonates indicate a predominant nosocomial transmission pattern which may complicate management of the disease in NICU setups

1728. Real world experience with cefiderocol for multi drug resistant gram negative bacterial infection in a tertiary hospital

Abstract Background Increasing rates of antibiotic resistance has emerged as one of the top concerns in health care. New resistance mechanisms such as carbapenemase production present a challenge in treating common infections. Novel antibiotics are being developed to tackle these pathogens, and one of the newest drugs in this arsenal is cefiderocol, a siderophore cephalosporin. Real-world data regarding the clinical use and outcomes of cefiderocol is limited. Methods A single-center, retrospective chart review was completed from July 2020 to March 2022 and included adult patients who received at least one dose of cefiderocol. Patient baseline characteristics were obtained along with other variables related to the primary infection. The primary outcome was clinical failure defined as 30 day mortality following cefiderocol administration. Secondary outcomes were development of resistance to cefiderocol, clearance of bacteremia, adverse drug reactions, and hospital length of stay. Results Cefiderocol was dispensed in 22 unique patient encounters. It was most often used to treat respiratory tract (45.4%, 10/22) and cardiac device infections (40.9%, 9/22). Most infections were polymicrobial, however, the most common organisms isolated were Pseudomonas aeruginosa and Klebsiella pneumoniae. The clinical failure rate was 22.7% (5/22). One patient developed resistance to cefiderocol throughout the hospitalization. Eight patients were treated for bacteremia, and 87.5% (7/8) demonstrated microbiological cure. Minimal side effects were observed in this cohort; two patients developed diarrhea, and one of whom had confirmed C. difficile colitis. The median length of stay was 24 days. Conclusion This cohort of patients demonstrated that cefiderocol can be used in a variety of multidrug resistant infections including bacteremia. Although limited by a small sample size, this study demonstrated a low prevalence of resistance. These results are promising in the ongoing and emerging issue of antibiotic resistance, and larger studies are needed to determine true efficacy and place in therapy for cefiderocol. Disclosures All Authors: No reported disclosures.

Low Dose Ruxolitinib in Indian Haplo-Identical Transplant Patients with Steroid Refractory Acute Graft Versus Host Disease

Background: The number of allogeneic bone marrow transplants done in India is increasing by year. Unlike west, this region had got its own unique complications in the post-transplant period especially multi drug resistant gram negative bacterial sepsis reflecting in a higher transplant related mortality. With increasing number of Haplo-identical transplants, the problem of steroid refractory Graft versus host disease (GVHD) is a major reason for post-transplant morbidity and mortality. As most available immunosuppressive agents worsen cytopenia and in turn sepsis, we need a relatively safer method of managing steroid refractory GVHD in our setting. In this observational study, we used a lower dose of 5mg per day Ruxolitinib for a period of minimum 60 days post-transplant to look into clinical benefit. This dose was chosen considering the recovering platelet count and financial considerations in our resource limited setting. Method: This observational study was conducted from July 2019 to January 2022. A total of 11 steroid refractory GVHD Haploidentical transplant patients were chosen for data analysis that completed the scheduled dosage regimen of Ruxolitinib. Nine out of 11 patients were Acute Myeloid leukemia and two were Acute Lymphoblastic leukemia. The standard conditioning regimen in all was Fludarabine / Busulfan & Cyclophosphamide based conditioning. Additional GVHD medications used were PTCy, Tacrolimus, MMF and steroids with oral budesonide. Tab Ruxolitinib was started around Day +5 post steroids in people who were fitting into steroid refractory acyte GVHD at a fixed dose of 5mg OD for a minimum period of ten weeks. Two cases had one week dose interruption due to cytopenia. Results : At start of Ruxolitinib seven patients had Grade III gut and liver GVHD and rest four had grade II GVHD. Skin was grade II in all patients. When evaluated at Day +58 of Ruxolitinib, all patients were alive and nine of them were on tapering doses of steroids. All continued Tacrolimus at therapeutic doses. No excess bleeding or cytopenia requiring transfusion support happened during this period. As mentioned earlier two cases Ruxolitinib was stopped in view of platelet counts going below 20,000. On evaluation at day +100 post transplant, two patients continued to have grade II Liver, gut and skin GVH and one patient expired on day +48 to sepsis. Rest of eight patients had a Grade I skin, liver and gut GVHD and were able to continue Ruxolitinib further. We started Ruxolitinib taper in cases where steroids were stopped. As case numbers were very low and it's a pilot observation study, we didn't do detailed statistical analysis which may be misleading and give false promises. Conclusion: In our setup, we were finding it a difficulty to use the prescribed 10mg BD Ruxolitinib dose in adults due to fear of worsening cytopenia /sepsis and financial toxicity. We tried to observe the benefit of a lower dose in steroid refractory GVHD post Haplo-identical bone marrow transplant. No pharmaco-genetic assays were done and it's a very small sample size. This observational study revealed a better efficacy and tolerability of this dosing regimen in Indian Haplo-identical patients with a considerable response without compromising blood counts. The results are looking more promising to conduct this study in a bigger scale to benefit our patients from resource limited settings. Reference Zhao Y, Shi J, Luo Y, Gao F, Tan Y, Lai X, Yu J, Wei G, Huang H. Calcineurin Inhibitors Replacement by Ruxolitinib as Graft-versus-Host Disease Prophylaxis for Patients after Allogeneic Stem Cell Transplantation. Biol Blood Marrow Transplant. 2020 May;26(5):e128-e133. doi: 10.1016/j.bbmt.2020.01.012. Epub 2020 Jan 23. PMID: 31982545.

ANTIBIOTICS RESISTANCE PATTERN AMONG GRAM POSITIVE COCCI ISOLATED FROM INTENSIVE CARE UNITS IN A TERTIARY CARE HOSPITAL

Multi drug resistant Gram positive cocci are implicated in life threatening infections, especially in critical care units. In the current study which was conducted from June 2018 to March 2019, the prevalence and antimicrobial resistance among gram positive cocci isolated from intensive care units of a tertiary care hospital has been detected . Various samples like urine, pus, body uids, sputum etc. were processed according to standard procedures. Total 81 gram positive cocci were isolated from 1650 samples. Maximum gram positive cocci were coagulase negative Staphylococcus followed by Staphylococcus aureus and enterococci. Coagulase negative Staphylococcus showed highest resistance to cotrimoxazole, erythromycin, ciprooxacin, and penicillin and maximum sensitivity to gentamicin. Staphylococcus aureus showed maximum resistance to penicillin, tetracycline, co - trimoxazole and ciprooxacin. All the isolates were sensitive to linezolid and vancomicin. Antimicrobial resistance among enterococci varied from 65% to 85%. Formation of antibiogram and strict adherence to hospital antibiotic policy is crucial to prevent spread of resistance among microorganisms.

Epidemiology of Multidrug Resistant Non-Fermentative Gram Negative Bacilli in Patients with Hospital Acquired Pneumonia: An Alarming Report from Somalia.

There is a lack of data concerning hospital-acquired pneumonia (HAP) due to multidrug resistant non-fermentative gram-negative bacilli (MDR-NFGNB) in Somalia, and this study will aim to analyze the epidemiology of MDR-NFGNB among HAP patients in tertiary care hospital in Somalia. This is a retrospective study which evaluated the presence of MDR-NFGNB among 2003 HAP patients between June 2017 and May 2022 in a tertiary care hospital in Somalia. NFGNB were cultured on blood agar and eosin methylene blue agar and were then used BBL crystal system andoxidase biochemical assays for identification. Antimicrobial sensitivity and resistance were assessed using Mueller-Hinton agar. A total of 160 confirmed cases of HAP due to NFGNB were examined. Among these, 114 (71%) were males compared to females (n = 46, 29%). The mean age was 49.49 ± 21.48. The overall prevalence rate of NFGNB among patients with HAP was 8%. It was more common in older patients and in patients with co-morbidities. Acinetobacter baumannii was the most common NFGNB with n = 93 (58%), followed by Pseudomonas aeruginosa (n = 55, 34%), and Stenotrophomonas maltophilia (n = 12, 8%). Regarding the antimicrobial sensitivity rate, Acinetobacter baumannii showed an overall resistance level of 82%, Stenotrophomonas maltophilia (81%), and Pseudomonas aeruginosa (62%). The rate of CR-NFGNB in our study was 79.4%. We found that 68.1% of NFGNB was multidrug resistant (MDR) pathogens. MDR patterns were more common in Acinetobacter baumannii infections (84%). MDR pathogens were strongly associated with ICU admissions (95% CI, 0.202-0.800, OR, 0.402, p value <0.009). Finally, the mortality rate of HAP caused by NFGNB in our study was 42.5%. The prevalence of NFGNB in HAP patients was 8%. These infections were more prevalent among men and the elderly. Acinetobacter baumannii and Stenotrophomonas maltophilia exhibited the highest antibiotic resistance rate. The MDR level of these pathogens was 68.1%.

Randomized control study of nebulized colistin as an adjunctive therapy in ventilator-associated pneumonia in pediatric postoperative cardiac surgical population.

Ventilator-associated pneumonia (VAP) with multidrug-resistant (MDR) gram negative organisms is a common problem in intensive care unit (ICU). Aerosolized antibiotics enhance the efficacy of systemic antibiotics when added as adjuvants. The primary objective of the study was to compare the clinical and bacteriological outcome of patients with VAP who were administered intravenous (IV) antibiotics alone with those patients who were treated with adjunctive nebulized colistin (NC) along with IV antibiotics. The secondary objective was to study the occurrence of any adverse events during colistin nebulization. The study was a prospective, randomized, double-blinded controlled study conducted at a tertiary-care teaching institution. Ninety-eight children from surgical ICU aged less than 12 years who were diagnosed with VAP due to gram negative bacteria following cardiac surgery were chosen and divided randomly into two groups. The experimental group (NC group) was treated with systemic antibiotics along with NC, whereas the control group (NS group) was administered systemic antibiotics with nebulized normal saline (NS). Clinical and bacteriological outcomes were noted. Statistical analysis was done using SPSS Version 20.0 software. The patient characteristics were compared using independent Student's t test and Chi-square test. There was a statistically significant reduction in the duration of mechanical ventilation, postoperative ICU and hospital stay (P &lt; 0.05) in the NC group compared with the NS group. Aerosolized colistin may be considered as an adjunct to systemic IV antibiotics in pediatric patients with VAP due to gram negative bacteria susceptible to colistin.

Susceptibility Pattern of Bacteria Isolated from Patients Admitted in Medical ICU with Pneumonia

Background: Patients in ICU presenting with severe disease suffer from multidrug-resistant bacteria, resuting in reduced therapeutic options which cover ICU pathogens spectrum. Identification of causative organism and its antibiotics sensitivity pattern has become cornerstone for antibiotic selection. β-Lactams i.e. cephalosporins, carbapenems, penicillins, and monobactams constitute 60% of total antibacterials used, however; with high usage of such antibiotics the emergence of resistance. Aim: To determine prevalence of local type of bacterial isolates and their antibiotic susceptibility. Study Design: Cross sectional study Place and Duration of Study: Medical ICU, Pakistan Ordinance Factories Hospital, Wah Cantt from 1st January 2020 till 31st July 2021. Methodology: One hundred patients were enrolled. Samples were taken from blood, sputum, bronchoalveolar lavage and tracheal secretions of the patient and sent to microbiologist for culture and sensitivity reports. Results: The mean age was 57.52+13.427 years, 47 females and 53 male patients. 72 patients were shifted out and remaining 28 died. In 57 patients, sputum specimen was taken, blood specimen of 24 patients, tracheal secretion of 8 patients and 11 patients bronchoalveolar lavage was taken. 33 were positive for Klebsiella pneumoniae, 29 patients were positive for acinetobacter baumanii. Pseudomonas aeuriginosa was positive in 18 patients, 05 patients for E. coli. In gram positive category; 05 patients were positive for staph aureus, 06 patients for MRCONS, 3 to MRSA and 1 patient for coagulase negative staphylococcus species. Conclusion: The high levels of antibiotic resistance are seen among both gram negative and gram positive isolates. Presence of elevated resistance to multiple drugs is an indicator for high prevalence of multi-drug resistant gram positive as well as gram negative organisms, so proper identification of organism in order to ascertain administration of emperical drugs most effective against the isolated organism is recommended in severe cases. Keywords: Antibiogram, Bacterial isolate, Pneumonia, Sensitivity

Antibacterial and antibiotic-potentiation activity of the constituents from aerial part of Donella welwitshii (Sapotaceae) against multidrug resistant phenotypes

BackgroundThe rise of multidrug-resistant (MDR) bacteria is a real public health problem worldwide and is responsible for the increase in hospital infections. Donella welwitschii is a liana or shrub belonging to the family Sapotaceae and traditionally used to cure coughs.ObjectiveThis study was conducted with the objective to validate the medicinal properties of this plant, the aerial part was studied for its phytochemical composition using column and PTLC chromatography and exploring its antibacterial and antibiotic-modifying activity as well as those of its phytochemicals.MethodsThe structures of the compounds were elucidated from their physical and spectroscopic data in conjunction with literature. The antibacterial activity of the isolated metabolites was performed toward a panel of MDR Gram negative and Gram-positive bacteria. The broth micro-dilution method was used to determine antibacterial activities, efflux pump effect using the efflux pump inhibitor (EPI) (phenylalanine-arginine-ß-naphthylamide (PAβN)), as well as the modulating activity of antibiotics. Monitoring the acidification of the bacterial growth medium was used to study the effects of the samples on the bacterial proton-ATPase pumps and cellular ATP production.ResultsEleven compounds were isolated including pentacyclic triterpenes, C-glucosyl benzophenones. With a MIC value < 10 μg/mL, diospyric acid (7) significantly inhibited the growth of Escherichia coli AG102, Enterobacter aerogenes ATCC13048, Klebsiella pneumoniae KP55, Providencia stuartii NEA16 and Staphylococcus aureus MRSA3. 28-hydroxy-β-amyrin (8) significantly impaired the growth of Enterobacter aerogenes EA27, Klebsiella pneumoniae ATCC11296 and Staphylococcus aureus MRSA6; and oleanolic acid (9) strongly impaired the growth of Escherichia coli AG 102, Enterobacter aerogenes EA27 and Providencia stuartii PS2636. Diospyric acid (7) and 28-hydroxy-β-amyrin (8) induced perturbation of H+-ATPase pump and inhibition of the cellular ATP production. Moreover, at MIC/2 and MIC/4, compounds 7, 8, and 9 strongly improved the antibacterial activity of norfloxacin, ciprofloxacin and doxycycline with antibiotic-modulating factors ranging between 2 and 64.ConclusionThe overall results of the current work demonstrate that diospyric acid (7), 28-hydroxy-β-amyrin (8) and oleanolic acid (9) are the major bioactive constituents of Donella welwitschia towards Gram-negative bacteria expressing MDR phenotypes.

Evaluation of novel immunological rapid test (K.N.I.V.O. Detection K-Set) for rapid detection of carbapenemase producers in multidrug-resistant gram negatives

The K.N.I.V.O Detection K-Set is a novel immunochromatographic test for detection of the 5 major carbapenemases (KPC, NDM, IMP, VIM, and OXA-48-like). This test is rapid, easy to perform, and shows a good specificity and sensitivity (96.8% and 100%, respectively), being suitable for microbiology laboratories together with biochemical rapid tests.

Detection of Extended Spectrum Beta-lactamase Gene (CTX-M) among Representative Multidrug-Resistant Gram Negative Bacterial Isolates from Patients with Urinary Tract Infection in Ekiti State, Nigeria

Urinary tract infection is huge public health burden and the emergence of extended spectrum beta lactamase producing bacterial pathogens increases the burden of infectious diseases in Nigeria. This study determined the current prevalence of cephalosporin resistance among Gram-negative bacteria isolated from patients with urinary tract infections between February 2018 and June 2018. Non-repetitive Gram–negative bacteria were recovered from 106 individuals with urinary tract infections who reported at two tertiary healthcare centers in Ekiti-State, Nigeria. A total of 106 bacterial isolates were obtained which included: Klebsiella pneumoniae 34 (29.1%), Klebsiella oxytoca 17 (16.0%), Proteus vulgaris 10 (9.4%), E. coli 24 (22.6%), Proteus mirabilis 18 (16.9%) and Pseudomonas aeruginosa 3 (2.8%). Sixty five of these organisms showed resistance to ceftazidime while 76 organisms showed resistance to ceftriaxone. Forty representative organisms were selected and tested for presence of extended spectrum beta-lactamase (ESBL) genes using primers specific for different ESBL genes. A total of eight (20.0%) organisms carried the blaCTX-M gene and other variants of the ESBL genes were not detected. The organisms carrying the blaCTX-M gene included E. coli 3 (37.5%), K. pneumoniae 1(12.5%), P. mirabilis 1(12.5%),) and K. oxytoca 3(37.5%). The high prevalence of cephalosporin resistant Gram-negative bacteria among patients with UTI is a serious threat to public health and efforts must be intensified to regulate the clinical use of the cephalosporins.

Phenotypic Identification and Molecular Detection of bla (NDM-1) Gene in Multidrug Resistant Gram Negative Bacilli in Sohag University Hospital

Phenotypic Identification and Molecular Detection of bla (NDM-1) Gene in Multidrug Resistant Gram Negative Bacilli in Sohag University Hospital

Febrile illness of bacterial etiology in a public fever hospital in Egypt: High burden of multidrug resistance and WHO priority Gram negative pathogens

Contemporary emergence of multidrug resistance (MDR) urges regular updates on circulating pathogens and their antimicrobial resistance profiles. We aimed to identify the burden of MDR and World Health Organization (WHO) priority Gram negative pathogens among patients admitted with febrile illness to Abbassia Fever Hospital, a major Public Fever Hospital in Egypt. The carbapenemase- and extended spectrum beta-lactamases (ESBLs)-encoding genes carried by the isolates were also identified. A total of 9602 clinical specimens were collected from febrile patients during 2018 and 2019. The recovered bacterial isolates were examined for antimicrobial susceptibility using disk diffusion test. Susceptibility to colistin was tested using E-test. ESBLs production was phenotypically and genotypically analyzed. A total of 790 bacterial isolates (612 Gram negative and 178 Gram positive) were recovered. A percentage of 77.6%, and 62.9% of the Gram negative and positive isolates showed MDR phenotype, respectively. WHO priority pathogens were abundant, including carbapenem-resistant (CR) Enterobacterales (105/187; 56.1%) and CR glucose non-fermenters (82/187; 43.8%) such as: A. baumannii (55; 29.4%), P. aeruginosa (27; 14.4%). Carbapenemase- and ESBLs-encoding genes were detected in 56.1% and 30.8% of Enterobacterales and in 43.8% and 46.3% of glucose non-fermenters, respectively. Antimicrobials such as fosfomycin and chloramphenicol retained good activities against MDR Gram negative pathogens. This study highlights the regional burden of MDR and priority Gram negative pathogens. The obtained data are of relevant medical importance for implementation of evidence-based antimicrobial stewardship programs and for tailoring the existing empirical treatment guidelines.

ChromagarTM Col-Apse for Detection of Colistin Resistance in Clinical Isolates of Multidrug Resistant Gram Negative Bacilli.

To determine the diagnostic accuracy of CHROMagarTM COL-APSE, for detection of colistin resistance in clinical isolates of multi drug resistant (MDR) gram negative bacilli (GNB). Cross-sectional validation study. Department of Microbiology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, Pakistan from February 2019 to August 2019. Total 96 MDR-GNB in clinical isolates were included. Isolates were identified using gram stain, catalase, oxidase, API 20E and API 20NE. After taking approval from Institutional Ethical Review Committee, colistin susceptibility was determined simultaneously by CHROMagarTM COL-APSE (using 1x105 CFU/ml inoculum) and Broth Micro Dilution (BMD) Minimum Inhibitory Concentration (MIC) method as per CLSI. EUCAST guidelines were followed for interpretation of susceptibility profile. Results were validated with gold standard test, i.e. BMD. Out of 96 MDR clinical isolates, the distribution was K. pneumoniae n=63, E. coli n=18, A. baumannii n=11, C. freundii n=3, and E. cloacae n=1. The sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy of CHROMagarTM COL-APSE for detection of colistin resistance, keeping BMD-MIC method as gold standard, was 97.96%, 97.87%, 97.96%, 97.87% and 97.92%, respectively. CHROMagarTM COL-APSE can be used as screening agar by direct streaking of specimen as well as diagnostic for detection of colistin resistance by the use of standardised inoculum. It can be used as a critical time saving method for colistin resistance detection in absence of expertise or manpower required for BMD as well as the cost required for genetic sequencing to detect MCR genes. Key Words: Multi drug resistant (MDR), Gram negative bacilli (GNB), Colistin resistance, CHROMagarTM COL-APSE.

Colistin Resistance in Monophasic Isolates of Salmonella enterica ST34 Collected From Meat-Derived Products in Spain, With or Without CMY-2 Co-production.

Colistin is a last-resort antibiotic in fighting severe infections caused by multidrug resistant Gram negative pathogens in hospitals. Zoonotic bacteria acquire colistin resistance in animal reservoirs and mediate its spread along the food chain. This is the case of non-typhoid serovars of Salmonella enterica. Colistin-resistant S. enterica in foods represents a threat to human health. Here, we assessed the prevalence of colistin-resistance in food-borne isolates of S. enterica (2014–2019; Asturias, Spain), and established the genetic basis and transferability of this resistance. Five out of 231 isolates tested (2.2%) were resistant to colistin. Four of them, belonging to the European monophasic ST34 clone of S. Typhimurium, were characterized in the present study. They were collected from pork or pork and beef meat-derived products, either in 2015 (three isolates) or 2019 (one isolate). Molecular typing with XbaI-PFGE and plasmid profiling revealed distinct patterns for each isolate, even though two of the 2015 isolates derived from the same sample. The MICs of colistin ranged from 8 to 16 mg/L. All isolates carried the mcr-1.1 gene located on conjugative plasmids of the incompatibility groups IncX4 (2015 isolates) or IncHI2 (2019 isolate). Apart from colistin resistance, the four isolates carried chromosomal genes conferring resistance to ampicillin, streptomycin, sulfonamides and tetracycline [blaTEM–1, strA-strB, sul2, and tet(B)] and heavy metals, including copper and silver (silESRCFBAGP and pcoGE1ABCDRSE2), arsenic (arsRSD2A2BCA1D1) ± mercury (merEDACPTR), which are characteristically associated with the European ST34 monophasic clone. The 2019 isolate was also resistant to other antibiotics, comprising third generation cephalosporins and cephamycins. The latter phenotype was conferred by the blaCMY–2 gene located on an IncI1-I(α)-ST2 plasmid. Results in the present study identified meat-derived products as a reservoir of a highly successful clone harboring transferable plasmids which confer resistance to colistin and other clinically important antibiotics. An important reduction in the number of food-borne S. enterica detected during the period of the study, together with the low frequency of colistin resistance, underlines the success of One Health initiatives, such as those implemented at the UE, to control zoonotic bacteria along the food chain and to halt the spread of antimicrobial resistance.

Pattern of Bacterial Infections among Children with Sickle Cell Disease in a Tertiary Care Hospital of Nagpur, Maharashtra, India

Introduction: In India Sickle Cell Disease (SCD) is associated with significant morbidity and represents a major health problem in central India. Patients with SCD are susceptible to a variety of bacterial infections, which are major cause of morbidity and mortality. The burden of disease caused by bacterial infections in patients with SCD is less studied and have long been neglected. Therefore there is scarcity of data for the association between SCD and invasive bacterial diseases. So it becomes necessary to have knowledge of distribution of these pathogens and their susceptibility to antibiotics. Aim: To identify the most common bacterial infections among children with SCD and to check antibiotic susceptibility patterns of all the clinical isolates. Materials and Methods: Present study was a descriptive observational study which was conducted in Department of Microbiology, Govt Medical College and Hospital Nagpur, Maharashtra, India, for two years from October 2013 to October 2015. Paediatric patients of age ≤12 years diagnosed as SCD and admitted in paediatric wards of tertiary care hospital, having body temperature ≥38.5°C were enrolled in the study. Clinical specimen i.e. blood, urine, pus, and body fluids such as Cerebrospinal Fluid (CSF), pleural fluids were collected aseptically and the bacteria causing infections were isolated and identified conventionally in accordance with the standard operative procedures. Antimicrobial susceptibility testing was performed as per the Clinical and Laboratory Standards Institute (CLSI) guidelines 2013 by modified Kirby-Bauer method. Data was presented as numbers and percentages. Results: A total of 824 samples including blood, urine, CSF, pus and pleural fluid were collected from 412 SCD patients. Prevalence of bacterial infections among patients with SCD was 17.35%. Bacteraemia was the most common infection among febrile children with SCD 84 (20.38%) followed by Urinary Tract Infection (UTI) 55 (15.02%), Osteomyelitis 2 (13.34%), and Meningitis 1 (5.56%). Total 143 organisms were isolated from different samples and gram negative bacilli 106 (74.12%) were found to be the commonest cause of bacterial infections among children with SCD. Imipenem was the best antibiotic for infections with multidrug resistant gram negative bacilli. Conclusion: To identify children with SCD and prevent bacterial infections in them should be a priority target for health research as these infections make a large contribution to the morbidity and mortality among children with SCD.