Growth Of Gram-positive Cocci Research Articles

A case report on the septic arthritis in a newborn: therapeutic approaches and major considerations

Introduction: The prevalence of septic arthritis (SA) is higher in underdeveloped countries than in developed countries, and the number of hospitalizations of children for this cause is decreasing and the most prevalent age group is between 0 and 4 years of age. Clinical history and detailed physical examination are essential to make an accurate diagnosis, but the neonatal period has certain limitations. Objective: To report a case of septic arthritis that occurred in a newborn admitted to the Neonatal Intensive Care Unit (ICU) due to its serious condition, morbidity, and unusual evolution of the pathology in the pediatric population, scarcity of epidemiological data on the subject, still aiming address aspects of the treatment of the disease. Case report: Male patient, born on 09/11/2020 at Santa Casa de Tupã, Sao Paulo, with a gestational age of 36 weeks and 6 days, through cesarean delivery and without complications, obtaining APGAR 9 and 10, and maintained in a room with the mother. The morning after birth, the newborn (NB) began to groan and have mild respiratory distress. The patient evolved with worsening of the breathing pattern and oxygen was concentrated at 6L/min in Hood's Halo, presenting hyperthermia, hyperemia, and edema in the right elbow. Laboratory tests showed an increase in C-Reactive Protein and worsened hematimetric indices of the blood count, and antibiotic therapy was then started with oxacillin 50 mg/kg/dose and amikacin 15 mg/kg/day. The ultrasound of the right elbow showed skin thickening, absence of collections, and no changes in vasculature, being suggestive of cellulite. A new ultrasound of the right elbow showed bulging, intra-articular collection, and discreet collection with extra-articular communication in the medial region, being suggestive of septic arthritis. Blood culture collection resulted in the growth of gram-positive cocci (Staphylococcus aureus) in a pair of samples. The patient received intravenous treatment with Vancomycin for 14 days with clinical and laboratory improvement of the infection and was discharged with the use of oral Clindamycin for another 14 days and physical therapy follow-up for rehabilitation of movement of the affected limb. Final considerations: Due to the unusual evolution of SA in the neonatal period, and the scarcity of literature on the pathology in this age group, the importance of further studies on the subject, and greater surveillance for neonatal diagnosis, in addition to early treatment to reduce of complications.

Identification of Bacterial Agents causing Meningitis in Adult Population using BACTEC FX40 System

Introduction: Bacterial meningitis among adults is a medical emergency and requires immediate diagnosis as well as immediate treatment. As clinical diagnosis is not always reliable, laboratory isolation along with antimicrobial susceptibility results are crucial. Conventional agar culture methods gives poor sensitivity and delayed results. Automated culture methods like BACTEC are suitable option for culture of sterile fluids beside blood culture. Aim: To determine the bacterial agents causing meningitis in this region, along with their antibiogram by using automated culture system BACTEC FX40. Materials and Methods: This cross-sectional, observational study was conducted between October 2019-September 2020 in Department of Microbiology, Rohilkhand Medical College and Hospital, Bareilly, Uttar Pradesh, India. Cereospinal Fluid (CSF) samples received from medicine Intensive Care Unit (ICU) and wards were processed by inoculating them on BD BACTEC Peds Plus/F bottle and then incubating them on BACTEC BD FX40 system. The bottles flagged and positive were subcultured and further processed according to standard laboratory procedures. The antibiotic sensitivity test was performed from isolated organisms by Kirby-Bauer Disc Diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: A total of 108 CSF were received for culture by automated methods from patients suspected of meningitis during the study period. Out of them 84 (77.8%) beeped as positive while 24 (22.2%) were sterile. From the positives, 77 showed growth of gram positive cocci, three were gram negative bacilli and four showed growth of environmental and skin contaminants that were excluded from the study. All gram positive cocci were identified as Staphylococcus aureus (S. aureus). The antibiotic sensitivity testing showed 100% sensitivity to vancomycin and linezolid. Only 25 isolates of S. aureus were sensitive to cefoxitin. Both the isolates of Escherichia coli (E. coli) were sensitive to cefazolin, ceftazidime, aztreonam. Pseudomonas aeruginosa (P. aeruginosa) was sensitive to amikacin and piperacillin/tazobactum. Conclusion: A shift in the trend was observed in the aetiology of bacterial meningitis with Methicillin Resistant Staphylococcus aureus (MRSA) as the most predominant isolate among adult population. BACTEC FX40 system was found more sensitive in detecting pathogens over the conventional methods with reduced time to positivity. Early detection of causative organism will facilitate early initiation of suitable antibiotic therapy, thereby reducing mortality and meningitis associated complications.

The profile of microorganisms that associate with acute tonsillitis in children and their antibiotics sensitivity pattern in Nigeria

Acute tonsillitis remains one of the common childhood diseases in developing countries. Prompt and appropriate treatment based on the knowledge of the causative microbiota and their antimicrobial susceptibility pattern will improve the treatment outcome and reduce time and resources spent on treatment. This study aims to determine the pattern of microbiota isolates and their susceptibility pattern. The study was a combination of the retrospective and cross-sectional method. The medical records of children treated for tonsillitis were retrieved, as well as those of children who presented with acute tonsillitis for the study. Interviewer-administered questionnaire was used to collect data, as well as document information retrieved from their medical record: presenting symptoms, treatments received. Swab sample was taken for culture and antibiotic susceptibility test. Out of the 72 swab cultures, 29 (40.3%) yielded insignificant growth of gram positive cocci. A total of nine (9) different isolates were obtained from all the significant swab cultures. Streptococcus species (13, 18.6%) and staphylococcus species (11, 15.3%) were the commonest isolates. Imipenem and levofloxacin revealed high level of susceptibility, while Ampicillin and Cefexine recorded high resistance rates. The proportion of the cultures that were positive of significant growth, the proportion of these positive isolates that were Streptococcus spp. and varied sensitivity pattern obtained underpinned the importance to advocate for culture isolates and susceptibility pattern guided treatment. These will not only an efficient approach to management of acute tonsillitis, but also a strong approach towards effective implementation of antibiotic stewardship.

Identifying gram-positive cocci in dermatoscopes and smartphone adapters using MALDI-TOF MS: a cross-sectional study

BackgroundThe increasingly frequent use of dermoscopy makes us think about the possibility of transfer of microorganisms, through the dermatoscope, between doctor and patients. ObjectivesTo identify the most frequent gram-positive cocci in dermatoscopes and smartphone adapters, as well as the resistance profile, and to evaluate the factors associated with a higher risk of bacterial contamination of the dermatoscopes. MethodsA cross-sectional study was carried out with 118 dermatologists from Porto Alegre/Brazil between September 2017 and July 2018. Gram-positive cocci were identified by MALDI-TOF MS and habits of use of the dermatoscope were evaluated through an anonymous questionnaire. ResultsOf the dermatoscopes analysed, 46.6% had growth of gram-positive cocci on the lens and 37.3% on the on/off button. The microorganisms most frequently found were S. epidermidis, S. hominis and S. warneri. Attending a hospital, using the dermatoscope at the hospital, with inpatients and in the intensive care unit were significantly associated with colonisation by gram-positive cocci. The highest resistance rates were observed for penicillin, erythromycin and sulfamethoxazole-trimethoprim. Study limitationsThe non-search of gram-negative bacilli, fungi and viruses. Moreover, the small number of adapters did not make it possible to better define if the frequency differences were statistically significant. ConclusionCoagulase-negative staphylococci were frequently identified. S. aureus was detected only on the lens.

Diagnostic accuracy and clinical impact of loop-mediated isothermal amplification for rapid detection of Staphylococcus aureus bacteremia: a retrospective observational study.

The aim of this study was to evaluate the diagnostic accuracy and the clinical impact of isothermal loop-mediated amplification (LAMP; eazyplex® MRSA kits) for rapid diagnosis of Staphylococcus aureus bacteremia (SAB) in comparison with conventional blood culture diagnostics. We performed a retrospective, single-center observational study over the period between November 2016 and December 2018 on patients (and blood cultures) with growth of Gram-positive cocci in clusters in their blood cultures. We quantified diagnostic accuracy with sensitivity and specificity for detection of S. aureus, methicillin-resistant S. aureus (MRSA), and the mecA/C resistance genes in 797 blood cultures. The clinical impact was assessed by time to result reporting, time to appropriate treatment, and length of stay in intensive care unit (ICU) in 190 SAB patients. We observed sensitivity and specificity above 90% for S. aureus detection (sensitivity (95% confidence interval (CI)), 99.57% (97.61%, 99.98%); specificity, 99.12% (97.95%, 99.71%)), for MRSA detection (sensitivity, 100% (89.11%, 100.00%); specificity, 99.72% (99.05, 99.96)), and for mecA/C detection (sensitivity, 94.71% (91.85%, 96.78%); specificity, 95.89% (93.58%, 97.54%)). LAMP testing was associated with shorter median time to result reporting (24.0h (first and third quartiles (Q1-Q3), 20.0-27.0h) vs 41.5h (36.0-46.0h); p < 0.001) and different distribution of time to appropriate treatment (2.0days (1.0-3.0) vs 2.0days (2.0-3.0); p = 0.004). No evidence for differences in length-of-stay in ICU was observed. Our analysis suggests for the application of LAMP (i) a high diagnostic accuracy for detection of S. aureus and the mecA/C genes in blood cultures, (ii) an earlier result reporting, and (iii) a shorter time to appropriate treatment.

Meningitis caused by micrococcus luteus: Case report and review of literature

Abstract Micrococcus Luteus is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family. They are considered as normal comensal of human skin and upper respiratory tract. Few cases have been reported as opportunistic infection or catheter/ shunt related infection. We here report a case of meningitis caused by Miccrococus luteus in previously healthy infant. Keywords: Micrococcus. Introduction Micro (small) Coccus(spherical) Luteus(Yellow) is a gram positive, non-motile, non-sporing cocci belonging to micrococcea family.[1],[2]Initially named as Micrococci lysodeikticus by Alexander flaming who discovered them in 1928, later Ganz et al renamed them as Micrococcus luteus in 2002. They are considered as normal commensal of human skin and upper respiratory tract and are considered harmless.[3],[4],[5],[6]Few cases have been reported as opportunistic infection or catheter/ shunt related infection.[7],[8],[9],[10],[11],[12],[13],[14],[15],[16],We here report a case of meningitis caused by Miccrococus luteus in previously healthy infant. Case Summary One year old male was brought to emergency with history of fever for 3 days and seizures from last 15 min, Baby was immunized as per age with normal developmental milestones. Child was managed with iv fluids and anibiotics. Investigations sent showed normal electrolytes, leucocytosis with raised CRP, CSF examination showed 350 cells (60% mononuclear cells). CSF culture showed growth of gram positive cocci, identified as Micrococcus luteus by Vitek 2, sensitive to all the commonly used antibiotics. IV antibiotics were continued as per the sensitivity profile (Ceftriaxone). Infant responded to the treatment, became afebrile, had no episode of seizure after admission, but had depressed consciousness and dystonia. MRI brain showed bilateral mid brain infarct. Child was discharged after completion of 10 days antibiotics with residual neurological sequeae. Discussion Bacter

Answer to October 2015 Photo Quiz

Answer: Streptococcus pyogenes. Figure 1 in the photo quiz demonstrates hypergranular, vacuolated neutrophils with intracellular cocci forming chains, resembling the genus Streptococcus. In light of the clinical features and blood film, a presumptive diagnosis of Streptococcus pyogenes toxic shock syndrome was made. Six hours following the blood film observation, growth of Gram-positive cocci in chains was detected in blood culture, which was subsequently confirmed to be S. pyogenes. As a result of the blood film findings, intravenous clindamycin was added; however, 19 h after presentation, the patient died of multiorgan failure. The published literature suggests that observation of bacteria on routine blood film is rare; case reports involving Capnocytophaga canimorsus (1), Citrobacter koseri (2), and Streptococcus pneumoniae (3) have been described, plus a case series of four neonates involving Staphylococcus aureus and coagulase-negative staphylococci (4). A larger case series, published in 1966, reported 26 bacteremic patients with bacteria observed within leukocytes on routine blood films (S. aureus, S. pneumoniae, Escherichia coli, Klebsiella pneumoniae, Neisseria meningitidis, Proteus species, viridans group Streptococcus, beta-hemolytic Streptococcus, and “paracolon bacillus”) (5). To the best of our knowledge, this is the first case of S. pyogenes observed in a routine blood film. The high fatality rate among these published cases (24 of 33 patients died) (1,–5) likely reflects the poor diagnostic sensitivity of blood films except in patients with very high bacterial loads observed during overwhelming septicemia, such as in our case. The use of buffy-coat smears may improve sensitivity (to a limit of detection of 104 CFU/ml) but is labor- and time-consuming, prohibiting its routine use (6). Bacteria observed during routine blood film examination should be discussed immediately with the treating physician, as the result may be used to predict a microbiological diagnosis and assist with choice of empirical antimicrobial and adjunctive therapy and portends a poor patient outcome. (See page 3109 in this issue [doi:10.1128/JCM.02824-13] for photo quiz case presentation.)

Evaluation of direct tube coagulase test in diagnosing staphylococcal bacteremia.

Blood Stream Infections (BSIs) are one of the most common nosocomial infections encountered in a hospital. It occurs 2 to 7 times more often in ICU patients than in ward patients and is associated with high rates of mortality and morbidity. Staphylococci is the most commonly encountered blood culture isolates for which, rapid and reliable methods for detection are warranted. To study the efficiency of direct tube coagulase test (DTC) in comparison with slide/tube coagulase test for early detection of Staphylococcal bacteraemia. This was a prospective study conducted in a tertiary care hospital, between January 2009 to December 2009. Two sets of blood cultures were obtained from each patient with a suspicion of Staphylococcal bacteremia. Both, direct tube coagulase test and conventional biochemical tests were done on the samples, to identify the species. Cohen's kappa coefficient of agreement was used to detect the differences between the two modes of detection. Ninety four samples (out of 460) yielded the growth of Gram positive cocci in clusters. The commonest isolates were Staphylococcus aureus (42.5%), Staphylococcus epidermidis (36.2%), Staphylococcus haemolyticus (8.6%), Staphylococcus cohnii (4.3%). Cohen's kappa coefficient which implies the extent of agreement of the results between the Direct Tube Coagulase test and conventional methods, was 95.6% along with sensitivity of 95.5% and specificity of 100%. Direct Tube Coagulase test (DTC) can be used for early detection of Staphylococcal bacteremia. It shows an almost perfect agreement with conventional coagulase test.

Prevalence of Multidrug resistant Staphylococcus Aureus isolates in Clinical specimens collected from local patients of Chittagong, Bangladesh

To observe the prevalence of Staphylococcus aureus a total of 1078 blood, pus and miscellaneous samples (sputum, swab and body fluids) were collected among which 478 samples exhibited bacterial growth on agar medium. Out of growth positive 300 pus samples, 116(38.67%) showed the growth of Gram positive cocci, of which 30(25.86%) were Staphylococcus aureus positive. In case of blood, the number of growth positive samples were 116(28.45%), of which 33(12.12%) showed the presence of Gram positive cocci including 4 Staphylococcus aureus. Among the 62 growth positive miscellaneous samples, the number of Gram positive cocci and Staphylococcus aureus positive samples were 22(35.48%) and 4(18.18%), respectively. From the antibiogram of 38 Staphylococcus aureus isolates, 26 were found multidrug resistant, showing resistance against two or more of the antibiotics used namely ampicillin, cefradine, gentamicin and ciprofloxacin, whereas all the isolates were sensitive to vancomycin. The MIC and MBC of these antibiotics against different Staphylococcus aureus isolates were determined, which were found to range from 2?g/ml to 8?g/ml and 4?g/ml to 16?g/ml, respectively, in case of vancomycin. The rate of resistance against ampicillin, cefradine, gentamicin and ciprofloxacin were 92.1%, 60%, 58.1% and 59.35%, respectively. DOI: http://dx.doi.org/10.3329/cujbs.v6i1-2.17242 The Chittagong Univ. J. B. Sci.,Vol. 6(1&amp;2):175-185, 2011

Reliability of Glass Bead Sterilization for Tried-in Orthodontic Bands

Objective: To evaluate the efficacy of two methods of sterilization for the decontamination of tried-in orthodontic bands after initial cleaning procedures. Materials and methods: A total of 68 molar bands were tried-in mouth of 17 patients. After try-in, 60 bands were cleaned in ultrasonic cleaning bath and then were equally divided into two groups (n = 30). Bands from group 1 were autoclaved in benchtop autoclave. Bands from group 2 were sterilized in glass bead sterilizer. After sterilization, the bands from both the groups were incubated in 50 ml brain heart infusion (BHI) broth. Eight remaining bands were neither cleaned nor sterilized. They served as positive control and were directly incubated in BHI broth. One bottle of BHI broth which did not contain any orthodontic band, served as negative control was incubated. Incubation of BHI broth for all the groups was done at 37°C for 5 days. After 5 days of incubation, BHI broth samples from all the groups were taken and incubated in blood agar culture medium. Results: All the groups except the positive control group did not show any turbidity in the BHI broth and growth in blood agar culture media. The broth containing positive control group had increase turbidity after 5 days and growth of gram-positive cocci in blood agar culture media. Conclusion: Glass bead sterilization is equally effective as benchtop autoclave for sterilization of orthodontic bands.

Evaluation of normal ocular bacterial flora with two different culture media.

The purpose of this study was to determine if the use of broth culture medium is efficient in investigating bacterial flora of the normal eyelid and conjunctiva. Samples from the conjunctiva and eyelid of healthy patients of various ages who were undergoing ocular surgeries were obtained and cultured at 3 periods: before topical antibiotic prophylaxis, in the postoperative period during topical antibiotic treatment, and 15 days after discontinuation of antibiotic use. Samples were inoculated into both brain heart infusion broth and blood agar plate, and the growth results of both media were analyzed. Brain heart infusion broth medium showed a significantly higher bacterial growth of gram-positive cocci in most periods. The solid blood agar medium had a higher recovery of gram-positive bacilli before prophylaxis only in the older patients. Our results show that a more complete analysis of eyelid and conjunctival flora can be obtained using both liquid and solid media to increase the chances of isolate recovery. The inclusion of liquid media in this analysis was even more relevant in the period of concomitant use of antibiotic treatment.

Streptococcus salivarius meningitis

A 7 1 -year-old man was admitted to the emergency room because of fever, progressive malaise, headache, and vomiting. His previous history was uneventful, and until the evening before admission, he had no complaints. His level of consciousness was decreased (Glasgow coma scale, 9/15). He had a stiff neck and a cluster of petechiae on the right chest. He had fever (temperature, 38°C) with blood pressure, 2 1 O/l 10 mmHg. Laboratory results showed an elevated leukocyte count of 11.6 x 109/1 (normal range, 3.5 to 9.8) with a neutrophilia of 90.4% (normal range, 37.9 to 70.2). The C-reactive protein (CRP) was slightly elevated at 1.6 mg/dl (normal range, < 1 mg/dl). Two blood cultures were obtained, and a lumbar puncture was performed. The cerebrospinal fluid (CSF) analysis revealed 5,820 leukocytes/mm3 (99% were polymorphonuclear cells) and 2,120 erythrocytes/mm3. A Gramstained smear revealed no organisms. Bacterial meningitis was suspected, and empirical antibiotic treatment was initiated with ceftriaxone, 2 x 2 g IV/d, and ampicillin, 4 x 2g IV/d. The next day, the patient was afebrile and alert but retained retrograde amnesia. The leukocyte count rose to 16.4 x 109/1 and CRP rose to 7.9 mg/dl. The CSF was cultured onto Columbia sheep blood agar (Becton Dickinson, Meylan Cedex, France) and chocolate agar with hemoglobin and IsoVitaleX (Becton Dickinson). After 15 h incubation at 35°C in 8% COZ, the chocolate agar showed poor growth of gram-positive cocci. The blood cultures obtained on admission were processed with the BacT/Alert 3D system (Organon Technika Corp., Durham, NC). After 36 h, three of four bottles were flagged positive. Gram-stained smears showed gram-positive cocci in chains. Overnight incubation of subcultures revealed small, non+-hemolytic colonies on sheep blood agar. A presumptive identification of Streptococcus salivarius was made based on the biochemical characteristics of the isolate (Table 1). The organism did not react with Streptococcus grouping antisera (Streptex kit; Murex Biotech Ltd., Dartford, England). It was not possible to identify the Streptococcus to the species level by using the API 20 Strep system (bio-Mtrieux, Marcy l’Etoile, France). Genetic analysis was performed with the MicroSeq 500 16s Ribosomal DNA bacterial sequencing kit (Applied Biosystems, Foster City, CA), and the obtained fragments were analyzed by the ABI PRISM 3 10 DNA sequencer (Applied Biosystems). The MicroSeq search resulted in a similarity of almost 99% with the S. salivarius sequence. Phenotypic methods and genetic analysis identified thejsolated strain in the CSF and blood as 5. salivarius. Antibiotic susceptibility testing was done by Etest (AB Biodisk, Solna, Sweden) and revealed susceptibility to ceftriaxone (MIC, 0.032 yg/ml), ampicillin (MIC, 0.094 pg/ml), and penicillin (MIC, 0.064 pg/ml). A transesophageal echocardiography was performed and showed no vegetations. Treatment with ampiciIlin was stopped on day 3. The patient improved clinically. Two weeks after admission, the intravenous ceftriaxone was stopped and the patient was discharged from the hospital in good condition.

The Diagnostic Value of Gram Stain for Initial Identification of the Etiologic Agent of Peritonitis in Capd Patients

To evaluate the effectiveness of the Gram stain in the initial diagnosis of the etiologic agent of peritonitis in continuous ambulatory peritoneal dialysis (CAPD). Retrospective study analyzing the sensitivity (S), specificity (SS), positive predictive value (+PV), and negative predictive value (-PV) of the Gram stain relating to the results of cultures in 149 episodes of peritonitis in CAPD. The data were analyzed in two studies. In the first, only the cases with detection of a single agent by Gram stain were taken (Study 1). In the second, only the cases with two agents in Gram stain were evaluated (Study 2). Dialysis Unit and Laboratory of Microbiology of a tertiary medical center. Sixty-three patients on regular CAPD who presented one or more episodes of peritonitis from May 1992 to May 1995. The positivity of Gram stain was 93.2% and the sensitivity was 95.7%. The values of S, SS, +PV, and -PV were respectively: 94.9%, 53.5%, 68.3%, and 90.9% for gram-positive cocci and 83.3%, 98.8%, 95.2%, and 95.6% for gram-negative bacilli. The association of gram-positive cocci plus gram-negative bacilli were predictive of growth of both in 6.8%, growth of gram-positive cocci in 13.7%, and growth of gram-negative bacilli in 72.5%. The Gram stain is a method of great value in the initial diagnosis of the etiologic agent of peritonitis in CAPD, especially for gram-negative bacilli.

Evaluation of a new agar in Uricult-Trio for rapid detection of Escherichia coli in urine

A new commercial agar (Uricult-Trio) with 8-hydroxyquinoline-beta-glucuronide was used to assess 2,536 uropathogens for beta-glucuronidase activity typical of Escherichia coli. Included in the study were 1,807 strains of the family Enterobacteriaceae, 284 strains of nonfermentative bacilli, 345 strains of gram-positive cocci, and 100 yeast strains. In identifying E. coli, the test agar gave a sensitivity of 95.5% and a specificity of 97.2%. Fifty E. coli isolates gave negative reactions; 31 non-E. coli strains produced black colonies characteristic of E. coli. No growth of gram-positive cocci and no false-positive reactions from yeasts were observed. The recovery rate for E. coli on this agar was at least 10% higher than that on blood agar.