Gram-stained Smears Research Articles

Prevalence and associated factors of bacterial vaginosis among pregnant women in Hue, Vietnam.

Bacterial vaginosis (BV) is the most frequent vaginal infection affecting women of childbearing age worldwide. It is associated with significant adverse healthcare outcomes, especially during pregnancy. Although screening for BV could reduce potential pregnancy-related obstetric complications, there is no routine screening of pregnant women for BV in Vietnam. We aimed to identify the prevalence of BV among pregnant women and the associated factors in two tertiary hospitals in Hue, Vietnam. This cross-sectional descriptive study included 885 pregnant women in third trimester, who received routine antenatal care in the Hue Central Hospital and Hue University Hospital of Medicine and Pharmacy, Hue city, Thua Thien Hue province, Vietnam. Gram-stained vaginal smears were used for calculating the Nugent score and recording the fungal elements. In total, 435 (49.1%) women had a normal BV score, 352 (39.8%) had intermediate vaginal microbiota, and 98 (11.1%) had BV. Among the 98 women with BV, 71 (72.4%) also had fungal infection. There was a significant association of BV with discharge (p = 0.004) and abnormal cervix (p = 0.014). BV was significantly more frequent among the women who reported previous abortion or miscarriage (p = 0.007). About a tenth of women in Thua Thien Hue province have BV in the third trimester of pregnancy being associated with previous adverse outcome. Discharge with fishy odour is still a characteristic feature among subtle clinical presentations of BV. Better awareness about this disease and routine test-and-treat management during pregnancy may improve pregnancy outcome.

Data augmentation for Gram-stain images based on Vector Quantized Variational AutoEncoder

Availability of large-scale datasets plays a significant role in segmentation and classification tasks using deep learning. However, domains such as healthcare inherently suffer from unavailability and inaccessibility of data. This leads to challenges in deploying CNN-based models for computer-aided diagnosis. This challenge extends to Gram-stain image analysis for detecting bacterial infections, which is a crucial task. The lack of datasets containing Gram-stained direct and culture smear images exacerbates the significant challenges in deep learning tasks. In this regard, we investigate a novel application of the Variational AutoEncoder. Specifically, the Vector Quantized Variational AutoEncoder model is trained to generate the Gram-stain images. Incorporating a novel loss function, where the quality loss (Lqu) is derived by integrating the LossSSIM, L1, and L2 losses with the VQ-VAE loss (Lossvq) for proposed approach for Gram-stained direct and culture smear images. This modification facilitates the creation of images closely resembling the original input, leading to notable SSIM scores of 0.92 for Gram-stained culture images and 0.88 for Gram-stained direct smear images. The current study compares the proposed method with state-of-the-art machine learning based and CNN based transformations. This work also demonstrates the classification process with and without image augmentation. It shows that the area under the curve in the case of augmentation is higher by an average of 20%.

Second trimester vaginal Candida colonization among pregnant women attending antenatal care in Bukavu, Democratic Republic of the Congo: prevalence, clinical correlates, risk factors and pregnancy outcomes

IntroductionVaginal Candida colonization (CC) can lead to vulvovaginal candidiasis, the second most prevalent vaginal condition worldwide, and has been associated with adverse birth outcomes. However, no data on CC in the Democratic Republic of the Congo are available. We investigated the prevalence, Candida species, clinical correlates, risk factors and pregnancy outcomes in women with CC in the second trimester of pregnancy.Material and methodsIn Bukavu, the Democratic Republic of the Congo, pregnant women were recruited during antenatal care between 16 and 20 weeks of gestation from January 2017 to October 2017 and followed until delivery. Sociodemographics, sexual behavioral, hygienic and clinical characteristics, microbiological data and pregnancy outcomes were collected. Candida detection and speciation was performed with microscopy (Gram-stained smears and wet-mount) and/or quantitative PCR. Multivariate regression models were used to estimate the different associations with CC.ResultsThe prevalence of CC by wet mount, microscopy of Gram-stain smears and qPCR was 27.9%, 28.1% and 38.2%, respectively. C. albicans was the most prevalent Candida species (91.0%). Previous genital infections, an intermediate vaginal microbiota, bacterial vaginosis, and the use of pit toilets were risk factors for CC. Clinically, CC was associated with itching only. Women with CC had twice the odds for preterm birth, if Candida concentration was high, the odds were four times higher.ConclusionsIn Bukavu, the Democratic Republic of the Congo, the prevalence of CC was high and associated with microbiological and modifiable risk factors. Screening and treatment for CC during antenatal care should be investigated as a possible strategy to reduce preterm birth.

Preliminary evaluation of in-Vivo and in-Vitro antifungal activity of Piper longum, Origanum majorana, Embelia ribes and Butea monosperma with Gas chromatography-mass spectrometry analysis of phytochemical property of plant extracts against Candida species causing dermatological

: Globally, fungi infections cause more than 1.5 million fatalities annually, mostly in those with weakened immune systems with Candida albicans found to be the chief pathogen. Approximately one to two billion people are affected by dermatological fungal infections involving skin and skin appendages, perhaps very difficult to estimate its exact prevalence due to under-reporting, this augments the requirements for more attention towards new molecules to combat resistance.: Evaluation of antifungal activity of Piper longum, Origanum majorana, Embelia ribes, Butea monosperma both in-vivo and in-vitro analysis against Candida species. This experimental study conducted for evaluating both in-vitro as well as in-vivo anti-fungal activity of Piper longum, Butea monosperma, Embelia ribes and Origanum majorana, and evaluation of phytoconstituent through Gas Chromatography and Mass Spectrometry analysis by GC-MS-QP-2010 plus system to identify the active bio-component. Fluconazole used as the standard control in-vivo and Itraconazole used as a standard drug for in-vitro infection and analysis. Ethical Committee approval was taken (IMS, BHU Dean/2018/CAEC/818). The antifungal activity of the extracts in-vitro was evaluated by Resazurin microtiter assay against Candida sp. Micro broth dilution methods. Rats were given an injection of 0.2 mL (intravenously/i.v) of a 10° UFC/mL inoculum made from a fresh 48-hour Candida albicans culture in sterile saline to cause infection. Induction of candidemia in rats was confirmed using qualitative Gram-stained smear. : The extracts of Butea monosperma (Alc.), Piper longum (Alc.), Origanum majorana (Aq.), and Piper longum (Aq.) showed effective antifungal activity against Candida sp. in-vitro research with a minimum inhibitory concentration (MIC) of less than 0.25 mg/mL. : This study is contributing towards the search for a natural drug candidate to combat resistance caused by the Candida genus, as it is one of the highest contributors to fungal dermatological/skin infections.

A Study of Bacteriological Profile of Bacterial Isolates in Sputum Samples in Azadi Teaching Hospital in Duhok City

The objective of this current study is to examine the bacteriological profile of microorganisms isolated in sputum samples collected from the patients in the Azadi Teaching Hospital in Duhok City. The current cross-sectional observational study was carried out in the Laboratory of Microbiology, Department of the Basic Science, College of Nursing at the University of Duhok, Kurdistan region, Iraq. A total of 150 patients with clinically verified bacterial infections were included in the study using a gram-staining and routine sampling procedure. Microscopic examination of the gram-stained sputum smears was done to assess the presence of epithelial cells, pus cells, or organisms. Consequently, 83 bacterial samples yielded positive results that indicated the presence of bacterial growth, while the remaining samples 67 did not exhibit any bacterial growth or normal flora. On assessing the organisms isolated from the study subjects, it was observed that out of 150 processed sputum samples, 44.7% (n=67) were non-bacterial samples and 55.3% (n=83) were pathogenic bacterial samples. It was seen that the most common isolated organism was Klebsiella pneumoniae in (44.6%; 37/83), followed by Pseudomonas aeruginosa in (35%; 29/83), and then Escherichia coli in (20.4%; 17/83), were the most frequently isolated microorganisms from sputum samples of patients with upper or lower respiratory infections. The current study comes to the conclusion that in order to obtain lower respiratory tract infection samples from subjects, high-quality sputum must be obtained, and initial sputum screening should be carried out.

Clinical screening of Nocardia in sputum smears based on neural networks.

Nocardia is clinically rare but highly pathogenic in clinical practice. Due to the lack of Nocardia screening methods, Nocardia is often missed in diagnosis, leading to worsening the condition. Therefore, this paper proposes a Nocardia screening method based on neural networks, aiming at quick Nocardia detection in sputum specimens with low costs and thereby reducing the missed diagnosis rate. Firstly, sputum specimens were collected from patients who were infected with Nocardia, and a part of the specimens were mixed with new sputum specimens from patients without Nocardia infection to enhance the data diversity. Secondly, the specimens were converted into smears with Gram staining. Images were captured under a microscope and subsequently annotated by experts, creating two datasets. Thirdly, each dataset was divided into three subsets: the training set, the validation set and the test set. The training and validation sets were used for training networks, while the test set was used for evaluating the effeteness of the trained networks. Finally, a neural network model was trained on this dataset, with an image of Gram-stained sputum smear as input, this model determines the presence and locations of Nocardia instances within the image. After training, the detection network was evaluated on two datasets, resulting in classification accuracies of 97.3% and 98.3%, respectively. This network can identify Nocardia instances in about 24 milliseconds per image on a personal computer. The detection metrics of mAP50 on both datasets were 0.780 and 0.841, respectively. The Nocardia screening method can accurately and efficiently determine whether Nocardia exists in the images of Gram-stained sputum smears. Additionally, it can precisely locate the Nocardia instances, assisting doctors in confirming the presence of Nocardia.

Multispectral in-line hologram reconstruction with aberration compensation applied to Gram-stained bacteria microscopy

In multispectral digital in-line holographic microscopy (DIHM), aberrations of the optical system affect the repeatability of the reconstruction of transmittance, phase and morphology of the objects of interest. Here we address this issue first by model fitting calibration using transparent beads inserted in the sample. This step estimates the aberrations of the optical system as a function of the lateral position in the field of view and at each wavelength. Second, we use a regularized inverse problem approach (IPA) to reconstruct the transmittance and phase of objects of interest. Our method accounts for shift-variant chromatic and geometrical aberrations in the forward model. The multi-wavelength holograms are jointly reconstructed by favouring the colocalization of the object edges. The method is applied to the case of bacteria imaging in Gram-stained blood smears. It shows our methodology evaluates aberrations with good repeatability. This improves the repeatability of the reconstructions and delivers more contrasted spectral signatures in transmittance and phase, which could benefit applications of microscopy, such as the analysis and classification of stained bacteria.

Comparison of Enhanced and Manual Urinalysis for Detecting Urinary Tract Infections in Children

Background: Urinary tract infections (UTIs) are among the most common bacterial infections in children. Urinalysis (UA) is a beneficial test for the preliminary diagnosis of UTIs. The presence of bacteriuria in UA can be determined by either an enhanced (using uncentrifuged Gram-stained specimens) or manual (using centrifuged specimens) technique. However, the diagnostic performance of enhanced UA is not well-established in childhood UTIs. Objectives: To assess the ability of enhanced and automated urinalysis to detect UTIs in children. Methods: This cross-sectional study was conducted on 191 children with the symptoms of UTI referred to Dr. Sheikh Hospital, Mashhad, Iran, from 2018 to 2019. Standard urinalysis, enhanced urinalysis, and quantitative urine culture were performed on specimens. A positive enhanced UA test was defined as ≥ 10 white blood cells per mL of urine and the presence of any bacteria per 10 high-power microscopic fields of a Gram-stained smear. A positive manual UA test was defined as ≥ 5 white blood cells per high-power field. The results of standard and enhanced UA were compared with urine culture findings to determine the accuracy of these two methods in detecting UTIs. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were determined for each test. Results: The results showed that the prevalence of UTI was 23%. Enhanced UA retrieved a sensitivity of 97.7%, specificity of 93.1%, PPV of 81.1%, and NPV of 99.3% for detecting UTIs. In standard UA, sensitivity, specificity, PPV, and NPV were 90.9%, 80.7%, 57.1%, and 96.6% for pyuria, 56.8%, 98.6%, 92.5%, and 88.4% for the nitrite test, 72.7%, 94.5%, 80%, and 92% for the leukocyte esterase test, respectively. Conclusions: Enhanced UA had higher sensitivity, specificity, PPV, and NPV than standard UA.

Role of rapid tests in diagnosis of bloodstream infections and their effect on promoting antibiotic stewardship among febrile neutropenic patients in Alexandria

Abstract Background Bloodstream infections (BSIs) in neutropenic patients significantly contribute to morbidity and mortality. The rapid identification and determination of antimicrobial susceptibility of pathogens causing BSIs can help in early initiation of targeted antimicrobial therapy and allow de-escalation of unnecessary empirical antimicrobials. Aim This study evaluated the rapidity and accuracy of gram-stained smear from blood culture bottles and direct VITEK 2 to identify and determine antimicrobial susceptibility of pathogens causing BSIs and assessed the effect of these tests on modification of empiric therapy. Patients and methods Blood cultures were collected from 100 febrile neutropenic patients from the Hematology Unit in Alexandria Main University Hospital. Gram-stained smears were prepared from blood culture bottles; blood from bottles with suggestive smears was directly inoculated into the VITEK 2 system, and results were compared with conventional methods. Results were instantly reported to clinicians. Empirical antimicrobials and their modification were recorded. Results BSI was detected in 30% of patients. Gram-negative bacteremia was predominant (24/30). Gram-stained smears showed complete accordance with conventional methods in 99 samples. VITEK 2 correctly identified all tested microorganisms, and the overall agreement in susceptibility testing was 100 and 95.2% for gram-positive and gram-negative bacteria, respectively. The effect of these rapid tests on empiric antimicrobial therapy was evident in 12 of 30 patients with documented BSIs (40%). Conclusion Gram-stained smear and direct VITEK 2 testing when integrated with instant reporting is a valid approach to guide targeted therapy in neutropenic patients in the context of extremely resistant pathogens.

Vitamin D supplementation in bacterial vaginosis

Research objectives: to evaluate the effect of vitamin D supplementation (Olidetrim) in the complex anti-relapse therapy of bacterial vaginosis (BV) in women of reproductive age.Materials and methods. The study included 63 women of reproductive age with BV and with a level of 25-hydroxyvitamin D below 75 nmol/l, who were divided into groups: the main group included 32 women who were prescribed standard therapy (metronidazole 500 mg twice a day for 7 days) and cholecalciferol (Olidetrim) 4000 IU/day with subsequent continuation of its intake in a dose 2000 IU/day for up to 6 months; the comparison group included 31 women, who were not intake vitamin D preparation during the standard therapy and during the next 6 months of observation.After treatment, women provided completed questionnaires and self-collected vaginal swabs 1, 3, and 6 months after beginning of the study or before BV relapse. Evaluation of Gram-stained smears was carried out by microscopy according to Hay/Ison criteria, where BV corresponded to type III. The level of 25-hydroxyvitamin D in blood serum was determined by the chemiluminescent immunoassay method before the start of thestudy and at 3 and 6 months of follow-up.The primary endpoint was BV recurrence at any post-treatment interval; the patients were offered a second treatment in this case.Results. Vitamin D deficiency at the beginning of the study was detected in 12 (37.5%) patients of the main group and in 13 (41.94%) women of the comparison group, an insufficient level – in 20 (62.5%) patients of the main group and in 58.06% women of the comparison group.Patients complained of pathological vaginal discharge, itching, burning and dyspareunia lasting from 1 week to 2 months. At the seven-day course of basic BV therapy all participants noted the normalization of discharge and the absence of vaginal discomfort. Positive dynamics were also observed according to the gynecological examination.One month after the treatment, 4 (12.5%) participants of the main and 6 (19.36%) women of the comparison group noted the recovery of symptoms. Absence of BV recurrence after 3 months was noted by 25 (78.12%) women of the main group and 16 (51.61%) women of the comparison group (p = 0.054), that was confirmed by the laboratory tests. At the same time, the level of 25-hydroxyvitamin D in the main group increased by 43.37% (p = 0.05), and in the comparison group it did not differ from the initial level. There were no complaints in 24 (75.0%) patients after 6 months, however, a laboratory-confirmed normal state of the vaginal microbiome was observed in 22 (70.96%) participants of the main group and in 14 (45.16%, p = 0.07) women of the comparison group. Continuation of vitamin D intake at a dose of 2000 IU/day for the next 3 months demonstrated maintenance of the 25-hydroxyvitamin D level within normal values (81.7 ± 6.23 nmol/l in the main group vs. 38.51 ± 5.43 nmol/l in comparison group, p = 0.05).Conclusions. A decrease in the frequency of BV recurrences in women during support a normal level of vitamin D was found, which allows recommending the inclusion of cholecalciferol supplementation (Olidetrim) in the standard treatment. In patients with recurrent BV a long-term monitoring of the serum level of vitamin D is advisable in order to timely prescribe supplementation.

Biocenosis of the vaginal tract in patients with fertility on the background of chronic endometritis

The objective: to determine the microbiota profile of vagina, cervical canal and endometrium by chronic endometritis (CE) in patients with a history of infertility.Materials and methods. A prospective study and clinical-laboratory analysis of the microbiocenosis of the vagina, cervical canal and uterine cavity in women with CE and infertility in history was conducted. The 1st group included 100 women of reproductive age with CE and infertility, the 2nd group – 40 women of reproductive age without impaired fertility.The scope of research is pH-metry of vaginal secretions, assessment of vaginal microbiocenosis based on Gram-stained smear microscopy, polymerase chain reaction method, cultural research of vaginal secretions, study of the microflora of the cervical canal and the uterine body.Results. In women with CE and a history of infertility, normobiocenosis of the vagina was found in only 32 patients (32.0±3.1%). In other cases, bacterial vaginosis, vulvovaginal candidiasis, and bacterial vulvovaginitis were detected. Microscopy and PCR-diagnostics of secretions from the cervical canal and uterine cavity in women with CE determined Streptococcus agalactiae, Enterococcus faecalis, Chlamidia trachomatis, Ureaplasma urealyticum, Mycoplasma genitalium, HPV, Gardnerella, HSV 1st or 2nd types and CMV.Conclusions. A characteristic feature of the vaginal flora in chronic endometritis is a high level of dysbiotic changes combined with high rates of insemination by opportunistic flora. The microbiota of the cervical canal and endometrium during inflammatory processes of the endometrium is characterized by the predominance of obligate anaerobic microorganisms in the form of associations with viral infection, which are significantly more often (more than 2 times) detected in the endocervix compared to the endometrium.

A Retrospective Study of Demographic Profile and Management Appraisal Non-Gonococcal Urethritis in Male Patients Attending a Tertiary Care Hospital in Odisha.

Male urethritis is primarily sexually transmitted in India and has also shown a high rate. Urethritis or inflammation of the urethra is a multifactorial condition. It is called gonococcal urethritis (GU) when Neisseria gonorrhoeae is detected in a urethral smear of the patient and non-gonococcal urethritis (NGU) when this organism cannot be visualized. To study the demographic profile and management pattern of NGU retrospectively in male patients. Urethritis cases were identified from the dermatology outpatient record. A retrospective study of data of patients diagnosed with NGU was done for 5 years from August 2015 to July 2020. We included only male patients more than 10 years of age. Chlamydia trachomatis (CT, 48%) was the most common causative organism found. The diagnosis was confirmed by demonstrating ≥5 polymorphonuclear lymphocytes (PMNLs) from the anterior urethra using a Gram-stained urethral smear followed by the polymerase chain reaction (PCR.) of urine. The most common age group affected was 21-30, (30, 46%) years with a mean ± standard deviation (SD) of 30.32 ± 10.80. Most patients were from low socioeconomic status, labourers (36, 55%). A history of heterosexual behaviour, (57, 88%) was the most common and it had been seen that sexual exposure was more common in unmarried patients (38, 59%). Associated features were seen in 18 patients. Among recommended initial therapies for NGU, doxycycline and azithromycin might be improved by the addition of antimicrobial coverage for Trichomonas vaginalis (T. vaginalis). Further, it was seen that response to doxycycline therapy was better than azithromycin. Though there is an increased incidence of Herpes infections, we found Chlamydia infection to be the most common cause of NGU. Doxycycline showed better results in NGU due to Chlamydia infection.

MICROSCOPIC AND DEMOGRAPHIC PROFILE OF SEXUALLY TRANSMITTED DISEASES AT A TERTIARY CARE CENTRE OF CENTRAL INDIA

Objective: To find out the prevalence of sexually transmitted diseases (STDS) among the patients attending the tertiary care center through microscopic screening. Methods: This Cross Sectional study was conducted from April 2018 to December 2019 wherein 500 samples were collected from patients attending the STD clinic of Skin and VD-OPD, and integrated counseling and testing center (ICTC), having cervical or urethral discharge along with signs and symptoms of sexually transmitted infections (STI) and were screened microscopically for the same. Results: This study reported only 2 cases suggestive of N. gonorrhoeae on microscopy, wherein the Gram-stained smear showed the presence of a particular arrangement of Gram-negative coffee bean shape cocci, both intracellular and extracellular, and plenty of pus cells. In Direct Microscopy findings, 49.2% of samples showed normal flora, 20.4% Gram-positive cocci, 15.8% Gram-negative cocci, 4% Clue cells, 13.8% with mixed flora, 3% Budding yeast-like cells and 0.4% showed Gram-negative cocci. Conclusion: Such studies involving the laboratory and demographic data should be conducted regularly, which can help in estimating the disease burden, strengthening the diagnostic capacity, and formulating the requisite strategy for tackling this problem through a syndromic approach.

Evaluation of the World Health Organization Global Invasive Bacterial Vaccine-Preventable Disease (IB-VPD) Surveillance Network's Laboratory External Quality Assessment Programme, 2014-2019.

Introduction. In 2009, the World Health Organization (WHO) established the Global Invasive Bacterial Vaccine Preventable Disease (IB-VPD) Surveillance Network (GISN) to monitor the global burden and aetiology of bacterial meningitis, pneumonia and sepsis caused by Haemophilus influenzae (Hi), Neisseria meningitidis (Nm) and Streptococcus pneumoniae (Sp).Hypothesis/Gap Statement. The GISN established an external quality assessment (EQA) programme for the characterization of Hi, Nm and Sp by culture and diagnostic PCR.Aim. To assess the performance of sentinel site laboratories (SSLs), national laboratories (NLs) and regional reference laboratories (RRLs) between 2014 and 2019 in the EQA programme.Methodology. Test samples consisted of bacterial smears for Gram-staining, viable isolates for identification and serotyping or serogrouping (ST/SG), plus simulated cerebrospinal fluid (CSF) samples for species detection and ST/SG by PCR. SSLs and NLs were only required to analyse the slides for Gram staining and identify the species of the live isolates. RRLs, and any SLs and NLs that had the additional laboratory capacity, were also required to ST/SG the viable isolates and analyse the simulated CSF samples.Results. Across the period, 69-112 SS/NL labs and eight or nine RRLs participated in the EQA exercise. Most participants correctly identified Nm and Sp in Gram-stained smears but were less successful with Hi and other species. SSLs/NLs identified the Hi, Nm and Sp cultures well and also submitted up to 56 % of Hi, 62 % of Nm and 33 % of Sp optional ST/SG results each year. There was an increasing trend in the proportion of correct results submitted over the 6 years for Nm and Sp. Some SSLs/NLs also performed the optional detection and ST/SG of the three organisms by PCR in simulated CSF from 2015 onwards; 89-100 % of the CSF samples were correctly identified and 76-93 % of Hi-, 90-100 % of Nm- and 75-100 % of Sp-positive samples were also correctly ST/SG across the distributions. The RRLs performed all parts of the EQA to a very high standard, with very few errors across all aspects of the EQA.Conclusion. The EQA has been an important tool in maintaining high standards of laboratory testing and building of laboratory capacity in the GISN.

The Importance of Gram-stained Smears in the Diagnosis of Nocardia Infections

Nocardia species are low virulence bacteria found in nature. They can be an infectious agent, especially in patients with risk factors such as underlying immunosuppression, chronic lung disease, and malignancy. They can be easily overlooked because they are not seen frequently and has no pathognomonic symptoms. With this study, it was aimed to draw attention to the importance of microscopic examination of Gram-stained smears in the diagnosis of Nocardia infections in routine microbiology laboratories. Cases in which Nocardia spp. were detected in their clinical samples between November 2014-December 2015 in Hacettepe University Medical Faculty Hospital were included in the study. In the direct microscopic examination of Gram-stained smears of the samples arriving to the laboratory, the incubation periods of the cultures of the samples compatible with Nocardia spp. were extended. Then relevant colonies were identified by conventional microbiological methods and also by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS, bioMerieux, France) automated system. Species-level identification of Nocardia isolates was performed by 16S rRNA gene sequence analysis. To demonstrate the genetic relationship between Nocardia isolates, pulsed-field gel electrophoresis (PFGE) was performed. In vitro susceptibility of the isolates against amoxicillin-clavulanate (AMC), linezolid, moxifloxacin, trimethoprim-sulfamethoxazole (TMP-SXT), amikacin, imipenem, clarithromycin, cefepime, cefotaxime, ceftriaxone, and ciprofloxacin was determined using the gradient strip method (E-test). A total of 19 Nocardia spp. strains were isolated from eight patients. Four cases exhibited repeated growth of Nocardia spp. up to a period of nine months. The most frequently isolated species was N.cyriacigeorgica, which was identified in four cases. Other species isolated from patients were N.asteroides, N.transvalensis, N.farcinicia, and N.asiatica/arthritidis. When the results obtained with DNA sequence analysis and MALDI-TOF MS were compared, 16 (84.2%) of 19 isolates were correctly identified to the genus level and 9 (47.4%) to the species level with MALDI-TOF MS, while three (15.8%) isolates could not be identified, and seven (36.8%) isolates were misidentified. According to the PFGE results, it was determined that the strains isolated from the same patient were genetically identical. All isolates were susceptible to amikacin, cefepime, cefotaxime, ceftriaxone, imipenem, linezolid, and except one isolate to TMP-SXT. Among the study isolates, the most common resistance was against ciprofloxacin (62.5%), followed by clarithromycin (37.5%). N.cyriacigeorgica was determined as the most frequently detected and the most resistant species to antibiotics in the study population. Direct microscopic examination of clinical specimens is one of the most valuable methods for the identification of Nocardia-type bacteria, which is difficult to isolate in microbiology laboratories. With this study, the importance of examining Gram-stained clinical samples was emphasized in the identification of Nocardia species, which can emerge with a wide variety of clinical forms and can be easily overlooked. In addition, antibiotic susceptibility profiles of the isolated bacteria were determinedto contribute to species-specific susceptibility profiles. Accurate identification of Nocardia species will contribute to clinical and epidemiological studies.

Comparison of BACTEC versus conventional enrichment medium forperitoneal dialysate fluid culture.

Bacterial peritonitis remains a significant cause of mortality in patients on peritoneal dialysis (PD). Early detection of causative organisms and targeted antimicrobial treatment allow for better clinical outcomes. This study compares bacterial growth results from peritoneal dialysate in the BACTEC blood culture system vs. conventional culture. We conducted a prospective study on 46 patients with 63 consecutive episodes of suspected PD peritonitis between August 2020 and August 2021. PD dialysate was simultaneously sent to the laboratory in both BACTEC and sterile bottles. BACTEC bottles were incubated in the BD BACTEC FX system for 5 days. PD effluent transported from the sterile bottles was centrifuged at 3,000 rpm for 15 minutes; the supernatant was inoculated into cooked-meat broth for enrichment. Both incubation methods were extended to 14 days if microorganisms were seen on the Gram-stained smear. Recovery of isolated micro-organisms and time to detection (TTD) were compared. 26 episodes of suspected PD peritonitis based on clinical criteria were identified during the study period. The sensitivity of the BACTEC and the conventional culture methods was 50% and 42.3%, respectively (p=0.45). Seven samples had partial concordance or discordant results. McNemar's χ2-test revealed no statistical difference between either method (p=0.45). TTD was 18.9 ± 24.4 hours via the BACTEC method vs. 37 ± 16.5 hours in conventional cultures (p=0.014). The comparable sensitivities and similar yield in identifying pathogens could be due to the enrichment medium and prolonged incubation period. The shorter TTD for the BACTEC method could facilitate earlier confirmation of bacteriological diagnosis and subsequent treatment strategies.

DESEMPENHO DO EXAME CITOPATOLÓGICO NO DIAGNÓSTICO DE VAGINOSE BACTERIANA

ABSTRACT Objective: to assess Pap smear performance in bacterial vaginosis diagnosis in women of childbearing age. Method: a cross-sectional, retrospective study carried out in a health unit in a city in the interior of São Paulo. The study included information from 1,173 women who underwent collection of Pap smear and Gram-stained vaginal smears (reference standard test for bacterial vaginosis diagnosis) and analysis based on the Nugent score, from January 2013 to June 2020. Secondary data collected from the information and internal control systems were used. Results: the prevalence of bacterial vaginosis in the sample was 31.8%. The degree of agreement between the two diagnostic methods by the Kappa index was 0.54, considered moderate, with a value <0.001. Sensitivity was 55.2% and specificity was 94.1%, with accuracy of 81.7% and positive predictive value of 81.4% and negative of 81.8%. Conclusion: Pap smear showed agreement with the reference standard of 0.77, however, does not replace the Gram-stained vaginal smear, but can be used as an auxiliary method for bacterial vaginosis diagnosis.

Filamentous Bacteria Resembling Fungal Hyphae: A Diagnostic Pitfall

Abstract Introduction/Objective When exposed to antibiotics or a host immune response, Gram-negative bacilli grow with inhibited septation producing long continuous cells; a phenomenon known as filamentation. Methods/Case Report We present a case of a 68-year-old man with quadriplegia, which was caused by the right vertebral artery dissection and subarachnoid hemorrhage requiring an emergent posterior inferior cerebellar artery direct bypass surgery. Lung lower lobe infiltrates were noted 3 days after the surgery. A bronchial wash culture was negative for growth, and a sputum specimen grew Serratia marcescens. The patient was treated with piperacillin/tazobactam, vancomycin, cefepime, cefazolin. After the surgery, he suddenly developed severe cerebral vasospasm of the posterior circulation with cerebellar strokes requiring emergent posterior fossa decompression. He remained on ventricular drainage by external ventricular drainage. After two days, the patient's cerebrospinal fluid (CSF) was sent for evaluation, on which a high segmented neutrophil count of 35% and Gram-negative filamentous organisms with occasional swellings were noted. Multi-drug resistant Serratia marcescens was isolated from the CSF specimen four days later. Neurosurgical procedures were reported to be highly associated with S. marcescens meningitis. The filamentous organisms noted on Gram-stained direct smear most likely represented Gram-negative S. marcescens with filamentation due to administration of sub-inhibitory concentrations of antibiotics. Results (if a Case Study enter NA) NA. Conclusion Gram-negative bacilli can develop abnormal morphology including elongation, swelling, and filamentation due to stress or sub-inhibitory antibiotic exposure, and it is important for microbiologists and pathologists to be aware of this phenomenon to avoid misinterpretation that may lead to inappropriate treatment.

Analysis Of Sputum Culture And Gram Staining In Subjects With Lower Respiratory Tract Infection: A Microbiological Assessment

Background: To diagnose the LRTIs (lower respiratory tract infections) in the microbiological laboratory, the most frequentlyemployed method is the microscopic examination of expectorated sputum samples. These sputum samples are generallycontaminated with the normal flora that is resident of the oropharynx. To manage and diagnose lower respiratory tract infections,sputum collection, culture, and microscopy are vital.Aim: The present study was conducted to analyse the sputum culture and gram staining in subjects with lower respiratory tractinfections.Materials & Methods: The study included 140 sputum samples where culture and gram-staining were done. Microscopic examinationof the gram-stained sputum smears was done to assess the presence of epithelial cells, pus cells, or organisms. Bartlett’s gradingsystem was used to assess the quality of the expectorated sputum samples. Bacterial isolates were identified using standard protocols.On Mueller Hinton agar, the Kirby Bauer disc diffusion method was used for testing antibiotic susceptibility.Results: In 140 processed sputum samples, it was seen that there were 55% (n=77) acceptable samples and 45% (n=63) nonacceptable samples. Total potential pathogens obtained from the study samples were 62.85% (n=88) samples were from acceptablesamples, in 76.13% (n=67) potential pathogens were isolated, whereas, from non-acceptable samples, potential pathogens wereisolated from 23.86% (n=21) samples. On assessing the organisms isolated from the study subjects, it was seen that the most commonisolated organism was Klebsiella pneumoniae isolated from 28.40% (n=25) samples followed by Pseudomonas aeruginosa in 14.77%(n=13) samples, Staphylococcus aureus in 13.63% (n=12) study samples, Escherichia coli in 12.5% (n=11) samples, Streptococcuspyogenes in 7.95% (n=7) samples, Klebsiella oxytoca in 6.81% (n=6) study samples, Streptococcus pneumoniae in 5.68% (n=5)study samples, Acinetobacter baumannii, Citrobacter koseri, and Enterobacter aerogenes in 3.40% (n=3) study samplesConclusion: The present study concludes that in subjects with Lower respiratory tract infections, good quality sputum must beobtained, and initial screening of the sputum should be done to obtain.

P223 First case of Candida auris candidemia in Manipur, Northeast India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjective Candida auris is known as an emerging ‘superbug’ because of its intrinsic resistance to one or more, sometimes to all available antifungal drugs and spreading globally. It has the ability to cause devastating nosocomial infections. In India, C. auris infection is on the rise with reports from north, south, central and eastern India. Here we present the first case of C. auris fungemia from a tertiary care hospital of Manipur in Northeast India.MethodsA 15-year-old Muslim girl was referred from a private hospital to Regional Institute of Medical Sciences (RIMS) hospital on November 19, 2021 with a history of burning epigastrium, headache, loss of appetite, shortness of breath, dry cough, fever, and generalized weakness for last 3 days. At the time of admission she was cyanotic. Family gave history of congenital heart disease and frequent visits to hospital. Echocardiogram revealed congenital cyanotic heart disease (Tetralogy of Fallot) showing large perimembranous VSD with bidirectional shunt. A complete hemogram showed neutrophilic leukocytosis with shift to left with band form, absolute monocytosis, and increased RBC count with mild anisocytosis. On November 24, 2021, 5 days after admission, her condition deteriorated and she was shifted to ICU. However, the condition of the patient deteriorated and she died on November 29, 2021 due to acute decompensated heart failure. Follow-up of other patients admitted in the same ward revealed no candidemia in next the few weeks.ResultsA single blood culture sent on November 29, 2021 was incubated in an automated blood culture system, BacT Alert and showed growth of budding yeast cells. Growth in SDA revealed it to be Candida sps. and Gram-stained smear examination revealed presence of budding yeast cells but no pseudohyphae. Germ tube test was negative. On CHROM agar, it produces pale yellow colonies at 24 h which progresses to light purple colonies around the rim at 48 h. Further processing in VITEK 2 (Biomerieux) identified it as C. auris. The isolate was sent to National Culture Collection of Pathogenic Fungi, WHO collaborating center, PGIMER and the isolate was confirmed as Candida auris by MALDI-TOF assay.Conclusion Candida auris is spreading irrespective of the level of health care. Blood culture before administration of antibiotics and in febrile sick patients cannot be underestimated. Rapid and accurate identification methods for timely diagnosis and stringent infection control measures with an emphasis on hand hygiene are important to prevent and control C. auris outbreaks.