Journal of Clinical Microbiology and Infectious Diseases

Gram stain evaluation in dermatovenereology clinic and microbiology laboratories: case series

Yolanda Pitra Kusumadewi — 2025-03-18

Introduction: The normal vaginal flora consists of aerobic and anaerobic bacteria; Lactobacillus sp. is the main microorganism in the vagina. In bacterial vaginosis, there is a decrease in Lactobacillus sp. and an increase in pathogenic bacteria. Direct gram staining of vaginal fluid is the standard microbiological method for diagnosing bacterial vaginosis. Gram staining allows us to distinguish between gram-positive and gram-negative bacteria based on differential staining with crystal violet iodine complex and safranin. This study aimed to evaluate and compare the gram stain results between the dermatovenereology clinic and microbiology laboratories.

Case description: Direct gram staining of vaginal or cervical swabs was carried out on women aged 19 years, women aged 29 years, and women aged 40 years. The gram staining was carried out at the dermatovenereology clinic and microbiology laboratory with different results between the two locations.

Conclusion: It is important to evaluate the process of taking vaginal fluid samples, making smears/slides, and gram staining so that the quality of the examination results is maintained

Appropriateness of antibiotic use and mortality risk factors in moderate to severe COVID-19 patients admitted to the intensive care unit of Bali Mandara Hospital in 2021

I Wayan Agus Gede Manik Saputra — 2025-04-30

Introduction: The COVID-19 pandemic has been closely linked to the overuse of antibiotics, particularly in hospitalized patients. Early clinical uncertainty and lack of definitive treatments led to widespread empiric use. This study assessed the appropriateness of antibiotic use and identified mortality risk factors among moderate to severe COVID-19 patients in Bali Mandara Hospital's ICU.

Methods: A retrospective cross-sectional study of 72 moderate to severe COVID-19 patients was conducted from 200 ICU admissions in 2021. Simple random sampling was used. Antibiotic appropriateness was assessed using the Gyssens method. Variables such as age, sex, severity, comorbidities (hypertension, diabetes), and culture results were analyzed using Chi-square tests.

Result: Among 322 antimicrobials, 226 were antibiotics. Gyssens' classification showed 40% appropriate use and 60% inappropriate use, mainly due to lack of indication (Category V). Age over 60, sex, and disease severity were significantly associated with mortality (p<0.05). Bacterial co-infection was confirmed in 30.6%, predominantly Enterobacteriaceae.

Conclusion: Antibiotic appropriateness remains low, with substantial overuse. Mortality correlated significantly with age, sex, and disease severity. These findings underscore the need for enhanced antimicrobial stewardship and targeted clinical management in cases of severe COVID-19.

The effect of prolonged exposure to the antibiotic meropenem on the resistance characteristics of Acinetobacter baumannii in vitro

Yossy I. Sutjipto — 2025-04-30

Introduction: To prove the effect of prolonged exposure to the antibiotic meropenem on the emergence of Acinetobacter baumannii resistance in vitro.

Methods: Meropenem-susceptible Acinetobacter baumannii isolates were exposed to 0.1 mg/ml meropenem and incubated for 8 days. The optical density and number of Acinetobacter baumannii bacterial colonies were observed every 24 hours. The average of optical density and the number of Acinetobacter baumannii bacterial colonies growing every 24 hours were analyzed using the Kruskal-Wallis test.

Results: The results showed a decrease in the average optical density value and the number of Acinetobacter baumannii bacterial colonies from day 1 to day 6 of incubation. However, there was an increase in the average optical density and the number of Acinetobacter baumannii bacterial colonies starting on day 7 and day 8.

Conclusions: Exposure to the antibiotic meropenem 0.1 mg/ml for 6 days might be associated with the emergence of Acinetobacter baumannii resistance in vitro.

Identification of Methicillin-Resistant Staphylococcus aureus (MRSA) carriers in the nasal passages of healthcare workers at Universitas Udayana Hospital

Made Illene Winaya — 2025-03-10

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a major pathogen responsible for healthcare-associated infections (HAIs), with growing global concern due to its resistance to multiple antibiotics, including beta-lactams. Healthcare workers (HCWs), particularly those in critical care settings, are at risk of being asymptomatic carriers, facilitating MRSA transmission. This study aimed to determine the prevalence and distribution of MRSA carriers among healthcare workers (HCWs) at Universitas Udayana Hospital, Bali, Indonesia.

Methods: A descriptive cross-sectional study was conducted from October to November 2024. A total of 44 nasal swab samples were collected from nurses across various hospital wards. Samples were cultured on Mannitol Salt Agar and underwent Gram staining, catalase, and coagulase testing. MRSA identification was confirmed using the cefoxitin disk diffusion method, with inhibition zones of ≤21 mm indicating methicillin resistance, as per CLSI guidelines. Data were analyzed descriptively.

Results: MRSA colonization was identified in 4 out of 44 samples (9.1%), with cases distributed in the NICU (2 cases), ICU (1 case), and Surgical Ward (1 case). In addition, 12 samples (27.3%) were positive for methicillin-sensitive Staphylococcus aureus (MSSA), while 28 samples (63.6%) contained coagulase-negative staphylococci (CNS), predominantly in the outpatient and emergency departments.

Conclusion: The presence of MRSA among HCWs, especially in critical units like NICU and ICU, emphasizes the importance of enhanced infection control measures. Routine screening, strict adherence to hand hygiene, appropriate use of personal protective equipment, and implementation of decolonization protocols, such as mupirocin application and chlorhexidine bathing, should be optimized to reduce MRSA transmission risks within healthcare facilities.

Trends in antimicrobial resistance of extended-spectrum beta-lactamase-producing Escherichia coli in urinary tract infections in ICU and non-ICU settings at Ngoerah Hospital, 2020-2022

Bryan Setiawan — 2025-03-26

Background: Urinary Tract Infection (UTI) is one of the most common nosocomial infections, with Escherichia coli (E. coli) as the primary pathogen. The misuse and overuse of antibiotics has led to the emergence of antibiotic resistance, particularly in Extended-Spectrum Beta-Lactamase (ESBL)-producing E. coli. This resistance presents a significant challenge in managing UTIs, especially among ICU and non-ICU ward patients. This study aims to analyze trends in antibiotic resistance of ESBL-producing E. coli in UTI patients in ICU and non-ICU settings at Ngoerah Hospital during 2020–2022.

Methods: A descriptive study with a cross-sectional approach was conducted. The inclusion criteria of this study were samples with identification results of E. coli bacteria with VITEK 2 Compact (bioMérieux, France), which showed E. coli with a probability greater than or equal to 90% and accompanied by sensitivity of E. coli bacteria to antibiotics.

Result: Among 789 samples meeting inclusion criteria, 90% (714) were from non-ICU wards, while 10% (75) were from ICU wards. ESBL-producing E. coli accounted for 53% (416), while non-ESBL strains constituted 47% (373). The prevalence of ESBL-producing E. coli in non-ICU wards increased from 43% (96 isolates) in 2020 to 47% (129 isolates) in 2021 and 51% (149 isolates) in 2022. In ICU wards, prevalence rose from 3% (6 isolates) in 2020 to 4% (12 isolates) in 2021 and 8% (24 isolates) in 2022. Tigecycline, meropenem, and ertapenem demonstrated high sensitivity (≥96%), while beta-lactam antibiotics like ampicillin, cefixime, and cefazolin showed 100% resistance.

Conclusion: The increasing resistance of ESBL-producing E. coli, particularly in non-ICU settings, highlights the need for strict antibiotic policies and monitoring to prevent further resistance.

Prevalence and antibiotic sensitivity patterns of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in pneumonia patients at Ngoerah Hospital from 2020 to 2022

Dewa Ayu Putu Aisaka Rajeshi Arikandini — 2025-03-18

Background: Pneumonia remains a major global cause of mortality, with Klebsiella pneumoniae recognized as a significant pathogen. The emergence of antibiotic resistance, particularly due to Extended-Spectrum Beta-Lactamase (ESBL) production by K. pneumoniae, complicates treatment efforts. This study aimed to determine the prevalence of ESBL-producing K. pneumoniae in pneumonia patients and assess its antibiotic sensitivity profile.

Methods: A descriptive cross-sectional study was conducted retrospectively using secondary data from VITEK 2 Compact (bioMérieux) laboratory results on sputum samples collected from pneumonia patients at Ngoerah Hospital from 2020 to 2022. A total of 515 samples met the inclusion criteria out of 1,350 tested.

Results: ESBL-producing K. pneumoniae was identified in 305 isolates (59.2%), with yearly prevalence rates of 63% in 2020, 52% in 2021, and 61% in 2022. Most patients were male (66.6%), aged 60 years or older (40%), and treated in non-ICU settings (69.2%), with expectorated sputum as the most common specimen type (63%). Antibiotic sensitivity testing revealed high susceptibility of ESBL-producing isolates to ertapenem (100%), meropenem (100%), amikacin (93%), and tigecycline (81%).

Conclusion: ESBL-producing K. pneumoniae accounted for over half of pneumonia cases, with fluctuating prevalence across the study period. The infection predominantly affected older male patients treated in non-ICU wards. Despite resistance to many beta-lactam antibiotics, high sensitivity to carbapenems and other specific agents highlights their continued relevance in treatment. Surveillance of antibiotic resistance patterns remains essential for effective clinical management.

Molecular characteristics of the gyrA gene among rifampicin-resistant Mycobacterium tuberculosis isolates

Febriana Aquaresta — 2025-04-28

Background: Drug-resistant tuberculosis (TB) remains a public health threat, especially during this pandemic. Meanwhile, fluoroquinolone is used as a second-line multidrug-resistant TB (MDR-TB) treatment since this drug was previously prescribed for respiratory, urinary, and genital tract infections. However, unregulated and excessive use of fluoroquinolones leads to resistance.

Methods: The design of this study is a descriptive observational study with a cross sectional approach. This study aims to determine the pattern of gyrA gene mutation in fluoroquinolone resistance among rifampicin-resistant Mycobacterium tuberculosis isolates during the COVID-19 pandemic in Sumatra, Indonesia. The Mycobacterium tuberculosis isolates were stored in the Palembang Health Center Laboratory as the referral laboratory in Sumatra from January to December 2020. Out of the 233 isolates that were tested phenotypically by BACTEC MGIT, 8 isolates of fluoroquinolone resistance (ofloxacin or moxifloxacin or both) were obtained and sequenced using an ABI PRISM 3730XL analyzer for Single Nucleotide Polymorphism analysis (SNP).

Results: Among the six fluoroquinolone-resistant Mycobacterium tuberculosis isolates, the gyrA mutations were identified in 5/6 isolates (84%), A90V (34%), D94A (16%), and D94G (34%), while 1/6 isolates (16%) had no mutation in gyrA gene among Mycobacterium tuberculosis that were fluoroquinolone resistance.

Conclusion: The gyrA gene mutation in fluoroquinolone resistance among rifampicin-resistant Mycobacterium tuberculosis was commonly present in codon 90 (2/6 isolates =32%) and 94 (3/6 isolates=68%).