Phytochemical Composition and Antimicrobial Efficacy of Salvadora persica Root Extracts Against Carbapenem-Resistant Acinetobacter baumannii.

Background Carbapenem-resistant Acinetobacter baumannii (CRAB) are difficult to eradicate from the environment and are virtually immune to all antibiotics. Consequently, CRAB may culminate in severe outbreaks and fatal infections among people attending hospitals and nursing homes. Salvadora persica has been used as an herbal remedy and chewing sticks for dental cleansing. Evaluating S. persica's efficacy against CRAB may provide an alternative approach to treating CRAB infections in healthcare environments, considering its traditional application in dental hygiene. Employing S. persica as an herbal remedy could be a part of a more sustainable approach to control CRAB infections. Aim To investigate the phytochemical composition of S. persica and evaluate its antimicrobial properties. Materials and methods The roots were extracted by Soxhlet apparatus using n-hexane, chloroform, and methanol. Each extract was analyzed using gas chromatography-mass spectrometry (GCMS) and characterized using WN908.L and National Institute of Standards and Technology (NIST) libraries. The antimicrobial activity of each extract against CRAB was evaluated using a broth microdilution assay to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Results The GCMS analysis of different solvent extracts of S. persica roots showed the presence of various phytochemical compounds such as steroids, phenolic compounds, fatty acids, alcohols, terpenoids, and vitamin E. Both chloroform and hexane extracts showed the most effective antimicrobial activity with a MIC value of 3.13 mg/mL and an MBC value of 12.50 mg/mL, respectively. Benzoic acid was the major phytochemical compound identified from S. persica extract. N-hexane, chloroform, and methanol extracts exhibited maximum antimicrobial activity due to the presence of active compounds in them. Conclusion Chloroform and hexane extracts showed the most potent antibacterial activities against CRAB.

Detection of Macrolide-Resistant Streptococcus pneumoniae Genes and Its Clinical Outcomes in a Tertiary Teaching Hospital in Malaysia.

Background: Streptococcus pneumoniae is one of the leading causes of mortality and morbidity worldwide. The dramatic increase in in-vitro resistance of antimicrobial agents, particularly beta-lactams and macrolides, makes pneumococcal infections difficult to treat. The aim of this study was to describe the drug resistance rate, assess the prevalence of macrolide-resistant genes and review the clinical complications of pneumococcal infections among patients presented to Hospital Universiti Sains Malaysia (HUSM), Kelantan, Malaysia. Methods: This is a descriptive cross-sectional study. All S. pneumoniae isolates collected from clinical specimens within a 1-year period were subjected to selected antimicrobial susceptibility testing using E-test strips. Polymerase chain reaction (PCR) analysis was conducted to detect macrolide-resistant determinants. The patient's clinical data were obtained from clinical notes. Results: A total of 113 patients with a positive growth of S. pneumoniae were included in the study. The most common predisposing factors among them were bronchopulmonary diseases (15.9%). The penicillin-resistant rate was 7.1%, with minimal inhibitory concentration (MIC) ranging between 0.012 µg/mL and >32 µg/mL, and the erythromycin-resistant rate was 26.5%, with a MIC range of 0.03 µg/mL-> 256 µg/mL. Most of the erythromycin-resistant isolates were found to have the mef(A) gene (50.4%) and the erm(B) gene (20%); 16.7% had a combination of genes mef(A) and erm(B), and 13.3% had none of the two genes. Community-acquired pneumonia is the predominant type of pneumococcal infection. There was no significant association between the presence of macrolide resistance determinants and mortality (P = 0.837) or complications (P > 0.999 for empyema and cardiac complication; P = 0.135 for subdural abscess). Conclusion: The majority of erythromycin-resistant isolates were found to have the mef(A) gene, followed by the erm(B) gene and a combination of genes mef(A) and erm(B).

Dysregulated expression of miR­367 in disease development and its prospects as a therapeutic target and diagnostic biomarker (Review).

MicroRNA (miR)-367 has a wide range of functions in gene regulation and as such plays a critical role in cell proliferation, differentiation and development, making it an essential molecule in various physiological processes. miR-367 belongs to the miR-302/367 cluster and is located in the intronic region of human chromosome 4 on the 4q25 locus. Dysregulation of miR-367 is associated with various disease conditions, including cancer, inflammation and cardiac conditions. Moreover, miR-367 has shown promise both as a tumor suppressor and a potential diagnostic biomarker for breast, gastric and prostate cancer. The elucidation of the essential role of miR-367 in inflammation, development and cardiac diseases emphasizes its versatility in regulating various physiological processes beyond cancer biology. However, further research is necessary to fully understand the complex regulatory mechanisms involving miR-367 in different physiological and pathological contexts. In conclusion, the versatility and significance of miR-367 makes it a promising candidate for further study and in the development of new diagnostic and therapeutic strategies.

Pediatric brain abscess with fatal outcome caused by hypervirulent Klebsiella pneumoniae, serotype K2-ST65.

Hypervirulent Klebsiella pneumoniae (hvKp) is an emerging pathotype in addition to classical Klebsiella pneumoniae, with its ability to cause life-threatening, community-acquired metastatic infections even in healthy individuals. We presented a case of cerebral abscess preceded by otitis media in a 10-year-old child caused by hvKp. The isolates from blood pus aspirate were later identified as K. pneumoniae capsular serotype K2 and closely related to sequence type (ST65), with multiple hypervirulent genes detected (rmpA, rmpA2, iucA and peg344). She succumbed to death despite surgical drainage and susceptible antibiotic therapy. Clinicians should be cognizant of the rising incidence of hvKp infections in pediatric populations.

Distribution of sasX, qacA/B and mupA genes and determination of genetic relatedness of methicillin-resistant Staphylococcus aureus among clinical isolates and nasal swab samples from the same patients in a hospital in Malaysia.

INTRODUCTION: This study determined the distribution of sasX, qacA/B and mupA genes from methicillin-resistant Staphylococcus aureus (MRSA) isolated from clinical samples and nasal swab samples of the same patients and analysed their genetic relatedness. METHODS: Polymerase chain reaction was used to detect the presence of sasX, qacA/B and mupA genes from 47 paired MRSA isolates. A paired isolate was defined as one nasal swab (colonising) isolate and clinical isolate that caused infection in the same patient. 22 selected paired isolates were subjected to multilocus sequence typing (MLST). The genetic relatedness among the isolates and association between the putative genes with epidemic sequence types (STs) were investigated. RESULTS: 7 (14.9%, n = 14) paired isolates were positive for the sasX gene. qacA/B genes were positive in 7.4% (n = 7) of the isolates, from three paired isolates and one clinical isolate whose paired colonising isolate was negative. The paired sample of three patients were positive for both genes. The mupA gene was not detected in all the isolates. MLST revealed two epidemic STs, ST22 and ST239, and a novel ST4649. sasX and qacA/B genes were found in ST239 in 29.5% (n = 13) and 13.6% (n = 6) of cases, respectively. Gene co-existence occurred in 13.6% (n = 6) of MRSA ST239 and 2.3% (n = 1) of MRSA ST4649. CONCLUSION: sasX and qacA/B genes were present in the MRSA isolates, while the mupA gene was undetected. ST22 and ST239 were the major MRSA clones. The circulating MRSA genotypes conferred different virulence and resistance determinants in our healthcare settings.

Seroprevalence of SARS-CoV-2 Antibodies in Africa: A Systematic Review and Meta-Analysis.

A reliable estimate of SARS-CoV-2-specific antibodies is increasingly important to track the spread of infection and define the true burden of the ongoing COVID-19 pandemic. A systematic review and a meta-analysis were conducted with the objective of estimating the seroprevalence of SARS-CoV-2 infection in Africa. A systematic search of the PubMed, Scopus, Web of Science and Google Scholar electronic databases was conducted. Thirty-five eligible studies were included. Using meta-analysis of proportions, the overall seroprevalence of anti-SARS-CoV-2 antibodies was calculated as 16% (95% CI 13.1-18.9%). Based on antibody isotypes, 14.6% (95% CI 12.2-17.1%) and 11.5% (95% CI 8.7-14.2%) were seropositive for SARS-CoV-2 IgG and IgM, respectively, while 6.6% (95% CI 4.9-8.3%) were tested positive for both IgM and IgG. Healthcare workers (16.3%) had higher seroprevalence than the general population (11.7%), blood donors (7.5%) and pregnant women (5.7%). The finding of this systematic review and meta-analysis (SRMA) may not accurately reflect the true seroprevalence status of SARS-CoV-2 infection in Africa, hence, further seroprevalence studies across Africa are required to assess and monitor the growing COVID-19 burden.

Evaluation of the current bacteriological profile and antibiotic sensitivity pattern in chronic suppurative otitis media.

OBJECTIVE: To determine the current microbiological profile of chronic suppurative otitis media (CSOM), their antimicrobial sensitivity, their resistance pattern to locally available antibiotics and the appropriate antibiotic against isolated microorganisms causing CSOM. METHODS: This cross-sectional study involved 91 ear swab specimens obtained from patients clinically diagnosed with active CSOM. Swabs were cultured for microbial identification according to a standard protocol. We performed antibiotic susceptibility testing, using the modified Kirby-Bauer disc diffusion method, and the diameter of the inhibition zone was interpreted based Clinical Laboratory Standards Institute guidelines. RESULTS: Microbial growth was seen in 85 (93.4%) samples, but 6 (6.6%) samples had no growth. Among the samples with growth, 63 (69.2%) were monomicrobial, 13 (14.3%) were polymicrobial, and 9 (9.9%) were of mixed growth with more than three microorganisms. The most common bacteria isolated was Pseudomonas aeruginosa (32.6%) followed by Staphylococcus aureus (16.9%) and Klebsiella spp. (5.6%). The most sensitive antibiotics against P aeruginosa were ceftazidime, meropenem, piperacillin-tazobactam, and cefepime. S aureus showed the highest sensitivity toward rifampin, cefoxitin, and fusidic acid. CONCLUSIONS: The bacteriological profile of CSOM showed a high prevalence of P aeruginosa, followed by S aureus and Klebsiella spp. with different distributions in different age groups. We observed a declining pattern of their antibiotic sensitivity. It is important to be aware of the current trend of the bacteriological profiles and to revise the antibiotic regime according to both the sensitivity and age groups.Level of Evidence: NA.

Design of InnoPrimers-Duplex Real-Time PCR for Detection and Treatment Response Prediction of EBV-Associated Nasopharyngeal Carcinoma Circulating Genetic Biomarker.

Nasopharyngeal carcinoma (NPC) is an epithelial tumor with high prevalence in southern China and Southeast Asia. NPC is well associated with the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) 30 bp deletion by having its vital role in increased tumorigenicity and decreased immune recognition of EBV-related tumors. This study developed an InnoPrimers-duplex qPCR for detection of NPC blood circulating LMP1 30 bp deletion genetic biomarker for early diagnosis and treatment response prediction of NPC patients. The analytical and diagnostic evaluation and treatment response prediction were conducted using NPC patients' whole blood (WB) and tissue samples and non-NPC cancer patients and healthy individuals' WB samples. The assay was able to detect as low as 20 ag DNA per reaction (equivalent to 173 copies) with high specificity against broad reference microorganisms and archive NPC biopsy tissue and FNA samples. The diagnostic sensitivity and specificity were 83.3% and 100%, respectively. The 30 bp deletion genetic biomarker was found to be a good prognostic biomarker associated with overall clinical outcome of NPC WHO type III patients. This sensitive and specific assay can help clinicians in early diagnosis and treatment response prediction of NPC patients, which will enhance treatment outcome and lead to better life-saving.

Prevalence and association of Panton-Valentine Leukocidin gene with the risk of sepsis in patients infected with Methicillin Resistant Staphylococcus aureus.

BACKGROUND: Panton-Valentine Leukocidin (PVL), is one of the virulence gene expressed by Methicillin Resistant Staphylococcus aureus (MRSA) and is known to be associated with severe form of community acquired MRSA infection. The aim of this study is to investigate its prevalence in our setting and patient's clinical outcome. METHODS: A cross sectional study involve retrospective record review were done involving 90 MRSA positive isolates between November 2016 and October 2017. Multiplex PCR was performed to detect femA, mecA and PVL genes. Clinical presentation and outcomes of patients were reviewed and presented as descriptive analysis. RESULTS: All of the 90 MRSA isolates included in this study were positive for femA and mecA genes following PCR. PVL gene was detected in 20% (n = 18) of the isolates of which 61.1% (n = 11) were community acquired infections and 38.8% (n = 7) were hospital acquired. Case distribution from community acquired infections include patients with skin and soft tissue infections (33.3%, n = 6), infected diabetic foot ulcers (16.7%, n = 3), and one patient each (5.5%, n = 1) for community acquired pneumonia and meningitis. Half of the PVL positive MRSA cases (50%, n = 9) were having sepsis and four of them succumbed to death due to severe infection. CONCLUSION: This study shows a high prevalence of PVL positive MRSA infection in our population. Skin and soft tissue infections accounting for the major sources. In addition, the presence of the PVL gene is associated with increased risk for developing sepsis.

The Significant Association between Polymicrobial Diabetic Foot Infection and Its Severity and Outcomes.

BACKGROUND: Foot infection is a major complication of diabetes mellitus (DM) and its agents are usually polymicrobial. This study aims to describe the agent and determine the association between polymicrobial infections and the severity of diabetic foot infections (DFI) and their outcomes. METHODS: This retrospective cohort study was conducted during one year and it involved 104 patients. Their records were reviewed and assessed. The causative agents and its sensitivity pattern were noted. The results were presented as descriptive statistic and analysed. RESULTS: A total of 133 microorganisms were isolated with 1.28 microorganisms per lesion. The microorganism isolated were 62% (n = 83) GN (Gram-negative) and 38% (n = 50) GP (Gram-positive). GN microorganisms include Pseudomonas spp (28%), Proteus spp (11%), Klebsiella spp (8%) and E. coli (4%). Staphylococcus aureus (54%) was predominant among GP, followed by Group B Streptococci (26%) and Enterococcus spp (6%). Thirty patients (28.8%) had polymicrobial infections. The association between the quantity of microorganisms and severity of DFI was significant. Among severe DFI cases, 77.8% with polymicrobial microorganisms underwent amputation compared to 33.3% with monomicrobial infection. CONCLUSION: GN microorganisms were predominantly isolated from DFIs and remained sensitive to widely used agents. Polymicrobial infections were associated with DFI severity.

High Prevalence of Toxigenic and Nontoxigenic Clostridium difficile Strains in Malaysia.

Accumulating evidence shows a high prevalence of Clostridium difficile in Southeast Asia associated with a range of clinical presentations. However, severe infections are rarely reported. We investigated C. difficile infection (CDI) across four hospitals in Kuala Lumpur and Kota Bharu, Malaysia. Enzyme immunoassays for glutamate dehydrogenase (GDH) and toxin A or B were performed on diarrheal stool specimens collected from patients in 2015 and 2016. Specimens were also cultured and isolates of C. difficile characterized by PCR ribotyping and detection of toxin genes. In total, 437 specimens were collected and fecal toxin was detected in 3.0%. A further 16.2% of specimens were GDH positive and toxin negative. After culture, toxigenic strains were isolated from 10.3% and nontoxigenic strains from 12.4% of specimens. The most prevalent PCR ribotypes (RTs) were RT 017 (20.0%) and RT 043 (10.0%). The high prevalence of RT 017 and nontoxigenic strains in Malaysia and in neighboring Thailand and Indonesia suggests that they localize to the region of Southeast Asia, with an implication that they may mediate the burden of CDI in the region.

Gut microbiota in early life and its influence on health and disease: A position paper by the Malaysian Working Group on Gastrointestinal Health.

The role of gut microbiota in early life and its impact on gut health and subsequent diseases remain unclear. There is a lack of research and awareness in this area, especially in the Asia-Pacific region, including Malaysia. This paper reports the position of a Malaysian Working Group on some key issues surrounding gut microbiota in early life and its role in gut health and diseases, as well as experts' stand on probiotics and prebiotics. The group reached a consensus that certain factors, including elective caesarean; premature deliveries; complementary feeding; use of antibiotics, prebiotics and/or probiotics; and exposure to the external environmental, have an impact on gut microbiota in early life. However, as evidence is lacking, especially from the Asia-Pacific region, further studies are needed to understand how gut microbiota in early life affects subsequent diseases, including allergy, inflammatory bowel disease, obesity and infantile colic. Lastly, although beneficial in acute diarrhoeal disease and probably allergic eczema, probiotics (and/or prebiotics) should be used cautiously in other gut dysbiotic conditions until more data are available.

A Combination of Trimethoprim-sulfamethoxazole and Ceftazidime Showed Good In Vitro Activity against Stenotrophomonas maltophilia.

BACKGROUND: Stenotrophomonas maltophilia has emerged as an important nosocomial pathogen, capable of causing a wide spectrum of infections. Treatment is difficult because it is resistant to many antimicrobial agents, thus reducing the treatment options. The aims of this study were to describe the antimicrobial susceptibility patterns and synergistic effect of selected antimicrobial combinations against S. maltophilia isolates. METHODS: This was a descriptive cross-sectional study undertaken in the Hospital Universiti Sains Malaysia from April 2011 to March 2012. S. maltophilia isolated from various clinical specimens were included in the study. Antimicrobial susceptibility testing was done using the epsilometer test (E-test) and interpreted according to the guidelines of the Clinical and Laboratory Standards Institute. In the synergy test, the isolates were tested against six different antimicrobial combinations. RESULTS: In total, 84 S. maltophilia isolates were collected and analysed. According to the E-test, the antimicrobial susceptibility of trimethoprim-sulfamethoxazole (TMP-SMX), tigecycline, and ciprofloxacin was 100%, 91.1%, and 88.9% respectively. The antimicrobial combination of TMP-SMX and ceftazidime showed the highest synergistic effect. CONCLUSION: TMP-SMX remains the antimicrobial of choice to treat S. maltophilia infection. TMP-SMX and ceftazidime was the most effective combination in vitro.

The significance of coagulase-negative staphylococci bacteremia in a low resource setting.

INTRODUCTION: Coagulase-negative staphylococci (CoNS) are a group of micro-organisms that are increasingly implicated as a cause of significant infection and the leading cause of bloodstream infection (BSI). One important predictor of true BSI is the isolation of CoNS from multiple blood cultures, presuming that the isolates represent the same species. Thus the objective of this study was to determine the significance of repeated CoNS isolated from blood cultures. METHODOLOGY: This was a prospective laboratory study which was initiated in June 2007 and lasted until July 2008. CoNS isolates were obtained from patients who had two positive blood cultures within a 14-day interval. CoNS were identified to the species level using an API-Staph, and antibiotics susceptibility testing was performed according to Clinical and Laboratory Standards Institute specifications. Strain relatedness was confirmed using pulsed-field gel electrophoresis. RESULTS: During the study period, 202 CoNS-positive samples were isolated from 101 patients. The most common species isolated was Staphylococcus epidermidis (59.0%), and 83.2% of the patients isolated the same species of CoNS from repeated blood cultures. Among the isolates of the same species, only 40.7% had the same antibiogram. CoNS with the same species and antibiogram had 93.3% probability of belonging to the same strain. Most (65.5%) of the patients were treated with antibiotics, primarily from the glycopeptides group. CONCLUSION: Speciation and antibiogram of CoNS from repeated blood cultures are adequate in determining the significance of repeated CoNS isolated from blood cultures.

Prevalence of Clostridium difficile toxin in diarhoeal stool samples of patients from a tertiary hospital in North Eastern Penisular Malaysia.

This study describes the prevalence of Clostridium difficile toxin (CDT) in loose stool samples from inpatients aged more than two years of a tertiary hospital. A total of 175 samples that had been examined were from stool samples that were sent to the Medical Microbiology & Parasitology Laboratory for various clinical indications. The toxin was detected by a commercial immunochromatograhic test, and the patients' demography, clinical features, treatment and outcomes were analyzed from their medical records. Clostridium difficile toxin was positive in 24 (13.7%) of the stool samples. Male and female were 11 (45.8%) and 13 (54.2%) respectively, with the majority of them aged more than 50 years. Most were from medical wards (n = 21, 87.5%), with the rest from surgical wards (n = 2, 8.3%) and intensive care units (n = 1, 3.4%). All the CDT positive patients had history of prior antibiotic usage within 6 weeks before the detection of the toxin. The mean duration of antibiotics usage was 17.75 (+/- 13.75) days, while the mean duration of diarrhea was 5.21((+/- 5.85) days. Eighteen patients had underlying medical illnesses that were diabetes mellitus, chronic renal disease, hypertension, ischaemic heart disease, cerebrovascular disease and malignancy; with seven of them being CDT positive while on chemotherapy. Stool occult blood test was positive in 15 patients whereas presence of pus cells in the CD positive stool samples were detected in 21 patients. The duration of hospitalization among the patients was 27.96 (+/- 23.22) days.

The antibacterial properties of Malaysian tualang honey against wound and enteric microorganisms in comparison to manuka honey.

BACKGROUND: Antibiotic resistance of bacteria is on the rise, thus the discovery of alternative therapeutic agents is urgently needed. Honey possesses therapeutic potential, including wound healing properties and antimicrobial activity. Although the antimicrobial activity of honey has been effectively established against an extensive spectrum of microorganisms, it differs depending on the type of honey. To date, no extensive studies of the antibacterial properties of tualang (Koompassia excelsa) honey on wound and enteric microorganisms have been conducted. The objectives of this study were to conduct such studies and to compare the antibacterial activity of tualang honey with that of manuka honey. METHODS: Using a broth dilution method, the antibacterial activity of tualang honey against 13 wound and enteric microorganisms was determined; manuka honey was used as the control. Different concentrations of honey [6.25-25% (w/v)] were tested against each type of microorganism. Briefly, two-fold dilutions of honey solutions were tested to determine the minimum inhibitory concentration (MIC) against each type of microorganism, followed by more assays within a narrower dilution range to obtain more precise MIC values. MICs were determined by both visual inspection and spectrophotometric assay at 620 nm. Minimum bactericidal concentration (MBC) also was determined by culturing on blood agar plates. RESULTS: By visual inspection, the MICs of tualang honey ranged from 8.75% to 25% compared to manuka honey (8.75-20%). Spectrophotometric readings of at least 95% inhibition yielded MIC values ranging between 10% and 25% for both types of honey. The lowest MBC for tualang honey was 20%, whereas that for manuka honey was 11.25% for the microorganisms tested. The lowest MIC value (8.75%) for both types of honey was against Stenotrophomonas maltophilia. Tualang honey had a lower MIC (11.25%) against Acinetobacter baumannii compared to manuka honey (12.5%). CONCLUSION: Tualang honey exhibited variable activities against different microorganisms, but they were within the same range as those for manuka honey. This result suggests that tualang honey could potentially be used as an alternative therapeutic agent against certain microorganisms, particularly A. baumannii and S. maltophilia.