Antimicrobial Susceptibility Patterns of Bacterial Isolates from Blood Culture of Pediatric Patients with Suspected Sepsis at a Tertiary Care Hospital in Mymensingh, Bangladesh.

Sepsis is a serious, life-threatening condition, occurring when an infectious agent invades the body, resulting in systemic inflammatory response syndrome (SIRS). Neonates and children are among the most vulnerable population groups of developing sepsis because of their weak immune barrier. Despite major advances in prevention, diagnosis and treatment of bacterial infections, invasive infections followed by sepsis remain one of the leading causes of childhood mortality. The aim of this study was to identify bacterial agents and antimicrobial resistance patterns of aerobic bacteria among children suspected of having sepsis. This cross-sectional descriptive type of observational study was conducted in the Department of Microbiology, Mymensingh Medical College, Bangladesh from March 2021 to February 2022. Blood samples were collected from pediatric patients, suspected of having sepsis referred from inpatient facility of department of Neonatology and Pediatrics, Mymensingh Medical College Hospital (MMCH). Blood samples were inoculated into BacT/ALERT PF Plus bottles followed by sub-culture of positive samples in blood agar, MacConkey agar and chocolate agar plates. Isolated bacteria were identified by routine biochemical tests. Antimicrobial resistance pattern of all isolated bacteria was seen by disk diffusion method. MIC of vancomycin by agar dilution method was determined for isolated S. aureus and Coagulase negative Staphylococci (CoNS). The prevalence of pediatric sepsis was 31.82% with highest isolation rate 35.55% among neonates. The isolation rate of gram-positive bacteria was 62.50% where S. aureus was the most common isolate 32.15% followed by CoNS 30.36%. Out of 21 gram-negative bacteria, Pseudomonas spp. was the most frequent isolate 7(33.33%), all of which were resistant to cefuroxime, ceftriaxone and ceftazidime along with all klebsiella and Acinetobacter isolates. Out of 18 S. aureus isolates, 94.44%, 88.89% and 66.67% were resistant to Azithromycin, Penicillin-G and Ciprofloxacin respectively. The MIC of Vancomycin by agar dilution method was observed <2µg/ml against all isolated S. aureus and CoNS. All the Gram-positive isolates were sensitive to Linezolid and Vancomycin. Early detection of bacteria followed by antimicrobial susceptibility test can help by selection of appropriate antibiotic and prevent spread of infection.

Detection of Oncogenic Human Papillomavirus (HPV-16 and HPV-18) from Bacterial Vaginosis Positive Patient Attending at Tertiary Care Hospital in Mymensingh.

Cervical cancer is the fourth most common cancer in women in the world and is the second leading malignancy among Bangladeshi women. Persistent infection with high risk human papillomavirus (HPV) is an important cause of development of cervical intraepithelial neoplasia (CIN) followed by cancer. Bacterial vaginosis (BV), a common treatable vaginal infection which can disrupt the balanced vaginal ecosystem and its innate protective mechanisms against infection, can play an essential role in the acquisition and persistence of high risk human papillomavirus (HR-HPV) infection. This cross sectional study was conducted to detect the HR-HPV (HPV-16 and HPV-18) infection among bacterial vaginosis positive patient in the Department of Microbiology, Mymensingh Medical College (MMC), Bangladesh, from March 2018 to February 2019. A total of 300 endocervical swabs and high vaginal swabs were collected from the VIA (Visual inspection with acetic acid) outdoor clinic of Obstetrics and Gynaecology Department of Mymensingh Medical college Hospital. HPV DNA was tested among all 300 cases by nested PCR. Typing of HPV 16 and HPV 18 was done among HPV DNA positive cases with BV and intermediate flora by multiplex PCR. BV was diagnosed according to Nugent criteria by using the gram stained smear of high vaginal swab. A total of 57/300 (19.0%) samples were positive for HPV DNA by nested PCR. Of the total 300 cases 78(26.0%) had BV, 38(13.0%) had intermediate flora and 184(61.0%) had normal vaginal flora. HPV DNA was more positive in patients having intermediate flora 08/38 (21.05%) followed by the patients having normal vaginal flora 37/184 (20.11%) and BV 12/78 (15.38%). Among the 12 BV patients who were also HPV DNA positive (83.33%) were belong to high risk HPV (type 16 and 18) group and among them 08(66.67%) were HPV-16 and 02(16.67%) were HPV-18. But among 08 HPV DNA positive intermediate flora containing patients only 01(12.5%) were belong to HR-HPV (type 16 and no type 18 was detected).

Ear Infections by Non albicans Candida Species with Isolation of Rare Drug Resistant Species in a Tertiary Care Hospital of Bangladesh.

Otomycosis, a fungal infection of external ear, is challenging for both patients and otolaryngologist as it requires long term treatment and follow up. Candida spp. is second common organism causing otomycosis with Aspergillus being first. Among Candida species, C. albicans is considered as most common but in recent years there is increasing incidence of Non albicans Candida (NAC) species with greater resistance and recurrence. This descriptive type of observational study was planned to determine the species distribution and antifungal susceptibility of Candida spp. causing otomycosis. From March 2021 to February 2022, 60 patients clinically suspected of Candida associated otomycosis at Mymensingh Medical College Hospital, Bangladesh were enrolled. Specimens were taken by an otorhinolaryngologist. After culture and microscopic examination, isolated Candida species were identified by phenotypic and genotypic method and antifungal susceptibility was determined at Department of Microbiology, Mymensingh Medical College. From 60 samples 18(30.0%) were positive for Candida on microscopy and culture. Of the isolates, C. albicans were 2(11.11%) and Non albicans Candida (NAC) 16(88.89%). Five different NAC species were identified of which C. parapsilosis was predominant 5(27.77%) followed by C. tropicalis 4(22.22%) and C. famata 3(16.67%). Rare species of C. ciferrii 2(11.11)%, Kodamaea ohmeri 2(11.11%) were isolated. Candida spp. showed highest resistance to Clotrimazole 8(44.0%) followed by Itraconazole 6(33.0%), Nystatin 4(22.0%) and Fluconazole 3(17.0%). C. ciferrii and Kodamaea ohmeri showed resistance to all antifungals except Nystatin. Outcomes from this study showed a different picture of species distribution, with isolation of rare and emerging drug resistant threatening species like C. ciferri and Kodamea ohmeri which necessitates more detailed survey.

Molecular Detection of MBL Encoding Genes in Acinetobacter baumannii strains Isolated from Various Samples at a Tertiary Care Hospital in Mymensingh.

MBL producing Acinetobacter baumannii is a major threat for therapeutic treatment of hospital acquired infections. The aim of this study was to determine the prevalence of metallo-ß-lactamase genes VIM, IMP & SIM genes amongst isolated A. baumannii. This cross sectional study conducted in the department of Microbiology Mymensingh Medical College from March 2019 to February 2020. 49 Acinetobacter spp. were isolated from different clinical samples including endotracheal aspirates, wound swab/pus, urine and blood. Among 380 clinical samples 130 organisms were isolated growth was 34.21%. Out of 130 isolated strains, 49(37.69%) were Acinetobacter spp identified by standard bacteriological method and resistance to different antibiotics was assessed with Kirby- Bauer Disc diffusion method. Among 49 Acinetobacter spp, 39(79.59%) were Acinetobacter baumannii which was identified by molecular method PCR directing OXA-51 like gene. Multiplex PCR was done to determine MBL genes existence VIM, IMP & SIM. Ceftriaxone (79.48%) showing higher resistance and colistin (12.82%) showing lower resistance. All the strains were sensitive to tigecycline. The distribution of MBLs genes such as VIM 20(51.28%), IMP 5(12.82%) and SIM 0 (0%). This study showed that high level of antibiotic resistance and VIM was the most prevalent MBL genes among A. baumannii highlighting the need for indigenous antibiotic usage plan & infection control measures to prevent the spread of these resistance genes.

Concurrent Infection of Orientia Tsutsugamushi with Rickettsia spp. Including Rickettsia felis in North Central Bangladesh.

Rickettsial diseases are one of the leading causes of treatable acute febrile illness in Asia pacific region. This cross-sectional descriptive study was conducted at Department of Microbiology, Mymensingh Medical College to diagnose scrub typhus by rapid Immunochromatographic Test (ICT) and Nested PCR followed by molecular identification of possible Rickettsial coinfection among suspected febrile patients in Mymensingh, Bangladesh from March 2019 to February 2020. Among the enrolled 402 patients, 89 samples (22.13%) were seropositive by Immunochromatographic Test (ICT) and 65 samples (16.16%) were positive for O. tsutsugamushi DNA by Nested PCR, targeting 47KDa gene. Therefore, 113/402 (28.10%) samples were positive for scrub typhus by PCR and/or ICT. All the scrub typhus positive samples were further subjected to Nested PCR targeting 17 KDa gene for identification of Rickettsial co-infection and 13/113 (11.50%) were documented as positive. Then 13 Rickettsial co-infected samples were undertaken to automate sequencing and all were genetically confirmed as Rickettsia felis. Findings of the study may help clinicians to expand their list of differential diagnoses for undifferentiated fever and detection of Rickettsial co-infection may guide them to prescribe effective antimicrobials.

Prevalence of qnr and aac(6')-Ib-cr Genes in Clinical Isolates of Proteus spp. at a Tertiary Care Hospital in Dhaka, Bangladesh.

Plasmid mediated quinolone resistance (PMQR) has been revealed to play not only a significant role in quinolone resistance but also this drug resistance can spread from one bacterium to another. There is limited data regarding the prevalence of PMQR are available from Bangladesh. So, the aim of this study was to detect the prevalence of qnr and aac(6')-Ib-cr genes among clinical isolates of ciprofloxacin resistant Proteus spp. This cross-sectional study was carried out in the Department of Microbiology of Dhaka Medical College, Dhaka, Bangladesh from July 2018 to June 2019. Fourty (40) Proteus spp. was isolated from 300 culture positive samples. Proteus mirabilis and Proteus vulgaris were identified by culture and biochemical test. Antibiotic susceptibility was performed by disc-diffusion technique. Quinolone resistance genes (qnrA, qnrB, qnrC, qnrD, qnrS and aac(6')-1b-cr) among ciprofloxacin resistant Proteus spp. were detected by PCR. Thirty (75%) ciprofloxacin resistant isolates were detected during disk-diffusion technique. Among them, quinolone resistance genes were found positive 11(36.67%) for aac(6')-Ib-cr, 6(20%) for qnrA, 5(16.67%) for qnrD, 4(13.33%) for qnrS and 3(10%) for qnrB genes. Co-existance of qnrA + aac(6')-Ib-cr and qnrD + qnrS were found in 3(10%) wound swab & pus and urine samples respectively followed by qnrA + qnrB in 2(6.67%) wound swab and pus and qnrA+qnrS in 1(3.33%) urine sample. The results of this study showed presence of high (66.67%) percentage of PMQR genes as well as high (30%) rate of co-carriage of the two genes among Proteus spp. isolates. The incidence of PMQR genes was found to be high which could be due to the increased prescription of fluoroquinolones. Thus, there is a need for rational usage of fluoroquinolones.

Socio-demographic and Clinico-epidemiological Study of Scrub Typhus in A Tertiary Care Hospital of Mymensingh, Bangladesh.

Scrub typhus is one of the leading causes of undifferentiated treatable febrile illness in Asia pacific region. It is grossly under diagnosed in many tropical countries of South Asia including Bangladesh, due to wide range of non-specific clinical presentations, low index of suspicion among clinicians, limited awareness and lack of accurate diagnostic facilities. This cross sectional observational study was conducted at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2020 enrolling 113 diagnosed cases of scrub typhus by Immunochromatographic test (ICT) and / or Nested PCR to characterize the socio-demographic and clinico-epidemiological features of scrub typhus in Mymensingh area. Majority of the scrub typhus cases came from rural areas (63.83%) and there was a slight female predominance (52.21%). The young (32.74%) and the young-adult age group (28.31%) were mostly affected. Most of the scrub typhus cases were housewives (30.98%), followed by farmers (23.89%) and students (21.23%). All the enrolled cases presented with fever. Other findings were myalgia (76.10%), headache (56.63%), cough (30.97%), vomiting (12.38%) and Respiratory distress (9.73%). Typical eschar of scrub typhus was present only in 9(7.96%) cases and 4(3.53%) patients had rashes on their skin. Few cases (3.53%) had jaundice and 15.96% cases were anaemic. Oliguria (7.96%) and neck rigidity (1.76%) were also documented. Most of the Nested PCR positive scrub typhus cases were documented during late rainy season and beginning of winter months. Findings of the study may offer increased awareness about high burden of scrub typhus as well as heightened suspicion among clinicians for early diagnosis, timely treatment and prevention of complications.

Detection of Quinolone Resistance Pattern and Presence of qnr Genes in Human Salmonella Isolates at Mymensingh, Bangladesh.

Among the quinolones, fluoroquinolones are broad spectrum antimicrobial agents used for treating many clinical infections including Salmonellosis. Although high level of resistance to fluoroquinolones remains low in Salmonella but reduced susceptibility is increasing worldwide. Plasmid-mediated quinolone resistance (PMQR) of qnr type (qnrA, B and S) has been identified now a day in several enterobacterial species including Salmonella spp. This cross-sectional study was held at department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh from March 2019 to February 2020. This study was conducted to determine the current quinolone resistance pattern and to detect the presence of qnrA, qnrB and qnrS genes among Salmonella isolates. Antimicrobial susceptibility test of 36 Salmonella isolates were done by disc diffusion method. MIC of ciprofloxacin was detected by agar dilution method. Then amplification with specific primers of qnrA, qnrB and qnrS genes were performed for all Salmonella isolates. The present study observed 80.5% resistance to nalidixic acid, 33.3% to ciprofloxacin and 19.4% to ofloxacin by disc diffusion method. qnr A gene was detected in 2(5.5%) isolates, where as qnrS was detected in 5 (13.8%) isolates. None of the isolates was positive for qnrB gene. All the qnrA positive isolates showed resistance to Ciprofloxacin (MIC=128µg/ml) and Ofloxacin. In conclusion, presence of qnr genes in the study isolates is alarming, because, rapid dissemination might occur due to conjugative plasmid mediated horizontal transfer.

Detection of Biocide Resistance Genes (qacE and qacΔE1) in Pseudomonas spp Isolated from Patients with CSOM at Mymensingh Medical College Hospital, Bangladesh.

Biocides, including disinfectants and antiseptics, are used for a variety of topical and hard surface applications in health care facilities. Biocides play a significant role for preventing and controlling nosocomial infections. However, failures in the antimicrobial activities of biocides have been reported. The resistance mechanism to disinfectants is usually determined by genes which are related to resistance to quaternary ammonium compounds, namely, qacE, qacΔE1 that are found in Gram-negative bacteria. The aim of this study is to detect the prevalence of Biocides resistance genes, qacE and qacΔE1, in clinical isolates of Pseudomonas spp. It was carried out from March 2017 to July 2018 in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. Samples were collected from Outpatient of ENT department, MMCH. In this study, 300 clinical samples of CSOM cases were tested by the PCR method. The present study shows detection of biocide resistance genes (qacE, qacΔE1) among 87 isolated Pseudomonas spp by uniplex PCR. Among 72 clinical isolates of Pseudomonas aeruginosa 67(93.05%) had the gene qacEΔ1 and 25(34.72%) had the gene qacE. In addition other 15 Pseudomonas spp 3(20%) isolates had the qacEΔ1 gene and 2(13.33%) isolates had the qacE gene. In this study there is a marked difference in detection of the qacEΔ1 gene between the MDR and non MDR P. aeruginosa isolates. The qacEΔ1 was identified in 50 of 54(92.59%) MDR isolates and 7 of 18(38.89%) non MDR strains respectively. While gene qacE was detect 25(46.29%) MDR isolates and did not show any qacEΔ1gene in non MDR isolates. This study shows that the genes, qacE, qacΔE1 are widespread among Pseudomonas aeruginosa, they are higher in MDR strains than non MDR strains.

Rapid Serologic and Molecular Diagnosis of Scrub Typhus among Suspected Febrile Patients Visiting Mymensingh Medical College Hospital.

Scrub typhus, caused by the bacterium- Orientia tsutsugamushi is one of the leading causes of undifferentiated treatable febrile illness in Asia pacific region. It is grossly under diagnosed in many tropical countries of South Asia including Bangladesh, due to wide range of non-specific clinical presentations, low index of suspicion among clinicians, limited awareness and lack of accurate diagnostic facilities. This cross-sectional descriptive study was conducted at Department of Microbiology, Mymensingh Medical College to diagnose scrub typhus by rapid Immunochromatographic test (ICT) as well as molecular detection of O. tsutsugamushi by Nested PCR and automated nucleotide sequencing among suspected febrile patients in Mymensingh, Bangladesh during 2019-20. Blood samples were collected from 402 febrile patients of suspected Rickettsial illness, referred from inpatient and outpatient departments of Medicine and Pediatrics, Mymensingh Medical College Hospital (MMCH). Among the enrolled 402 patients, 89 samples (22.13%) were seropositive by Immunochromatographic test (ICT) and 65 samples (16.16%) were positive for O. tsutsugamushi DNA by Nested PCR, targeting 47KDa gene. Therefore, 113/402 (28.10%) samples were positive for scrub typhus by PCR and/ or ICT. Highest number of patients was detected positive by nested PCR during the first 5-10 days of fever but only 2 cases were positive after 20 days. In case of ICT, highest positivity for only IgM (8.13%) and both antibodies (2.43%) were documented in first 5-10 days of fever, but IgG positivity was highest (41.66) in >20 days of fever. From 65 PCR positive samples, automated nucleotide sequencing was performed on 20 randomly selected samples and all were genetically confirmed to be O. tsutsugamushi.

Early and Rapid Detection of Typhoid Fever by Nested PCR in Blood.

Typhoid fever caused by Salmonella typhi is one of the major health problems in developing countries including Bangladesh. Still now blood culture is gold standard method for diagnosing typhoid fever, but this method is laborious, requires several days and detection rate is low. Failure of early laboratory diagnosis often leads to increased morbidity and mortality. This study was intended to apply a nested PCR in blood for early diagnosis of typhoid fever. In this cross sectional study blood samples were collected from 200 suspected typhoid fever patients attending Mymensingh Medical College Hospital, Bangladesh. Nested Polymerase Chain Reaction (n PCR) of flagellin gene was done in all the blood samples. At the same time all blood samples were subjected to culture by lytic centrifugation method. Culture positive isolates were identified as S. typhi by biochemical tests. Among the 200 blood samples, 57 (28.5%) were positive for S. typhi on nested PCR where as blood culture was positive for S. typhi in 16 (8%) samples. Among the 57 PCR positive samples, only 15 (26.3%) samples were culture positive for S. typhi and rest 42 (73.7%) were culture negative. So, in culture negative cases PCR can be used as a rapid diagnostic test for diagnosing typhoid fever. Considering time requirement, PCR takes one day, whereas blood culture takes 3 or more days to confirm diagnosis.

Prevalence of ESBL Encoding Genes in Acinetobacter baumannii Strains Isolated from Various Samples of a Tertiary Care Hospital in Mymensingh.

The aim of this study was to find the prevalence of ESBL genes among A. baumannii isolates. In this cross sectional study, 49 Acinetobacter spp. were isolated from various clinical samples from March 2019 to February 2020 conducted in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. Clinical samples including endotracheal aspirates, wound swab/pus, urine and blood. A total of 380 samples were analyzed. Growth was obtained in 34.21% of the samples yielding 130 organisms. Out of 130 organisms, 49(37.69%) were Acinetobacter spp. Among 49 Acinetobacter spp, 39(79.59%) were Acinetobacter baumannii which was identified by PCR targeting OXA-51 like gene. Amplification of the ESBL encoding genes, namely CTX-M, TEM, SHV done by molecular technique PCR. The most antibacterial resistance was against ceftriaxone (79.48%) and lower resistance only showed in colistin (12.82%). All the isolates were sensitive to tigecycline. The distribution of ESBLs genes such as TEM 20(51.28%), CTX-M 16(41.02%) and SHV 0(0%). The high resistance to most of the antibiotics among the studied strains and also a high prevalence of TEM gene in A. baumannii strains found in our study gives alarming sign towards the treatment complexity of these strains.

Clinical Features of Covid-19 Infection: a Study of SARS-CoV-2 Positive Patients from Mymensingh Region, in Bangladesh.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spread to almost every country on the globe and it is considered by World Health Organization as a pandemic. SARS-CoV-2 causes the Coronavirus disease 2019 (COVID-19). Many of country are reporting the symptomatic characteristics of their cases to give better observations into the various clinical presentations of SARS-CoV-2 infection. However, the symptomatic literature is limited in Bangladesh. The aim of the study is to analyze the symptomatic characteristics of patients having the SARS-CoV-2 positive by real time reverse transcription polymerase chain reaction (rRT -PCR) test. Here, the data of 146 patients who were positive for the SARS-CoV-2 virus and were residents of different districts of Mymensingh region were analyzed. Patients' demographics, symptoms, history of co-morbidities condition like DM, HTN, Hypothyroid etc, travel and contact were collected from MMC Daily Reported data from April 1, 2020 to April 30, 2020. Among the total 3184 patients' nasopharyngeal samples, we have got 146 (4.58%) positive for SARS-CoV-2. Of the 146 positive patients most of the patients were male 95(65%), the majority 80(54.8%) were the 21 to 40year age group. Most of the patients 61(41.78%) were residents of Mymensingh include Mymensingh Sadar, Valuka, Trishal and Ishhorganj. Among the patients 94(64.4%) were symptomatic and 52(35.6%) were asymptomatic. The symptomatic patients presented mostly were with fever 45(30.82%), cough 33(22.6%) and breathlessness 9(6.16%). The majority of patients 54(36.9%) had a history of contact with SARS-CoV-2 patients and 16(11%) had a travel history within 14 days of their rRT-PCR test positive. The only 3(2%) patients had history of comorbidities condition like DM, HTN, Hypothyroid etc. The number of SARS-CoV-2 cases is rapidly increasing in our country. The education of the population about the most common symptoms of the virus infection is needed mostly; therefore, individuals may able to recognize these symptoms. So, that people might get themselves tested.

Detection of Antimicrobial Resistance Pattern and ESBL Production among Clinical Isolates of Salmonella Species in Mymensingh, Bangladesh.

The occurrence of antimicrobial resistance in Salmonella enterica serovars (both typhoidal and non-typhoidal Salmonellae) is a major public health problem especially in developing countries, which have been associated with treatment failures. Therefore, the study was undertaken to determine the current antimicrobial resistance pattern and extended spectrum ß-lactamase (ESBL) production among clinical isolates of Salmonella spp. during 2019-2020 in Mymensingh, Bangladesh. In this cross sectional study, 36 Salmonella enterica isolates were obtained from blood and stool culture of suspected 200 enteric fever and 100 gastroenteritis patients attending at Mymensingh Medical College Hospital, Mymensingh, Bangladesh. Isolated Salmonella species were identified by biochemical tests and Polymerase Chain Reaction (PCR). Disk diffusion test was performed by modified Kirby Bauer method. Minimum Inhibitory Concentration (MIC) of ceftriaxone was detected by agar dilution method. Double disk synergy test was used as a screening test for ESBL production. PCR was done for detection of blaTEM, blaSHV and blaCTX-MU genes. The isolates showed 25% resistance to Ceftriaxone and 58.3% to Azithromycin. The highest sensitivity rates were 88.9% to Meropenem and 83.3% to Amikacin. Whereas 6(16.7%) isolates were Multi Drug Resistant (MDR). Eight (8) isolates were confirmed as ESBL producer by DDST. The marked increase in MIC was observed between 8->512µg/ml to ceftriaxone. blaTEM, blaSHV and blaCTX-MU genes were detected in 3, 5 and 8 isolates respectively. In conclusion, the current study observed, higher level of resistance to ceftriaxone and azithromycin. At the same times 22.2% isolates showed ESBL production, which is a cause for concern as it may lead to treatment failure. On the other hand the study also showed the re-emergence of chloramphenicol and Sulfamethoxazole-Trimethoprim sensitivity.

Predominance of Leptospira wolffii in north-central Bangladesh, 2019.

Leptospira was detected in 48.9% of blood samples from 182 febrile patients in north-central Bangladesh in 2019. Most Leptospira were classified as L. wolffii (93%) on the basis of phylogenetic analysis of 16S ribosomal RNA genes, while others were assigned to L. borgpetersenii and L. meyeri.