Pulmonary Paragonimiasis: A Case Series.

Paragonimiasis contributes to significant foodborne zoonosis worldwide. The major mode of transmission in humans is by consumption of uncooked or undercooked crabs and crayfish harbouring Paragonimus metacercariae. It begins with symptoms like fever and lower respiratory involvement from a few months to a year, mimicking those of tuberculosis and leading to diagnostic delay. Here, we report two cases of paragonimiasis during a period of nine months. Both cases presented with symptoms of productive cough with rusty sputum, chest pain, along with eosinophilia, and pleural effusion and had a history of consumption of smoked crab from the local river. The diagnosis was established by microscopic demonstration of Paragonimus ova in the sputum. They were treated with praziquantel and recovered. Indeed, it is challenging to diagnose paragonimiasis due to the lack of its specific symptoms but should be considered in the differential diagnosis of eosinophilia and pleural effusion in such lung diseases. Keywords: case reports; eosinophilia; paragonimiasis; pleural effusion.

Metallo-ß Lactamase Producing Non-Fermentative Gram-Negative Bacilli from Various Clinical Isolates in a Tertiary Care Hospital: A Descriptive Cross-sectional Study.

INTRODUCTION: Non-fermentative gram-negative bacilli are common causes of human infections especially nosocomial infections. These organisms are usually resistant to multiple antimicrobial agents including carbapenems. The study aimed to find out the prevalence of metallo-ß-lactamase producing non-fermentative gram-negative bacilli among the samples which yielded growth of bacteria in a tertiary care hospital. METHODS: This is a descriptive cross-sectional study conducted in a tertiary care hospital from February 2017 to May 2017. Convenience sampling method was used. Bacterial identification, characterization and antimicrobial susceptibility testing were done by following standard microbiological guidelines. Metallo-ß-lactamase production was detected by using combined disk diffusion test and double-disc synergy test. Data were analyzed by using Statistical Package of Social Science software version 16. Point estimate at 95% confidence interval was calculated along with frequency and proportion for binary data. RESULTS: Among 628 samples which yielded growth of bacteria, 118 (18.79%) at 95% Confidence Interval (15.74-21.84) were metallo-ß-lactamase producing non-fermentative gram-negative bacilli. Among them, 54 (45.76%) were Pseudomonas aeruginosa and 64 (54.24%) were Acinetobacter baumannii. CONCLUSIONS: A high prevalence of metallo-ß-lactamase production was observed among the nonfermentative gram-negative bacilli than the study done in similar settings. It is mandatory to perform routine monitoring of metallo-ß-lactamase producing isolates in clinical laboratories in order to help the clinicians prescribe proper antibiotics.

Urinary Calculi: A Microbiological and Biochemical Analysis at a Tertiary Care Hospital in Eastern Nepal.

BACKGROUND: The occurrence of urinary tract infection in presence of urolithiasis is frequently noted; however, microbial agents of urolithiasis and their antimicrobial susceptibility patterns remain underinvestigated. This study aimed to identify the microorganisms isolated from urine and stone matrices to determine their antimicrobial susceptibility, to find the association between the pathogens of urine and stone matrices, and to perform the biochemical analysis of stones. METHODS: A total of 88 cases of urolithiasis admitted for elective stone removal at Department of surgery, B.P. Koirala Institute of Health Sciences (BPKIHS), were enrolled. Preoperative urine culture and postoperative stone culture were performed. Isolation, identification, and AST were done by the standard microbiological technique. Further qualitative biochemical analysis of stones was also attempted. RESULT: Among 88 stone formers recruited, culture of urine, whole stone, and nidus yielded the growth of bacteria 44, 32, and 30, respectively. Bacteria isolated from urine culture correlated with those from stone matrices with a sensitivity of 90%, specificity of 79.69%, PPV of 63.64%, and NPV of 95.45%. Escherichia coli (46.7%) was the most common bacteria followed by Klebsiella pneumoniae (16.7%) and Proteus mirabilis (13.3%) from urine and stone cultures. Almost all the uropathogens isolated were susceptible to commonly used antibiotics. Calcium oxalate (84.1%) was common biochemical constituent found in stone formers followed by calcium oxalate + phosphate (8%). CONCLUSIONS: The association of microorganism isolated from urine and nidus culture was significant that can predict the source of infective stone; however, in some cases, microorganisms and the antimicrobial susceptibility pattern from urine and nidus were different. This study emphasizes the use of appropriate antimicrobial agents to prevent the regrowth of residual stones and minimize the risk of infectious complications after surgical removal of stones.

Virulence factors of uropathogenic Escherichia coli (UPEC) and correlation with antimicrobial resistance.

BACKGROUND: Escherichia coli has found to be the predominant uropathogen (50-90%) in uncomplicated, community acquired urinary tract infection (UTI). Uropathogenic Escherichia coli (UPEC) express a multitude of virulence factors, which enable the bacteria to establish UTI. The objective of this study was to evaluate the presence of different phenotypic virulence markers in UPEC isolates and determine their correlation with antibiotic resistance pattern. RESULTS: Out of 105 patients, 56 (53%) were females and 49 (47%) were males. The age of the patients in the study ranged from 18 years to 87 years and majority of the patients belonged to the age group 20-29 years. Virulence factor was observed in 65% (n = 69) of UPEC and 20% (n = 22) of control isolates (P = 0.0001). Haemolysin production was observed in 34(32.3%) of uroisolates and 12 (11.4%) of control strain. Similarly, 62% of UPEC and 1% of control produced biofilm (P = 0.0001). The expression of Mannose-resistant hemagglutinin (MRHA) and mannose-sensitive hemagglutinin (MSHA) in uroisolates were 52.3% (n = 55) and 5.7% (n = 6) respectively, whereas in faecal isolates, 8.5% (n = 9) expressed MRHA and none produced MSHA. Antimicrobial resistance showed a high degree of resistance towards ampicillin, cotrimoxazole and norfloxacin. The resistance was observed in significant higher degree in biofilm formers as compared to non-formers. MDR and ESBL was observed in 51 and 46% of test strains and 9 and 7.6% of control strains (P = 0.0001). CONCLUSION: A significant association between virulence factors of UPEC and antimicrobial resistance in UPEC was present. Routine testing of these factors and co-relation with AMR is recommended. These findings will certainly help understand the pathogenicity and proper management of UTI patients, thus decreasing the improper use of antibiotics.

Characterization of Clinical Isolates of Enterococci with Special Reference to Glycopeptide Susceptibility at a Tertiary Care Center of Eastern Nepal.

BACKGROUND: Enterococci, once considered as a harmless commensal of intestine, have now emerged as medically important pathogens and are associated with both community-acquired and nosocomial infections. They bear the potential to exhibit resistance against all commonly used antibiotics either by inherent or acquired mechanism, posing a therapeutic challenge. OBJECTIVES: This study aimed to characterize enterococci up to the species level and study their antibiogram with special regard to vancomycin. METHODS: A descriptive cross-sectional study was conducted in the Department of Microbiology, B.P. Koirala Institute of Health Sciences, Dharan, Nepal, from February to May 2017. A total of 91 enterococcal isolates recovered from clinical specimens were investigated in this study. Their identification and speciation were done according to standard microbiological guidelines. Kirby-Bauer disc diffusion technique was used to study antimicrobial susceptibility pattern, whereas minimum inhibitory concentration of vancomycin was determined by the agar dilution method, with reference to Clinical and Laboratory Standards Institute guidelines. RESULTS: Seven different species of enterococci were isolated, E. faecalis and E. faecium accounting about 45% each. The other species encountered were E. avium, E. cecorum, E. dispar, E. durans, and E. CONCLUSIONS: Enterococcus faecalis and E. faecium were the predominant species in causing enterococcal infections. The alarming rise in prevalence of vancomycin and multidrug resistance strains warrants immediate, adequate, and efficient surveillance program to prevent and control its spread.

Comparative study of antimicrobial resistance and biofilm formation among Gram-positive uropathogens isolated from community-acquired urinary tract infections and catheter-associated urinary tract infections.

Background: Gram-positive cocci have emerged to be an important cause of urinary tract infection (UTI) both in community-acquired UTI (Com-UTI) and catheter-associated urinary tract infection (CA-UTI). The objective of this study was to investigate the frequency of Gram-positive cocci urinary tract infections, their susceptibility patterns to commonly used antimicrobial agents and the biofilm forming property with respect to catheter-associated UTI and community-acquired UTI. Methods: A total of 1,360 urine samples from indwelling catheter and 10,423 from mid-stream urine were obtained during a 6-month period and processed following standard microbiological guidelines. Biofilm formation was detected using congo red agar (CRA), tube method (TM) and tissue culture plate (TCP) method. Chi-square test and independent sample t-test were employed to calculate the significance. Statistical significance was set at P-value ≤0.05. Results: The infection rate was significantly higher in CA-UTI as compared to Com-UTI (25% vs 18%, p=0.0001). Among 2,216 organisms isolated, 471 were Gram-positive cocci; 401 were obtained from Com-UTI while 70 were from CA-UTI. Enterococcus faecalis was the most common organism isolated from Com-UTI, while Staphylococcus aureus was commonest among CA-UTI. Multi-drug resistance, methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococci were also significantly higher in CA-UTI as compared to Com-UTI. Biofilm-forming property was significantly higher in CA-UTI than Com-UTI. The sensitivity of congo red agar method and tube method was 79% and 81.9% respectively and specificity was 98.5% each. Antimicrobial resistance was significantly higher in biofilm-formers as compared to non-formers. Conclusion: Gram-positive bacteria are a significant cause of both CA-UTI and Com-UTI with Enterococcus faecalis and Staphylococcus aureus as common pathogen. Biofilm formation and multi-drug resistance is significantly higher in CA-UTI than Com-UTI. Routine surveillance of antimicrobial resistance and biofilm formation is necessary in all cases of UTI to ensure the proper management of patients.

Clinical, etiological and antimicrobial susceptibility profile of pediatric urinary tract infections in a tertiary care hospital of Nepal.

BACKGROUND: Urinary tract infection (UTI) is one of most common pediatric infections. The study was designed to assess the clinical profile, common bacterial microorganisms causing UTI and their antimicrobial susceptibility patterns at B. P. Koirala Institute of Health Sciences (BPKIHS) hospital. METHODS: This is a prospective cross-sectional study conducted at Department of Microbiology and Infectious Diseases for 6 months (January to June 2018). A total of 1962 non-repetitive urine specimens (midstream, nappy pad, catheter aspirated) of pediatric patients (0-14 years age) suspected of UTI were obtained in the Microbiology laboratory. Clinical data was obtained from requisition form and hospital software. Culture and bacterial identification was done by using standard microbiological guidelines. Antimicrobial susceptibility testing was done by Kirby-Bauer disc diffusion method following clinical and laboratory standards institute (CLSI) guidelines. Resistance to methicillin and vancomycin were confirmed by calculating minimum inhibitory concentration using broth dilution method. RESULTS: Among 1962 samples, 314 (16%) were positive for bacterial infection. Fever, irritability and poor feeding was the most common symptoms in neonates while older children presented with fever and urinary symptoms. E. coli was reported the most common etiological agent (53%), followed by Enterococcus faecalis (22%), Klebsiella pneumoniae (7%) and Staphylococcus aureus (7%). Multidrug resistant (MDR) isolates accounted for 32% of isolates, while 5% were extensively drug resistant (XDR). Fourty percentage of gram-negative bacilli were ESBL producer, 38% of S. aureus were methicillin resistant Staphylococcus aureus (MRSA) and 5% E. faecalis were vacomycin resistant enterococci (VRE). E coli was highly resistant to Ampicillin (87%), Ceftriaxone (62%) and Ofloxacin (62%). Amikacin (11% resistance) and Nitrofurantoin (5% resistance) are the most effective drugs for gram-negative bacilli (GNB) while vancomycin and linezolid are functional against gram-positive cocci. CONCLUSIONS: High-level antimicrobial resistance was observed in pediatric UTI with alarming incidence superbugs like MDR, XDR, ESBL and MRSA. Regular surveillance should be carried out to determine the local prevalence of organisms and antimicrobial susceptibilities in order to guide the proper management of children.

Study of biofilm formation and antibiotic resistance pattern of gram-negative Bacilli among the clinical isolates at BPKIHS, Dharan.

OBJECTIVES: Gram-negative bacilli are the common causative agents for community-acquired, nosocomial and opportunistic infections. The recent upsurge of biofilm, as well as beta-lactamases producing strains, have synergistically led to the extensive dissemination of multi-drug resistant gram-negative bacilli. This study was carried out with an intention to detect the biofilm formation by gram-negative bacilli and determine their antibiogram along with the detection of extended-spectrum beta-lactamases (ESBLs) and metallo-beta-lactamases (MBLs) production. RESULTS: Among 314 isolates, Escherichia coli (38%) were the predominant isolates followed by Acinetobacter spp. (20%), Klebsiella spp. (16%), and Pseudomonas spp. (12%). Overall, 197 (62.73%) of isolates were biofilm positive. 84 (26.75%) and 51 (16.24%) were confirmed as ESBL and MBL producers respectively. The association between MBL production and biofilm formation was statistically significant (χ2 = 10.20, P value= 0.002) whereas it was insignificant between ESBL and biofilm production (χ2 = 0.006, P-value= 0.937). Most of the biofilm and MBL producing strains were multi-drug resistant.

Incidence and outcomes of multidrug-resistant gram-negative bacteria infections in intensive care unit from Nepal- a prospective cohort study.

Background: Infections caused by multi-drug resistant gram-negative bacterial infections are the principle threats to the critically ill patients of intensive care units. Increasing reports of these infections from the Nepalese intensive care unit underline the clinical importance of these pathogens. However, the impact of these infections on the patient's clinical outcome has not yet been clearly evaluated. The objective of our study was to determine the incidence and associated clinical outcome of multi-drug resistant gram-negative bacterial infections in intensive care unit from a tertiary care center of Nepal. Methods: A prospective cohort study was conducted among adult patients admitted in intensive care unit of B. P Koirala Institute of Health Sciences from July to December 2017. Patients infected with multi-drug resistant gram-negative bacteria, non-multi-drug resistant gram-negative bacteria and those without infection were included. Identification of gram-negative bacteria and their antibiotic susceptibility pattern was performed with standard microbiological methods. Demographic, clinical profiles and outcomes (in-hospital-mortality, intensive care unit and hospital length of stay) were documented. Results: The incidence rate of multi-drug resistant gram-negative bacteria infections was 47 per 100 admitted patients (64/137) with 128 episodes. Acinetobacter species (41%, 52/128) was the commonest followed by Klebsiella pneumoniae (28%, 36/128) and Pseudomonas spp (21%, 27/128). Patients with multi-drug resistant gram-negative bacteria in comparison to non-multi-drug resistant gram-negative bacteria had high healthcare-associated infections (95%, 61/64 versus 20%, 2/10; p = < 0.001). In-hospital-mortality was 38% (24/64), 20% (2/10) and 10% (4/41) in multi-drug resistant, non-multi-drug resistant and uninfected group respectively (p = 0.007). After adjustment for independent risk factors, compared to uninfected patients, the odds ratio (CI) for in-hospital-mortality in multi-drug resistant and non-multi-drug resistant group was (4.7[1.4-15.5], p = 0.01) and 2.60 [0.38-17.8], p = 0.32) respectively. Multi-drug resistant patients also had longer intensive care unit and hospital stay, however, it was statistically insignificant. Conclusion: The incidence of multi-drug resistant gram-negative bacterial infections was remarkably high in our intensive care unit and showed a significant association with healthcare-associated infections and in-hospital-mortality.

Stenotrophomonas maltophilia causing blood stream infection in neonates and infants: a cause for concern.

Stenotrophomonas maltophilia ( S. maltophilia) is an important Gram-negative, non-fermentative, multidrug resistant (MDR) nosocomial organism. We evaluated the isolation of S. maltophilia from the seven blood culture specimens received from the Paediatric Emergency Department (PED) of BP Koirala Institute of Health Sciences (BPKIHS) over the duration of two weeks. The suspicion of a possible outbreak was raised and the hospital infection control team investigated the source and found the hand of one healthcare provider harbouring a similar organism. All six steps of hand hygiene were subsequently strictly enforced after which the same bacteria were no longer isolated. Infection control measures should be rigorously adopted for the control of such nosocomial bacteria.

A study of prevalence of intestinal parasites and associated risk factors among the school children of Itahari, Eastern Region of Nepal.

INTRODUCTION: Intestinal parasitic infestation is a major public health problem in children of developing countries Because of poor socio-economic conditions and lack of good hygienic living. The aims of this study were to measure the prevalence of intestinal parasitic infestations and to identify risk factors associated with parasitic infestations among the school children of Itahari Municipality. MATERIALS AND METHODS: The cross-sectional study was conducted in Grade VI, VII and VIII in Government and private schools of Itahari Municipality. Stratified random sampling method was applied to choose the schools and the study subjects. Semi-structured questionnaire was administered to the study subjects and microscopic examination of stool was done. The Chi-square test was used to measure the association of risk factors and parasitic infestation. RESULTS: Overall intestinal parasitic infestation was found to be 31.5%. Around 13% of the study population was found to be infested with helminthes and 18.5% of the study population was protozoa infected. Not using soap after defecation, not wearing sandals, habit of nail biting and thumb sucking were found to be significantly associated with parasitic infection. CONCLUSIONS: The prevalence of intestinal parasitic infestation was found to be high in school children of Itahari. Poor sanitary condition, lack of clean drinking water supply and education is supposed to play an important role in establishing intestinal parasitic infections.

Asymptomatic bacteriuria in pregnant women.

INTRODUCTION: Asymptomatic bacteriuria is the significant presence of bacteria in urine of an individual without symptoms. The aim of the study is to determine the prevalence of asymptomatic bacteriuria in pregnant women. METHODS: This study was a prospective study conducted in the department of Obstetrics and Gynaecology at B. P. Koirala Institute of Health Sciences. The duration of the study was six months from January to June 2012. A total of 600 pregnant women were enrolled. All women were clinically identified to have no signs and symptoms of UTI. Clean catch midstream urine sample was collected from each patient into a sterile vial. The urine samples were examined for microscopic and culture sensitivity test. RESULTS: Out of 600 pregnant women, 52 were positive for significant bacteriuria with a prevalence rate of 8.7%. There was a significant difference in prevalence of asymptomatic bacteriuria with respect to trimester (p=0.005). Age did not show any significant difference in the prevalence of asymptomatic bacteriuria (p=0.807). There was not any significant difference in the prevalence of asymptomatic bacteriuria with respect to parity (p=0.864) and booking status (p=0.397). Escherichia coli (35%), Acinetobacter species (15%), Enterococcus species (12%) and Klebsiella pneumoniae (10%) were the common isolates. Most of the isolates were sensitive either to Nitrofurantoin, Norfloxacin or Amikacin. CONCLUSIONS: Asymptomatic bacteriuria is common in pregnancy. Urine culture sensitivity should be carried out routinely on all pregnant patients in order to prevent the dangerous complications associated with it.