Safeguarding health on every plate: Intestinal parasitic infection survey of food handlers in a tertiary care centre.

Food borne infections pose significant public health problem, especially in developing countries of the world. A continuous surveillance to ensure the health of the personnel involved in preparation of the hospital food is important as they can be a source of spreading the infections and possible outbreaks. We analysed the data of the prevalence of intestinal parasitic infections in food handlers in our tertiary care centre from 2018 to 2022 and 6.8% were observed to harbour intestinal parasites during the period. This signifies the importance of routine screening, and the need of awareness and education of the food handlers in hospitals.

Modified trichrome stain for faster and improved detection of intestinal protozoan parasites.

Permanent stains such as trichrome have better sensitivity but are time-consuming and the fixative includes toxic mercuric chloride. Thus, a newer modification was tested and found to be a superior, faster and safer staining technique for intestinal parasitic detection. Our study lasted 9 months and a single stool sample was collected from each enrolled patient. We evaluated classical trichrome (T1 - using Schaudinn fixative) with newer modifications, which involved different fixatives with mordant combinations (T2 - acetic acid + hydrated aluminium sulphate, T3 - citric acid + copper sulphate hydrate). Conventional PCR targeting Entamoeba histolytica, Giardia lamblia and Cryptosporidium spp. was taken as the reference. Out of 175 stool samples, 25.1% protozoa were identified by wet mount, 24% by each T1 and T2, 25.7% by T3. Statistically, T3 and T2 had higher sensitivity as compared to T1 and wet mount when PCR was used as reference.

Comparison of B1 and RE 529 gene targets by real time PCR and LAMP assay for diagnosis of toxoplasmosis in pregnant females.

PURPOSE: The aim of this study is to accurately diagnose the presence of toxoplasmosis in pregnant women. In this study we evaluated two gene targets B1 and RE-529 using two different molecular methods i.e., real-time PCR and LAMP. PROCEDURE: A total of 150 blood samples were collected from pregnant women attending the PGIMER outpatient clinic. The serum and Buffy layer were extracted and various serological (ELISA) and molecular tests (qPCR and LAMP) targeting B1 and RE-529 were carried out. FINDING: Out of 150 patients, 32 were seropositive. Amongst which for the RE-529 gene, 18 were LAMP positive and 16 were qPCR positive, while for the B1 gene, 14 were LAMP positive and 13 were qPCR positive. CONCLUSIONS: Molecular methods were more sensitive than serological tests to diagnose congenital toxoplasmosis in antenatal females. Few seronegative patients were reported positive using molecular methods. In addition, LAMP targeting the RE-529 gene is more sensitive than qPCR, and LAMP targets the B1 gene.

Novel diagnostic approach for amoebic liver abscess using cell free (cf) DNA: a prospective study.

BACKGROUND: Amoebic liver abscess (ALA) is commonly seen in tropical countries and diagnosis of ALA relies mainly on non-specific serological and imaging techniques as well as PCR from pus. OBJECTIVE: This study evaluated the potential of using cell free DNA (cfDNA) from serum and urine for diagnosing ALA. METHODS: We prospectively evaluated quantitative PCR (qPCR) for detection of cf DNA in serum and urine sample in all liver abscess patients. The samples were collected from patients reporting to emergency ward of Postgraduate Institute of Medical Education and Research, Chandigarh, India with symptoms suggestive of liver abscess. Real time PCR was done to detect cf DNA in serum and urine by targeting 99-bp unit of small subunit rRNA of Entamoeba histolytica and conventional PCR for pus. RESULTS: A total 113 samples (serum and urine) and 100 pus samples were analysed. A total of 62 ALA patients were confirmed; with maximum 57 patients detected by qPCR for cfDNA in the serum, 55 patients by PCR on pus aspirate and 50 ALA patients by qPCR for cfDNA in urine sample. Therefore, the sensitivity of qPCR for detection of cf DNA in serum was 91.94% and for urine was 80.65%. CONCLUSION: A total of 11.2% of ALA patients were diagnosed only through detection of E. histolytica cf DNA in their serum and urine. Detection of cfDNA from serum, urine of ALA has a potential role in future especially for developing countries as it is a rapid, sensitive and patient friendly diagnostic approach.

Seroprevalence of human cystic echinococcosis: A study from a tertiary care center of North India.

Present study retrospectively analysed the serological data of patients suspected of cystic echinococcosis (CE) attending the outpatient clinics or admitted in our hospital. An enzyme-linked immunoassay was used to analyse anti-CE antibodies in serum samples of 3680 patients. Microscopy of aspirated cystic fluid was performed on 170 cases only. CE seropositive cases were 595 (16.2%), of which 293 (49.2%) were males and 302 (50.8%) were females. A higher percentage of seropositivity was found in adults within age range of 21-40 years of age. There has been a decrease in seropositivity in the study years (2016-2021) in comparison to previous years (1999-2015).

Molecular Diagnosis of Encephalitis/Meningoencephalitis Caused by Free-Living Amoebae from a Tertiary Center in India.

BACKGROUND: Pathogenic free-living amoeba (FLA) such as Acanthamoeba spp., Naegleria fowleri, and Balamuthia mandrillaris are causative agents of fatal amoebic encephalitis/meningoencephalitis. The diagnosis of such infections is challenging due to a lack of clinical suspicion and expertise in microscopic identification. We evaluated the performance of molecular assays for the timely and accurate detection of FLA-causing central nervous system (CNS) afflictions. METHODS: This study included samples from 156 patients with suspected encephalitis/meningoencephalitis, including 149 cerebrospinal fluid (CSF) samples, 5 brain tissue biopsies, and 2 brain abscess samples. All the samples were subjected to PCR-based detection of Acanthamoeba spp., N. fowleri, and B. mandrillaris. The diagnostic characteristics and the inter-rater reliability scores were evaluated for parasite-specific polymerase chain reaction (PCR) using culture on non-nutrient agar (NNA)/microscopy or histopathological examination as a confirmatory test for Acanthamoeba spp. and N. fowleri and histopathology for B. mandrillaris. RESULTS: We detected 11 samples positive for FLA, including 6 Acanthamoeba spp., 3 B. mandrillaris, and 2 N. fowleri. Furthermore, all 11 samples were positive according to the confirmatory tests, i.e., culture on NNA/microscopy/histopathology in the case of Acanthamoeba spp. and N. fowleri and histopathology of tissue biopsies for B. mandrillaris. The inter-rater reliability between the PCRs and the confirmatory tests for the detection of Acanthamoeba spp., N. fowleri, and B. mandrillaris was 100%. CONCLUSIONS: The PCR-based detection of FLA in patients suspected of encephalitis/meningoencephalitis was found to be fast, efficient, and reliable in our study. We suggest the use of these PCRs in laboratories to obtain additional data on their efficiency in diagnosing FLA infections of the CNS. The present study was conducted with a small sample size of 156 patient samples, and we found only six Acanthamoeba spp., three B. mandrillaris, and two N. fowleri. The present study should be conducted on a larger sample size for better evaluation of the primer pairs.

Bilateral Pulmonary Hydatid Cyst in a Young Child: A Rare Case Report from North India.

Echinococcosis or hydatid disease is caused by the larval stage of the dog tapeworm, that is, Echinococcus granulosus , E. multilocularis , E. vogeli , or E. oligarthrus . Echinococcus granulosus causes cystic echinococcosis, which has a worldwide distribution. Liver is the most common site, affecting approximately two-third of the patients, whereas lung involvement is seen in approximately 25% of cases. This case is a very rare scenario of bilateral pulmonary hydatid cysts in a young child having exposure to a pet dog with negative hydatid serology and normal eosinophil count.

Cyclosporiasis in an immunocompromised patient who had undergone renal allograft transplant-A rare case report.

Cyclospora spp. is an important cause of traveler's diarrhea or water and food-borne diarrhoeal diseases. We present an interesting but rare case report of cyclosporiasis in a 51-year-old male who had undergone renal allograft transplant six years ago. He also had a past history of tuberculosis, cytomegalovirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and hepatitis C infection and was being treated with immunosuppressants. The patient had a prolonged history of gastrointestinal manifestations with recent acute onset of watery diarrhea associated with abdominal cramps. Stool examination after modified Ziehl-Neelsen staining revealed oocysts of Cyclospora spp. The patient was successfully treated with cotrimoxazole.

Bacteriological Surveillance of Ambulance Vehicles from a Tertiary Care Hospital of North India.

Objective An ambulance is a medically equipped vehicle which is used in case of any medical emergency for the transport of patients to treatment facilities. The ambulances help in the transportation of thousands of patients per year, and such patients may carry infectious microorganisms which pose a major threat to the treatment of such patients. In this study, we analyzed the extent of bacterial contamination in our ambulance vehicles and measured the degree of antimicrobial resistance among isolated pathogens. Material and Method This study included five ambulances of our tertiary care hospital and different random sites were swabbed in each vehicle. These were selected based on their well-known high frequency of contact by emergency personnel and patients. Swabs were inserted into sterile test tubes containing normal saline and immediately transferred to our microbiology laboratory to identify bacterial contaminants utilizing standard microbiological procedures. Result A total of 198 swab samples were collected from all the five ambulances, out of which 170 (85.8%) swabs were sterile and 28 (14.2%) swabs yielded potentially pathogenic bacterial isolates. The highest contamination rate with pathogenic bacteria was detected in the oxygen flow meter knob (60%), suction machine tubing (60%), and stethoscope (40%). Staphylococcus aureus (32%) was the most frequently detected microorganism. Conclusion Our study showed low prevalence of bacterial contamination in ambulances because of good infection control policy of our hospital, however, some areas still need improvement and require proper standard operating procedures of disinfection policies of these emergency vehicles.

A Prospective Study to Assess the Optimal Incubation Times for Culture and Aerobic Bacterial Profile in Prosthetic Joint Infections.

Introduction With an increase in the number of total joint arthroplasties, the count of prosthetic joint infections (PJIs) is also increasing and has become a nightmare for an orthopaedic surgeon. Microbiological diagnosis is important for administering definitive antimicrobial treatment. Negative culture reports hamper patient management and prolonged incubation periods have increased the culture yield but at the risk of culture contamination in conventional microbiology settings. Thus, we aimed to optimize the best incubation time for culture and the aerobic bacterial profile of PJIs. Material and Methods Over a year, samples from clinically suspected PJI patients were collected and processed for culture using standard techniques. The samples were incubated for up to 10 days with daily subculturing on to solid media. The bacterial isolates were identified and antibiotic susceptibility was performed. Results Out of 200 patients, 105 were included in the study and samples were collected in triplicate. In 70 cases culture was positive and 35 were culture negative PJIs. Mean incubation days to culture positivity was 3.6 days with 97.14% culture positivity seen by the seventh day of incubation. Twenty-four percent methicillin-resistant Staphylococcus aureus , and more than 50% extended spectrum ß lactamase producing Klebsiella pneumoniae and Escherichia coli were isolated. Discussion Culture positivity in PJIs provides definitive evidence of infection and guides the treatment. Increasing the incubation times can help in maximizing the culture yield, and we found that 97.14% pathogens grew within 7 days of incubation. Prolonging it further would not provide an added advantage especially in a resource-constraint setting.

Clinical and laboratory profile of trichinellosis from a non-endemic country.

Trichinellosis is a zoonotic parasitic infection caused by nematodes of the genus Trichinella. Humans mostly get infected by eating raw/under-cooked pork. In India, it has been reported mostly as sporadic cases especially from North Eastern and Eastern part of country. In this study, we retrospectively analyzed the socio-demographic pattern, clinical presentation, laboratory profile and treatment response in Trichinella-infected patients visiting our tertiary care center which mainly caters to patients from North India. For this retrospective laboratory-based analysis, patients diagnosed on the basis of positive anti-Trichinella IgG antibodies between 1st June 2008, and 31st May 2019 were included. A total of 11 positive trichinellosis cases were detected, of whom majority were children who presented with history of fever, gastrointestinal symptoms, myalgia, eosinophilia along with hepatomegaly and pulmonary manifestations. No CNS involvement was seen in any of the patients although it is commonly associated with Trichinella infection. All patients recovered uneventfully after antihelminthic treatment.

Epidemiology and clinical outcome of ventilator-associated events at a tertiary care hospital from North India.

The aim of our study was to determine the incidence, microbiological profile, risk factors and outcomes of patients diagnosed with ventilator-associated events in our tertiary care hospital. In this prospective study, intensive care patients put on mechanical ventilation for >48 h were enrolled and monitored daily for ventilator-associated event according to Disease Centre Control guidelines. A ventilator-associated event developed in 33/250 (13.2%); its incidence was 3.5/100 mechanical ventilation days. The device utilisation rate was 0.86, 36.4% of patients had early and 63.6% late-onset ventilator-associated pneumonia whose most common causative pathogen was Acinetobacter sp. (63.6%). Various factors were significantly associated with a ventilator-associated event: male gender, COPD, smoking, >2 underlying diseases, chronic kidney disease and elevated acute physiological and chronic health evaluation II scores. Therefore, stringent implementation of infection control measures is necessary to control ventilator-associated pneumonia in critical care units.

Characterization of Bacteriological Isolates from Patients and Environment Samples of Burn Ward: A Study from a Tertiary Care Hospital of India.

OBJECTIVE: Source of infection in a burn patient is from the patient's flora, contaminated environmental surfaces and transmitted from health care workers. Insufficiently disinfected hospital environmental surface provides a niche for multidrug resistant bacteria. This study was carried out to assess the bacteriological profile of the pathogens from burn wounds and the surrounding environmental areas. METHODS: During 6 months, wound swabs from burn patients were collected on admission (after 48 hours of admission), on day 5 and then weekly. Environmental samples were also collected from burn ward and studied for the bacteriological and anti-microgram profiles. RESULTS: Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii were the major bacterial isolates from the wound swabs and the environmental samples. ESBL was detected in 56.6% of our Enterobacteriaceae isolates. The environmental sites from which these bacterial isolates were found were nursing counter, sink, dressing trolley, medicine locker and patient's bed. The percentage of MRSA decreased from 50 to 5% and there was an increased role of Enterococci species causing infections (13.63%). CONCLUSION: In this study, there appears that the colonizers of the environment may play a role in the causation of infection in burn patients. In burns ward, rigorous implementation of infection control program should be warranted, which includes and hygiene and use of personal protective equipment, environmental disinfection, cohort nursing care and antibiotics stewardship programme.

Corynebacterium Striatum, an Emerging Nosocomial Pathogen: Case Reports.

BACKGROUND: Corynebacterium striatum is an emerging nosocomial pathogen, capable of causing a variety of infections in immunocompromised and hospitalized patients. ; Case Presentation: We describe three cases of infection by C. striatum that were initially considered as contamination. Clinical suspicion in the wake of predisposing factors and accurate identification, using the Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS), were key to implicate this commensal bacterium as a cause of infection.

Increasing Resistance to Reserve Antibiotics: The Experience of a Tertiary Level Neonatal Intensive Care Unit.

BACKGROUND: Blood stream infections are considered as a major cause of morbidity and mortality in neonates. Recent trend shows increasing resistance to commonly used antibiotics. AIMS AND OBJECTIVES: The aim of this study is to find the antibiotic susceptibility pattern of various bacteria from blood samples in neonates and associated risk factors. METHODS: All consecutive cases of intramural neonatal sepsis were enrolled for >12 months. Before starting or changing antibiotic, blood sample under all aseptic precautions was taken for culture. Clinical and demographic details were recorded to analyze risk factors for sepsis. Antibiotic sensitivity tests were done as per CLSI 2019 guidelines. RESULTS: Of the 898 participants, 107 showed culture positivity. Klebsiella pneumoniae (25.2%) and Coagulase-negative Staphylococcus (23.3%). The blood culture positivity rate was 11.9%. Approximately 79% of isolates were multidrug-resistant: extended-spectrum beta-lactamase 90%, carbapenemase-resistant Enterobacteriaceae 27.7% and MRSA 43%. The risk factors found to be associated with sepsis were period of gestation ≤37 weeks, meconium-stained liquor, birth weight <1500 g, mechanical ventilation, partial exchange transfusion, duration of antibiotics for >10 days and duration of both NICU stay and hospital stay for >10 days. The case fatality rate (CFR) was more due to K. pneumoniae (19.2%) and the relative risk of death was 2.53 in culture-positive cases with an attributable risk of 60% and the population attributable risk of 15.4%. CONCLUSION: Increase in antibiotic resistance organisms can lead to an increase in the neonatal CFR, so regular surveillance is needed.

Detection of various beta-Lactamases in Escherichia coli and Klebsiella sp.: A study from Tertiary Care Centre of North India.

Objective: The emergence of carbapenem-resistant Escherichia coli and Klebsiella species is a global threat. We aimed to compare two phenotypic methods and evaluate the genotypic method for the detection of beta-lactamases produced by E. coli and Klebsiella spp. Materials and Methods: One hundred and twenty-six E. coli and Klebsiella isolates were examined for phenotypic production of beta-lactamases by using disc diffusion, combined disc test (CDT) and modified carbapenem inactivation method (mCIM). All strains were also studied for the presence of various genes by polymerase chain reaction. Results: Out of 126 isolates, 96% of the isolates were extended-spectrum ß-lactamase (ESBL) producers based on the presence of various ESBL genes. CDT method showed higher number of total (89%) carbapenemases in comparison to mCIM (81%). Among carbapenemases none of the isolates were Klebsiella pneumoniae carbapenemase producer by CDT, while 69% isolates were metallo-beta-lactamase (MBL) producers. Another method, mCIM/ethylene diamine tetraacetic acid mCIM showed 100% agreement for MBL detection. As regards, AmpC and class D carbapenemases; 0.04% and 16% positivity was detected, respectively, based on CDT method. Molecular analysis revealed 91% of the isolates harbouring carbapenemase genes. blaNDMwas the most common gene detected followed byblaOXA-48. Nine of the blaNDM-positive isolates also possessed blaOXA-48gene. Conclusion: Our finding shows high percentages of ESBL and carbapenemases in E. coli and Klebsiella spp. Among phenotypic methods, CDT seems to be a better choice as prevalence of carbapenemases shows lots of variation in our country. For Class B enzymes, both CDT and mCIM/eCIM can be used in the routine laboratories.

Burkholderia Cepacia Complex Causing Pneumonia in an Immunocompetent Non-Cystic Fibrosis Patient: Case Report and Review of Literature.

BACKGROUND: Burkholderia cepacia complex is widespread in the environment and has been recognized as a cause of opportunistic pulmonary infections, particularly in patients with Cystic Fibrosis (CF). The natural ecology of the bacteria as part of plant growth-promoting rhizosphere provides stark contrast to its infectious potential. Its preponderance as a nosocomial pathogen may be due to its ability to survive in antiseptic solutions, contaminate equipments and intrinsic antimicrobial resistance. CASE: An elderly, diabetic male was evaluated for hemoptysis, fever and cough. Chest computed tomography showed a thick walled cavity in the left lung and hilar lymphadenopathy. Sputum examination showed Gram negative bacilli and no acid fast bacilli. Sputum culture yielded growth of non-fermentative Gram negative bacilli on two occasions, but blood culture was sterile. The isolate was identified as B. cepacia by Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS). The patient's general condition remained poor and in spite of initiation of antibiotics, the patient expired after an episode of massive hemoptysis. CONCLUSION: This report raises concerns regarding the spread and severity of B. cepacia infection in non-compromised patients in the community and the need to suspect and identify it. Since the organism is inherently resistant to antipseudomonal penicillins, aminoglycosides and polymyxin B, differentiation from Pseudomonas spp. and determining antimicrobial susceptibility is paramount for treatment.

Post-Traumatic Endophthalmitis Due to Acinetobacter radioresistens: Virulence Potential of Commensal Bacteria.

Acinetobacter species are widely distributed in soil, water and hospital environment. In addition to A. calcoaceticus-baumannii complex, the clinically most relevant species, there are other genomospecies which are less frequently identified, mostly due to lack of accurate methods for routine identification. We describe the first case of post-traumatic endophthalmitis caused by otherwise non-pathogenic A. radioresistens, in India. Gram-negative coccobacilli were observed on Gram stain and culture of vitreous fluid specimen, and identified biochemically as Acinetobacter spp. The species was identified by Matrix Assisted Laser Desorption Ionization- Time of Flight Mass Spectrometry (MALDI-TOF MS). The pathogenic potential of 'commensal' A. radioresistens and its role in dissemination of carbapenem resistance genes underlines the importance of species-level identification in Acinetobacter infections.

Next-generation strategy for treating drug resistant bacteria: Antibiotic hybrids.

Resistance against nearly all antibiotics used clinically have been documented in bacteria. There is an ever-increasing danger caused by multidrug-resistant Gram-negative bacteria in both hospital and community settings. In Gram-negative bacteria, intrinsic resistance to currently available antibiotics is mainly due to overexpressed efflux pumps which are constitutively present and also presence of protective outer membrane. Combination therapy, i.e., use of two or more antibiotics, was thought to be an effective strategy because it took advantage of the additive effects of multiple antimicrobial mechanisms, lower risk of resistance development and lower mortality and improved clinical outcome. However, none of the benefits were seen in in vivo studies. Antibiotic hybrids are being used to challenge the growing drug resistance threat and increase the usefulness of current antibiotic arsenal. Antibiotic hybrids are synthetic constructs of two molecules which are covalently linked. These could be two antibiotics or antibiotic with an adjuvant (efflux pump inhibitor, siderophore, etc.) which increases the access of the antibiotics to the target. The concepts, developments and challenges in the future use of antibiotic hybrids are discussed here. Majority of the studies have been conducted on fluoroquinolones and aminoglycosides molecules. The antibiotic tobramycin has the property to enhance the action of antimicrobial agents against which the multidrug-resistant Gram-negative bacteria were earlier resistant, and thus potentiating the action of legacy antibiotics. Antibiotic hybrids may have a role as the silver bullet in Gram-negative bacteria to overcome drug resistance as well as extend the spectrum of existing antibiotics.

Evaluation of various risk factors associated with multidrug-resistant organisms isolated from diabetic foot ulcer patients.

AIMS: Diabetic foot ulcer is a dreaded complication of diabetes. Diabetic foot ulcer patients are often infected with multidrug resistant organism (MDRO) due to chronic course of the wound, inappropriate antibiotics treatment, frequent hospital admission, neuropathy, nephropathy, and peripheral vascular disease. MATERIALS AND METHODS: This prospective study was conducted in our 750 bedded hospital for a period of 6 months. The present study was undertaken to isolate various MDRO methicillin resistant Staphylococcus aureus; Gram-negative bacteria producing enzymes such as extended spectrum beta-lactamases (ESBL), Amp C, Carbapenamases; Pseudomonas and Acinetobacter species producing metallo-beta-lactamases (MBL). In addition we attempted to identify risk factors for association of diabetic foot ulcer and MDRO. RESULTS: A total of 149 bacterial isolates were identified. Of the total isolates 73.2% were Gram-negative and remaining 26.8% were Gram-positive bacteria. Among Enterobacteriaceae 59% were ESBL producers and 48% were Amp C producers. In addition, 41.5% of the isolates produced both ESBL and Amp C and 13.4% were carbapenem resistant Enterobacteriaceae. Among 20 Pseudomonas and Acinetobacter isolates, 5 were MBL producers (25%). Furthermore, in the study, 56% of patients with diabetic foot ulcer harbored MDRO. The risk of multidrug-resistant infection is significantly more in patients having diabetes duration >20 years and size of ulcer more than 4 cm2. CONCLUSION: The detection of MDRO in patients of diabetic foot ulcer changes the treatment strategies limits the antimicrobial options and causes higher complications among them.