Monkeypox scenario in India: a review study.

The monkeypox virus, which causes the viral zoonotic disease, is known as the most significant orthopoxvirus infection following the elimination of smallpox. The monkeypox virus, which was previously exclusive to West and Central African nations and caused endemic diseases in monkeys and people, has recently been linked to human infections in non-endemic areas including the United States of America (USA) and more than 30 additional countries. Guidelines for the diagnosis and treatment of monkeypox have also recently been made available by the Ministry of Health and Family Welfare of India and the Indian Government. The monkeypox outbreak continues to be a worldwide health emergency, the highest degree of alert recognised by the World Health Organization. The Centers for Disease Control and Prevention (CDC) advises vaccination for those who have been exposed to the disease as well as those who may be at higher risk of contracting it, such as those who have been identified by public health officials as a contact of someone who has the disease.

Prevalence of Hepatitis B Virus, Genotypes, and Mutants in HBsAg-Positive Patients in Meerut, India

Background: Genetic changeability of hepatitis B virus (HBV) signifies a challenge for the sensitivity of immunologic and molecular diagnostics. Therefore, knowing the spread of HBV genotypes (GENs) and mutation has considerable impacts on treatment strategies, vaccination program, diagnosis, and prevention. The present study aimed to detect HBV GENs and mutants in HBsAg-positive patients. Methods: The study conducted on 4927 patients in Meerut, India, between March 2013 and April 2017. The blood specimens were analyzed for HBsAg using an ELISA kit, then the blood samples from HBsAg-positive patients were subjected to HBeAg assay and DNA isolation. Amplification of the HBV DNA of pre-S gene and pre-core or basal core promoter region were performed by RT-PCR and sequenced to analyze both GEN and mutation. Results: According to the results, 245 cases were positive for HBsAg, and 55 were HBeAg-positive. With regard to HBV DNA levels, 16 samples were found positive in PCR assay with 7 (43.8%) less than 2000 IU/mL, 4 (25%) between >2000 and 20,000 IU/mL, and 5 (3.25%) >20,000 IU/mL. No mutations were detected in GENs B and A. The prevalence of HBV GENs B and A were 68.8% (n = 11) and 31.25% (n = 6), respectively. Conclusion: GEN-B was more prevalent in comparison to GEN-A. The genetic diversity of HBV and distribution of its GENs and mutation improve the current knowledge of epidemiological, clinical and virological patterns of hepatitis B in this region, which help physicians to prescribe proper antiviral/interferon therapy according to current genotyping pattern.

Linezolid and Vancomycin Resistant Enterococci: A Therapeutic Problem.

INTRODUCTION: Enterococci are recognized as opportunistic pathogens, as well as commensals in both humans and animals. They are an important cause of nosocomial infections, difficult to treat, as the organism have intrinsic and acquired resistance to many antibiotics. AIM: To isolate and identify clinically relevant Enterococcus up to species level from all the clinical samples processed in the microbiology laboratory and also to study their resistance pattern. MATERIALS AND METHODS: A prospective study was carried out for a period of one year from May 2014 to April 2015 at the Department of Microbiology, Subharti Medical College, Meerut, Uttar Pradesh, India. A total of 200 isolates of Enterococcus species from 15342 clinical samples obtained from IPD/OPD patients irrespective of age, having suspicion of bacterial infection were processed in the microbiology laboratory. Identification was done with standard biochemical methods. Antibiotic susceptibility testing was done on Muller Hilton agar plate by Kirby Bauer disc diffusion method. High-Level Gentamycin Resistance (HLGR) and High-Level Streptomycin Resistance (HLSR) were further confirmed by Agar dilution method and Broth microdilution method. Vancomycin and linezolid resistance was further confirmed by Agar dilution method and MIC was calculated by using VITEK 2, Biomerieux. All methodology was followed as per Clinical and Laboratory Standards Institute (CLSI) M100-S 24, 2014 guidelines. RESULTS: E. faecalis (n=169, 84.5%) was the predominant species isolated, followed by E. faecium (n=27, 13.5%) and E. casseliflavus (n=4, 2%). A total of 25 (12.5%) isolates were HLGR, 13 (6.5%) isolates were HLSR and 62 (31%) isolates were HLGR+HLSR. Vancomycin resistance was found in 14 (7%) isolates of which 11 (78.5%) were Van A and 3 (21.4%) were Van B, detected phenotypically as per relative MIC of vancomycin and teicoplanin. Linezolid resistance was seen in 4 (2%) of isolates which were vancomycin as well as high-level gentamycin and high-level streptomycin resistant. CONCLUSION: Isolation of Enterococcus species resistant to most of the higher antibiotics like vancomycin and linezolid, with high prevalence of High-Level Aminoglycoside Resistance (HLAR), from hospitalized patients is a major concern as such isolates have limited or no therapeutic option.

Bio-Medical Waste Managment in a Tertiary Care Hospital: An Overview.

INTRODUCTION: Bio-Medical Waste (BMW) management is of utmost importance as its improper management poses serious threat to health care workers, waste handlers, patients, care givers, community and finally the environment. Simultaneously, the health care providers should know the quantity of waste generated in their facility and try to reduce the waste generation in day-to-day work because lesser amount of BMW means a lesser burden on waste disposal work and cost saving. AIM: To have an overview of management of BMW in a tertiary care teaching hospital so that effective interventions and implementations can be carried out for better outcome. MATERIALS AND METHODS: The observational study was carried out over a period of five months from January 2016 to May 2016 in Chhatrapati Shivaji Subharti Hospital, Meerut by the Infection Control Team (ICT). Assessment of knowledge was carried out by asking set of questions individually and practice regarding awareness of BMW Management among the Health Care Personnel (HCP) was carried out by direct observation in the workplace. Further, the total BMW generated from the present setup in kilogram per bed per day was calculated by dividing the mean waste generated per day by the number of occupied beds. RESULTS: Segregation of BMW was being done at the site of generation in almost all the areas of the hospital in color coded polythene bags as per the hospital protocol. The different types of waste being collected were infectious solid waste in red bag, soiled infectious waste in yellow bag and sharp waste in puncture proof container and blue bag. Though awareness (knowledge) about segregation of BMW was seen in 90% of the HCP, 30%-35% did not practice. Out of the total waste generated (57912 kg.), 8686.8 kg. (15%) was infectious waste. Average infectious waste generated was 0.341 Kg per bed per day. The transport, treatment and disposal of each collected waste were outsourced and carried out by 'Synergy' waste management Pvt. Ltd. CONCLUSION: The practice of BMW Management was lacking in 30-35% HCP which may lead to mixing of the 15% infectious waste with the remaining non-infectious. Therefore, training courses and awareness programs about BMW management will be carried out every month targeting smaller groups.

Methicillin Resistant Staphylococcus aureus: Inconsistencies in Vancomycin Susceptibility Testing Methods, Limitations and Advantages of each Method.

BACKGROUND: Vancomycin may be ineffective against an increasing proportion of methicillin resistant Staphylococcus aureus (MRSA) with minimum inhibitory concentrations (MICs) well within the susceptible range. On the other hand it is common knowledge that determination of vancomycin MICs is method dependent. Therefore, given the apparent variability in vancomycin MIC results obtained with the different methods, the use of the vancomycin MIC to predict the outcome of serious S. aureus infections needs to take into account the method used and the results of studies using that particular method. AIM: Comparative study was carried out to evaluate the MICs obtained by BMD method, E-test, and Vitek 2 method and to detect inconsistencies in these vancomycin for 66 MRSA isolates obtained from various samples of patients attending the OPDs & IPDs within a period of one year. MATERIALS AND METHODS: A comparative study was carried out to evaluate the MICs obtained by BMD method, E-test, and Vitek 2 method to detect vancomycin susceptibility in 66 clinical isolates of MRSA obtained from various samples of patients attending the OPDs & IPDs within a period of one year. The study was conducted in Department of Microbiology, Subharti Medical College, Meerut from January to December 2012. RESULTS: On determination of MICs for vancomycin for the MRSA isolates, all were identified as VSSA by BMD, E-Test & Vitek 2 methods. However, the vancomycin MIC values obtained by E-test correlated better with BMD method (correlation factor= 0.6727) than Vitek 2 (correlation factor=0.5316), indicating E-Test to be a better method for determination of vancomycin MICs as compared to Vitek 2. CONCLUSION: MRSA isolates with higher vancomycin MICs, even within the susceptibility range, are being observed more frequently which result in treatment failures with vancomycin. Because of the discrepancy that exists in vancomycin MIC results from different methods, the prediction of outcome of serious S.aureus infections should take into account the method used & results of studies using that particular method.

Comparison of biofilm formation in clinical isolates of Candida species in a tertiary care center, North India.

BACKGROUND AND OBJECTIVES: Biofilms are colonies of microbial cells encased in a self-produced organic polymeric matrix. The biofilm production is more important for nonalbicans Candida (NAC); as C. albicans possess many other mechanisms to establish infections. Correct identification of Candida species has gained importance due to persistent rise in infections caused by NAC. We sought to isolate, identify Candida species in clinical isolates and study biofilm formation. MATERIALS AND METHODS: Modified microtiter plate method was performed to study biofilm formation by isolates in Sabouraud's dextrose broth. It was then quantitatively assessed using a spectrophotometer. Biofilm formation was graded as negative, +1, +2, +3 and + 4 on the basis of percentage absorbance. RESULTS: Biofilm formation was observed in 16 of 40 (40.0%) isolates of C. albicans as compared to 39 of 78 (50.0%) of isolates of NAC. Strong (+4) biofilm production was seen in maximum biofilm producers in C. tropicalis (12 of 27) followed by C. albicans (8 of 16). Total biofilm producers were significantly more among high vaginal swab isolates 63.2% (12 of 19) and urine isolates 59.2% (29 of 49), when compared to blood isolates 34.2% (13 of 38) as well as other isolates 27.5% (11 of 40). INTERPRETATION AND CONCLUSIONS: NAC species are qualitatively and quantitatively superior biofilm producers than C. albicans. Biofilm production is the most important virulence factor of NAC species and compared to other lesions, it is more significantly associated with luminal infections.

Recurrent hydatid cyst of liver with asymptomatic concomitant hydatid cyst of lung: an unusual presentation-case report.

A 40-year-old male patient presented to us with complaints of pain in abdomen for the past 2 weeks and fever for 3 days. The patient gave history of being previously operated for hydatid cyst of liver 15 years back. His chest radiograph and computed tomography scan revealed a cystic lesion in the right lobe of liver and a cyst in left lung. The patient was managed surgically. Aspirate from the cyst fluid showed plenty of hooklets and scolices of Echinococcus granulosus. An intact brood capsule was also seen. Diagnosis of hydatidosis was further confirmed by histopathological examination. Post-operative the patient had a good recovery.

Evaluation of phenotypic tests for detection of Klebsiella pneumoniae carbapenemase and metallo-beta-lactamase in clinical isolates of Escherichia coli and Klebsiella species.

CONTEXT: Carbapenemase production is an important mechanism responsible for carbapenem resistance. AIMS: Phenotypic detection and differentiation of types of carbapenemase in carbapenem resistant Enterobacteriaceae is important for proper infection control and appropriate patient management. SETTINGS AND DESIGN: We planned a study to determine the occurrence of Class A Klebsiella pneumoniae carbapenemase (KPC type) and Class B Metallo-ß-lactamase (MBL type) carbapenemase in hospital and community. MATERIALS AND METHODS: Clinical isolates of Escherichia coli and Klebsiella species and simultaneously evaluate different phenotypic methods for detection of carbapenemases. RESULTS: It was observed that 20.72% clinical isolates of E. coli and Klebsiella spp. were resistant to carbapenem on screening of which, 14.64% were E. coli and 29.69% were Klebsiella spp. Using phenotypic confirmatory tests the occurrence of carbapenemase production was found to be 87.01% in E. coli and 91.51% in Klebsiella spp. using both modified Hodge test (MHT) and combined disk test (CDT) using imipenem-ethylenediaminetetraacetic acid. CONCLUSIONS: Both MBL and KPC type carbapenemases were seen among clinical isolates of E. coli and Klebsiella spp. CDT is simple, rapid and technically less demanding procedure, which can be used in all clinical laboratories. Supplementing MHT with CDT is reliable phenotypic tests to identify the class A and class B carbapenemase producers.

Coordination mode of pentadentate ligand derivative of 5-amino-1,3,4-thiadiazole-2-thiol with nickel(II) and copper(II) metal ions: synthesis, spectroscopic characterization, molecular modeling and fungicidal study.

Complexes of nickel(II), and copper(II) were synthesized with pantadentate ligand i.e. 3,3'-thiodipropionicacid-bis(5-amino-1,3,4-thiadiazole-2-thiol) (L). The ligand was synthesized by the condensation of thiodipropionic acid and 5-amino-1,3,4-thiadiazole-2-thiol in 1:2 ratio, respectively. Synthesized ligand was characterized by elemental analysis, mass, (1)H NMR, IR, and molecular modeling. All the complexes were characterized by elemental analysis, molar conductance, magnetic moment, IR, electronic spectra, ESR, and molecular modeling. The newly synthesized complexes possessed general composition [M(L)X2] where M = Ni(II), Cu(II), L = pantadentate ligand and X = Cl(-), CH3COO(-). The IR spectral data indicated that the ligand behaved as a pantadentate ligand and coordinated to the metal ion through N2S3 donor atoms. The molar conductance value of Ni(II), and Cu(II) complexes in DMSO corresponded to their electrolytic behavior. On the basis of spectral study, octahedral and tetragonal geometry was assigned for Ni(II) and Cu(II) complexes, respectively. In vitro fungicidal study of ligand and its complexes was investigated against fungi Candida albicans, Candida parapsilosis, Candidia krusei, and Candida tropicalis by means of well diffusion method.

Genotype MTBDR plus assay for molecular detection of rifampicin and isoniazid resistance in Mycobacterium tuberculosis.

AIM: This study was performed for the rapid identification of Mycobacterium tuberculosis complex and its resistance to rifampicin and isoniazid, directly from the sputum samples of pulmonary tuberculosis patients. MATERIALS AND METHODS: A commercially available genotype MTBDR plus assay was used for the identification and detection of mutations in Mycobacterial isolates. A total of 100 sputum samples of pulmonary tuberculosis patients were analyzed by using the genotype MTBDR plus assay. The MTBDR plus assay is designed to detect the mutations in the hotspot region of rpoB gene, katG and regulatory region of inhA gene. RESULTS: The genotype MTBDR plus assay detected 22% multidrug resistant (MDR), 2% rifampicin (RMP) monoresistant and 1% isoniazid (INH) monoresistant isolates. In 22 MDR isolates, the codons most frequently involved in RMP-associated mutations were codon 531 (54.55%), 516 (31.82%) and 526 (13.63%), and 90.90% of MDR isolates showed KatG S315T mutations and 9.1% showed inhA C-15T mutations associated with INH resistance. CONCLUSION: The new genotype MTBDR plus assay represents a rapid, reliable tool for the detection of MDR-TB, wherein results are obtained in 5 h allowing early and appropriate treatment, which is essential to cut the transmission path and reduce the spread of MDR-TB. The genotype MTBDR plus assay can readily be included in a routine laboratory work for the early diagnosis and control of MDR-TB.

Extended spectrum beta lactamase producing Proteus penneri: a rare missed pathogen?

Indole negative Proteus species are invariably incorrectly identified as Proteus mirabilis, often missing out isolates of Proteus penneri. We report a case of extended spectrum beta lactamase producing and multidrug-resistant P. penneri isolated from pus from pressure sore of a patient of road traffic accident. Correct and rapid isolation and identification of such resistant pathogen are important as they are significant nosocomial threat.

A study of cervical intraepithelial neoplasia in pregnancy.

OBJECTIVE: To find the incidence of human papillomavirus (HPV) infection and cervical intraepithelial neoplasia (CIN) in pregnant women and compare Pap smear with the HPV DNA test in detecting HPV infection. MATERIALS AND METHODS: Hundred antenatal women, irrespective of gestational age, were enrolled as subjects in this prospective pilot study for blood investigations, wet mount examination of cervical discharge, Pap smear, and high-risk HPV DNA detection of cervical scrape by PCR. Women showing abnormality in Pap smear and/or those who were high-risk HPV DNA positive were subjected to colposcopy. RESULTS: The incidence of HPV-positive pregnant women was 18 %. Koilocytosis on Pap smear was observed in six women. Three high-risk HPV DNA-positive women showed changes consistent with CIN 1 on colposcopy. CONCLUSIONS: The HPV DNA test is the most sensitive and reliable in detecting HPV infection as compared to Pap smear, but considering the cost of PCR, Pap smear screening of all antenatal women was recommended.

Effect of addition of 2% chlorhexidine or 10% doxycycline on antimicrobial activity of biodentine.

AIM: The purpose of this in vitro study was to determine whether the addition of 2% chlorhexidine gluconate or 10% doxycycline would enhance the antimicrobial activity of Biodentine against Staphylococcus aureus (ATCC-25923), Enterococcus faecalis (ATCC-29212), Candida albicans (ATCC-90028), and Streptococcus mutans (MTCC-497). MATERIALS AND METHODS: Three wells of 4 mm diameter and 4 mm depth on each plate were prepared on the agar medium with standardized suspensions of each microorganism. Biodentine powder mixed with 2% chlorhexidine (0.06 g) or 10% doxycycline (0.30 g) in its liquid or liquid alone was placed to fill each well. Plates were incubated at 37°C as required for microbial growth. A blinded, independent observer measured zones of inhibition. The data were analyzed using independent "t" test to compare the differences among the three cement preparations for different micro-organisms. RESULTS: All Biodentine samples inhibited microbial growth. The highest mean diameters of zone of inhibition for all the micro-organisms were found around Biodentine/chlorhexidine (13.417) followed by Biodentine alone (12.236) and Biodentine/doxycycline (11.25). CONCLUSION: In conclusion, adding 2% chlorhexidine gluconate in liquid of Biodentine enhanced the antimicrobial activity of Biodentine against all the tested micro-organisms except Candida albicans, while addition of 10% doxycycline decreased the antimicrobial activity of Biodentine. The differences were significant statistically (P < 0.05).

Micropathological changes in the sub-epidermal zone of normal appearing skin in leprosy.

Leprosy is a chronic inflammatory disease caused by Mycobacterium leprae, which affects not only the peripheral nerves and skin but also various internal viscera through the hematogenous spread, especially in lepromatous cases. The micropathological changes in epidermis, nerves and skin appendages from lesioned skin reported by various authors but reports of involvement of apparently normal sites are few. We investigated skin biopsy material taken from 130 patients with clinically diagnosed leprosy. Biopsies were taken at least 10 cm away from site of lesion. Hematoxylin and Eosin staining and Harada's modified Allochrome method for acid-fast bacilli were applied for histological investigations. The pattern of leprosy among the patients were indeterminate in 53 cases (40.8%), tuberculoid in 29 cases (22.3%), borderline tuberculoid in 14 cases (10.8%), borderline leprosy in ten cases (7.7%), borderline lepromatous in nine cases (6.9%) and lepromatous leprosy in 15 cases (11.5%). The changes were seen in sub-epidermal zone of normal appearing skin in all type of leprosy, but involvement was greater at the lepromatous end of the spectrum compared to tuberculoid end. Acid-fast bacillus (AFB) was seen in subepidermal zone of normal appearing site. Presence of AFB is significant as far as dissemination and transmission of disease is concerned.

Microbial keratitis in Gujarat, Western India: findings from 200 cases.

INTRODUCTION: The objective of this study was to study the epidemiological characteristics and the microbiological profile of patients suspected with microbial keratitis in Gujarat. METHODS: Corneal scraping was collected from 200 consecutive cases of suspected microbial keratitis and was subjected to direct examination and culture. RESULTS: Of the 200 ulcers 55% were culture positive, 26.5% were bacterial ulcers of which 47% were due to Staphylococcus spp. Pure fungal growth was seen in 22% while 6% were mixed ulcers. Fusarium spp. (30%) was the most common fungus followed by Aspergillus spp. (21%). Only one case of Acanthamoeba keratitis was encountered. Patients were mainly from rural areas (61.5%) with male preponderance (61.5%). Corneal injury was seen in 78.5% cases of which 53% had injury with vegetative matter. Prior treatment was seen in 58% of which 5% had been treated by village healers. Nineteen patients (9.5%) also used some kind of traditional topical treatment. Increased incidence was seen from August to December. Five case of fugal ulcers lead to perforation of which three were due to Fusarium spp. whereas perforation was seen in only two cases of bacterial ulcers due to Pseudomonas aeruginosa. CONCLUSION: Staphylococcus and Fusarium spp. were the most common etiological agents in our region. Predominant outdoor agricultural activity is the principal causative factor for corneal injury. Corneal ulcers complicated due to treatment by village healers are another important concern. The information regarding regional etiology will help empirical management as many eye clinics do not have microbiological facilities.

Maxillary osteomyelitis by mucormycosis: report of four cases.

Mucormycosis is a fungal infection commonly affecting structures in the head and neck, such as the air sinuses, orbits, and the brain. Common predisposing factors include diabetes mellitus and immunosuppression. We describe our clinical experience with four cases of mucormycosis of the maxillary antrum associated with uncontrolled diabetes mellitus managed at our centre. Early diagnosis and prompt treatment can significantly reduce the mortality and morbidity of this lethal fungal infection.

Physician accessories: doctor, what you carry is every patient's worry?

BACKGROUND: Nosocomial infections are on the rise worldwide and many a times they are carried by the health care personnel. Accessories used by physicians and healthcare personnel can be a potential source of nosocomial infection. MATERIALS AND METHODS: We designed a survey with the aim to investigate the prevalence of microbial flora of accessories such as pens, stethoscopes, cell phones and white coat used by the physicians working in a tertiary care hospital. OBSERVATIONS: It was observed that 66% of the pens, 55% of the stethoscopes, 47.61% of the cell phones and 28.46% of the white coats used by the doctors were colonized with various microorganisms. Staphylococcus spp. was the predominant isolate followed by Escherichia coli. Methicillin resistance in Staphylococcus aureus was also found, which was a matter of concern. CONCLUSIONS: Awareness of appropriate hand hygiene is important in order to prevent potential transmission to patients.

External ophthalmomyiasis caused by Oestrus ovis: a rare case report from India.

Myiasis of different organs has been reported off and on from various regions in the world. We report a human case of external ophthalmomyiasis caused by the larvae of a sheep nasal botfly, Oestrus ovis, for the first time from Meerut city in Western Uttar Pradesh, India. A 25-year-old farmer presented with severe symptoms of conjunctivitis. The larvae, 3 in number, were observed in the bulbar conjunctiva, and following removal the symptoms of eye inflammation improved within a few hours.