Diverse aminoglycoside phosphotransferase types conferring aminoglycoside resistance in Enterobacteriaceae: A single-centre study from Northeast India

The present study investigates the molecular basis of aph-mediated aminoglycoside resistance and their transmission dynamics in a tertiary care hospital of Northeast India. Two hundred forty one isolates (230 Escherichia coli and 11 Klebsiella pneumoniae) were collected and screened for aminoglycoside resistance genes. Various aph types were amplified using polymerase chain reaction (PCR) assay. Plasmid incompatibilty, horizontal transferability and ERIC-PCR based typing were carried out for all the positive isolates. Among them, 67 isolates showed the presence of aph gene. Aph (3“)-IIIa and aph (3')-Via were predominant and horizontally transferable. All the plasmids were of incompatibility I1 group. Twenty-eight different haplotypes of E. coli were found harbouring aph gene types. This study was able to identify diverse aph types in a single centre and their corresponding phenotypic trait.

Sub-inhibitory concentration of ertapenem induces overexpression of regulator of antibiotic resistance A in Escherichia coli

AcrAB-TolC is a tripartite efflux pump system constitutively expressed which functions as an intrinsic-resistant mechanism found to be responsible for conferring resistance towards dyes, detergents and different compounds including various classes of antibiotics. One global regulator belonging to AraC-type regulator family, regulator of antibiotic resistance A (RarA) up-regulates the expression of AcrAB-TolC encoded in Klebsiella pneumoniae, Enterobacter sp. 638, Serratia proteamaculans 568 and Enterobacter cloacae resulting in multidrug-resistant phenotypes. The present work was initiated to find out the transcriptional response of RarA in clinical isolates of Escherichia coli against concentration gradient carbapenem stress. A total of 22 clinical isolates of E. coli and expression level of regulators were analysed via quantitative real-time polymerase chain reaction with and without carbapenem stress. As a result, a strong correlation between the expressional levels of RarA in AcrAB overexpressed isolates of E. coli and elevated expression was observed when exposed under concentration gradient ertapenem stress. The clones containing pRar showed reduction in the zone of inhibition towards carbapenem, indicating the active participation of RarA in AcrAB overexpressed isolates of E. coli conferring resistance towards carbapenems.

High-sensitivity detection of human malaria parasites by the use of rapid diagnostic tests and nested polymerase chain reaction in burdened communities of North East India

Background and Objectives: The study aimed to evaluate the diagnostic performance of malaria through microscopy and rapid diagnostic test (RDT) analysis performed locally and the accuracy evaluated by nested polymerase chain reaction (PCR) for diagnosis of Plasmodium falciparum from hotspot regions of North East (NE) India. Materials and Methods: One thousand one hundred and seventy-three blood samples were collected for identification of P. falciparum infection using microscopy and RDT analysis. DNA was extracted from whole blood using QIAamp DNA blood mini kit, and nested PCR was performed to confirm P. falciparum for evaluating sensitivity and specificity from various epidemiological surveys and geographical areas of NE India. Results: Of 1173 symptomatic malaria suspected patients, 15.6% (183/1173) patients were diagnosed as malaria positive by RDT and 67.94% cases (53/78) with microscopy. Of 183 malaria-positive patients, 42.62% (78/183) were diagnosed with P. falciparum and 84.61% (66/78) further confirmed to be P. falciparum positive by nested PCR. High sensitivity (97.9%) and low specificity (2.03%) of the RDT and high sensitivity (99.1%) and low specificity (0.9%) in microscopy against nested PCR results was statistically significant (P < 0.05). Epidemiological comparisons expressed highest incidences in Manipur (51.11%) followed by Meghalaya (48.93%) and Assam (35.16%). Overall incidence rate among the genders was observed to be higher in males than in females. Conclusions: Our findings suggest that PCR, RDT and microscopy can potentially determine hotspots at moderate transmission intensities, but PCR testing has a diagnostic advantage as transmission intensity falls. Therefore, malaria control programs should consider PCR testing when the prevalence of infection is low.

Transcriptional response of AcrEF-TolC against fluoroquinolone and carbapenem in Escherichia coli of clinical origin

Introduction: Efflux pump systems constitute a major means of intrinsic resistance in Escherichia coli. AcrEF-TolC pump is known to exhibit higher expression level in quinolone resistant isolates. However, the transcriptional response of this pump is yet to be known when exposed to quinolone and other group of antibiotics. Objective: The present study analyses the transcriptional response of AcrEF-TolC in the presence of quinolones and carbapenems. Methodology: A total of 167 non-duplicate clinical isolates from Silchar medical college and Hospital, Silchar, India were included in this study. Of which 27 were devoid of any carbapenemase activity and among them 13 isolates showed overexpression of AcrE and AcrF gene. Transcriptional response of AcrE was directly proportional to increasing concentration of levofloxacin and ofloxacin. However, the response of AcrE and AcrF was inconsistent with carbapenems. Result: The study isolates showed susceptibility towards amikacin (68.4%), gentamicin (59.6%), cefepime (52.7%) and pipercillin/tazobactam (48.3%). The present investigation highlights that apart from qnr genes and mutational changes in gyr region, AcrEF-TolC plays a major role in fluoroquinolone resistance in this part of the world. Conclusion: Upregulation of AcrE in the presence of levofloxacin and ofloxacin warrants further investigation to establish their active role in efflux of this drug.

Occurrence of II and V staphylococcal cassette chromosome mec types among coagulase-negative staphylococci from Northeastern part of India

Introduction: Coagulase Negative Staphylococci, the most commonly isolated pathogen are becoming emerging threats to the community as well as to the nosocomial environment. The present study underscores the distribution of Staphylococcal cassette chromosome mec (SCCmec) types among Methicillin resistant Coagulase Negative Staphylococci from the environmental origin. Methods and Materials: Environmental and food sample (n = 460) from different location of northeastern region of India were collected for a period of one year and were phenotypically and genotypically screened using cefoxitin disc and PCR techniques for mecA and mecC gene detection. All the MR-CoNS isolates possessing mecA gene were subjected to 16srDNA sequencing for species identification. SCCmec typing was determined by evaluating using primer sets from type I to type V. Antibiotic susceptibility testing was performed for all the isolates. Statistical analysis with chi-square test using SPSS-21 statistical software. Results: Methicillin resistance shown by one hundred forty three isolates were carried out for molecular analysis, among them 53.84% serves as mecA carrier. Distribution of Staphylococcus haemolyticus was more frequent and was found that SCCmec types II and V were predominant among the study isolates. Linezolid was the drug of choice for the CoNS isolates. Statistical analysis showed an insignificant result for the tested antibiotics and SCCmec types. Conclusion: This study therefore interprets the relative importance of SCCmec types among MR-CoNS isolates.

Association of glycerol kinase gene with class 3 integrons: A novel cassette array within Escherichia coli

Background: Integrons are genetic elements which are known for their role in capturing and spreading of antibiotic resistance determinants among Gram-negative bacilli. So far, there is no study regarding Class 3 integron and their genetic organisation in India. Objective: This study investigates the occurrence of Class 3 integron and their gene cassette array among Escherichia coli. Materials and Methods: In this study, a total of 200 E. coli isolates were collected from indoor and outdoor patients from Silchar Medical College and Hospital during September 2015 to February 2016. Detection of the integrase genes and gene cassettes within the Class 3 integron was performed by polymerase chain reaction which was further analysed by sequencing. Results: Twenty-seven isolates were found to harbour Class 3 integron. Sequencing of the gene cassettes and whole Class 3 integron revealed the presence of nine different types of cassettes array, out of which the arrangement with glycerol kinase gene cassette was found to be the most prevalent. Arrangement with blaCTX-Mgene cassette was also detected in few isolates. Conclusion: This study provides epidemiological profiling of Class 3 integrons in this geographical area. The data generated in this study are helpful in infection control programme, anti-infective research and search for epidemiological markers.

The quality of life of women volunteering as live-related kidney donors in India.

Background. Women outnumber men 6:1 as live-related donors in our renal transplant programme. Women donors in developing regions are often illiterate and unemployed. This study was done to assess the change in quality of life of women who donate kidneys. Methods. We prospectively studied 73 consecutive women volunteering as live-related kidney donors over a 6-month period using the World Health Organization Quality of Life Brief (WHO QoL Bref) Questionnaire and Hospital Anxiety and Depression Scale (HADS). Each woman was interviewed 2 weeks before and 6 months after kidney donation. Results. There was a significant improvement in all the domains, namely physical (p=0.0001), psychological (p<0.0001), social relationship (p=0.037) and environment (p<0.0001) of the WHO QoL Bref questionnaire. Donors who were mothers had a greater improvement in all 4 domains than donors with other relationships. There was a significant decrease in the depression score (p<0.0001), but no change in the anxiety scores (p=0.065) following kidney donation. All donors would donate again, if possible. Conclusion. In live-related women kidney donors, quality of life improves and depression scores decline after kidney donation.

Public health strategies to stem the tide of chronic kidney disease in India.

It is estimated that 10-15% of the population has chronic kidney disease (CKD), resulting in significant health expenditure, which is largely met by out-of-pocket by the patient in India. However, the seriousness of this public health problem has remained largely under-recognized so far. Luckily the preventive measures are simple and not difficult to implement. Public health strategies are essential to control the burgeoning problem. Lifestyle modifications can reduce the incidence of obesity, hypertension and diabetes. These diseases account for a significant proportion of CKD cases. Active involvement of the primary care physicians is vital for early detection of CKD with retardation of its progress, since nephrologists do not see the patients in the early stages. The role of community health specialists is essential to organize health education programs and screening camps, form active patient support groups; and incorporate the prevention program in the various tiers of the health-care system. Availability of optimal nephrology services in peripheral hospitals can rehabilitate most cases of end-stage renal disease, and also prevent the illegal organ trade, which keeps rearing its ugly head at regular intervals in India.

Rare presentation of a common disease of tropics.

A 19 years male presented with fever, oliguria and purpuric lesions involving both hands. The patient was diagnosed as a case of purpura fulminans with disseminated intravascular coagulation due to complicated falciparum malaria. The case is presented to sensitize the physicians to keep malaria as a differential in cases of fever with purpura fulminans.

Purpura fulminans in a complicated Falciparum malaria.

A 19-year-old male presented with fever, oliguria and purpuric lesions involving both hands. The patient was diagnosed as a case of purpura fulminans with disseminated intravascular coagulation due to complicated Falciparum malaria. The case is presented to sensitize the physicians to keep malaria as a differential in cases of fever with purpura fulminans.

The first successful simultaneous pancreas-kidney transplant in India.

Insulin-dependent diabetes mellitus is associated with renal failure, diabetic retinopathy, neuropathy and vasculopathy. We report the first successful simultaneous pancreas-kidney transplant in India in a young diabetic with renal failure. The dual transplant has cured his diabetes and renal failure and has had a beneficial effect on his neuropathy, retinopathy and quality of life. Obstacles to dual transplant in India include a lack of suitable recipients and a cadaver donor programme that is still in its infancy.

Pulmonary mucormycosis in a diabetic renal transplant patient.

A 50-year-old male, diabetic, post transplant patient had recurrent pneumonia. The first episode responded well to antibiotics, while on the second occasion he had a necrotising pneumonia, which developed into a thick-walled cavity. Despite antibiotics, his condition rapidly deteriorated precluding bronchoscopy or percutaneous biopsy. Post-mortem lung biopsy revealed typical hyphae of mucormycosis.

Pneumocystis carinii pneumonia: role of high resolution computed tomography.

Pneumocystis carinii (PC) is a common opportunistic infection in renal transplant recipients and requires an early diagnosis for its successful treatment. For its definitive diagnosis PC needs to be demonstrated in bronchoalveolar lavage (BAL) fluid. At times BAL may not be possible or get delayed. In such conditions typical appearances seen on high resolution CT of the chest help in early diagnosis of PC pneumonia (PCP). This easily performed procedure helped in early diagnosis of PCP in two patients.

Results of renal transplantation on conventional immunosuppression in second decade in India: a single centre experience.

BACKGROUND: In-spite of many renal transplant (RT) centres in the country, there is paucity of data on the survival results in India. Furthermore, there is hardly any data of results in second decade. In this study we present the results of RT in second decade at our centre. METHODS: RT at our hospital is being done since 1972. First eight years saw occasional RT and complete data is not available in these patients. So, we have excluded these patients from the study. We have included rest all 144 patients who had RT between Jan. 1981 to Dec. 1989, so as to have at least 10 years of follow-up or more. All these patients were on conventional immunosuppression. RESULTS: Of the 144 patients, 126 (87.5%) were males and the mean age was 29.5 +/- 8.5 years (range 14-54). Basic disease was presumed chronic glomenulonephritis (CGN) in 79.9%, vesicouretenic reflux (VUR) in 8% and diabetic nephropathy in 1.4% patients. Mean number of haemodialysis (HD) and blood transfusion before RT were 44 +/- 24.7 and 8.9 +/- 4.7 (range 0-25), respectively. Mean donor age was 40.9 +/- 10.1 year (range 18-62). HLA matching was haploidentical in 74.5% cases, HLA identical in 3% cases and 11.5% were less than two-antigen match. Of the 63% patients, who had at least one acute rejection (AR) during their follow-up, 78.8% cases had this AR within first three months after the renal transplant. During 10 years of follow up, 65 patients died (23 with normal graft function and 42 with chronic rejection) and 17 lost to follow-up. Eight patients lost their graft but were surviving at 10 years and 54 had functioning graft till the time of analysis. With mean follow-up of 83.9 +/- 61.2 (range 1-216) months, 10-year actuarial patients and graft survival was 53% and 47%, respectively. Graft half-life in these patients is 92 months. In multivariate analysis of the donor age, number of blood transfusion (BT), degree of HLA matching and AR, only AR was found to be statistically correlated with the graft outcome (p=0.004). CONCLUSION: Our study concludes that in our set-up during eighties, on conventional immunosuppression, actuarial patient and graft survival at 10 year is 53% and 47%, respectively with graft half-life being 92 months. Only acute rejection has been found to affect the graft outcome in these patients.