Heavy metal, salinity and azo dye tolerant, Cr (VI) reducing, plant growth-promoting Pseudomonas aeruginosa R32 reverses Cr (VI) biotoxic effects in Vigna mungo

Hexavalent chromium [Cr (VI)], derived from various industries, including fly ash from coal-based Thermal Power Plants, can be a source of toxic pollution of land and water bodies. This study aimed to bioremediation of such pollutant dump sites using bacteria capable of both Cr(VI) reduction and plant growth-enhancing substance production. The bacteria were isolated from the rhizospheric fly ash of a Thermal Power Plant, Kanpur. One of the rhizospheric isolate, Pseudomonas aeruginosa R32 showed high minimum inhibitory concentration (MIC) for Cr(VI) (1250 µg/ml), heavy metal tolerance (ZnCl2, CdCl2, Pb(NO3)2) up to 100 µg/ml, Acid Red 249 (AR) tolerance and halotolerance (6% NaCl). The isolate R32 also produces plant growth-promoting (PGP) hormones in the absence or presence of Cr (VI). R32 could completely reduce Cr(VI) at a tested dose of 100 and 500 μg/ml after 24h and 72h, respectively. However, decolorization of AR was observed after 48 hours at an initial concentration of 100 µg/ml and confirmed by Fourier transform infrared spectroscopy analysis. Vigna mungo seed inoculation with isolate R32 showed increased rootling growth compared to shoot after 7 d treatment with 0, 100, 500, and 1000 μg/ml of Cr(VI) concentrations, respectively. Root length tolerance index in Cr(VI) treated V. mungo seedlings was reduced to 56%, 35%, and 29%, respectively, when treated with 100, 500, and 1000 μg/ml Cr(VI) in comparison to control. Cr(VI) sub-MIC concentrations can affect the plant growth-promoting properties of rhizospheric bacteria. Herein, we report the isolation of rhizospheric bacteria P. aeruginosa R32 showing concurrent PGP substance production and Cr(VI) bioreduction capabilities in the presence of PGP inhibitory Cr(VI) concentrations.

Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus

Azadirachta indica Juss (Neem) is well documented for its antimicrobial activity. The effect of varying concentrations (0.1 to 50% v/v) of Azadirachta indica derived neem seed oil (NSO), neem seed oil with tween 20 and neem bark extract was evaluated on planktonic, biofilm formation and mature biofilms of multiple drug resistant Pseudomonas aeruginosa ATCC 15442 and Staphylococcus aureus ATCC 25923 using the crystal violet assay and scanning electron microscopy. NSO showed antimicrobial activity at 25% v/v for P. aeruginosa but not S. aureus in zone of inhibition assay. Neem bark extract on the contrary showed antimicrobial activity against both the isolates at 50% v/v concentrations. Interestingly, in biofilm formation assay, low concentrations of NSO (3.5 to 0.2% v/v) induced biofilm formation while inhibition of both planktonic and biofilm was seen in concentration dependent manner from 12.5% v/v onwards. Complex of NSO and tween in comparison of NSO alone caused low induction in S.aureus biofilm formation, while inhibiting biofilm formation of P. aeruginosa at all the concentrations. In biofilm eradication assay, NSO induced biofilm of both P. aeruginosa (50 to 0.1%v/v) and S. aureus (50 to 3.13%v/v). Eradication effect of neem bark extract was found on P. aeruginosa biofilm in a dose dependent fashion from 50 to 20% v/v followed by 0.2 to 0.1%v/v concentration respectively. S. aureus biofilm were eradicated at 50 to 25%v/v concentrations. At low concentrations, both the neem derivatives induced biofilm mediated growth of the pathogenic organisms. The data also indicate that neem seed oil was more effective against Gram negative P. aeruginosa while neem bark extract was effective against Gram positive S. aureus. This study highlights the crucial but variable effects of concentration dependent effect of phytochemicals and their composition on biofilm induction as well as eradication, the primary growth form in clinical settings. This challenges the notion that all herbal products are safe as antimicrobial activities differ as per microbial growth modes. Hence, concentration dependent effect of medicinal plant derived products requires thorough investigation prior to their use as antimicrobial agents

MOLECULAR DETECTION OF HUMAN RHINOVIRUS IN RESPIRATORY SAMPLES OF SWINE FLU NEGATIVE NORTH INDIAN CHILDREN WITH FLU-LIKE ILLNESS

Objectives: Flu-like illness may also be caused by different respiratory viruses other than influenza. Human rhinovirus (HRV) shows almost flu-likesymptoms. The purpose of this study is the molecular detection of HRV in throat swab of swine flu negative North Indian children during the years2012 and 2013. Reverse transcriptase (RT) - polymerase chain reaction (PCR) amplification of 5'non-coding region (NCR) was used for HRV detectionfollowed by cell culture isolation of HRV.Methods: PCR confirmed swine flu negative throat swab samples were collected from the Department of Microbiology, Sanjay Gandhi Post GraduateInstitute of Medical Sciences, Lucknow, Uttar Pradesh, India. The RNA isolation of samples was done using the QIAampViral RNA Mini Kit (Qiagen),followed by single step RT-PCR amplification (AgPath-ID, Life Technologies). All PCR positive HRV samples were cell cultured in HeLa and HEp-2 celllines for viral isolation.®Results: 135 swine flu negative throat swab samples were examined. Out of which 34 samples (25.2%) were found HRV positive by RT-PCR, while onlyfour samples (11.8%) were culture positive on HeLa cell line. Younger children (0-4 year) were found more susceptible to HRV infection. This studyindicated the highest prevalence of HRV (37.0%) during the months (September-October) of the Autumn season in 2012 and 57% in Winter-springseason (February-March) during 2013.Conclusion: HRV may be a cause of flu-like symptoms in swine flu suspected North Indian children with a higher rate during Autumn and Springseason. Molecular detection of HRV using RT-PCR is more sensitive than cell culture assay.Keywords: Human rhinovirus, Swine flu, Influenza-like illness, Lower respiratory tract infections

A study of factors influencing decisions on organ donation among patient attendees in a Tertiary Care Hospital in North India

Introduction: Organ transplantation is the most preferred treatment modality for end-stage organ disease and organ failures as it improves the quality of life. Patients deemed fit for transplantation by the transplant team often wait expectantly for a donor organ. Each year thousands of individuals wait for organs to be donated for transplantation. There remains, therefore, a great need for increasing organ donation and harnessing of donor organs. Aims and Objectives: To determine knowledge, attitude, and practices of the general population for organ donation. To determine the factors which facilitate or inhibit general population from organ donation. Methodology: A cross-sectional survey was conducted among a total of 120 patient attendants attending outpatient department in the age group of 18–65 years at Sir Ganga Ram Hospital, New Delhi. Results: One hundred and twenty participants were enrolled in the study, but only 119 participated. The mean age of males was 37.7 ± 14.7 years while that of females was 32.7 ± 11.5. Maximum awareness was found to be for kidney donation, least awareness for ear drums, and intestine donation. About 49.1% males and 51.6% females were aware of the legislation related to organ transplantation. The majority of female respondents (73.3%) showed their willingness to donate their organs in contrast to males only 16.7% were ready to donate in any situation. Females (60.7%) outnumbered males (45.8%) in their readiness to sign an organ donor card. About 73.3% females and 64.4% males were ready to participate in any promotion campaign related to organ donation. Conclusion: The magnitude of organ retrieval for the execution of a successful donor program is heavily dependent upon the level of knowledge and attitudes of the general public, so the need of hour is to increase the awareness among masses

INHIBITION OF DIFFERENT STAGES OF BIOFILM OF PSEUDOMONAS AERUGINOSA PA01 BY ISOLATED BACTERIOPHAGE P2

 Biofilm mediated protection of pathogens from environmental stresses are recognized as one of the major threat worldwide, which encourage theresearcher to search for other alternative approach. Bacteriophage therapy is one such effective and controlled strategy that undertakes the utilizationof tailored phage mediated biofilm inhibition. Hence, in this study, P2 phage, specific to Pseudomonas aeruginosa PA01 was isolated and its potentialto inhibit biofilm formation was studied. Important findings are summarized in the manuscript.Keywords: Pseudomonas aeruginosa, Biofilm, Bacteriophage and Transmission electron microscopy, Antibiotic resistance

High Prevalence of Multiple Drug Resistant and Biofilm Forming Staphylococcus aureus among HIVInfected Patients with Suspected Pneumonia

Staphylococcus aureus is one of the major causes of life threatening pneumonia, especially in immunocompromised population. In HIV positive patients, S. aureus associated pneumonia can be either health care associated or community acquired and responsible for high rate of mortality. In this study total 102 throat swab samples of HIVInfected Patients with suspected pneumonia were collected during 2014-2016, out of them 46 samples (45.1%) were found positive for S. aureus by biochemical tests. 38 (82.6%) isolates were found multiple drug resistant while 9 (19.6%) strains showed resistance to cefoxitin antibiotic, were considered as methicillin resistant Staphylococcus aureus (MRSA). Only one strain (2%) was found vancomycin intermediate (VISA), remaining 98% isolates were sensitive to vancomycin antibiotic. In PCR test, all cefoxitin resistant strains were found positive for the presence of MecA gene. Biofilm former S. aureus were screened by tissue culture plat (TCP) methods. In TCP assay, 21 (45.6 %) isolates were confirmed as high biofilm formers (OD value > 0.250), 16 (34.8 %) were moderate biofilm formers (OD values- between 0.150 to 0.250), while 9 (19.6 %) were low biofilm formers (OD value < 0.150). A significant association was found among multiple drug resistance and high biofilm formation (p value < 0.05). High prevalence of biofilm forming MDR isolates in airways of pneumonia suspected HIV patientsis matter of great concern as poor antibiotic response may cause more severe diseases with increasing cost and duration of treatment. The MecA gene might be a cause of methicillin resistance among MRSA isolates