Indian Journal of Microbiology Research

83-88

Authors: Tina Chaturvedi, Amit Bhattacharya

Background: Several diseases are caused by microbes present in the environment. The respiratory tract is the most common site for infection by pathogens. In this study Aspergillus species were isolated from clinical specimen which is the most prevalent fungi causing Aspergillosis in immuno compromised patients. Drug susceptibility test against the isolated fungi was determined. Method: The specimens were collected and cultured on Sabouraud’s Dextrose Agar (SDA) slants for the isolation of fungi. Antifungal drug susceptibility test was performed using E-test strip on RPMI-1640 media plates. Results: Out of 280 patients 7 patients were found infected with Aspergillus species. Voriconazole was observed to have least MIC value as compared to other drugs. Conclusion: The study reveals that the inhalation of spores of Aspergillus can cause health risk and pulmonary infection (Aspergillosis). This infection usually occurs in those individuals who have weakened immune system. Our study has showed that Voriconazole is a better treatment against Aspergillus infections.

95-105

Authors: V.Y. Shukla, H. G. Pansuriya

Background: Wastewater from various industrial plants such as paper and pulp mills, pigment and textiles can contain various substances that tend to increase chemical oxygen demand (COD) of wastewater. Various agencies like Central Pollution Control Board (CPCB) have placed limits on allowable levels of COD in industrial wastewater effluents. It is desirable to develop a process suitable for treating wastewater to meet the regulatory limits. Objective: The present study was attempted to solve the problem of high COD values as well as concerned heavy metals like Cu+2 and Cr+6 in industrial wastewater using native bacteria. Methods: The organic matter that contributed to high COD in wastewater was degraded by native bacterial consortium under aerobic conditions using batch process. Flask treatment experiment was carried out to optimize inoculum size, temperature, pH and HRT using consortium for effective COD, Cu+2 and Cr+6 removals. Lab scale reactor study was carried out in which bacteria were allowed to attach to a supporting matrix to prevent slough off during treatment. Results: Under the experimental conditions, the reduction efficiencies of COD, Cu+2 and Cr+6 were found to be as high as 57%, 97% and 93% respectively with 72h HRT in fixed film bioreactor. Conclusion: Use of native biological population for the treatment of industrial wastewater is a permissible and cost effective technique.

111-114

Authors: Gayatri Patel

Background: Enzymes are the most important substances used in so many areas either in research, medicine or commonly in industries. Starch degrading bacteria are mostly important in food, texture, fermentation and paper industries. Amylase production from bacteria is economical because the enzyme production rate is higher in bacteria as compared to other microorganism. The isolation and manipulation of pure culture of starch degrading microorganisms from soil have a great importance on biotechnology as well as microbiology field. Methods: Present study involves the isolation by dilution method, characterization and identification by bergey’s menual of systematic bacteriology. Enzyme assays by DNS a method. Optimization of enzymes at different parameters. Results: In this research the bacterial four strains A1, A2, A3 and A4 were isolated from soil at area of college campus Ganpat University, Gujarat, India. Isolated bacteria have great amylase activity at 37˚Ctemperature, the highest hydrolysis zone of starch using with concentration of 1.5% amylase activity at different pH in highest growth of bacteria at pH 6.7 and amylase activity at different time duration at 48hrs with high activity 23.5U/ml in isolate A4 by DNS a method. Conclusions: In the presence study, the bacteria were isolated from the garden soil from the college campus. This area also consisting of starchy materials we found starch degrading bacteria have capacity to grow in physical condition pH, temperature and concentration of starch. As per morphological character, colony character and biochemical testing isolated bacteria similar to genus bacillus.

120-125

Authors: Sarika Kombade, Gopal N. Agrawal

Background and objectives: Nonfermenting gram-negative bacilli (NFGNB) are ubiquitous in the nature. They are considered as environmental contaminants and most of them have emerged as important nosocomial pathogens. So this study was undertaken to identify the various NFGNB with its antimicrobial susceptibility pattern of specimens from patients admitted to Intensive care unit (ICU) in our hospital Methods: Total 107 NFGNB were identified during the study period of September 2009-October 2011. All isolates were identified using a standard conventional method like motility, oxidase production; oxidation-fermentation test etc. and failing to acidify butt in triple sugar iron medium. Antibiotic susceptibility testing was performed with the help of the Kirby-Bauer disc diffusion method and MBL detection done by DDST and Modified Hodge test. Result: P. aeruginosa (16%) was the commonest isolate obtained followed by A. baumannii (13.5%). NFGNB were maximum susceptible to imipenem (80.4%), piperacillin tazobactum (71.9%) and amikacin(52.3%). P.aeruginosa and A. baumannii were 90% and 60.6% sensitive imipenem and 85% and 54.5% to piperacilli-tazobactum. 19.6% MBL producers were obtained in all nonfermenters. Maximum MBL production was obtained in Acinetobacter spp. (39.4%) followed by S. maltophilia (20%) and P. aeruginosa (10.0%) Interpretation & conclusion: Multidrug resistance infection in ICU is emerging; we also obtained 19.6% MBL producers in all NFGNB. Local antibiotic policy should be generated to prevent the emerging MDR in NFGNB.

128-132

Authors: Ranjana Hawaldar, Sadhna Sodani

Bacteremia due to Stenotrophomonas maltophilia – a series of three cases. Stenotrophomonas maltophilia is an aerobic ubiquitous gram-negative bacillus that has been isolated from humans, animals, soil, food, and pharmaceuticals. Because of its limited invasiveness and low level of pathogenicity, it is rarely responsible for community-acquired serious infections. In recent years, S. maltophilia has been increasingly reported as a cause of life-threatening infections, in particular in immune compromised patients. Antimicrobial pressure, indwelling venous catheterization, long-term hospitalization, aggressive. Cytoreductive treatments for malignancy and neutropenia seem to be contributory factors. Management is problematic because of resistance to commonly used antibiotics including those of carbapenem class. In vitro susceptibility testing specific guidelines are not available for management. The development of rapid, inexpensive and reproducible culture systems such as Vitek II has made possible the recognition of this as a cause of bacteremia. In particular more information on nosocomial reservoirs and routes of transmission of the bacterium is needed for effective infection control strategies. Here we report a series of three cases of Stenotrophomonas maltophilia bacteremia from patients admitted in a paediatric ICU.

7-13

Authors: G. S Kocher, Nishu Joshi

Background: The present study was carried out to use potato peel which is available in plenty from food processing industries as a substrate for alkaline protease production using B. circulans MTCC 7906. Method: The potential of potato peel as substrate for alkaline protease has been studied for its production and purification was studied by Bacillus circulansMTCC 7906. In order to make the process economically viable the casein was replaced by Potato peel. The results were analyzed using factorial CRD. Results: Alkaline protease activity was increased by2.74fold and was obtained in 96 h of fermentation. Ammonium sulphate precipitation followed dialysis of the fraction 60-90% saturation with maximum specific activity was partially purified using DEAE column chromatography. Purification profile was studied and the fraction with highest specific and enzyme activity were pooled to achieve specific activity of 18.761 U/mg with 6.185 fold purification. The concentration of enzyme was done using PEG had the specific activity of 69.332 U/mg and 22.859 fold purification. Conclusion: The concentrated partially purified enzyme was subjected to SDS PAGE followed by silver staining for both the substrates casein and potato peel showed the protein band for alkaline protease at 40 kDa revealing the purity of the enzyme. Experiments revealed successful utilization of potato peel as an economical substrate for enhanced production of alkaline protease by B. circulans MTCC 7906. Projects which are inclined to reduce global wastage should be encouraged and this is one of the same type.

20-26

Authors: Sandip Bandopadhyay

Purpose: Purpose of the present study is to check the potency of the strains Bacillus thuringiensis A5-BRSC and Bacillus megaterium ATCC 9885 for promoting growth of different plants without hazardous chemical fertilizers. Methods: Four non-leguminous seasonal plants like Amaranthus; Ocimum, Abelmoschus and Capsicum are used to study combined growth promoting effects of two bacteria viz. Bacillus thuringiensis A5-BRSC and Bacillus megaterium ATCC 9885 under pot condition. Different morphological parameters like shoot height, leaf area, fresh weight and dry weights of the plants; as well as biochemical parameters like leaf chlorophyll content, catalase and peroxidae enzyme activities of both culture treated and untreated plants were investigated. Results: All the biofertilizer treated plants showed remarkably higher results in all parameters, except chlorophyll content, than control plants. Conclusion: The strains proved as potential strains for plant growth promotion in pot study and hence may be used as biofertizer under field condition for sustainable agriculture.

30-39

Authors: Animesh Samanta, Shrabani Pradhan, Arpita Mandal, Arpita Patra, Suchismita Roy, Shreya Mandal,

Background: Curd is an important fermented food which is generally consumed by the community people in large scale and becomes a functional upon incorporating probiotics-live microorganisms due to a great variety of positive health effects. Methods: In this study, pure cultures of Lactobacillus casei (MTCC1408), L fermentum (MTCC 903), L rhamnosus (MTCC1462), L ingluviei ADK10 (GenBank Accession No-JQ395039), Enterococcus faecium ADK18 (KF032592.1), E durans ADK14 (KF032593.1) and Streptococcus thermophillus (MTCC 1938) were inoculated to pasteurized Amul Milk separately to produce curd. Different physiochemical and antimicrobial characteristic of curd against some pathogen were analyzed to assay the quality of curd. Results: The results of the study revealed that the moisture content of the curd samples ranged between 86.36%- 88.71% and the pH values of the samples ranged between 3.96-5.43, which were reasonably suitable for curd processing industries in tropical countries. Titratable acidity (0.81-1.71), protein (range 1.4g-9.6g) and fat (2.2g-6.8 g) content also varied with respect to different lactic acid bacterial strains. All seven lactic acid bacteria (LAB) strains showed their antagonistic activity (zone of inhibition 5-10 mm) against four enteropathogenic bacterial strains (Escherichia coli ATCC 25922, Klebsiella pneumonia ATCC 15380, Streptococcus mutans ATCC 25175, Bacillus subtilis ATCC 6633). L ingluviei ADK10 and L. rhamnosus strains showed wide inhibitory spectrum against all the tested strains. Conclusions: It can be conclude that the physiochemical parameters were varies according to their starter culture with antimicrobial activity against some entero-pathogens and has been fruitful starter culture for the preparation of curd having better health promoting effects.

46-49

Authors: Madhurendra Singh Rajput, Pradeep Kumar, Ramesh Chandra thanna

Background: Psudomonas aeruginosa is a major cause of nosocomial infections. Despite advances in sanitation facilities and the introduction of wide variety of antimicrobial agents with antipseudomonal activities, life threatening infections caused by this agent continue to cause devastations in the hospitals. Aims & objective: To study the Resistance Pattern of Pseudomonas Aeruginosa Isolates From surgical wounds. Material & Method: This study was a retrospective study done in Department of Microbiology, Index Medical College hospital and research center, Indore, m.p, india from March 2014 to december 2014. A total of 300 isolates of Pseudomonas aeruginosa both from indoor patients and patients attending outpatient department who were having surgical wound infections, were included in the study. Each isolate was evaluated for susceptibility to nine different antibiotics i.e cefotaxime, ceftriaxone, ceftazidime, cefdinir, amikacin, gentamicin, ciprofloxacin, piperacillin and imipenem. ESBL production was detected by double disc potentiation method by applying disc of cefoperazone [75μ] and combination of cefoperazone-sulbactam [75/30μ]. The results were interpreted according to Clinical Laboratory Standard Institute (CLSI) guidelines. Result: Maximum resistance was seen to third generation cephalosporins-68.5 % to cefotaxime, 80.23% to ceftriaxone, 75.5% to ceftazidime, 93% to cefdinir. Amikacin showed resistance in 42.5% and Gentamicin in 81% of the isolates. Ciprofloxacin resistance was seen in 72.5% isolates while piperacillin resistance was seen in 43% of the isolates. Minimum resistance was seen to imipenem -2.5% Conclusion: The present study highlights that the Pseudomonas aeruginosa remains an important cause of nosocomial wound infections. Is study thus gives the alarming signal for the future, making the therapeutic options more difficult. Strict infection control measures are to be followed to contain the so called water and soil organism as Pseudomonas aeruginosa.

55-59

Authors: Varjani Sunita Patel Kinjal

Methane (CH4), a potent greenhouse gas (GHG) with a global warming potential (GWP) ~23 times higher than that of carbon dioxide (CO2) is produced by Landfills. There is urgent need to prevent methane emissions from large landfills. Methanotrophs are a group of bacteria that consume methane. In cover soil methanotrophs prevent fugitive methane emissions. In present scenario two general research techniques are performed to prevent methane emissions. First, a dimensionless number method developed based on Michaelis-Menten kinetics. Second, effects of nutrient amendments on methane oxidation and nitrous oxide production were examined by constructing soil microcosms using landfill cover soils. Mehanotrophic activity and community structure can be differentially affected by both landfill gas composition and amendments thus provides insights for best manipulation of methanotrophic processes to better mitigation of GHG emission.

64-71

Authors: Annu Goel, Priyanka Batra, Leela Wati

Ebola virus disease (EVD) is a rare severe disease, often fatal, caused by the Ebola virus. It is transmitted through direct contact with blood or other bodily fluids (e.g. saliva, urine) from infected people, dead or alive. This includes unprotected sexual contact with patients up to seven weeks after they have recovered. Ebola virus does not transmit through the air as influenza does. After two days and up to 21 days following exposure to the virus, disease may start suddenly with fever, muscle aches, weakness, headache and sore throat. The next stage of disease is characterised by vomiting, diarrhoea, rash and failure of the liver and kidneys. Some patients also have heavy internal and external bleeding and multi-organ failure. There is no specific vaccine or treatment for the disease, but World Health Organization and other regulatory partners are currently working to identify potential viable treatments. Key Words: Virus, Ebola, Diagnosis, Therapy

4-13

Authors: Vijay N. Charde1,*, Shweta M. Bhiwankar2, Pratik M. Bezalwar3

Background: Pseudomonas aeruginosa has become an important cause of infection, especially in patients with compromised host defense mechanisms. Thus, it is a frequent cause of nosocomial infections. Development of drug resistance in Pseudomonas is worldwide problem and tough challenge to medical practitioners; Methods: Present study involves the isolation of Pseudomonas from the patients suffering from the ailments like respiratory tract infection, UTI, etc. and studying the antibiotic resistant pattern of these isolates and synergistic action of cold and hot water and methanol extracts of Cinnamomum zeylanicum on the activity of antibiotics by agar diffusion method; Results: Out of 28 isolates of Pseudomonas isolates five isolates exhibiting wide resistance pattern were selected for study. The results showed synergistic effect of Cinnamomum zeylanicum on the antibiotics which are ineffective against the Pseudomonas isolates. Cold methanol and cold water extract showed good synergistic effect with antibiotics AK, CTX, CAZ, PI, G, CPZ, CPM, CB, NET, TCC, CTR, IPM, AT & AZ against Isolate P-19 making it susceptible to these antibiotics. Hot extract of both methanol and water showed no synergistic effect against Isolate P-19. All the extracts of Cinnamomum zeylanicum have shown no significant synergistic effect on the antibiotics against Isolate P-3, P-4, P-20 and P-26; Conclusions: Cinnamom extract is effective in combination with antibiotic in dealing the drug resistance problem of Pseudomonas and such synergy is important in exploring the new options for drug resistance problem of many pathogens.

37-45

Authors: Sodani Sadhna, Hawaldar Ranjana.

Background: Growing multiple drug resistance in microorganisms is of great importance and so is the knowledge about the current drug susceptibility pattern for deciding the empirical usage of antibiotics. E.coli is one of the commensals in the human intestinal tract and contributes to the maintenance of Health of the person. However when E.coli enters into unnatural sites, it can cause variety of infections, bacteremia, meningitis and other soft tissue infections. The emergence of antibiotic resistance in Ecoli is threatening the effectiveness of many antibiotics which has resulted in increased hospital stay of patients, which in turn causes economic burden. Extended spectrum Beta lactamase (ESBL) producing E.coli has tremendously increased worldwide and it is one of the most common causes of morbidity and mortality associated with hospital acquired infections. Objective: The present retrospective study highlights the susceptibility pattern of E.coli in clinical specimens obtained at Sampurna Sodani Daignostic Clinic Indore (MP) Microbiology department for the year 2014 from January to July. Materials and methods: This was a retrospective study of ESBL producing E.coli in different clinical specimens including urine, blood, pus, vaginal swab, stool, semen, aural swab, BAL fluid and conjuctival swab obtained from out patients at Sampurna Sodani Diagnostic Clinic Microbiology department located in Indore (Madhya Pradesh) from January to July 2014 and reported in fully automated Microscan (Siemens) according to CLSI guidelines. A total of 542 samples were reviewed. A total of 29 antibiotics were assessed for the resistance pattern in E.coli isolates. Results: Out of 542 E.coli isolates grown in the lab from urine, blood, pus, vaginal swab, stool, aural swab, BAL fluid, and conjunctival swab. 420 isolates showed sensitivity to imipenem, amikacin, meropenem, and piperacillin/tazobactum in more than 70 % cases. Conclusion: ESBLs are enzymes that mediate resistance to extended spectrum Beta lactamase (third generation) cephalosporins (eg. Ceftazidime, cefotaxime, & ceftriaxone) and monobactums (eg. Aztreonam) but do not affect cephamycins (eg.cefoxitin) or carbapenems (eg.meropenem or imipenem). 40. 95% isolates of E.coli were ESBL. Among oral antibiotics highest sensitivity was observed to nitrofurantoin (68.33 %) and Amoxy/ Clavulinic acid (44.8%) which can be the drug of choice for E.coli infections specially in UTI. Injectable drugs like Imipenem (90.4%) and Amikacin showed sensitivity in 84.87% isolates but being injectable drugs, should be used with caution.

52-55

Authors: C. Sundaravadivelan, P. Kumar, J. Anburaj, T. Kuberan, S. Sivasubramanian.

Tea is one of the major nonalcoholic beverages in the world. Being a perennial crop, it provides a stable environment for a number of pests and diseases. Pests are important factors limiting the productivity and quality of processed tea. Among the pests, the shot hole borer (SHB), Euwallacea fornicatus (Eichhoff) is a serious pest of tea in south India. These beetles have a symbiotic relationship with the fungus Fusarium bugnicourtii. The fungal spores are carried by the beetles in special organs called mycangia located in the buccal cavity of head. Spores of the ambrosia fungus borne by the female beetles adhere to the walls of the stem galleries. On germination of spores, the grubs and adults feed on the fungus. Hence a novel approach has been attempted to find out the efficacy of pesticides on the fungus, F. bugnicourtii. Results showed 50 % reduction of fungi while tested with Dicofol, Endosulfan, Quinalphos and Lambdacyhalothrin. The high significance was observed in Dicofol and Endosulfan, and no significance was recorded in Lime sulphur, Ethion and control treatments at 0.05 %.

60-71

Authors: Souvik Nath and Abul Kalam.

Biological diversity - or biodiversity - is a term we use to describe the variety of life on Earth. Microbes are one of the dominant life forms in the earth. Their contribution to the earth and human being is beyond the imagination. Their science is concerned with their form, structure, reproduction, physiology, metabolism and classification. It also includes their distribution in nature, their relationship with each other and other living organisms, their effects on human beings, other animals and plants. Biological diversity (biodiversity) encompasses the variety of life forms occurring in nature, from the ecosystem to the genetic level, as a result of evolutionary history (Wilson 1992). Microbial Diversity is an integral part of biodiversity which includes bacteria, archaea, fungi, algae, protozoa and protests. Fungi constitute a major portion of natural resources likely to provide innovative applications useful to man. Fungi are one of the major sources of antimicrobial agents and produce a wide range of other important medicinal compounds, industrially important biomolecules, novel enzymes, insecticides the microbial level is beginning to be recognized, but this richness of diversity amongst bacterial, fungal and virus species has yet to be catalogued particularly in West Bengal. The East Kolkata Wetland (EKW) is situated at 880 20’ E - 880 35’ E and200 25’ N -200 35’ N. Climate here is more or less sub-tropical with the annual mean rainfall around 200 cm. The maximum temperature during summer rises around 390 C. while minimum temperature during winter is around 100 C. The average temperature during most part of the year is around 300 C during day time with a fall in temperature of 50-60C at night. East Kolkata Wetland shows an immense diversity of flora and fauna both at the macro and micro level. Microbial richness of a region is its unseen asset that needs to be explored and conserved. Soil samples collected from East Calcutta Wetland shows the presence of various new strains of microbes which are not only ecologically important but also have commercial value. Isolation, characterization, documentation and conservation of these resources are important considering their strategic importance for future generation as well as complimentary economic growth and prosperity. In this present work several fungi were isolated and purified from diverse area of East Calcutta Wetland out of which about 10 organisms was identified by microscopic studies. Among the isolates it is expected that one or two new genus may obtained.

77-83

Authors: Supriya Christopher, P M Giridhara Upadyaya.

Background: Brucella sps are Gram negative, facultative intracellular bacteria causing brucellosis in humans and animals. Since it is a multi-organ disease and symptoms are nonspecific the diagnosis by clinical finding is difficult and can be missed .Many serological tests have been used form the diagnosis of human brucellosis. This study compared the diagnostic value of in-house ELISA and IFA using lipopolysaccharide heat extracted antigen (LPS-HE) and Lipopolysaccharide sonicated extracted antigen (LPS- SE) with STT. Methods: The present study was carried out to evaluate the two different antigenic preparations from smooth strains of Brucella abortus S99 for standardising ELISA and IFA as the alternative test for STT. Standard Tube test antigen and Standard anti brucella serum was obtained from IVRI, Izatnagar were used as controls for standardizing ELISA and IFA. 81 human sera (cases) were collected from people working in organized farms including veterinary staff, 32 animal sera from organized farms mentioned above, 100 human sera (controls) collected from KIMS, Bangalore Blood bank. 100 WIDAL positive samples from Department of Microbiology used to check for cross reactivity. All the serum samples (cases, controls and WIDAL positive samples) were tested with STT. Result: Out of 81 human sera 8 (9.87%) was found to be positive with SAT showing a titre of ≥ 1:80. Whereas by ELISA, 10 (12.34%) & 9 (11.11%) cases showed positive in LPS –SE and LPS –HE coated plates respectively. The sensitivity and specificity for IFA was 77 and 87.5. Conclusion: It was found that ELISA was considered to be better test over IFA and STT. It is cheap and reproducible and a specific assay for the diagnosis of Brucellosis.

91-94

Authors: Vijay Prabha, Harpreet Vander, Sonia Gupta.

Contraception is a well-known conventional method for stabilizing the world population. Traditional contraceptive techniques that have been used for several years are associated with disadvantages and side effects. There is urgent need of improvements in prevailing birth control methods available to us. Various vaginal contraceptive agents marketed today are chemical or detergent based that are accompanied by numerous negative effects on vaginal flora and epithelium. Therefore, novel mode for birth control that is attaining limelight at the present time is the use of microbial products.