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Cellulolytic enzymes have gained considerable importance as potential candidates for the efficient bioconversion of agricultural biomass into second-generation biofuel. This study aimed to improve cellulase production from a bacterial strain isolated from cow dung. The biochemical and molecular identification was performed, and the capacity of the isolated bacterium to hydrolyze sugarcane bagasse was also determined. Response surface methodology (RSM) and one-variable-at-a-time (OVAT) designs were employed to maximize the cellulase production. The bacterium identified as Bacillus subtilis gave the highest cellulase yield at temperature of 45 °C, pH-7.0, and 72 h of incubation time. Optimization by the Plackett–Burman design consisting of 12 experimental runs at two levels of nine independent variables displayed the significant contribution of galactose (22%) and sodium nitrate (16%) in cellulase production. The responses in the form of contour and 3D plots (generated by Box-Behnken design) predicted maximum cellulase production with an activity of 0.8 U/mL/min in a medium containing 3.0 g/L carboxymethyl cellulose, 3.0 g/L galactose, 0.5 g/L sodium nitrate, 0.75 g/L KH2PO4, and 0.375 g/L MgSO4·7H2O. The experimentally deduced activity value was 0.698 U/mL/min, showing 90% validity of the predicted value and gave a threefold increase in cellulase production. The process adequately represented by the quadratic model (p < 0.0343) for all the responses was significant. Results obtained show that optimization of fermentation medium favored the increased production of cellulase by the bacterium. The bacterial isolate B. subtilis CD001 was found to be a potential candidate to hydrolyze the lignocellulosic feedstock, sugarcane bagasse.

Background: Aspergillosis is one of the most common fungal infection among both general patients and immunocompromised individuals. Methods: The study was carried out on 2 groups of patients: immunocompetent and immunocompromised comprising of a total of 90 patients. Microscopy, culture, identification of isolates was done and some specialised tests on serum and BAL for antigen detection were performed. Result: Most of the patients i.e., 47 (31.3%) were between 31-40 years with a mean age of 32.5 years. The male to female ratio was 1.8:1. Galactomannan antigen was positive in 13 (28.9%) BAL samples and in 15 (33.3%) serum samples wherein the 45 BAL samples represented immunocompetent cases and 45 serum samples represented immunocompromised cases. Sensitivity and specificity of GM assay in BAL samples were found to be 100% and 97% respectively. On the other hand, sensitivity and specificity in serum samples were found to be 94.1% and 100% respectively. All patients were categorized into 4 categories as proven IPA, probable IPA, possible IPA and non-IPA. 22 (24.4%) patients were of proven invasive pulmonary aspergillosis (IPA), 7 (7.8%) of probable IPA, 2 (2.2%) belonged to possible IPA while 59 (65.5%) were of non IPA. All of the cases of proven IPA were positive on culture. Conclusion: GM assay in BAL is a very sensitive and specific marker for Aspergillosis while in case of serum samples it is somewhat lesser sensitive.

Background: Scrub typhus is lesser known cause of fever of unknown origin in India. Even if there have been reports documenting the prevalence of scrub typhus in different parts of India, it is still an unknown entity, and clinicians usually do not consider it as differential diagnosis. The present study was performed to document the prevalence of scrub typhus among febrile patients in western part of Uttar Pradesh and to assess the clinical profile of infected patients on the one hand and knowledge, attitude, and practices among clinicians on the other. Materials and Methods: A total of 357 adult patients with fever of more than 5-day duration were recruited. All patients underwent complete physical examination, and detailed clinical history was elicited as per predesigned pro forma. After primary screening to rule out malaria, enteric fever, and leptospirosis infection, secondary screening for scrub typhus was done by rapid screen test and IgM ELISA. Results: Scrub typhus infection was positive in 91 (25.5%) cases. The most common symptoms among the patients were fever (100%), pain in abdomen (79.1%), pedal edema 56 (61.5%), rash 44 (48.3%), headache 44 (48.3%), vomiting 42 (46.1%), constipation 33 (36.2%), cough 28 (30.7%), and lymphadenopathy 20 (21.9%). The median values of interleukin-8, interferon-gamma, and tumor necrosis factor-alpha in healthy controls were 15.54 pg/ml, 7.77 pg/ml, and 54.1 pg/ml, respectively, while the median values of these cytokines in scrub typhus-positive patients were 21.04 pg/ml, 8.74 pg/ml, and 73.8 pg/ml, respectively. Conclusion: Our results highlight that scrub typhus infection is an important cause of pyrexia of unknown origin, and active surveillance is necessary to assess the exact magnitude and distribution of the disease.

Introduction: Invasive fungal infections are increasingly common in the nosocomial setting. Materials and Methods: The patients were divided into two groups immunocompetent and immunocompromised that is, patients with significant neutropenia

The occurrence of microbial communities harboring antibiotic resistance bacteria and antibiotic resistance genes in the drinking water distribution system pose a significant threat to the aquatic ecosystem and to public health, especially in developing countries. In this study, we have used next-generation sequencing technology to explore bacterial community diversity and the abundance of antibiotic resistance genes in biofilms collected from the drinking water distribution system of Peshawar, the capital city of the Khyber Pakhtunkhwa province of Pakistan. The results showed that Proteobacteria were the most abundant phyla (89.79%) in all biofilm samples, followed by Bacteroidetes (3.48%) and Actinobacteria (2.79%). At genus level, Pseudomonas was the most common (22.45%) in all biofilm samples. Overall, bacterial diversity and richness was higher in biofilm samples collected from the consumer end than the source site. Bacterial diversity was also dependent on the piping material (GI vs. PVC) and water supply (direct vs. indirect). Functional annotation reveals a differential abundance of common metabolic pathways at source and consumer end. Resistome analysis revealed a prevalence of resistance genes against 12 classes of antibiotics in all samples with macrolides resistance being the commonest at the consumer end (42.1%) and fluoroquinolone resistance at the source end (24%). To our knowledge, this is the first study that provides new insight and evidence into the microbial community diversity and antibiotic resistance in the drinking water supply system of Peshawar. These findings may ultimately help the authorities to design and implement effective strategies for controlling biofilms and ensuring a continuous supply of safe drinking water to the community.

Nanoparticles (NPs) prepared through safer, eco-friendly and natural-based products can be used various applications including antimicrobial and biomedical applications. Quorum sensing (QS) is a complex system for intercellular communication that regulates the expression of many virulence determinants, in the opportunistic Pseudomonas aeruginosa. Thus, the disruption of QS-mediated communication system could be an alternative strategy to combat infection caused by drug resistant P. aeruginosa. In this study, zinc oxide nanoparticles (ZnO NPs) were synthesized by using seed extract of Butea monosperma and further characterized by XRD, SEM and TEM. The biosynthesized ZnO NPs were then subjected to the investigation of their antibacterial, antibiofilm activities as well as the anti-quorum sensing potentialities for biomedical applications. The minimum inhibitory concentration (MIC) value against P. aeruginosa was found 1600 µg/ml, and it was found that ZnO NPs effectively reduced virulence factors Viz. swarming and swimming of P. aeruginosa by 62.8 and 45%, respectively. Exopolysaccharides production was also affected by ZnO NPs which indicate the disruption of biofilm formation. Further, the in silico docking analysis displayed that ZnO NPs efficiently interacted within QS (Las/Rhl) mechanism of P. aeruginosa by binding to the catalytic cleft of LasI synthase (Gly-116-ZnO = 2.8 A), RhlI synthase (Gly-180-ZnO = 3.7 A) and transcription receptor protein LasR receptor (Asn-141-ZnO = 2.9 A), RhlR receptor (Tyr-72-ZnO = 2.6 A). Thus, it was concluded that the effective interaction of ZnO NPs with Las and Rhl system led to the inactivation of autoinducers such as N-acyl homoserine lactones (AHL) molecules which finally led to the inactivation of QS and down regulation of virulence determinants. Consequently, the present in vitro and in silico results obtained suggest that the synthesized ZnO NPs can be efficiently used as potent antimicrobial agents to prevent the colonization, biofilm formation as well as anti-QS-mediated determinants caused by pathogenic drug resistant bacteria.

INTRODUCTION: This study was conducted to assess the prevalence of metallo-beta-lactamases (MBLs) in general and blaNDM-1 in particular. It also aimed at evaluating clinical characteristics and outcome in patients infected with MBLs. MATERIALS AND METHODS: A total of 116 carbapenem-resistant Gram-negative bacilli (CRGNB) were evaluated in the study. These CRGNB were tested for MBL production both phenotypically for MBLs and genotypically for blaNDM-1 gene by polymerase chain reaction (PCR). Representative stains of NDM-1 isolates were further sequenced by Triyat Scientific Co., (Nagpur, India). RESULTS: Among 116 CRGNB Citrobacter species 28 (24.13%) was the most common pathogen. Phenotypically, MHT, imipenem-EDTA (IPM-EDTA) double‑disk synergy test and IPM-EDTA combined disk synergy test (CDST) detected MBL production in 105 (90.51%), 96 (81.03%), and 87 (75%) CRGNB, respectively. However, blaNDM-1 genes were detected in 66 (56.89%) isolates. The prevalence of blaNDM-1 gene was highest among Escherichia coli 26 (100%). Considering PCR as gold standard, it was observed that IMP-EDTA CDST was most specific (78.38%) while MHT was most sensitive (97.47%). Results of blaNDM-1 gene by PCR were further confirmed by sequencing (Triyat genomics, Nagpur). All the 11 representative strains were confirmed to be an NDM-1 gene. The presence of MBLs in our group of patients (non-Intensive Care Unit patients) is a cause for concern. However, on tracing their outcome, it was interesting to note that while the duration of stay lengthened in a large number of patients 112 (96.5%), mortality was relatively low 5 (4.31%). CONCLUSION: The results of this study provide insight into the prevalence of MBLs, including blaNDM-1, in a tertiary care hospital. Antibiotic stewardship implemented in all seriousness may to a great extent stave off the impending pan-drug resistance. The surprising outcome of our patients suggests either that the bacteria trade off virulence for drug resistance or the relatively robust immune response of non ICU patients fights back.

Background: Cefotaximases (CTX-M), temoniera (TEM), and sulfhydryl variable (SHV) constitute a rapidly growing cluster of enzymes that have disseminated geographically. They are spreading to species other than Enterobacteriaceae and might be responsible for the presence of blaCTX-M,blaTEM, and blaSHVgenes in Pseudomonas and Acinetobacter spp. The present study was designed to characterize CTX-M, TEM, and SHV phenotypically and genotypically in Pseudomonas and Acinetobacter spp. Materials and Methods: A total of 90 isolates (73 Pseudomonas and 17 Acinetobacter spp.), resistant to any of the third-generation cephalosporins, were randomly selected from clinical samples. Results: Of 90 isolates, 64 (71.11%) were tested positive for extended-spectrum β-lactamase (ESBL) production. Among phenotypically tested ESBL producers, forty isolates were randomly selected for molecular characterization. The prevalence of CTX-M, TEM, and SHV was found to be 57.5%, 15%, and 75%, respectively. Multiplex polymerase chain reaction assay categorized blaCTX-Mgenes into Groups 1 and 26 where Group 1 was present in only 5 isolates and Group 25 was present in rest of the 18 isolates. Conclusion: This is among the premier systemic reports from India documenting phenotypic and molecular characterization of CTX-M, TEM, and SHV β-lactamases in Pseudomonas and Acinetobacter spp. With judicious use of antibiotics and strict infection control procedures, it may be possible to limit the effects of these newer β-lactamases.

Introduction: Urinary tract infection (UTI) continues to be the most common infection diagnosed in outpatients as well as in hospitalized patients. Citrobacter spp. is an emerging urinary pathogen. The present study assessed the prevalence and antibiotic sensitivity pattern of Citrobacter spp. in patients admitted to or attending outpatient departments with Complain of UTI in a tertiary care hospital. Materials and Methods: A total of 36,250 urine samples were included in the study. UTI was confirmed in 7099 samples. Of these 246 (3.46%) had UTI due to Citrobacter spp. Identification was by conventional biochemical methods. Antimicrobial susceptibility testing was performed using Kirby-Bauer disc diffusion method as recommended by Clinical and Laboratory Standards Institute M2-A9. Multidrug-resistant (MDR) was defined as resistance to more than two groups of drugs. Extended spectrum beta-lactamase (ESBL), AmpC, and metallo beta-lactamase (MBL) were detected by phenotypic methods. Results: Females predominated in the study 193 (78.4%) with 53 (21.5%) males. Most of the isolates were in the age group 21-30 and 31-40. 49 (19.9%) patients were inpatients, and 197 (80.08%) were outpatients. Aminoglycosides had a better spectrum of antimicrobial sensitivity (70.2%) with 85.2% isolates sensitive to amikacin. Fluoroquinolones had a poor activity against Citrobacter (46% sensitive). Ureidopenicillins had poor efficacy (15.4%). Out of the 246 strains, 129 (52.4%) were MDR, 61 (24.7%) of which were ESBL producers and 65 (26.4%) were AmpC producers. Most of the ESBL producing strains were isolated from inpatients. No MBL were identified in this study. Conclusion: Citrobacter though not uncommon isolate is posing a problem due to its MDR character. Infection control practices should be observed strictly, and any type of unnecessary instrumentation should be avoided.

Development of multidrug resistance among pathogens has become a global problem for chemotherapy of bacterial infections. Extended-spectrum β-lactamase- (ESβL-) producing enteric bacteria and methicillin-resistant Staphylococcus aureus (MRSA) are the two major groups of problematic MDR bacteria that have evolved rapidly in the recent past. In this study, the aqueous extract of Murraya koenigii leaves was used for synthesis of silver nanoparticles. The synthesized MK-AgNPs were characterized using UV-vis spectroscopy, FTIR, XRD, SEM, and TEM, and their antibacterial potential was evaluated on multiple ESβL-producing enteric bacteria and MRSA. The nanoparticles were predominantly found to be spheroidal with particle size distribution in the range of 5–20 nm. There was 60.86% silver content in MK-AgNPs. Evaluation of antibacterial activity by the disc-diffusion assay revealed that MK-AgNPs effectively inhibited the growth of test pathogens with varying sized zones of inhibition. The MICs of MK-AgNPs against both MRSA and methicillin-sensitive S. aureus (MSSA) strains were 32 μg/ml, while for ESβL-producing E. coli, it ranged from 32 to 64 μg/ml. The control strain of E. coli (ECS) was relatively more sensitive with an MIC of 16 μg/ml. The MBCs were in accordance with the respective MICs. Analysis of growth kinetics revealed that the growth of all tested S. aureus strains was inhibited (∼90%) in presence of 32 μg/ml of MK-AgNPs. The sensitive strain of E. coli (ECS) showed least resistance to MK-AgNPs with >81% inhibition at 16 μg/ml. The present investigation revealed an encouraging result on in vitro efficacy of green synthesized MK-AgNPs and needed further in vivo assessment for its therapeutic efficacy against MDR bacteria.

Eco-friendly and cost effective Al2O3 NPs were synthesized by a green method using a leaf extract of Cymbopogon citratus. The characterization of the synthesized NPs was performed by XRD, HR-TEM, EDS, AFM, DLS, and zeta potential. To our knowledge, for the first time, the antifungal activity of Al2O3 NPs against various Candida spp isolated from oropharyngeal mucosa of HIV+ patients has been investigated. The antifungal potential of Al2O3 NPs has been assessed by determining minimum inhibitory concentrations (MIC), minimum fungicidal concentration (MFC), growth kinetics, and well diffusion methods. The minimum concentration at which Al2O3 NPs inhibit the growth of Candida spp were 250–500 μg ml−1 and more or less no growth has been observed at higher concentration (i.e., 1500 μg ml−1) when incubated for 48 h. The mechanisms of Al2O3 NPs–Candida cell biointerface interaction and its intracellular localization were further characterized by SEM and HR-TEM. The electron microscopic analysis shows that Al2O3 NPs not only attach to the surface but also penetrate inside the Candida cells and disrupt the morphological as well as physiological activity that led to the death of the cells. Due to the scavenging potential of Al2O3 NPs, the antifungal activity of Al2O3 NPs is mild towards tested yeast spp and that is why these NPs suppressed the growth of yeast cells at higher concentration. The result obtained in this study shows that Al2O3 NPs inhibit the growth of fluconazole susceptible and fluconazole resistant C. albicans and C. dubliniensis in a similar manner regardless of their drug resistant mechanisms. Thus, the present study revealed that Al2O3 NPs may be a promising remedy for the development of anticandidal agents and play an important role for sustained controlled drug delivery in medical implants and for therapeutic coatings on implantable devices used in orthopedics and dentistry.

Background: Urinary tract infection (UTI) is one of the most common bacterial infections, affecting 150 million people each year worldwide with substantial clinical and financial burden. With upcoming multi drug resistance (MDR) and carbepenem resistance among uropathogens there is urgent need to explore other new or old treatment options like nitrofurantoin and fosfomycin trometamol.Methods: This is a cross-sectional (descriptive study) conducted over 6 month’s period from October 2019 to March 2020. Out of 9045 urine samples, 1788 (19.8%) were positive (1721 samples with single organism and 67 samples with 2 organisms). Total 1855 isolates were identified and antimicrobial susceptibility was performed by Kirby-Bauer method and VITEK 2 system. Methicillin‑resistant Staphylococcus aureus (MRSA), vancomycin resistant enterococci (VRE), multi drug resistance (MDR) and metallo‑beta‑lactamases (MBL) production was detected.Results: E. coli 41.8% was found commonest followed by enterococcus species (21.6%). Methicillin resistance was 66% while 1.8% were VRE. 429 (34.5%) were CRE (carbapenem resistant enterobacteriales) out of which, 154 (36%) were MBL while 188 (44%) were detected as serine carbapenemase producers via modified carbapenem inactivation method (mCIM) and EDTA-modified carbapenem inactivation method (eCIM) testing. Among 742 (40%) MDR, fosfomycin was effective in 611 (82.3%) while 331 (77.1%) of the CRE isolates were susceptible to fosfomycin.Conclusions: Fosfomycin should be reserved for MDR and nitrofurantoin should be used cautiously otherwise resistance will increase to these drugs in the coming days.

Objectives: Detection and comparison of metallo-β-lactamase (MBL) production in clinical isolates by phenotypic and genotypic measures. The objective of this study is to evaluate clinical characteristics and risk factors in patients infected with MBLs. Materials and Methods: Study was conducted by the Department of Microbiology, Jawaharlal Nehru Medical College from February 2014 to December 2015. Bacterial culture, identification, and antibiotic susceptibility were carried out according to standard guidelines. MBL production was detected both phenotypically (Modified Hodge test [MHT], imipenem-ethylene diamine tetraacetic acid double disk potentiation test [IMP-EDTA DDPT], IMP-EDTA combined disk synergy test [IMP-EDTA CDST]), and genotypically (bla NDM-1, bla VIM and bla IMP). Results: Among 116 carbapenem-resistant Gram-negative Bacilli (CRGNB), Citrobacter species 28 (24.1%) was the most common pathogen. Phenotypically, MHT, IMP-EDTA DDPT, and IMP-EDTA CDST detected MBL production in 105 (90.5%), 96 (81%), and 87 (75%) CRGNB, respectively. Bla NDM-1 genes were detected in 6 6 (56.8%) isolates, however, very few bla VIM (16, 15.2%) and bla IMP (1, 1.2%) were identified. Considering polymerase chain reaction (PCR) as the gold standard, it was observed that IMP-EDTA CDST was most specific (78.3%) while MHT was most sensitive (97.4%). Results of bla NDM-1 gene by PCR were further confirmed by sequencing (Triyat genomics, Nagpur). All the 11 representative strains were confirmed to be NDM-1 gene. Major risk factors in patients infected with MBLs were in-dwelling devices (68%), prolonged hospital stay (72%) and prior antibiotic treatment (86%). However, on tracing their outcome, it was interesting to note that mortality was relatively low 5 (4.3%). Conclusion: The present study shows a rising trend of bla NDM-1 in CRGNB, an ominous sign heralding the post antibiotic era. It is essential to assess the prevalence of various MBLs so that infection control measures can be reinforced. We recommend three phenotypic tests in tandem for the detection of MBL. While phenotypic tests are easy and cost-effective to perform, quick, effective molecular diagnostic techniques can tailor treatment guidelines to optimize patient's management.

The objective of the present study was one step extracellular biosynthesis of silver nanoparticles (AgNPs) using supernatant of Candida glabrata isolated from oropharyngeal mucosa of human immunodeficiency virus (HIV) patients and evaluation of their antibacterial and antifungal potential against human pathogenic bacteria and fungi. The mycosynthesized AgNPs were characterized by color visualization, ultraviolet-visible (UV) spectroscopy, fourier transform infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The FTIR spectra revealed the binding and stabilization of nanoparticles with protein. The TEM analysis showed that nanoparticles were well dispersed and predominantly spherical in shape within the size range of 2&ndash, 15 nm. The antibacterial and antifungal potential of AgNPs were characterized by determining minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC)/ minimum fungicidal concentration (MFC), and well diffusion methods. The MBC and MFC were found in the range of 62.5&ndash, 250 &mu, g/mL and 125&ndash, 500 &mu, g/mL, which revealed that bacterial strains were more susceptible to AgNPs than fungal strains. These differences in bactericidal and fungicidal concentrations of the AgNPs were due to the differences in the cell structure and organization of bacteria and yeast cells. The interaction of AgNPs with C. albicans analyzed by TEM showed the penetration of nanoparticles inside the Candida cells, which led the formation of &ldquo, pits&rdquo, and &ldquo, pores&rdquo, that result from the rupturing of the cell wall and membrane. Further, TEM analysis showed that Candida cells treated with AgNPs were highly deformed and the cells had shrunken to a greater extent because of their interaction with the fungal cell wall and membrane, which disrupted the structure of the cell membrane and inhibited the normal budding process due to the destruction and loss of membrane integrity and formation of pores that may led to the cell death.

Introduction: Tuberculosis (TB) remains a significant public health problem with an estimated onethird of the world’s population being infected. Cytokines play a major role in protection against Mycobacterium tuberculosis infection and regulate the immune responses at cellular level. Most studies on cytokines during TB are from ‘in vitro’-stimulated lymphoid cells with few reports on in vivo plasma levels. This study was aimed to evaluate the levels of TNF-α & IL4 in new, underOriginal Research Article Fatima et al.; BMRJ, 7(4): 159-166, 2015; Article no.BMRJ.2015.107 160 treatment (UT) and multidrug resistant (MDR) pulmonary and extra-pulmonary cases. Methodology: The study was conducted at the Department Of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, India. Levels of TNF-α and IL-4 were measured in 76 serum samples from TB patients by ELISA kit (Diaclone France) along with 10 BCG vaccinated control. A complete clinical, radiological & treatment data was collected on questionnaire forms of each patients. Results: TNF-α levels were elevated in new (P 0.05). TNF-α and IL-4 levels showed no significant variations according to site of involvement in pulmonary vs. extra-pulmonary TB cases. Conclusion: An understanding of the development of this response may lead to insight into pathogenesis and novel therapies for TB.

Article history: Received on: 07/07/2015 Revised on: 21/07/2015 Accepted on: 09/08/2015 Available online: 28/10/2015 Emergence of AmpC beta-lactamases in isolates of Pseudomonas and Acinetobacter species, is a threatening condition as they mediate resistance to a wide variety of β-lactam drugs, including α-methoxy-β-lactams, such as cefoxitin, narrow-, expanded- and broad-spectrum cephalosporins, aztreonam and are poorly inhibited by βlactam inhibitor combinations. The present study was conducted to determine the occurrence of blaampC genes in these pathogenic non-fermenters for their rapid and accurate detection. Monoplex PCR was done to detect blaampC genes in 40 non-duplicate clinical Pseudomonas and Acinetobacter isolates, that were found resistant to any of the third-generation cephalosporin and cefoxitin. Multiplex PCR assay was carried out to identify family-specific AmpC beta-lactamase genes within Pseudomonas and Acinetobacter spp. PCR detected blaampC in 43.24% of Pseudomonas and 33.33% of Acinetobacter isolates. Overall 42.50% of the total isolates were found to harbour blaampC genes by PCR. By multiplex PCR, total eight (20%) isolates yielded a positive amplicon with AmpCspecific primers. High prevalence of blaampC genes in cefoxitin-resistant isolates of Pseudomonas and Acinetobacter isolates emphasizes that molecular detection methods should be carried out to know the exact prevalence of beta-lactamases.

Background: Chronic Suppurative Otitis Media (CSOM) is an important cause of preventable hearing loss. Global emergence of resistant strains is of great concern. The aim of the present study was t o determine the etiology and antibiotic sensitivity pattern of bacterial isolates from CSOM cases with special emphasis on ESBL (Extended Spectrum Beta- Lactamases) and AmpC beta lactamases. Methods: Patients with sign and symptoms suggestive of CSOM, ESBL (Extended Spectrum Beta-Lactamases), AmpC beta lactamases and MBLs (Metallo beta lactamases) were included. Two ear swabs were taken from all the patients and cultured on blood agar and MacConkey agar. Bacterial identification of isolates was done using standard biochemicals. Antimicrobial susceptibility was performed by Kirby-Bauer's disc diffusion method as per the Clinical Laboratory Standards Institute (CLSI) guidelines using antibiotic discs (HI MEDIA). Results: Out of 130 patients, 110(84.62%) had bacterial growth. The common pathogenic species were Pseudomonas aeruginosa 36(37.89%), Staphylococcus aureus 31(32.63%), Citrobacter koseri 9(9.47%) and Proteus vulgaris 6(6.32%). P. aeruginosa showed maximum sensitivity to colistin (94.4%), polymixin-B (91.3%) and imipenem (91.3%) . Gram positive cocci showed maximum sensitivity to vancomycin (99%). MRSA (Methicillin Resistant Staphylococcus aureus) and HLAR (High Level Aminoglycoside Resistance) were detected in 9(29%) S. aureus and 1(50%) Enterococcus faecalis respectively. ESBL and AmpC were detected in 11 (18.3%) and 12(20%) Gram negative bacteria, respectively and MBL producer was not detected. Conclusion: P. aeruginosa was found to be the most common isolate in CSOM cases and colistin, polymixin-B and imipenem was found to be most effective antibiotics.

Objective To evaluate the role of Toxoplasma gondii in women with bad obstetric (BOH) history and in women with primary and secondary infertility. Methods This study was conducted in the Department of Microbiology, Jawaharlal Nehru Medical College and Hospital for a period of 5 years from January 2004 to December 2009. Quantitative determination of IgM antibodies to Toxoplasma gondii infection was done by IgM capture ELISA in patients with history of BOH or infertility. Results Out of a total of 441 subjects included in the study, 417 (94.6%) had a BOH and 24 (5.4%) subjects had infertility. Toxoplasma was found to be more common in females with two or more abortions 52 (76.5%). Similarly in patients with infertility due to Toxoplasma , secondary infertility (66.7%) was more common than primary infertility. About 40.3% patients with BOH and 20% patients with infertility had healthy live issues after treatment with spiramycin. Conclusions Toxoplasmosis is thus, an easily treatable cause of abortions and infertility. All antenatal females and females with infertility should be screened for toxoplasmosis.

Clinical isolates (n = 55) of Pseudomonas aeruginosa were screened for the extended spectrum β-lactamases and metallo-β-lactamases activities and biofilm forming capability. The aim of the study was to demonstrate the antibiofilm efficacy of gum arabic capped-silver nanoparticles (GA-AgNPs) against the multi-drug resistant (MDR) biofilm forming P. aeruginosa. The GA-AgNPs were characterized by UV-spectroscopy, X-ray diffraction, and high resolution-transmission electron microscopy analysis. The isolates were screened for their biofilm forming ability, using the Congo red agar, tube method and tissue culture plate assays. The biofilm forming ability was further validated and its inhibition by GA-AgNPs was demonstrated by performing the scanning electron microscopy (SEM) and confocal laser scanning microscopy. SEM analysis of GA-AgNPs treated bacteria revealed severely deformed and damaged cells. Double fluorescent staining with propidium iodide and concanavalin A-fluorescein isothiocyanate concurrently detected the bacterial cells and exopolysaccharides (EPS) matrix. The CLSM results exhibited the GA-AgNPs concentration dependent inhibition of bacterial growth and EPS matrix of the biofilm colonizers on the surface of plastic catheters. Treatment of catheters with GA-AgNPs at 50 µg ml(-1) has resulted in 95% inhibition of bacterial colonization. This study elucidated the significance of GA-AgNPs, as the next generation antimicrobials, in protection against the biofilm mediated infections caused by MDR P. aeruginosa. It is suggested that application of GA-AgNPs, as a surface coating material for dispensing antibacterial attributes to surgical implants and implements, could be a viable approach for controlling MDR pathogens after adequate validations in clinical settings.

Objective To study the prevalence of toxoplasmosis in pregnant females with history of habitual abortions.

Objective: To estimate superoxide dismutase (SOD) activity in erythrocytes infected with Plasmodium falciparum in the cases of cerebral malaria. Methods: The diagnosis of cerebral malaria was made clinically and by Giemsa stained peripheral blood smear examination, quantitative buffy coat (QBC) examination and rapid antigen detection test (RDT). Parasitemia per micro litre of blood was evaluated by counting 200 white blood corpuscles and used to calculate parasite density considering 8000 white blood corpuscles per micro litre. SOD activity was estimated by the method given by Joe M. McCord and Irwin Fridovich spectrophotometrically. Statistical analysis was performed by using SPSS software version 17. Results: The SOD activity in the cases was found to be (1.06 ± 0.50) nmol/mL and that in the controls was (3.55 ± 0.07) nmol/mL. The SOD activity in the cases was significantly decreased (P < 0.05) as compared to the controls. The Pearson’s coefficient of correlation between SOD activity and parasitemia was found to be -0.93 showing strong negative relationship. Conclusions: There is severe oxidative stress in falciparum malaria due to reactive oxygen species and supplementation of antioxidants may modify the course and outcome of the disease.

BACKGROUND: Cryptosporidium has emerged as one of the major parasitic agents as a cause of diarrhoea in children. Various epidemiological factors have been described by different workers. Aims: This study was done to determine different epidemiological factors incriminated in cryptosporidiosis. Methods: Stool samples from 240 children with diarrhoea were examined for presence of Cryptosporidium. Wet mount examination, modified Ziehl- Neelsen (Z.N.) and Safranine-methylene blue staining methods were performed. For 177 samples, ELISA was also done. Detailed history of patients regarding their socioeconomic status and various sociodemographic factors was taken. Statistical Analysis: Chi-square and z tests were used to compare differences between the groups. A p value of ≤0.05 was considered significant. Results: Majority of patients were in Class IV socioeconomic group. Top-feeding, use of insanitary wells for drinking purposes, close association with animals, field defaecation and residence in rural areas were different factors that contributed to the spread of infection. Oocysts were present in 21 children on different staining procedures and 23 were positive by ELISA. Conclusion: Different sociodemographic factors like improper sanitation practices, drinking contaminated water, early withdrawal of breast feeding and close intimacy with animals are various factors that can enhance the spread of infection in community. Preventive measures are of great importance in control of spread of infection as there is no specific therapy for cryptosporidiosis.

Introduction: Klebsiella pneumoniae is a ubiquitous environmental organism and a common cause of serious gram-negative infections in humans. This study was conducted to examine the association between seasonal variation and the incidence rate of Klebsiella pneumoniae blood stream infection. Material and methods: The retrospective study was conducted in the Department of Microbiology, JN Medical College AMU Aligarh for a period of 5 years from January 2011 to December 2015. Samples were received for blood culture in brain heart infusion broth. Cultures showing growth of Klebsiella pneumoniae were identified using standard biochemical procedures. Antimicrobial susceptibility testing was done on Mueller Hinton’s agar by Kirby Bauer Disc diffusion method as per the CLSI guidelines. Detection of ESBL, AmpC and MBL production was done. Results: 495 (30.0%) isolates were found to be positive for Klebsiella pnuemoniae during the five year study period. Prevalence of Klebsiella pneumoniae isolates in blood stream infection for the warmest four months (July- September) was found to be 61.8% as compared to 38.2% in the remaining year (January-June and October- December). Maximum number of Klebsiella pneumoniae were isolated the month of August 86 (17.3%) followed by July 75 (15.1%) while minimum isolates were found in the month of February 14 (2.8%) and January 19(3.8%). ESBL producing isolates were 44 (8.8%) and 318 (64.20%) isolates were AmpC producers. 36 (7.2%) isolates were found to be MBL producer. Conclusion: Our study suggests that K. pneumoniae is an important pathogen in hospitals worldwide and that rates of K. pneumoniae blood stream infection vary seasonally.

Cefoxitin-resistant Escherichia coli ( n = 109) and Klebsiella pneumoniae ( n = 16) isolates collected from patients in India in 2009 to 2010 were screened for bla ampC families and mobilizing elements ( ISE cp1, IS 26, IS CR1, and sul -1-type class 1 integrons) and their association with bla ampC and for the occurrence of class A beta-lactamases (BLs) (CTX-M, TEM, and SHV). The concurrent occurrences of two distinct AmpC families ( bla CIT and bla EBC ) and of class A with class C beta-lactamase were observed. All but one of the isolates harboring CTX-M extended-spectrum BLs (ESBLs) were carrying bla CTX-M genogroup 1; the remaining isolate carried bla CTX-M genogroup 9. The mobilizing elements occurred in different combinations in the study isolates.

Objectives: To study the origin and spread of the chloroquine-resistant Plasmodium falciparum population in the Indian subcontinent. Methods: Fourteen microsatellites spanning a ∼120 kb region, flanking the P. falciparum chloroquine resistance transporter (pfcrt) gene, were analysed in 185 parasite isolates. Results: The Indian P. falciparum population exhibited a selective valley of reduced genetic variation in the flanking microsatellites of the mutant pfcrt alleles (up to ±29 kb) as compared with the wild-type allele. This valley is much narrower than the ±200 kb valley reported from African and South-East Asian countries. The majority of the isolates showed asymmetry in the selective valley, where upstream microsatellites showed less genetic variation than the downstream microsatellites. Regional variation in the width and symmetry of the selective valley was noticed, which seems to be related to the number of pfcrt alleles present in the parasite population of a region. Forty-six different microsatellite haplotypes were observed among the P. falciparum isolates containing mutant pfcrt alleles. Parasite populations from different regions of mainland India shared microsatellite haplotypes between them, but they shared none with the isolates from the Andaman and Nicobar Islands, and vice versa. Indian isolates shared microsatellite haplotypes with the isolates from Papua New Guinea and Thailand. Conclusions: With regard to chloroquine there is regional variation in the selection pressure on the P. falciparum population in India. These findings will help the regional implementation of drug policy in India's malaria control programme.

The use of successive generations of beta-lactams has selected successive generations of beta-lactamases including CTX-M ESBLs, AmpC beta-lactamases, and KPC carbapenamases in Enterobacteriaceae. Moreover, this cephalosporin resistance, along with rising resistance to fluoroquinolones, is now driving the use of carbapenems and unfortunately the carbapenem resistance has emerged markedly, especially in Acinetobacter spp. due to OXA- and metallo-carbapenemases. The industry responded to the challenge of rising resistance and recently developed some novel beta-lactams such as ceftobiprole, ceftaroline etc. and many beta-lactam compounds, including beta-lactamase-inhibitors, such as BMS-247243, S-3578, RWJ-54428, CS-023, SMP-601, NXL 104, BAL 30376, LK 157, and so on are under trials. This review provides the comprehensive accounts of the developments in penicillins, cephalosporins, carbapenems, and beta-lactamase-inhibitors, and the insight about medicinal chemistry, mechanism(s) of action and resistance, potential strategies to overcome resistance due to beta-lactamases, and also the recent advancements in the development of newer beta-lactam compounds; some of which are still under trials and yet to be classified. This review will fill the gap since previously published reviews and will serve as a comprehensive update on the current topic.

Background: Trichomonas vaginalis causes a common sexually transmitted disease trichomoniasis, which may lead to increased risk of transmission of human immunodeficiency virus infection and other pelvic inflammatory diseases. Wet mount examination is the most common test for diagnosis, but it has low sensitivity. Acridine orange staining can be used for diagnosis, but it requires special microscopic facility. Culture is considered as the gold standard, but it takes a long time for diagnosis. OSOM Trichomonas Rapid Test is a recently introduced rapid method based on immunochromatographic assay of trichomonal protein antigens. Hence, the present study was done to compare these four diagnostic techniques for detection of trichomoniasis in females with vaginal discharge. Materials and Methods: Vaginal swabs were taken from 835 female patients and wet mount examination, acridine orange staining, culture in Kupferberg medium, and OSOM Trichomonas Rapid Test, were performed. Results: Out of 835 patients included in our study, 68 (8.1%) positive cases of trichomoniasis were detected by culture. OSOM Trichomonas Rapid Test detected 63 (7.5%) cases, acridine orange staining detected 53 (6.3%) cases, whereas, wet mount examination detected only 45 (5.4%) positive cases. OSOM Trichomonas Rapid Test performed well and showed high sensitivity and specificity of 88.2% and 99.6%, respectively. Conclusion: As OSOM Trichomonas Rapid Test is a point of care test and gave better results than both wet mount examination and acridine orange staining; it can be used as a routine test in peripheral areas lacking laboratory facilities.

PURPOSE: To find out the prevalence of Klebsiella in hospital acquired neonatal infections in a tertiary care set up and to evaluate the role of klebocin typing and protein profile by SDS-PAGE in epidemiological typing of the isolates. METHODS: Hospital born neonates transferred to the neonatal unit after birth and available in the unit 48 hours later comprised the study group. Two hundred and three neonates were found eligible for inclusion in the study. Repeated blood cultures, other relevant clinical specimens and environmental samples were collected and identified according to the standard techniques. Isolated clinical and environmental Klebsiella pneumoniae strains were subjected to klebocin typing and protein profiling by SDS-PAGE at regular intervals. RESULTS: Multi drug resistant K. pneumoniae were the commonest organism isolated in 30 neonates leading to the incidence of Klebsiella nosocomial infection to be 14.7%. Klebocin typing of the K. pneumoniae isolates showed four patterns with type 312 being the commonest (43.4%). Whole cell protein analysis by SDS-PAGE of K. pneumoniae isolates revealed four types of banding pattern. Analysis of the typing method showed that the typeability and reproducibility of klebocin was 83.3% and 73.3% respectively whereas typeability and reproducibility of SDS-PAGE was 100%. CONCLUSIONS: Based on the present study it is concluded that SDS-PAGE typing method is better than klebocin typing in neonatal nosocomial infection. It is also suggested that protein profile by SDS-PAGE may be used as a tool for epidemiological typing of Klebsiella pneumoniae isolates in laboratories where genomic based molecular typing technique is not available.

About 50% of malaria infections in India are attributed toPlasmodium falciparumbut relatively little is known about the genetic structure of the parasite populations. The molecular genotyping of the parasite populations by merozoite surface protein(msp1 and msp2)and glutamate-rich protein (glurp) genes identifies the existing parasite population in the regions which help in understanding the molecular mechanisms involved in the parasite’s drive for survival. This study reveals the genetic profile of the parasite population in selected regions across the country with varying degree of endemicity among them. We also report the prevalence ofPfcrtmutations in this parasite population to evaluate the pattern of drug resistance development in them.

The high prevalence of extended-spectrum β-lactamases (76.3 %) and metallo-β-lactamases (7.3 %) amongst the bacteria Pseudomonas aeruginosa is a critical problem that has set forth an enormous therapeutic challenge. The suggested role of nanoparticles as next generation antibiotics, and inadequate information on antibacterial activity of aluminium oxide nanoparticles has led us to investigate the green synthesis of aluminium oxide nanoparticles (Al2O3 NPs) using leaf extracts of lemongrass and its antibacterial activity against extended-spectrum β-lactamases and metallo-β-lactamases clinical isolates of P. aeruginosa. The synthesized Al2O3-NPs were characterized by scanning electron microcopy, high resolution-transmission electron microscopy, atomic force microscopy, X-ray diffraction, Zeta potential, and differential light scattering techniques. The X-ray diffraction data revealed the average size of the spherical Al2O3-NPs as 34.5 nm. The hydrodynamic size in Milli Q water and Zeta potential were determined to be 254 nm and +52.2 mV, respectively. The minimal inhibitory concentration of Al2O3-NPs was found to be in the range of 1,600–3,200 µg/ml. Treatment at concentrations >2,000 µg/ml, resulted in complete growth inhibition of extended-spectrum β-lactamases and metallo-β-lactamases isolates. Scanning electron microcopy analysis revealed the clusters of nanoparticles attached to the bacterial cell surface, causing structural deformities in treated cells. High resolution-transmission electron microscopy analysis confirmed that nanoparticles crossed the cell membrane to become intracellular. The interaction of nanoparticles with the cell membrane eventually triggered the loss of membrane integrity, most likely due to intracellular oxidative stress. The data explicitly suggested that the synthesized Al2O3-NPs can be exploited as an effective bactericidal agent against extended-spectrum β-lactamases, non-extended-spectrum β-lactamases and metallo-β-lactamases strains of P. aeruginosa, regardless of their drug resistance patterns and mechanisms. The results elucidated the clinical significance of Al2O3-NPs in developing an effective antibacterial therapeutic regimen against the multi-drug resistant bacterial infections. The use of leaf extract of lemongrass for the synthesis of Al2O3-NPs appears to be cost effective, nontoxic, eco-friendly and its strong antibacterial activity against multi-drug resistant strains of P. aeruginosa offers compatibility for pharmaceutical and other biomedical applications.

Natural products of plant origin are being used to cure infections since ancient times. However, the real breakthrough in the field of antimicrobial agents was after the advent of a chemical substance produced by a strain of the mold Penicillium, by Alexander Fleming, that he named penicillin in 1928. Since that time, the fungi and higher plants have been searched for antibacterial compounds, including cephalosporins and aminoglycosides. In some of our previous articles published during years 2009–2010 we reviewed the literature on plant natural products harboring the antibacterial and antifungal properties. In this article we will explore the very recent developments in the said field with an emphasis on antibacterial, antifungal, anti-parasitic and antiviral potentials (especially against Human Immunodeficiency virus and Influenza virus) and some of the other interesting recent developments.

Introduction: Mycobacterium tuberculosis (Mtb) infects one third of the world population. Despite advance in therapy, tuberculosis (TB) remains responsible for 2-3 million deaths annually. During TB, cytokines play a role in host defense. Many cytokines are produced during TB with a predominance of Th1 cytokines during the early stage and Th2 cytokines in the later stages of the infection. The present study aims to screen tumor necrosis factor-alpha (TNF-α), interferon gamma (IFN-γ), interleukin (IL)-12p70, IL-4, IL-6, and IL-1β among TB patients and controls. Materials and Methods: The study was conducted at the Department Of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, India. Different cytokine levels were measured in 65 samples of TB patients of whom 30 (46.1%) were new TB cases and 35 (53.8%) were suspected multidrug-resistant (MDR)-TB cases along with 15 bacille Calmette-Guerin (BCG) vaccinated healthy controls by enzyme-linked immunosorbent assay (ELISA) (Diaclone, France). Complete clinical, radiological, and treatment data were collected. Informed consent was taken from all subjects. Study was approved by the Institutional Bioethical Committee, Jawahar Lal Nehru Medical College. Statistical analysis had been performed by using Sigma plot (10.0). Results: The serum concentration of TNF-α, IFN-γ, IL-12p 70 , IL-1β, and IL-6 in patients with active TB and MDR-TB was elevated than in patients after anti-TB treatment (P < 0.001), in contacts (P < 0.001) and in controls (P < 0.001). It was found that IL-6 seems critical in resistance to TB. IL-4 concentration increased only in MDR-TB patients (P < 0.001) as compared to controls. All these cytokines showed no significant variations according to the site of involvement in pulmonary versus extrapulmonary TB cases. Conclusions: Measuring the serum levels of several cytokines may be useful for evaluating the activity of TB disease and monitoring the clinical effects of anti-tubercular treatment (ATT). The serum TNF-α, IFN-γ, IL-12p 70 , IL-1β and IL-6 level can be a useful marker to diagnose the effectiveness of therapy in the patients.

Chloroquine (CQ) has been the mainstay of treatment of malaria for decades. This cost-effective and safe drug has become ineffective for treatment of falciparum malaria in many parts of the world due to development of resistance by the parasite. In addition CQ is not gametocytocidal for P. falciparum and thus cannot block transmission. The extent of problem of chloroquine resistance in P. falciparum is increasing every year. The study was done in period of 2 years. A total of 5653 specimens were examined for malarial infection by employing different diagnostic modalities. Four hundred and thirty-five were found to be positive for P. falciparum by using different diagnostic techniques. All positive specimens were cultured on RPMI 1640 medium; only 108 were found to be culture positive. Sensitivity of isolates to chloroquine was done using Mark III WHO sensitivity plates. The prevalence of malaria infection was found 9.54% in 2010. There were schizont formation at 8 pmol/liter or more of chloroquine concentration in 26 isolates. The emergence of chloroquine (CQ) resistance pattern in Aligarh isolates increases. Antimalarial agents should be used with caution; monotherapies should be avoided.

Background: Enteric parasitic protozoan of genus cryptosporidium has become an important cause of diarrhoea in developing countries. It is a cause of acute and persistent diarrhoea in immunocompromised and immunocompetent subjects. Methodology: This study was conducted to determine the prevalence of Cryptosporidium spp. as an enteric pathogen causing diarrhoea in children. 250 stool specimens of children with diarrhoea were examined using staining procedures like modified Ziehl-Neelsen (Z.N.) staining, safranine-methylene blue staining after concentration of stool specimens by formol-ether concentration technique and antigen detection ELISA was performed for diagnosis. Results: Modified Z. N. staining and safranine-methylene blue staining were positive in 21 cases while 29 cases were positive by ELISA. Prevalence of Cryptosporidium in children with diarrhoea was found to be 11.6%. Conclusion: High prevalence as noted in this study advocates need to screen for this parasite as a routine diagnostic measure in patients of diarrhoea.

Introduction: Immunity forms the backbone of our very existence and opportunistic infections have plagued the immunosuppressed since long. A significant morbidity and mortality is attributable to intestinal parasitosis in the immunocompromised population. Objectives: This study aims to study the prevalence of intestinal protozoa in various immunosuppressed groups and to correlate degree of immunosuppression with protozoal infections. Material and Methods: The study was conducted on 400 patients divided into 4 groups: Group I (HIV patients), Group II (patients on chemotherapy/chemotherapy and radiotherapy for various malignancies), and Group III (diabetes patients) and Group IV included children presenting with diarrhea. Group I, II and III included patients with and without gastrointestinal symptoms. Stool samples were investigated microscopically for cysts/trophozoites after concentration with formol-ether method. Iodine wet mount and Modified acid fast staining methods were used. Result: Intestinal protozoa were detected in 40.75% (163/400); more commonly in patients with higher degree of immunosuppression (CD4 count 7: 95.3% in diabetics and moderate to severe malnutrition: 61.8%. Most common protozoa associated was Cryptosporidium in 66 patients (40.5%) followed by, Entamoeba histolytica in 48 (29.4%), Giardia lamblia in 35 (21.5%), Isosporabelliin 9 (5.5%), Blastocystis hominis in 4 (2.5%), Cyclospora cayetenensis in 1 (0.61%). Conclusion: High prevalence of intestinal protozoa was seen in all groups of immunosuppressed patients and a significant association seen between degree of immune suppression and protozoal infection. Focus needs to be on routine screening of all immunosuppressed patients and their immune reconstitution.

ObjectiveTo determine the occurrence of class A and class C β-lactamase genes and their co-occurrence in Indian Enterobacteriaceae.Methods52 third generation cephalosporin resistant isolates were phenotypically detected by combination disk method and screened by PCR to identify class A and class C type β-lactamase genes.ResultsOf the 52 isolates, 94.2% (49) were found harboring any of the blaESBLs. blaCTX-M, blaSHV and blaTEM were present in 82.6% (43/52), 59.6% (31/52), and 42.3% (22/52) isolates, respectively. Of the 49 ESBL positive isolates 57.1% (28/49) showed co-occurrence of blaampC with blaESBLs. On the contrary, the collection from 2009 showed their co-occurrence in 81.4% isolates.ConclusionsThe comparative study shows a downward trend for co-existence of blaESBLs with blaampC from 2009 to 2010. Further large scale studies are needed to address the co-occurrence of class A and class C β-lactamases in India and the resistance trend occurring over a period of time.

Histamine, a biogenic amine derived from the decarboxylation of amino acid histidine by an enzyme histidine decarboxylase. It involves the local immune responses as well as regulating physiological functions. As part of an immune response to foreign pathogen (including malarial parasite infection), histamine is produced by basophils and by mast cells found in nearby connective tissue. Elevation in immune mediators such as IL-1, IL-6, IL-8, TNF-α, NO and histamine have been associated with disease severity in malarial infection. Histamine releasing factor (HRF) is a peptide described in mice and humans, causes the release of histamine from basophils. HRF belongs to a class of protein called translationally controlled tumour protein (TCTP) homologs. Recently a Plasmodium falciparum TCTP is identified. This protein has a high homology to human HRF. The central nervous system signs and symptoms such as drowsiness, coma, multiple seizures, destruction of blood brain barrier etc are supposed to be due to histamine secretion in CNS during Plasmodium falciparum infection. In this chapter we will discuss the pathophysiological effects of histamine in severe malaria infection.

Introduction: Tuberculosis (TB) remains a significant public health problem, with an estimated one-third of the world's population being infected. Cytokines play a major role in protection against Mycobacterium tuberculosis ( M. tuberculosis ) infection and regulate the immune responses at a cellular level. Most studies on cytokines during TB are from in vitro -stimulated lymphoid cells with few reports on in vivo plasma levels. The aim of this study was to evaluate the levels of tumor necrosis factor-α (TNF-α) in new, undertreatment (UT), and multidrug-resistant (MDR) pulmonary and extrapulmonary cases. Materials and Methods: The study was conducted in the Department of Microbiology, Jawaharlal Nehru Medical College (J.N.M.C.), Aligarh Muslim University (A.M.U.), Aligarh, Uttar Pradesh, India. Results: The levels of TNF-α were measured in 76 serum samples from TB patients by an enzyme linked immunosorbent assay (ELISA) kit (Diaclone Sas, 1 BD, A Flemming Besancon Cedax, France), along with 10 healthy controls. Complete clinical, radiological and treatment data were collected. The TNF-α levels were elevated in new cases ( P P P > 0.05). Conclusions: An understanding of this response may lead to an insight into the pathogenesis and novel therapies for TB.

Mycobacterium tuberculosis (Mtb) infects approximately one third of world population. Many cytokines are produced during tuberculosis, with predominance of Th1 cytokines during the early stage and Th2 cytokines in the later stages of the infection. Most studies on cytokines during TB are from ‘in vitro’-stimulated lymphoid cells with few reports on in vivo plasma levels. The high expression of IL-4 has been implicated as a virulence factor. In the present study we examined the level of IL-4 in the serum of active tuberculosis in new cases, during anti-tubercular therapy and MDR cases. This Study was conducted at Department Of Microbiology, JNMC, AMU, Aligarh. A total 76 blood samples of Pulmonary & Extra-Pulmonary tuberculosis patients were tested by ELISA (Diaclone France) along with 10 healthy controls. The test was carried out according to the manufacturer instructions. O.D. was taken at 450 nm. There was no stastical difference in age and sex in our study. The level of IL-4 show significant changes in MDR TB cases (P 0.05). An understanding of the development of the cellular immune response may lead to insight into pathogenesis and novel therapies for TB. Further studies are needed to address the role of cytokines in immunity to TB under natural conditions.

A few key papers which have recently been published on the characterization of amoeba antigens are reviewed. Immunofluorescence tests and immunoelectron microscopy have demonstrated the localization of certain surface antigens on axenically cultured trophozoites. Most of the surface antigens have largely been shown to elicit a humoral response. The elicitation of cellular response has not been well illustrated. The localization of a large number of antigens in cytoplasmic vacuoles and plasma membrane indicates that a greater stimulus to the host would be provided by intracellular antigens than by those located on the surface of amoeba trophozoites. In a few inoculation studies, amoeba antigens, in combination with several adjuvants, have been successfully employed for inducing protective immunity in various animal model systems. These and other results clearly demonstrate that amoeba antigens are fully capable of generating humoral and as well as CMI responses. A combination of these two effector limbs of immunity can be fully exploited through effective use of future vaccines.

The ability of bacteria to develop antibiotic resistance and colonize abiotic surfaces by forming biofilms is a major cause of medical implant-associated infections and results in prolonged hospitalization periods and patient mortality. Different approaches have been used for preventing biofilm-related infections in health care settings. Many of these methods have their own demerits that include chemical-based complications; emergent antibiotic-resistant strains, and so on. Silver nanoparticles (AgNPs) are renowned for their influential antimicrobial activity. We demonstrate the biofilm formation by extended spectrum β-lactamases-producing Escherichia coli and Klebsiella spp. by direct visualization applying tissue culture plate, tube, and Congo red agar methods. Double fluorescent staining for confocal laser scanning microscopy (CLSM) consisted of propidium iodide staining to detect bacterial cells and concanavalin A-fluorescein isothiocyanate staining to detect the exopolysaccharides matrix were used. Scanning electron microscopy observations clearly indicate that AgNPs reduced the surface coverage by E. coli and Klebsiella spp. thus prevent the biofilm formations. Double-staining technique using CLSM provides the visual evidence that AgNPs arrested the bacterial growth and prevent the exopolysaccharides formation. The AgNPs-coated surfaces effectively restricted biofilm formation of the tested bacteria. In our study, we could demonstrate the complete antibiofilm activity AgNPs at a concentration as low as 50 μg/ml. Our findings suggested that AgNPs can be exploited towards the development of potential antibacterial coatings for various biomedical and environmental applications. These formulations can be used for the treatment of drug-resistant bacterial infections caused by biofilms, at much lower nanosilver loading with higher efficiency.

Objective To determine the prevalence of malaria in Aligarh and analyze species dominance in different years over a decade. Methods Diagnosis of malaria was done using microscopy as gold standard, rapid antigen detection assays and quantitative buffy coat (QBC) assays. Giemsa stained blood smear examination was done, thick and thin films were examined for presence of different Plasmodium spp. Rapid antigen detection assays employing detection of HRP-2 and parasite lactate dehydrogenase antigen (pLDH) by immunochromatography was done in patients whose blood smear found to be negative by conventional Giemsa slide examination. QBC was done in cases where there is strong clinical suspicion of malaria with blood smear negative, in patients with chronic malaria, splenomegaly, or in those patients who had inadequate treatment and for post-treatment follow up. Results Plasmodium vivax and Plasmodium falciparum were only species detected in our hospital. Overall prevalence of malaria in Aligarh was found to be 8.8%. The maximum prevalence of 20.1% was observed in year 2008 and lowest 2.3% in 2002. Conclusions High prevalence of malaria is observed in this part of country with dominance of both species particularly Plasmodium falciparum should be monitored and factors accounting for occurrence should be studied to employ effective control measures.

Golden hamsters were immunized with various antigen fractions of Leishmania donovani promastigotes. Beta 1,3-glucan was used as an adjuvant in these vaccination experiments. The results indicate that immunization of animals with the microsomal fraction (subcellular fraction III) in combination with glucan confers considerable immune protection against L. donovani infection. The immune protection was confirmed by correspondingly lower parasite burden in the livers and spleens of test animals compared to controls. Additionally, the vaccinated animals showed positive skin test responsiveness after challenge, along with increased antibody titres. Immunization of animals with whole and particulate antigen fractions was also found to afford a high degree of resistance. The other subcellular and soluble antigen fractions conferred very little protection. In these experiments, glucan was found to be a potent adjuvant when injected, intraperitoneally, with Leishmania antigens. Similar doses of parasite extracts given without an adjuvant were able to confer only very little or no protection.