Purpose: To study the demographic characteristics, associated factors, causative agents, of infectious keratitis and develop a diagnostic tool to aid easy diagnosis of keratitis.Methods: Corneal scrapes were collected and subjected to culture, microscopy, considering age, occupation, geographical, Frequency of predisposing ocular conditions, antibiotic susceptibility test, resistance patterns test, drug sensitivity and 16s r-DNAas well as 18s r-DNA based identification was performed and submitted to data bank with accession number. The 16S rDNA sequences of the individual bacteria and fungi were used for a universal primer design and thereby multiplex PCR can be performed.Results: A total of 250 consecutive patients with infective keratitis were evaluated, of which 77 (30.8%) were found to be of bacterial, 67 (26.8%) were fungal, 16 (6.4%) were both fungal and bacterial, and the remaining 90 (36%) were found to be culture negative. Contact lens wear was the main risk factor (80.8%). Ocular surface disease (23.6%), ocular trauma (14.8%), corneal surgery (4.4%) and corneal suture (6.4%) of cases were found in corneal ulcers. Most community acquired bacterial and fungal ulcers resolve with appropriate treatment. 64% of the infections involved positive cultures and 36% involved negative cultures, were found in polymicrobial mode of infection. Fusarium spp. (32.75%) was the most predominant species followed by Aspergillus sp. (20.68%) was found in fungal corneal ulcers and also Staphylococcus sp. was the most common bacteria found in bacterial cultures. The 16S rDNA sequence of the bacteria and fungi cultured from the isolates of the corneal scrapping were performed and the genes were submitted in Genbank.The primers designed for bacteria and fungi gave good results in the multiplex PCR carried out and we suggest this can be used as a diagnostic tool for keratitis.
Rapid identification of dermatophyte species by 28S rDNA Polymerase Chain Rea...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Study of Bacterial isolates with antibiogram from clinically suspected cases ...Saptadeep Roy
The document summarizes a study on bacterial isolates from clinically suspected neonatal sepsis cases at Burdwan Medical College & Hospital in West Bengal, India. Blood cultures were collected from 64 neonates showing signs of sepsis and bacteria were isolated from 26 samples. The most common isolates were Klebsiella pneumoniae (35%) and Acinetobacter baumannii (23%). Late onset sepsis was more prevalent than early onset. Antibiotic susceptibility testing found high resistance to cephalosporins in gram-negative bacteria and 22% of gram-positive isolates were methicillin-resistant Staphylococcus aureus. 17.64% of total isolates were multidrug-resistant.
power point conventional bacteria identification.pptxDebasish Sahoo
This document discusses conventional methods for bacterial identification from clinical samples. It describes the importance of microscopic examination of direct mounts or stained smears to provide a presumptive diagnosis before culturing. A variety of culture media, stains, and techniques are discussed for the isolation and identification of bacteria from different anatomic sites like the throat, lower respiratory tract, urine, genital tract, and gastrointestinal tract. Selective media are recommended to recover specific pathogenic bacteria while inhibiting normal flora for each sample type.
This document discusses oral fungal infections and their microbiological laboratory diagnosis. It begins by introducing common fungal pathogens found in the oral cavity such as Candida species. It then covers topics such as sample collection and processing, direct examination techniques including various staining methods, culture techniques using different media, non-culture diagnostic methods, serodiagnosis, and molecular methods for diagnosis. The role of oral health care professionals in the diagnosis and management of oral fungal infections is also emphasized.
This document describes the isolation and characterization of a new giant virus called Cedratvirus. Key points:
- Cedratvirus was isolated from an environmental sample in Algeria using Acanthamoeba castellanii.
- It has an ovoid shape with a cork structure at each end, resembling Pithovirus sibericum but with a unique double cork feature.
- The 589kb genome is most closely related to the pithovirus genomes, sharing over 100 genes, but with only 21% of genes involved in best reciprocal hits, indicating genetic distance from known pithoviruses.
PREVALENCE AND CHARACTERIZATION OF VIRULENCE PROPERTIES OF PSEUDOMONAS AERUGI...SUS GROUP OF INSTITUTIONS
Pseudomonas aeruginosa is the epitome of an opportunistic pathogen of humans that cause urinary tract infections, respiratory system infection, particularly in victim of severe burns, cancer and AIDS patient who are immunocompromised. Most Pseudomonas infections are both invasive and toxigenic. The particular bacterial determinants of virulence mediate different stages of infection and are ultimately responsible for the characteristic syndromes that accompany the disease. In the present study P. aeruginosa was found to be more prevalent in burn patients (100%) followed by urinary tract infection samples (71%), sputum samples (66%) and wound samples (59%). 85% isolates recovered from clinical samples were mucoid. A total of 35% isolates were strong siderophore producers, 19% isolates were strong protease producers while 52% were strong phospholipase producers. Isolates from burns, sputum and environment sample were strong rhamnolipid producers. Elevated level of hemolysin production was observed in burn, urine and wound isolates. The prominence of haemagglutination ability in environmental isolates followed by burns isolates provided evidence for its being a nosocomial pathogen. The association between virulence determinants and disease can indicate the precise role played by the determinant in estabilishing the disease. Isolates were maximally sensitive towards lactam antibiotics.
This study evaluated the use of multiplex PCR (M-PCR) to detect leprosy among close contacts of leprosy patients in India. M-PCR detected leprosy in nasal swab samples from 10.9% of multibacillary contacts and 1.3% of paucibacillary contacts. During a 2-year follow up, 2 contacts who had initially tested positive by M-PCR later developed clinical leprosy. The results suggest that M-PCR could be a more effective tool than conventional methods for early detection of leprosy among contacts, helping to prevent further transmission.
Genotyping and subgenotyping of Trichophyton rubrum isolated from dermatophyt...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Rapid identification of dermatophyte species by 28S rDNA Polymerase Chain Rea...iosrjce
IOSR Journal of Dental and Medical Sciences is one of the speciality Journal in Dental Science and Medical Science published by International Organization of Scientific Research (IOSR). The Journal publishes papers of the highest scientific merit and widest possible scope work in all areas related to medical and dental science. The Journal welcome review articles, leading medical and clinical research articles, technical notes, case reports and others.
Study of Bacterial isolates with antibiogram from clinically suspected cases ...Saptadeep Roy
The document summarizes a study on bacterial isolates from clinically suspected neonatal sepsis cases at Burdwan Medical College & Hospital in West Bengal, India. Blood cultures were collected from 64 neonates showing signs of sepsis and bacteria were isolated from 26 samples. The most common isolates were Klebsiella pneumoniae (35%) and Acinetobacter baumannii (23%). Late onset sepsis was more prevalent than early onset. Antibiotic susceptibility testing found high resistance to cephalosporins in gram-negative bacteria and 22% of gram-positive isolates were methicillin-resistant Staphylococcus aureus. 17.64% of total isolates were multidrug-resistant.
power point conventional bacteria identification.pptxDebasish Sahoo
This document discusses conventional methods for bacterial identification from clinical samples. It describes the importance of microscopic examination of direct mounts or stained smears to provide a presumptive diagnosis before culturing. A variety of culture media, stains, and techniques are discussed for the isolation and identification of bacteria from different anatomic sites like the throat, lower respiratory tract, urine, genital tract, and gastrointestinal tract. Selective media are recommended to recover specific pathogenic bacteria while inhibiting normal flora for each sample type.
This document discusses oral fungal infections and their microbiological laboratory diagnosis. It begins by introducing common fungal pathogens found in the oral cavity such as Candida species. It then covers topics such as sample collection and processing, direct examination techniques including various staining methods, culture techniques using different media, non-culture diagnostic methods, serodiagnosis, and molecular methods for diagnosis. The role of oral health care professionals in the diagnosis and management of oral fungal infections is also emphasized.
This document describes the isolation and characterization of a new giant virus called Cedratvirus. Key points:
- Cedratvirus was isolated from an environmental sample in Algeria using Acanthamoeba castellanii.
- It has an ovoid shape with a cork structure at each end, resembling Pithovirus sibericum but with a unique double cork feature.
- The 589kb genome is most closely related to the pithovirus genomes, sharing over 100 genes, but with only 21% of genes involved in best reciprocal hits, indicating genetic distance from known pithoviruses.
PREVALENCE AND CHARACTERIZATION OF VIRULENCE PROPERTIES OF PSEUDOMONAS AERUGI...SUS GROUP OF INSTITUTIONS
Pseudomonas aeruginosa is the epitome of an opportunistic pathogen of humans that cause urinary tract infections, respiratory system infection, particularly in victim of severe burns, cancer and AIDS patient who are immunocompromised. Most Pseudomonas infections are both invasive and toxigenic. The particular bacterial determinants of virulence mediate different stages of infection and are ultimately responsible for the characteristic syndromes that accompany the disease. In the present study P. aeruginosa was found to be more prevalent in burn patients (100%) followed by urinary tract infection samples (71%), sputum samples (66%) and wound samples (59%). 85% isolates recovered from clinical samples were mucoid. A total of 35% isolates were strong siderophore producers, 19% isolates were strong protease producers while 52% were strong phospholipase producers. Isolates from burns, sputum and environment sample were strong rhamnolipid producers. Elevated level of hemolysin production was observed in burn, urine and wound isolates. The prominence of haemagglutination ability in environmental isolates followed by burns isolates provided evidence for its being a nosocomial pathogen. The association between virulence determinants and disease can indicate the precise role played by the determinant in estabilishing the disease. Isolates were maximally sensitive towards lactam antibiotics.
This study evaluated the use of multiplex PCR (M-PCR) to detect leprosy among close contacts of leprosy patients in India. M-PCR detected leprosy in nasal swab samples from 10.9% of multibacillary contacts and 1.3% of paucibacillary contacts. During a 2-year follow up, 2 contacts who had initially tested positive by M-PCR later developed clinical leprosy. The results suggest that M-PCR could be a more effective tool than conventional methods for early detection of leprosy among contacts, helping to prevent further transmission.
Genotyping and subgenotyping of Trichophyton rubrum isolated from dermatophyt...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Antibiotic resistance and virulence genes in enterococcusMohamed Hassan
This document summarizes a study on antibiotic resistance and virulence genes in Enterococcus strains isolated from hospitals in Saudi Arabia. 89 bacterial isolates were obtained from hospital patients, of which 12 were Enterococcus species. These Enterococcus isolates were tested for antibiotic resistance, identified using specific gene primers, and characterized using repetitive sequence-based PCR (Rep-PCR). The results showed that 58.3% of isolates were Enterococcus faecium, 16.6% were Enterococcus durans, and 25.1% were other Enterococcus species. 67% of isolates showed multi-drug resistance. Virulence genes were also detected in some isolates. 16S rDNA sequencing was used to further identify the isolates. The isolates exhibited genetic diversity
Development of Dot-blot Hybridization Based on 522 bp Repetitive Sequence (R5...Tenri Ashari Wanahari
Toxoplasmosis, arising from infection by Toxoplasma gondii, is one of the most common parasitic diseases in humans and other warm-blooded animals. In humans, infections are usually asymptomatic but severe disease can occur in immunocompromized individuals and newborns. Due to the importance of the disease and in order to take suitable measures, an early diagnosis of the disease is essential, particularly for pregnant women and in industry of domestic animals. The genome of T. gondii contains repeat sequences B1 and R522 which constitute ideal targets for genome-based detection methods. The 522 base pairs repeat sequences R522 are the most promising due to the high copy number, evaluated to be 200 to 300 units within the genome. We developed a simple dot-blot hybridization based on R522 sequences. The method is simple and does not require sophisticated devices. The test of the method, using cloned R522 as target, showed that the parasite detection method was sensitive and proved to be promising for use in routine health controls as well as for the survey of Toxoplasma infections.
Key words: DIG-probe, dot blot hybridization, repeat sequences, R522, Toxoplasma gondii.
ABSTRACT- This study was an attempt to estimate the prevalence of fungal isolates in superficial mycoses cases attending IPD and OPD of IIMS&R, Lucknow, Uttar Pradesh.A prospective study over a period of six (6) months was conducted from January 2015 to June 2015.The suspected cases of superficial mycoses were subjected to mycological examination with direct microscopy using 10%-40% KOH depending on the types of samples (skin, nail, hair) processed and culture on Sabouraud’s dextrose agar with chloramphenicol and cycloheximide (SDCCA) and also on Potato dextrose agar (PDA) medium. Causative agents were identified macroscopically and microscopically from the growth obtained on SDCCA and PDA. Direct microscopy revealed fungal elements in 78 (66.1%) cases whereas 54 (45.7%) were positive on culture. Out of 54 (45.7%) culture positive samples 6 (15%) were negative on microscopy (KOH mount). Tineacorporis 38 (32.2%) was the most common clinical types and male is to female ratio in relation to clinical types was 2.2:1. Commonest age group affected were 21-30 years with 41 (34.7%) cases. Males were predominantly affected 41 (75.9%) and male to female ratio being 3.1:1. 60% of the patients came from the rural background. College students formed a major chunk of the cases 29 (24.6%) followed by housewives 18 (15.3%) and unskilled workers 16 (13.6%). Trichophyton mentagrophytes 20 (37%) was the predominat isolate followed by T. tonsurans15 (27.7%), T. rubrum3 (5.5%), M. audouinii 3 (5.5%) and T. schoenleinii 2 (3.7%) with no Epidermophyton species. A non-pigmented variant of T. rubrum was identified in this study. Both SDCCA and PDA were found equally effective in isolating fungal isolates from clinical samples in our study. We are reporting change in frequency of dermatophytes isolated from superficial mycoses cases in our region.
Key words- Superficial mycoses, Non-pigmented variants, Dermatophytes
Antifungal activity of a toothpaste containing Ganoderma lucidum against Cand...Róbert Končal
The document discusses a study that tested the antifungal properties of a toothpaste containing Ganoderma lucidum against Candida albicans. Through a serial broth dilution method, the researchers found that the toothpaste had antifungal activity against C. albicans and determined its minimum inhibitory concentration (MIC) to be less than 2 mg/ml. Ganoderma lucidum has been used in traditional Chinese medicine and has antimicrobial and antiviral properties. The low MIC suggests the toothpaste could help treat or prevent oral candidiasis. However, further clinical studies are needed to confirm the laboratory findings.
Ganoderma lucidum against Candida albicansCsikós Ilona
Ganoderma lucidum has been in use in Traditional Chinese Medicine for years. Literature supports the use of this Ganoderma lucidum as a medicinal mushroom for its antimicrobial, antiviral properties. Candida albicans is the most common oral fungus associated with oral candidial infections. More: http://www.ganodermacoffee.dxnnet.com
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
A novel coronavirus from patients with pneumonia in china, 2019MANUELPERALTA33
- In December 2019, a cluster of pneumonia cases of unknown cause emerged in Wuhan, China and was linked to a seafood market.
- Researchers isolated a novel coronavirus (2019-nCoV) from bronchoalveolar lavage fluid of patients with pneumonia.
- The virus was able to infect and replicate in human airway epithelial cells in vitro, causing cytopathic effects. Electron microscopy images showed spherical virus particles around 60-140nm in diameter with distinctive spikes.
Background and study aim: During last two decades, there has been a world-wide trend in increasing occurrence of enterococcal infections in the hospitals. The aim of present study was to determine the spectrum of enterococcal infections, species prevalence, antimicrobial and characteristics of vancomycin resistant enterococci (VRE) in a tertiary care hospital, Eastern India.
Patients and Methods: Between January 2013 and July 2014, 152 Enterococcus species were obtained from clinical samples. Enterococci were identified using standard biochemical tests. Antimicrobial susceptibility was tested by Kirby-Bauer disk diffusion according to Clinical resistance
& Laboratory Standards Institute (CLSI) guidelines.VRE agar base was used to screen VRE isolates. Minimum inhibitory concentration (MIC) values of VRE isolates were determined using Epsilometer-test. VRE isolates were also examined by PCR to detect vanA gene.
Results: From 1602 clinical samples, 961 (60%) were culture positive and 152 (15.8%) enterococcal isolates were obtained. Most common species isolated was E. faecalis (63.8%) followed by E. faecium (35.5%). Majority of enterococcal infections were detected from ICUs and surgical wards and clinically presented as UTIs. Disk diffusion method showed 67.1% were resistant to penicillin, 61.2% ampicillin, 58.5% ciprofloxacin, 46.7% high-level gentamicin, 42. 8% high-level streptomycin, 7.9% teicoplanin and none to linezolid. Twenty (13.2%) enterococcal isolates were vancomycin resistant in VRE screen and disk diffusion method. Epsilometer-test of VRE isolates showed 8 (40%) isolates were resistant and 9 (45%) were intermediately resistant. From 20 VRE isolates, six showed VanA and two VanB phenotypes and all six VanA phenotypes had vanA gene cluster.
Conclusion: More accurate and reliable MIC determination tests should be performed in all suspected VRE isolates. Confirmatory PCR is required for identifying resistant gene cluster.
Key words: Enterococci, E. faecalis, E. faecium, VRE, vanA gene
The document summarizes a study on slime production by Staphylococcus epidermidis isolated from patients with corneal infections and healthy controls. A total of 57 S. epidermidis isolates were tested: 37 from patients with keratitis and 20 from healthy eyes. 52.63% of all isolates were positive for slime production, with 62.16% of patient isolates and 35% of control isolates positive. Positive slime producers exhibited higher antibiotic resistance than negative producers. Cephalothin and ciprofloxacin were the most effective antibiotics against both patient and control isolates. The results indicate slime production contributes to S. epidermidis antibiotic resistance and virulence in corneal infections.
This document outlines procedures for detecting various contaminants in human cell cultures, including microbes, fungi, viruses, and mycoplasma. It describes microscopic and macroscopic checks for initial detection of bacterial or fungal contamination. Detection of viral contamination involves using PCR or other techniques to test for pathogens like HIV, HBV, and EBV. Mycoplasma contamination is detected using one of two methods - a PCR-based test or a luminometric assay that detects mycoplasma enzyme activity. Periodic screening is important to ensure cell cultures are free of contamination.
Disinfectants play an important role in health careassociated
infection control by either minimizing or preventing
microorganism dissemination. This article to study the
morphological changes which may be related to the lose of
antibiotic resistance after disinfectant exposure using SEM.
Showed all isolates resistant to ampicillin, amoxicillin, cloxacillin,
cephalexin, tetracycline, doxycycline, rifampin, chloramphenicol,
trimethoprim cefotaxime and erythromycin, while one of burn
isolates was susceptible for gentamicin, chloramphenicol and
trimethoprim, and 15 of burn, 6 of wound, 5 of ear, and all urine
isolates were susceptible to gentamicin using Kirby-Bauer
method.
The MICs of four common in use disinfectants (Hexatane,
Dettol, Savlon and Povidone – Iodine) were determined for all
isolates. The results showed that the MICs of Hexatane ranged
from (64–512) µg/ml, Dettol (2048–16384) µg/ml,
Savlon (4096:40960)–(32768:327680) µg/ml and for Povidone –
Iodine MICs were (8192–32768) µg/ml. It has been found that
burn and urine isolates were more resistant to disinfectants than
wound and ear isolates. According to the effect of subMICs of
disinfectants at different exposure patterns on antibiotic
resistance, the results showed lose of resistance to tetracycline,
doxycycline, rifampin, chloramphenicol, cefotaxime and
trimethoprim in %72, %72, %68, %22, %28 and %36 of isolates,
respectively. The results of SEM micrograph showed normal
morphology and small sized bacteria with nub formation on some
of them when exposed to dettol, and shape changes in cells with
bulging in exposed to Povidone-iodine, while elongation and
deformation were recorded in some cells in exposed to
Savlon(chlorohexidine/ cetrimide) and Hexatane (chlorohexidine/
gluconate), respectively.
This document describes a study that identified several host defense peptide (HDP) mimetic compounds that exhibited potent antifungal activity against Candida species both in vitro and in vivo. In screening tests, several HDP mimetics were found to be broadly active against C. albicans and other Candida species, rapidly fungicidal against yeast and hyphal forms, and exhibited low cytotoxicity to mammalian cells. Importantly, the compounds showed specificity for Candida over commensal oral bacteria. In mouse models of oral candidiasis, topical application of three HDP mimetics significantly reduced the Candida burden in the tongues of infected animals.
Detection of Integrons in Multidrug Resistant Wound Isolatesijtsrd
Integrons are mobile genetic structures that carry genes responsible for resistance to different classes of antibiotics. These genetic platforms are disseminated easily among bacteria through horizontal transfer. This makes it possible for bacteria infecting parts of the body including wounds to harbor integrons resulting to poor therapeutic outcomes. This study was conducted to detect the presence of integrons in multidrug resistance isolates from wounds. Three hundred and sixty chronic wound patients were sampled using sterile cotton tipped swab sticks. The specimens were cultured according to standard microbiological procedures. The isolates were characterized by standard biochemical tests. The genomic DNA of the isolates was extracted by boiling method and was sequenced using the Big Dye kit on 3510 ABI sequencer. Antimicrobial susceptibility test was done using disc diffusion method. Multiplex Polymerase Chain Reaction was carried out on The DNA extracts using Class 1 and Class 11 Integron primers. The result shows that all 360 wound swab specimens yielded single bacteria isolate each. Pseudomonas aeruginosa was the most prevalent isolate 44.2 . The antimicrobial susceptibility test indicates that 42 isolates 11.7 were multidrug resistant MDR . Streptomycin attracted the highest resistance of 88.89 . The least resistance was to Imipenem 35.71 . The gel electrophoresis of the Multiplex PCR product indicates that 90.5 of the MDR isolates possess Class 1 Integron, 33.33 possess Class 11 Integron and 23.8 possess both Integron 1 and Integron 11. In conclusion, this study reports high prevalence of Pseudomonas aeruginosa in chronic wound swabs and 11.7 multidrug resistance among all isolates. The study also reports high prevalence of Class 1 Integron in multidrug resistance isolates. It is therefore recommended that stringent infection control measures be adopted to prevent the spread of bacteria harbouring antibiotic resistance genetic structures. Also rational antibiotic policy is recommended to avoid selection of drug resistance under antibiotic pressure. Ere, Justus Ejike | Enwuru, Chika Paulinus | Wachukwu, C. K "Detection of Integrons in Multidrug Resistant Wound Isolates" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-2 , February 2022, URL: https://www.ijtsrd.com/papers/ijtsrd49409.pdf Paper URL: https://www.ijtsrd.com/other-scientific-research-area/other/49409/detection-of-integrons-in-multidrug-resistant-wound-isolates/ere-justus-ejike
This document summarizes a study on the isolation and molecular characterization of human adenovirus. The study found that out of 83 samples collected from eye secretions, 69 (83.13%) tested positive for human adenovirus using rapid tests and PCR. The highest rate of infection was found in individuals aged 16-30 years old (55.04%) and males had a higher rate of infection than females. Human adenovirus was successfully isolated by inoculating samples on chicken embryo fibroblast cell cultures and embryonated eggs, where cytopathic effects were observed. Molecular characterization was also conducted to identify the adenovirus strains present.
A novel coronavirus from patients with pneumonia in china, 2019Juan Rubio
- In late December 2019, a cluster of patients with pneumonia of unknown cause was linked to a seafood market in Wuhan, China. Through testing samples from these patients, a novel coronavirus was discovered and named 2019-nCoV.
- Using samples from the pneumonia patients, researchers were able to isolate and culture the novel coronavirus (2019-nCoV) using human airway epithelial cells. Electron microscopy of the cultured cells showed coronavirus particles.
- Genomic sequencing of samples from the patients identified the virus as a new strain of coronavirus within the subgenus sarbecovirus, most closely related to SARS-CoV and MERS-CoV but distinct from them.
Infective endocarditis is a life-threatening disease caused by bacterial infection of the endothelium and cardiac valves, either native or prosthetic. In the present work the role of the new microbiological techniques (techniques of detection and amplification of the subunit 16 ribosomal sRNA by means of the chain reaction of the polymerase in blood or tissue, fluorescent in situ hybridization, and matrix-assisted laser is reviewed desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF MS) in the diagnosis of infective endocarditis.
This study investigated the prevalence of Toxoplasma gondii infection in 197 pets and stray cats in 4 districts of Khyber Pakhtunkhwa, Pakistan using serological (ELISA) and molecular (PCR) techniques. The results showed that T. gondii infection was significantly higher in stray cats (74.6%) compared to pet cats (25.4%). Infection rates also varied significantly between districts and were highest in Kohat (95.5%). Older cats (>4 years) had significantly higher infection rates (91.66%) than younger cats. Chronic and reactivated chronic infections (58.37%) were more common than acute infections. This research suggests T. gondii is widely
This randomized double-blinded clinical trial compared the antibacterial efficacy of a final rinse with MTAD (BioPure MTAD) and intracanal medication with 2% chlorhexidine gel (CHX) to a saline rinse in teeth with apical periodontitis. Bacterial samples were taken from the root canals at several time points and analyzed using epifluorescence microscopy and colony-forming unit counts. The results showed that bacterial counts were significantly reduced after canal preparation with NaOCl. However, the final rinse with MTAD and medication with CHX did not further reduce bacterial counts beyond the levels achieved with canal preparation alone.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
The document summarizes a study conducted over six months to identify common bacterial contaminants in a microbiology laboratory. Samples were collected from surfaces, air, hands, and clothing of laboratory personnel using culture methods and identified using biochemical and molecular tests. The most common contaminants isolated were Micrococcus (52.94% of cultures) and Bacillus subtilis (23.52% of cultures), primarily from surfaces and air, respectively. The study concludes that proper disinfection, sterilization, and personal hygiene are needed to reduce laboratory contaminants and prevent false positive culture reports and laboratory-acquired infections.
This document summarizes a study that evaluated the effects of various antiseptics on nosocomial fungi isolated from a hospital in Iraq. Eighteen fungal species were tested against Celavex-c, Povidine-iodine, Asl-slidex, and Clorox solution at different concentrations using a gradient plate technique. The study found that 0.5% Celavex-c and 2% Povidine-iodine were the most effective at inhibiting fungal growth. Asl-slidex showed the least effectiveness against the fungi even at high concentrations. In conclusion, Celavex-c, Povidine-iodine, and Clorox solution were found to be effective antise
Tobacco smoke contains mutagenic chemicals that are in the “probably carcinogenic” or “possibly carcinogenic” categories. In addition to free radicals, cigarette smoke is also rich in combustion toxic gases that can reach a very high concentration and become involved in more radical formation. Tobacco smoke contains a mixture of chemicals, including a host of reactive oxygen species (ROS), among others, that can damage cellular and sub-cellular targets, such as lipids, proteins, and nucleic acids. A growing body of evidence supports a key role for smoking-induced ROS and the resulting oxidative stress in inflammation and carcinogenesis. Smoking is one of the causes of the incidence and mortality of cancer in the world. This study aimed to review the relationship between smoking and especially the use of cigarettes with common cancers of various organs of the body. In addition to free radicals, cigarette smoke is also rich in combustion toxic gases that can reach a very high concentration and become involved in more radical formation. Smoking increases the risk of cancers of the lungs, bladder, cervix, kidney, larynx (voice box), pharynx (upper throat), nose, mouth, oesophagus (foodpipe), pancreas, stomach, liver and some types of leukaemia. Within this review article we will focus on the correlation between smoking and oxidative stress and the role of smoking in increasing the risk of cancer.
Hyper IgE syndrome especially the autosomal recessive genotype caused by DOCK8 mutation is not rare immunodeficiency disorder in Saudi Arabia due to high consanguinity. DOCK8 disruption will alter the normal immune function leading to higher susceptibility to viral infection and allergic diseases. Chronic diarrhea in AR-HIES is extremely rare; thus, we are reporting a 5-year old DOCK8 mutation who had chronic severe diarrhea explained only by cytomegalovirus infection and died despite intensive management.
More Related Content
Similar to Diagnostic tool development using advanced techniques in biotechnology for microbial keratitis.pdf
Antibiotic resistance and virulence genes in enterococcusMohamed Hassan
This document summarizes a study on antibiotic resistance and virulence genes in Enterococcus strains isolated from hospitals in Saudi Arabia. 89 bacterial isolates were obtained from hospital patients, of which 12 were Enterococcus species. These Enterococcus isolates were tested for antibiotic resistance, identified using specific gene primers, and characterized using repetitive sequence-based PCR (Rep-PCR). The results showed that 58.3% of isolates were Enterococcus faecium, 16.6% were Enterococcus durans, and 25.1% were other Enterococcus species. 67% of isolates showed multi-drug resistance. Virulence genes were also detected in some isolates. 16S rDNA sequencing was used to further identify the isolates. The isolates exhibited genetic diversity
Development of Dot-blot Hybridization Based on 522 bp Repetitive Sequence (R5...Tenri Ashari Wanahari
Toxoplasmosis, arising from infection by Toxoplasma gondii, is one of the most common parasitic diseases in humans and other warm-blooded animals. In humans, infections are usually asymptomatic but severe disease can occur in immunocompromized individuals and newborns. Due to the importance of the disease and in order to take suitable measures, an early diagnosis of the disease is essential, particularly for pregnant women and in industry of domestic animals. The genome of T. gondii contains repeat sequences B1 and R522 which constitute ideal targets for genome-based detection methods. The 522 base pairs repeat sequences R522 are the most promising due to the high copy number, evaluated to be 200 to 300 units within the genome. We developed a simple dot-blot hybridization based on R522 sequences. The method is simple and does not require sophisticated devices. The test of the method, using cloned R522 as target, showed that the parasite detection method was sensitive and proved to be promising for use in routine health controls as well as for the survey of Toxoplasma infections.
Key words: DIG-probe, dot blot hybridization, repeat sequences, R522, Toxoplasma gondii.
ABSTRACT- This study was an attempt to estimate the prevalence of fungal isolates in superficial mycoses cases attending IPD and OPD of IIMS&R, Lucknow, Uttar Pradesh.A prospective study over a period of six (6) months was conducted from January 2015 to June 2015.The suspected cases of superficial mycoses were subjected to mycological examination with direct microscopy using 10%-40% KOH depending on the types of samples (skin, nail, hair) processed and culture on Sabouraud’s dextrose agar with chloramphenicol and cycloheximide (SDCCA) and also on Potato dextrose agar (PDA) medium. Causative agents were identified macroscopically and microscopically from the growth obtained on SDCCA and PDA. Direct microscopy revealed fungal elements in 78 (66.1%) cases whereas 54 (45.7%) were positive on culture. Out of 54 (45.7%) culture positive samples 6 (15%) were negative on microscopy (KOH mount). Tineacorporis 38 (32.2%) was the most common clinical types and male is to female ratio in relation to clinical types was 2.2:1. Commonest age group affected were 21-30 years with 41 (34.7%) cases. Males were predominantly affected 41 (75.9%) and male to female ratio being 3.1:1. 60% of the patients came from the rural background. College students formed a major chunk of the cases 29 (24.6%) followed by housewives 18 (15.3%) and unskilled workers 16 (13.6%). Trichophyton mentagrophytes 20 (37%) was the predominat isolate followed by T. tonsurans15 (27.7%), T. rubrum3 (5.5%), M. audouinii 3 (5.5%) and T. schoenleinii 2 (3.7%) with no Epidermophyton species. A non-pigmented variant of T. rubrum was identified in this study. Both SDCCA and PDA were found equally effective in isolating fungal isolates from clinical samples in our study. We are reporting change in frequency of dermatophytes isolated from superficial mycoses cases in our region.
Key words- Superficial mycoses, Non-pigmented variants, Dermatophytes
Antifungal activity of a toothpaste containing Ganoderma lucidum against Cand...Róbert Končal
The document discusses a study that tested the antifungal properties of a toothpaste containing Ganoderma lucidum against Candida albicans. Through a serial broth dilution method, the researchers found that the toothpaste had antifungal activity against C. albicans and determined its minimum inhibitory concentration (MIC) to be less than 2 mg/ml. Ganoderma lucidum has been used in traditional Chinese medicine and has antimicrobial and antiviral properties. The low MIC suggests the toothpaste could help treat or prevent oral candidiasis. However, further clinical studies are needed to confirm the laboratory findings.
Ganoderma lucidum against Candida albicansCsikós Ilona
Ganoderma lucidum has been in use in Traditional Chinese Medicine for years. Literature supports the use of this Ganoderma lucidum as a medicinal mushroom for its antimicrobial, antiviral properties. Candida albicans is the most common oral fungus associated with oral candidial infections. More: http://www.ganodermacoffee.dxnnet.com
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
A novel coronavirus from patients with pneumonia in china, 2019MANUELPERALTA33
- In December 2019, a cluster of pneumonia cases of unknown cause emerged in Wuhan, China and was linked to a seafood market.
- Researchers isolated a novel coronavirus (2019-nCoV) from bronchoalveolar lavage fluid of patients with pneumonia.
- The virus was able to infect and replicate in human airway epithelial cells in vitro, causing cytopathic effects. Electron microscopy images showed spherical virus particles around 60-140nm in diameter with distinctive spikes.
Background and study aim: During last two decades, there has been a world-wide trend in increasing occurrence of enterococcal infections in the hospitals. The aim of present study was to determine the spectrum of enterococcal infections, species prevalence, antimicrobial and characteristics of vancomycin resistant enterococci (VRE) in a tertiary care hospital, Eastern India.
Patients and Methods: Between January 2013 and July 2014, 152 Enterococcus species were obtained from clinical samples. Enterococci were identified using standard biochemical tests. Antimicrobial susceptibility was tested by Kirby-Bauer disk diffusion according to Clinical resistance
& Laboratory Standards Institute (CLSI) guidelines.VRE agar base was used to screen VRE isolates. Minimum inhibitory concentration (MIC) values of VRE isolates were determined using Epsilometer-test. VRE isolates were also examined by PCR to detect vanA gene.
Results: From 1602 clinical samples, 961 (60%) were culture positive and 152 (15.8%) enterococcal isolates were obtained. Most common species isolated was E. faecalis (63.8%) followed by E. faecium (35.5%). Majority of enterococcal infections were detected from ICUs and surgical wards and clinically presented as UTIs. Disk diffusion method showed 67.1% were resistant to penicillin, 61.2% ampicillin, 58.5% ciprofloxacin, 46.7% high-level gentamicin, 42. 8% high-level streptomycin, 7.9% teicoplanin and none to linezolid. Twenty (13.2%) enterococcal isolates were vancomycin resistant in VRE screen and disk diffusion method. Epsilometer-test of VRE isolates showed 8 (40%) isolates were resistant and 9 (45%) were intermediately resistant. From 20 VRE isolates, six showed VanA and two VanB phenotypes and all six VanA phenotypes had vanA gene cluster.
Conclusion: More accurate and reliable MIC determination tests should be performed in all suspected VRE isolates. Confirmatory PCR is required for identifying resistant gene cluster.
Key words: Enterococci, E. faecalis, E. faecium, VRE, vanA gene
The document summarizes a study on slime production by Staphylococcus epidermidis isolated from patients with corneal infections and healthy controls. A total of 57 S. epidermidis isolates were tested: 37 from patients with keratitis and 20 from healthy eyes. 52.63% of all isolates were positive for slime production, with 62.16% of patient isolates and 35% of control isolates positive. Positive slime producers exhibited higher antibiotic resistance than negative producers. Cephalothin and ciprofloxacin were the most effective antibiotics against both patient and control isolates. The results indicate slime production contributes to S. epidermidis antibiotic resistance and virulence in corneal infections.
This document outlines procedures for detecting various contaminants in human cell cultures, including microbes, fungi, viruses, and mycoplasma. It describes microscopic and macroscopic checks for initial detection of bacterial or fungal contamination. Detection of viral contamination involves using PCR or other techniques to test for pathogens like HIV, HBV, and EBV. Mycoplasma contamination is detected using one of two methods - a PCR-based test or a luminometric assay that detects mycoplasma enzyme activity. Periodic screening is important to ensure cell cultures are free of contamination.
Disinfectants play an important role in health careassociated
infection control by either minimizing or preventing
microorganism dissemination. This article to study the
morphological changes which may be related to the lose of
antibiotic resistance after disinfectant exposure using SEM.
Showed all isolates resistant to ampicillin, amoxicillin, cloxacillin,
cephalexin, tetracycline, doxycycline, rifampin, chloramphenicol,
trimethoprim cefotaxime and erythromycin, while one of burn
isolates was susceptible for gentamicin, chloramphenicol and
trimethoprim, and 15 of burn, 6 of wound, 5 of ear, and all urine
isolates were susceptible to gentamicin using Kirby-Bauer
method.
The MICs of four common in use disinfectants (Hexatane,
Dettol, Savlon and Povidone – Iodine) were determined for all
isolates. The results showed that the MICs of Hexatane ranged
from (64–512) µg/ml, Dettol (2048–16384) µg/ml,
Savlon (4096:40960)–(32768:327680) µg/ml and for Povidone –
Iodine MICs were (8192–32768) µg/ml. It has been found that
burn and urine isolates were more resistant to disinfectants than
wound and ear isolates. According to the effect of subMICs of
disinfectants at different exposure patterns on antibiotic
resistance, the results showed lose of resistance to tetracycline,
doxycycline, rifampin, chloramphenicol, cefotaxime and
trimethoprim in %72, %72, %68, %22, %28 and %36 of isolates,
respectively. The results of SEM micrograph showed normal
morphology and small sized bacteria with nub formation on some
of them when exposed to dettol, and shape changes in cells with
bulging in exposed to Povidone-iodine, while elongation and
deformation were recorded in some cells in exposed to
Savlon(chlorohexidine/ cetrimide) and Hexatane (chlorohexidine/
gluconate), respectively.
This document describes a study that identified several host defense peptide (HDP) mimetic compounds that exhibited potent antifungal activity against Candida species both in vitro and in vivo. In screening tests, several HDP mimetics were found to be broadly active against C. albicans and other Candida species, rapidly fungicidal against yeast and hyphal forms, and exhibited low cytotoxicity to mammalian cells. Importantly, the compounds showed specificity for Candida over commensal oral bacteria. In mouse models of oral candidiasis, topical application of three HDP mimetics significantly reduced the Candida burden in the tongues of infected animals.
Detection of Integrons in Multidrug Resistant Wound Isolatesijtsrd
Integrons are mobile genetic structures that carry genes responsible for resistance to different classes of antibiotics. These genetic platforms are disseminated easily among bacteria through horizontal transfer. This makes it possible for bacteria infecting parts of the body including wounds to harbor integrons resulting to poor therapeutic outcomes. This study was conducted to detect the presence of integrons in multidrug resistance isolates from wounds. Three hundred and sixty chronic wound patients were sampled using sterile cotton tipped swab sticks. The specimens were cultured according to standard microbiological procedures. The isolates were characterized by standard biochemical tests. The genomic DNA of the isolates was extracted by boiling method and was sequenced using the Big Dye kit on 3510 ABI sequencer. Antimicrobial susceptibility test was done using disc diffusion method. Multiplex Polymerase Chain Reaction was carried out on The DNA extracts using Class 1 and Class 11 Integron primers. The result shows that all 360 wound swab specimens yielded single bacteria isolate each. Pseudomonas aeruginosa was the most prevalent isolate 44.2 . The antimicrobial susceptibility test indicates that 42 isolates 11.7 were multidrug resistant MDR . Streptomycin attracted the highest resistance of 88.89 . The least resistance was to Imipenem 35.71 . The gel electrophoresis of the Multiplex PCR product indicates that 90.5 of the MDR isolates possess Class 1 Integron, 33.33 possess Class 11 Integron and 23.8 possess both Integron 1 and Integron 11. In conclusion, this study reports high prevalence of Pseudomonas aeruginosa in chronic wound swabs and 11.7 multidrug resistance among all isolates. The study also reports high prevalence of Class 1 Integron in multidrug resistance isolates. It is therefore recommended that stringent infection control measures be adopted to prevent the spread of bacteria harbouring antibiotic resistance genetic structures. Also rational antibiotic policy is recommended to avoid selection of drug resistance under antibiotic pressure. Ere, Justus Ejike | Enwuru, Chika Paulinus | Wachukwu, C. K "Detection of Integrons in Multidrug Resistant Wound Isolates" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-2 , February 2022, URL: https://www.ijtsrd.com/papers/ijtsrd49409.pdf Paper URL: https://www.ijtsrd.com/other-scientific-research-area/other/49409/detection-of-integrons-in-multidrug-resistant-wound-isolates/ere-justus-ejike
This document summarizes a study on the isolation and molecular characterization of human adenovirus. The study found that out of 83 samples collected from eye secretions, 69 (83.13%) tested positive for human adenovirus using rapid tests and PCR. The highest rate of infection was found in individuals aged 16-30 years old (55.04%) and males had a higher rate of infection than females. Human adenovirus was successfully isolated by inoculating samples on chicken embryo fibroblast cell cultures and embryonated eggs, where cytopathic effects were observed. Molecular characterization was also conducted to identify the adenovirus strains present.
A novel coronavirus from patients with pneumonia in china, 2019Juan Rubio
- In late December 2019, a cluster of patients with pneumonia of unknown cause was linked to a seafood market in Wuhan, China. Through testing samples from these patients, a novel coronavirus was discovered and named 2019-nCoV.
- Using samples from the pneumonia patients, researchers were able to isolate and culture the novel coronavirus (2019-nCoV) using human airway epithelial cells. Electron microscopy of the cultured cells showed coronavirus particles.
- Genomic sequencing of samples from the patients identified the virus as a new strain of coronavirus within the subgenus sarbecovirus, most closely related to SARS-CoV and MERS-CoV but distinct from them.
Infective endocarditis is a life-threatening disease caused by bacterial infection of the endothelium and cardiac valves, either native or prosthetic. In the present work the role of the new microbiological techniques (techniques of detection and amplification of the subunit 16 ribosomal sRNA by means of the chain reaction of the polymerase in blood or tissue, fluorescent in situ hybridization, and matrix-assisted laser is reviewed desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF MS) in the diagnosis of infective endocarditis.
This study investigated the prevalence of Toxoplasma gondii infection in 197 pets and stray cats in 4 districts of Khyber Pakhtunkhwa, Pakistan using serological (ELISA) and molecular (PCR) techniques. The results showed that T. gondii infection was significantly higher in stray cats (74.6%) compared to pet cats (25.4%). Infection rates also varied significantly between districts and were highest in Kohat (95.5%). Older cats (>4 years) had significantly higher infection rates (91.66%) than younger cats. Chronic and reactivated chronic infections (58.37%) were more common than acute infections. This research suggests T. gondii is widely
This randomized double-blinded clinical trial compared the antibacterial efficacy of a final rinse with MTAD (BioPure MTAD) and intracanal medication with 2% chlorhexidine gel (CHX) to a saline rinse in teeth with apical periodontitis. Bacterial samples were taken from the root canals at several time points and analyzed using epifluorescence microscopy and colony-forming unit counts. The results showed that bacterial counts were significantly reduced after canal preparation with NaOCl. However, the final rinse with MTAD and medication with CHX did not further reduce bacterial counts beyond the levels achieved with canal preparation alone.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
The document summarizes a study conducted over six months to identify common bacterial contaminants in a microbiology laboratory. Samples were collected from surfaces, air, hands, and clothing of laboratory personnel using culture methods and identified using biochemical and molecular tests. The most common contaminants isolated were Micrococcus (52.94% of cultures) and Bacillus subtilis (23.52% of cultures), primarily from surfaces and air, respectively. The study concludes that proper disinfection, sterilization, and personal hygiene are needed to reduce laboratory contaminants and prevent false positive culture reports and laboratory-acquired infections.
This document summarizes a study that evaluated the effects of various antiseptics on nosocomial fungi isolated from a hospital in Iraq. Eighteen fungal species were tested against Celavex-c, Povidine-iodine, Asl-slidex, and Clorox solution at different concentrations using a gradient plate technique. The study found that 0.5% Celavex-c and 2% Povidine-iodine were the most effective at inhibiting fungal growth. Asl-slidex showed the least effectiveness against the fungi even at high concentrations. In conclusion, Celavex-c, Povidine-iodine, and Clorox solution were found to be effective antise
Similar to Diagnostic tool development using advanced techniques in biotechnology for microbial keratitis.pdf (20)
Tobacco smoke contains mutagenic chemicals that are in the “probably carcinogenic” or “possibly carcinogenic” categories. In addition to free radicals, cigarette smoke is also rich in combustion toxic gases that can reach a very high concentration and become involved in more radical formation. Tobacco smoke contains a mixture of chemicals, including a host of reactive oxygen species (ROS), among others, that can damage cellular and sub-cellular targets, such as lipids, proteins, and nucleic acids. A growing body of evidence supports a key role for smoking-induced ROS and the resulting oxidative stress in inflammation and carcinogenesis. Smoking is one of the causes of the incidence and mortality of cancer in the world. This study aimed to review the relationship between smoking and especially the use of cigarettes with common cancers of various organs of the body. In addition to free radicals, cigarette smoke is also rich in combustion toxic gases that can reach a very high concentration and become involved in more radical formation. Smoking increases the risk of cancers of the lungs, bladder, cervix, kidney, larynx (voice box), pharynx (upper throat), nose, mouth, oesophagus (foodpipe), pancreas, stomach, liver and some types of leukaemia. Within this review article we will focus on the correlation between smoking and oxidative stress and the role of smoking in increasing the risk of cancer.
Hyper IgE syndrome especially the autosomal recessive genotype caused by DOCK8 mutation is not rare immunodeficiency disorder in Saudi Arabia due to high consanguinity. DOCK8 disruption will alter the normal immune function leading to higher susceptibility to viral infection and allergic diseases. Chronic diarrhea in AR-HIES is extremely rare; thus, we are reporting a 5-year old DOCK8 mutation who had chronic severe diarrhea explained only by cytomegalovirus infection and died despite intensive management.
AIM AND OBJECTIVE: • To compare and assess the clinical features and outcome of pneumonia occurring in infants with traditional child rearing practices and in infants without traditional child rearing practices. • To assess the influence of individual traditional child rearing practices with the outcome of pneumonia occurring in infants. MATERIALS AND METHODS: : For this study detailed clinical history was taken. Infants with clinical and radiological evidence of pneumonia were selected as per the selection criteria The study. After eliciting necessary history some children were excluded using exclusion criteria. Detailed questionnaires were asked to mother / care taker which included details regarding various traditional child rearing practices. . RESULTS: On comparing and analyzing the clinical parameters and outcome of pneumonia in infants between traditional child rearing practices and without traditional child rearing practice. it is found that increased morbidity pattern of pneumonia in infants associated with traditional child rearing practices is high while comparing non traditional child rearing infants CONCLUSION: In my study increased morbidity in the infants is attributed to traditional child rearing practice, mortality was very less probably due to increased vaccination status and also due to increasing literacy rate in mothers.
The aim of the study is to compare intubating conditions using rocuronium with gold standard relaxant suxamethonium and to evaluate intubating conditions with rocuronium using the timing principle. This study was carried out in the department of Anaesthesiology, Patients posted for elective general surgical procedure were included in study. In this study, 96.7% patients in group I (Sch) exhibited excellent intubating conditions. In group II only 63.3% patients exhibited excellent intubating conditions, where timing principle is not followed. In group III and group IV 100% and 96.7% patients exhibited excellent intubating conditions respectively. Good intubating conditions were found in 3.3%, 36.6% and 3.3% patients in group I (Sch), group II and group IV respectively. The excellent and good intubating conditions were considered acceptable whereas the fair and poor intubating conditions were unacceptable. To conclude rocuronium bromide can be considered as an alternative to suxamethonium even if rapid sequence intubation is required.
Background &Objective: Klebsiella pneumonia causes different serious nosocomial infections for human and several strains became multiple drug resistance .This study was conducted to describe the epidemiology and molecular typing of Klebsiella pneumonia with the extended spectrum of B lactamase enzyme in Gaza strip .Methods :A cross-sectional survey was conducted during the period of December 2008 to November2009. One hundred and fifty clinical specimens were collected from patients admitted in different wards . Results : Sixty six percentage of the isolates were K.pneumonia .These were isolated from different infected sites : urine 24% , sputum 14%, wound 11% , stool11% , blood14% , cerebrospinal fluid 11% , skin16% . The ESBLs was detected in 67% of the strains ,53% strains were resistant for more than eight antibiotics , PCR demonstrated different patterns for the presence of SHV(80%) , TEM(60%) enzyme and CTX-M(20%), PFGE Showed 10 clusters of genetically unrelated strains with high prevalence of polyclonal strains of Klebsiella pneumonia. Antibiotic resistance was found against Cephalothin(95.0%),Cefotaxime(82.0%),Ceftazidime(59.0%),Ceftriaxone(86.0%),Gentamicin(56.0%),Trimethoprim/sulphamethoxazole(47.0%)..Chloramphenicol(42%),Amikacin(33%),Aztreonam (32%) and Imipenem(0%). Interpretation, Conclusion : our findings showed that genetically-related isolates of K. pneumoniae producing SHV and TEM and CTX-M were present in Gaza Strip. Larger studies need to be done to better define the molecular epidemiology of ESBL producing K. pneumoniae and its clinical implications
Introduction Psoriasis is a chronic inflammatory & most common dermatologic diseases. It is related to several diseases but most common is cardiovascular disease. Lipid abnormalities are observed in psoriasis patients at the earliest stages of the disease and so it may be genetically determined.Hence there is a great need to study lipid abnormalities in psoriatic patients, which will help us to evaluate the level of risk individuals may experience for developing atherosclerosis and vascular obstructive disorders, as well as associated morbidity and mortality. Aim: To estimate lipoproteins levels & related risk of cardiovascular diseases in psoriasis patients Materials and Methods: This is a community-based cross sectional analytical study. The study population was derived from the healthy ambulatory person in and around Pimpri area & was conducted in Dr. D.Y.Patil Medical College & Hospital, Pimpri Pune. Study group consist of 40 psoriatic patients with mean age of 50.68 ± 8.86 years and control group consisted of 40 healthy individual of either sex with mean age of 50.15 ± 9.6 years. 55 % males & 45 % females were present in control group while test group comprises of 59 % males & 41 % females. estimation of Serum total cholesterol, Serum triglyceride, Serum High-Density Lipoprotein-cholesterol & Serum Low density Lipoprotein levels were done the same day. Estimation of lipoproteins levels done by Enzymatic End Point Method. Tha data was analysed by calculating mean, standard deviation, p-value and Chi-square test. Results : Findings of this show that, total cholesterol ,VLDL was significantly increased in psoriasis compared to control(p<0.05).Difference between psoriasis cases(6.8 ± 1.49) and control (3.08 ± 1.3) in findings of serum TGs were highly significant(p<0.001).However, difference in between serum LDL level in psoriasis cases and control was not significant. There were no significant changes shown in serum HDL level in both psoriasis cases and control . LDL/HDL ration has not shown any significant changes in both psoriasis case and control, while in TC/HDL ratio, difference was statistically significant(p<0.05). Conclusion: Present study suggests that patients with psoriasis were associated with hyperlipidaemia. So have high risk of developing the cardiovascular diseases. We suggest that screening of serum lipid profile in psoriatic patients will help in evaluating risk at early stage. Impromptu treatment in these patients will prevent the risk of cardiovascular diseases.
This document provides instructions for authors submitting manuscripts to the International Journal of Biological and Medical Research (IJBMR). The journal publishes original research papers across all areas of contemporary biological and medical fields. Manuscripts should be organized with sections including title page, abstract, keywords, introduction, materials and methods, results, discussion, conclusions, acknowledgements, and references. Authors are instructed to submit manuscripts online through the journal website and provide all relevant publication details and formatting according to the instructions. The submission and review process is also outlined.
Over the years, I have noticed that my hands would react vigorously to hand-sanitizers. The anti-bacterial gel could cause my skin to burn with a slight redness, but the degree to which this happened varied. This led me to wonder why this burning sensation was different for different sanitizers, or why some sanitizers were more reactive than others. With research, I understood that this difference was due to differences in the effect on live bacteria. During my internship at the Shri Baidyanath Research Institute, an ayurvedic pharmaceutical institute, I learned in detail the manner in which the effectiveness of their antibacterial medicines was experimentally tested. Using the breadth of knowledge I gained from my time there, I decided to investigate, through experimentation, the effectiveness of different hand-sanitizers against bacteria, or more specifically, E. coli strain K-12.
Pregnancy with portal hypertension with splenectomy is an uncommon condition.It is seen that maternal mortality is 2-18% ,hematemesis is seen in 20-30% and perinatal mortality is 11-18% [1]. A number of patients with Extra hepatic portal venous obstruction (EHPVO) and non cirrhotic portal fibrosis (NCPF) are surviving to adult life. In patients with cirrhosis as long as liver function is relatively preserved pregnancy is possible.
INTRODUCTION: In India maxillofacial injuries are among the common ones that present to emergency department. Maxillofacial injury can be defined as injury to the facial soft tissue, skeleton and other associated structures resulting deformity or destruction of jaw and eyes. MATERIAL & METHOD: The cases were documented as per proforma, noting name, age, sex, education, religion, socioeconomic status, occupations, mode of trauma and clinical history were recorded and also noted the past H/o of different complaints of ear, nose, throat and face. X-rays and CT scan were done to identify fractures of different maxillofacial region. RESULTS: Results of this study showed that among 150 patients, 122 (81.33%) were males, maximum 62 (41.33%) patients were in 21-30 years of age group, the face was the commonest site of maxillofacial injuries (73.33%), followed by nose (67.33). The nasal bleeding (78.67%) was commonest noted symptom and assaults were the most common cause (48%) of facial injury and nasal bone was the commonest fractured bone (32.66%), CONCLUSION: The incidence of maxillofacial injuries were maximum in age group of 21-30 years and male were more prone for maxillofacial trauma. The most common symptom in present series was epistaxis. Nasal bone was the most commonly fractured bone and assault was the commonest cause of facial injury as compared to road traffic accident. This study showed that increase number of interpersonal violence responsible for facial injury.
The aim of the study was to evaluate the antibacterial evaluation of root extracts of Juglans regia against Extended Spectrum Beta Lactamase (ESBL) producing E. coli and Klebsiella pneumonia in Bombay Hospital and Research Centre Jabalpur. The antibacterial activity of, ethyl acetate and methanol root extracts of Juglans regia was determined by disk diffusion method. The antibacterial activity was calculated based on the minimum inhibitory concentration using Mueller–Hinton broth in a tube-dilution method. The best antibacterial activity, calculated as minimum inhibitory concentration values, against ESBL was shown by the methanol root extract Juglans regia (25 mg/mL) for both isolated organisms and ethyl acetate (25mg/mL) against E. coli. The methanol extract showed zone of inhibition in the range of 17-26mm as compared to ethyl acetate extract which showed zone of inhibition in the range of 11-16mm against the uropathogens. The zone of inhibition ranged from 17 mm to 26 mm and MIC was 25mg/ml. This effect is comparable to antibiotics. The results obtained in the present study suggest that Juglans regia have the potential to be developed as antibacterial agents against ESBL producing UTI bacteria strain. Further investigations are needed to identify the active compounds and their mechanism of action
Introduction: Snake bite is considered to be one of the common, neglected and potential life-threatening public health issues in tropical and sub-tropical countries. According to World Health Organization (WHO), 4.5- 5.4 million people get bitten by snakes annually. Acute kidney injury is one of the major health issues due to snake envenomation and can be managed with appropriate dose of Anti-snake venom and other symptomatic approach. Methods: a prospective observational study, all the patients who were admitted to tertiary care hospital between October 2019 to September 2020 and developed Acute kidney injury were included. All the relevant data were collected from case reports and case sheets and patient interaction. Patients with the definitive history of renal disease, exposure to nephrotoxic drugs were excluded. All the statistical data were analyzed using descriptive statistics. Results: a total of 40 patients were included with a mean age of 40.07 (±10.01) years. This study shows a male predominance of 72.5% (n=29). The clinical manifestations like Cellulitis, local swelling, hypotension was commonly seen. Severe damage to kidney was seen in patients who received native medicine before admitting to hospital. Conclusion: this study concludes that acute kidney injury is one of the major causes of morbidity, there is need of education on snake envenomation and its treatment and first aid and effect of native medicine
Objective: To study the effects of plant growth promoter fungicide & plant hormone Abscisic acid on the growth and biochemical properties of the plant. In our study we have taken Basella alba (Malabar Spinach) as a model system for our study. Methodology: It was an experimental approach. Wines with 3-6 nodes were selected and planted separately in 20 pots. Difenoconazole and ABA at a specific concentration were used. The plants were irrigated with tap water. The treatments were given on 25, 50 and 75 days after planting (DAP) by soil drenching, The plants were taken randomly on 45, 65 and 85 DAP for analyses and separated into root, stem and leaves used for determining growth and biochemical changes. Results: The total height of the plant increased with ABA treatment but decreased with Difenoconazole whereas there was increase in rootlenghth in both the treatments. The total leaf area of the plant decreased in both Difenoconazole and Abscisic acid treatment. We found significant increase in Chlorophyll, Cartoneoid & Xanthophyll content of the plant against both the treatments. In the same manner remarkable increase in the total protein content, amino acid content and phenol content was also observed after both the treatments. Conclusion: This study aims to reveal the efficacy of the growth regulating fungicide and Abscisic acid in the growth and biochemical properties of the plant. Conclusion: It was concluded that Difenoconazole and Abscisic acid are effective in overall plant development though Difenoconazole is least effective in increasing the total leaf area and height of the plant.
Paratyphoid fever has been emerged as a global public health problem, especially in developing countries. There should be a consistent monitoring of the isolation throughout the countries to analyze the spectrum of the Salmonella enteric serovar Paratyphi. This study examined current isolation pattern of Salmonella paratyphi A, B and C over a period of 6 years at National Salmonella and Escherichia Centre (NSEC), Central Research Institute (C.R.I), Kasauli. Miscellaneous suspected cultures of Salmonella had been recieved from various regions of India during six years span of January 2010- December 2015. These samples were characterized by biotyping as well as serotyping at NSEC situated at Central Research Institute, Kasauli. Isolates were serotyped on the basis of somatic O and phase 1 and phase 2 flagellar antigens by agglutination tests with antisera according to the Kauffmann White scheme. Out of 71 isolates of Salmonella enterica, 51 (71.830%) were S. Paratyphi A, 16 (22.532%) were S. Paratyphi B and 4 (5.633%) were S. Paratyphi C. Among total 71 samples of Salmonella Paratyphi, 11.267% were obtained from North India while 88.732% cases were from South India. 81.96 % samples were isolated from blood while rest of samples were from feces, urine, pus etc. Increasing rates of antibiotic resistance among S. enterica, particularly in S. Paratyphi A strains, is of concern, as S. Paratyphi A infection is becoming increasingly common and is not prevented by current vaccinations. This study caters the Salmonella Paratyphi A, B, C characterization by biotyping and serotyping status in various regions of India irrespective of their co-relation to the region of isolation, source of sample isolation, types of isolates including their age, gender and season during period of 2010 to 2015 across India.
This article discusses compound heterozygosity and its role in the pathogenesis of hypertrophic cardiomyopathy (HCM). It analyzes 100 HCM cases and 100 controls to study compound heterozygous variations, especially in the MYH7 and MYBPC3 genes. 22 cases were found to have at least one mutation in a sarcomeric gene as well as other polymorphisms. These cases exhibited genetic anticipation, with the disease manifesting at an earlier age in each generation. The article focuses on several cases in detail, analyzing how compound heterozygous mutations affect splicing, mRNA structure, codon usage bias, and protein structure. It suggests that creation or loss of splicing sites and cryptic splice sites, as well as effects on
This article analyzes the parasporin (anti-cancer) proteins of Bacillus thuringiensis through in silico methods. Physico-chemical properties, secondary structure, 3D structure, motifs, and phylogenetic relationships were analyzed for 19 parasporin proteins. The findings help characterize the proteins and predict their cancer cell killing mechanisms. Secondary structure analysis showed predominance of random coil. 3D structure prediction identified domains and structural quality. Motif analysis found endotoxin motifs common to most proteins. Understanding these proteins could help develop them as anti-cancer agents.
Rasamanikya is a excellent preparation in the field of Rasashastra, it is used in various Kushtha Roga, Shwasa, Vicharchika, Bhagandara, Vatarakta, and Phiranga Roga. In this article Preparation& Comparative analytical profile for both Formulationon i.e Rasamanikya prepared by Kushmanda swarasa & Churnodhaka Shodita Haratala. The study aims to provide insights into the comparative efficacy and analytical aspects of these formulations for enhanced therapeutic outcomes.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Cell Therapy Expansion and Challenges in Autoimmune DiseaseHealth Advances
There is increasing confidence that cell therapies will soon play a role in the treatment of autoimmune disorders, but the extent of this impact remains to be seen. Early readouts on autologous CAR-Ts in lupus are encouraging, but manufacturing and cost limitations are likely to restrict access to highly refractory patients. Allogeneic CAR-Ts have the potential to broaden access to earlier lines of treatment due to their inherent cost benefits, however they will need to demonstrate comparable or improved efficacy to established modalities.
In addition to infrastructure and capacity constraints, CAR-Ts face a very different risk-benefit dynamic in autoimmune compared to oncology, highlighting the need for tolerable therapies with low adverse event risk. CAR-NK and Treg-based therapies are also being developed in certain autoimmune disorders and may demonstrate favorable safety profiles. Several novel non-cell therapies such as bispecific antibodies, nanobodies, and RNAi drugs, may also offer future alternative competitive solutions with variable value propositions.
Widespread adoption of cell therapies will not only require strong efficacy and safety data, but also adapted pricing and access strategies. At oncology-based price points, CAR-Ts are unlikely to achieve broad market access in autoimmune disorders, with eligible patient populations that are potentially orders of magnitude greater than the number of currently addressable cancer patients. Developers have made strides towards reducing cell therapy COGS while improving manufacturing efficiency, but payors will inevitably restrict access until more sustainable pricing is achieved.
Despite these headwinds, industry leaders and investors remain confident that cell therapies are poised to address significant unmet need in patients suffering from autoimmune disorders. However, the extent of this impact on the treatment landscape remains to be seen, as the industry rapidly approaches an inflection point.
Clinic ^%[+27633867063*Abortion Pills For Sale In Tembisa Central19various
Clinic ^%[+27633867063*Abortion Pills For Sale In Tembisa Central Clinic ^%[+27633867063*Abortion Pills For Sale In Tembisa CentralClinic ^%[+27633867063*Abortion Pills For Sale In Tembisa CentralClinic ^%[+27633867063*Abortion Pills For Sale In Tembisa CentralClinic ^%[+27633867063*Abortion Pills For Sale In Tembisa Central
Osteoporosis - Definition , Evaluation and Management .pdfJim Jacob Roy
Osteoporosis is an increasing cause of morbidity among the elderly.
In this document , a brief outline of osteoporosis is given , including the risk factors of osteoporosis fractures , the indications for testing bone mineral density and the management of osteoporosis
TEST BANK For Community Health Nursing A Canadian Perspective, 5th Edition by...Donc Test
TEST BANK For Community Health Nursing A Canadian Perspective, 5th Edition by Stamler, Verified Chapters 1 - 33, Complete Newest Version Community Health Nursing A Canadian Perspective, 5th Edition by Stamler, Verified Chapters 1 - 33, Complete Newest Version Community Health Nursing A Canadian Perspective, 5th Edition by Stamler Community Health Nursing A Canadian Perspective, 5th Edition TEST BANK by Stamler Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Pdf Chapters Download Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Pdf Download Stuvia Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Study Guide Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Ebook Download Stuvia Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Questions and Answers Quizlet Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Studocu Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Quizlet Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Pdf Chapters Download Community Health Nursing A Canadian Perspective, 5th Edition Pdf Download Course Hero Community Health Nursing A Canadian Perspective, 5th Edition Answers Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Ebook Download Course hero Community Health Nursing A Canadian Perspective, 5th Edition Questions and Answers Community Health Nursing A Canadian Perspective, 5th Edition Studocu Community Health Nursing A Canadian Perspective, 5th Edition Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Pdf Chapters Download Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Pdf Download Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Study Guide Questions and Answers Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Ebook Download Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Questions Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Studocu Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Stuvia
Adhd Medication Shortage Uk - trinexpharmacy.comreignlana06
The UK is currently facing a Adhd Medication Shortage Uk, which has left many patients and their families grappling with uncertainty and frustration. ADHD, or Attention Deficit Hyperactivity Disorder, is a chronic condition that requires consistent medication to manage effectively. This shortage has highlighted the critical role these medications play in the daily lives of those affected by ADHD. Contact : +1 (747) 209 – 3649 E-mail : sales@trinexpharmacy.com
Integrating Ayurveda into Parkinson’s Management: A Holistic ApproachAyurveda ForAll
Explore the benefits of combining Ayurveda with conventional Parkinson's treatments. Learn how a holistic approach can manage symptoms, enhance well-being, and balance body energies. Discover the steps to safely integrate Ayurvedic practices into your Parkinson’s care plan, including expert guidance on diet, herbal remedies, and lifestyle modifications.
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Diagnostic tool development using advanced techniques in biotechnology for microbial keratitis.pdf
1. BioMedSciDirect
Publications
Int J Biol Med Res.2023 ;14(2):7575- 7581
Contents lists available at BioMedSciDirect Publications
Journal homepage: www.biomedscidirect.com
International Journal of Biological & Medical Research
International Journal of
BIOLOGICAL AND MEDICAL RESEARCH
www.biomedscidirect.com
Int J Biol Med Res
Volume 14, Issue 2, April 2023
Copyright 2010 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685. All rights reserved.
c
ARTICLE INFO ABSTRACT
Keywords:
Ocular infection
keratitis
bacterial keratitis
mycotic diseases
multiplex PCR
Introduction
Cornealinfectionisoneoftheleadingcauseofvisualloss[1]an
estimated 6.8 million people suffer from keratitis in India and the
number is expected to increase up to 10.6 million by 2020 [2].
Bacterial keratitis is the most common form of suppurative
corneal ulceration and a variety of fungi are also implicated in this
infection. Infective keratitis especially of mycotic origin has been
reported from diverse climatic zones, it appears to be more
common in subtropical and tropical regions. A global increase in
the incidence of this disease has been noted in recent decades,
mainly due to the topical use of antibacterial and steroidal
preparations [3] which compromise the cornea to opportunistic
fungalinfections.
While many organisms are capable of causing ocular infection
microbiologic examination of clinical specimens is required for
diagnosis. Microbiology tests are successful in identifying the
causative organism; however, Multiplex PCR has been shown to be
especially suited for detecting small amounts of microbial DNA
present in ocular specimens [4-10]. This is particularly true for
virusesthatsetupinfectionoftheeye.Whileonlyalimitednumber
of viruses especially, cytomegalovirus, herpes simplex virus and
zosterviruswhicharetheetiologicagents.
A large number of bacterial and fungal pathogens are
commonly encountered during diagnosis of keratitis. The 16S
subunit of rDNA of bacteria and 18S subunit (ITS regions) of the
fungi are the most conserved region that can different one species
from another [11]. Our study aims at using PCR amplification and
sequence analysis of 16S rDNA to detect bacterial and 18S rDNA to
detect fungal pathogens in subjects with keratitis. The amplified
sequences were deposited in the GenBank. A unique primer was
designed for each organism and efforts are underway to design a
universalprimerforbacteriaandfungiwhichwillenableusto
Purpose: To study the demographic characteristics, associated factors, causative agents, of
infectious keratitis and develop a diagnostic tool to aid easy diagnosis of keratitis.Methods:
Corneal scrapes were collected and subjected to culture, microscopy, considering age,
occupation, geographical, Frequency of predisposing ocular conditions, antibiotic
susceptibility test, resistance patterns test, drug sensitivity and 16s r-DNAas well as 18s r-DNA
based identification was performed and submitted to data bank with accession number. The
16S rDNA sequences of the individual bacteria and fungi were used for a universal primer
designandtherebymultiplexPCRcanbeperformed.Results:Atotalof250consecutivepatients
with infective keratitis were evaluated, of which 77 (30.8%) were found to be of bacterial, 67
(26.8%) were fungal, 16 (6.4%) were both fungal and bacterial, and the remaining 90 (36%)
were found to be culture negative. Contact lens wear was the main risk factor (80.8%). Ocular
surface disease (23.6%), ocular trauma (14.8%), corneal surgery (4.4%) and corneal suture
(6.4%) of cases were found in corneal ulcers. Most community acquired bacterial and fungal
ulcers resolve with appropriate treatment. 64% of the infections involved positive cultures and
36% involved negative cultures, were found in polymicrobial mode of infection. Fusarium spp.
(32.75%)wasthemostpredominantspeciesfollowedbyAspergillussp.(20.68%)wasfoundin
fungal corneal ulcers and also Staphylococcus sp. was the most common bacteria found in
bacterial cultures. The 16S rDNA sequence of the bacteria and fungi cultured from the isolates
of the corneal scrapping were performed and the genes were submitted in Genbank.The
primers designed for bacteria and fungi gave good results in the multiplex PCR carried out and
wesuggestthiscanbeusedasadiagnostictoolforkeratitis.
Original article
Diagnostic tool development using advanced techniques in biotechnology for
microbial keratitis
V.Nithya,* Anusha Bhaskar
Assistant Professor in Microbiology, Urumu Dhanalakshmi College, Kattur, Tiruchirappalli, Tamil Nadu, India.
* Corresponding Author :
Assistant Professor in Microbiology, Urumu Dhanalakshmi College,
Kattur, Tiruchirappalli, Tamil Nadu, India.
E-mail: nithyavaradharaj20@gmail.com
*Dr. V.Nithya
Copyright 2011. CurrentSciDirect Publications. IJBMR - All rights reserved.
c
2. develop a diagnostic tool for easy diagnosis of the nature of
keratitis by multiplex PCR. This analysis will ascertain the
organism(s) responsible for the infection and also help in the
diagnosisofmixedinfections.
MATERIALSANDMETHODS
Studypopulationandsamplecollection
In this study 250 consecutive outpatients 143 males and 107
females were with corneal ulcerations were examined in the Eye
ClinicoftheGeneralHospitalofTiruchirappallifromJanuary2009
to February 2012. This study was approved by the Research Ethics
Committee, of the Hospital. An ophthalmic history, including risk
factors for corneal ulceration (ocular surface disease, previous
ocular surgery, contact lens use of trauma) and use of antibiotics
or steroids was obtained from each patient. All patients were
examinedunderaslit-lampbiomicroscopebyanophthalmologist.
Corneal scrapings were collected after instillation of 4%
lignocaine without preservative under aseptic conditions from
each ulcer by an ophthalmologist using a sterile Bard Parker blade
(No 15). Scrapings were performed under magnification of slit-
lamp or operating microscope. Leading edge and base of each
ulcer were scraped initially and the material obtained were
directly inoculated onto the surface of solid media such as sheep
blood agar, chocolate agar and Sabouraud Dextrose Agar (SDA) in
a row of C- shaped streaks and also deep inoculation in the liquid
media such as brain heart infusion (BHI) broth without
gentamycin sulphate and thioglycollate medium. Subsequent
scrapings were spread onto labeled slides in a thin, even manner
for 10 % potassium hydroxide (KOH) wet mount and Gram
staining. Control patients were defined as patients who had
epithelial defects of noninfectious etiologies or infective keratitis.
Thecontrolsubjectswereagematched.
PrimaryScreening
All bacterial cultures were incubated aerobically. Cultures on
chocolate agar were evaluated at 24 - 48 hours and then discarded
if no growth was seen. All media were incubated at 35°C (±1)
except SDA, which are incubated at 27°C (±1) in BOD incubator.
Cultures inoculated in BHI broth were examined for turbidity and
subsequently sub cultured and gram stained for identification.
Determination of positive culture samples were done according to
methodsbyBharathiet.al[12].
SecondaryScreening(Culturebased)
Thespecificidentificationofbacterialpathogenswasbasedon
microscopic morphology, staining characteristics, and
biochemical properties using standard laboratory criteria. Fungi
were identified by their colony characteristics on SDA and by their
microscopicappearanceinLactophenolcottonblue.
Antibacterialsensitivitytest
Antibiotic susceptibility testing of each isolate was done using
routine antibiotics discs Gentamycin; Ampicillin; Tetracycline; Co-
trimaxazole; Ceftriaxon; Cephalotin; Cefotaxime; Ceftriaxon;
Kanamycin according to disc diffusion technique by Kirby Bauer
method on Muller Hinton agar (MHA) and were evaluated by as
perthestandards[13].
Anti-fungaldrugsensitivitytest
Various antimicrobials were incorporated in the study to
check the sensitivity of the filamentous isolates. The diffusion
methodhasbeenemployedusingtabletsofamphotericinB,
clotrimazole, econazole, flucytosine, miconazole, and nystatin.
These diffusion tablet tests were performed as recommended by
Casals[14].Forallcases,postdiagnosisantimicrobialtherapywas
given. Subsequent treatment was tailored according to the
microbiological diagnosis and sensitivity results. The final visual
acuity was defined as the visual acuity on discharge from the ward
(Notdocumentedinthiswork).
MolecularIdentification
In this study aliquots from positive cultures were obtained
from the keratitis.. For most sequencing experiments a 'universal'
primer was used, this being one that was complementary to the
partofthevectorDNAimmediately.
In most cases, agarose gel electrophoresis is used for the
separationofDNAfragmentsrangingfrom10kb~0.2kb,avertical
polyacrylamide gel is more appropriate henece used. The
amplification of 16S r-DNA of any bacterial species is possible
without prior cultivation when broad-range PCR primers targeted
to highly conserved regions are applied[15]. Identified bacteria in
corneal scrapings and in vitreous samples of patients who had
keratitis and endophthalmitis by amplification and subsequent
direct sequencing of 16S r-DNA were performed by designing
specificprimersfordesignatedbacterialandfungalisolates.
PrimerdesignformultiplexPCR
The primer for individual bacteria and fungi were designed
using Primer 3 software. Then using these individual primers a
commonprimerforbacteriaandfungiweredeveloped.
Reactionvolume
Take five 0.2ml or 0.5 ml PCR tubes and added 0.5 μl of each
DNA sample, Distilled water 18.3µl, Taq DNA polymerase buffer
(10x) 2.5µl, dNTP mix (2mM) 2.5µl, 2 Universal Primers (10µM)
0.5µl for each, DNA polymerase enzyme (5U/µl) 0.2µl, Total
ReactionVolume25.0µltotherespectivePCRtube
MultiplexPCR
Multiplex PCR amplification was performed in a Gene Cycle
under the following conditions: first heat initial denaturation
94ºC for 5 minutes, cycle denaturation 94ºC for 45 seconds for 34
cycles, annealing 50ºC for 45 seconds and extension 72ºC for 1
minute. This was followed by incubation at 72 ºC for 10 minutes
and cooling at 4 ºC. Negative control reaction mixtures contained
sterile distilled water in place of template DNA. Finally resolve the
PCR product onto 1% agarose gel electrophoresis was used to
visualize multiplex PCR DNA product and photographed under UV
(Progen Biotech, Salem, Tamil Nadu). The universal primers were
synthesizedatSigma,Bangalore.
Results
Epidemiologicalcharacteristics
Out of 250 patients 143 (52.7 %) were males and 107 (42.8 %)
were females. There were 153 (61.5%) rural residents and
97(38.5%)urbanresidents.Patientsabovetheageof50years115
(46 %) were significantly less than patients below 50 years 135
(54 %) were shown in Figure 1. Non-agricultural workers (Figure
2)weresignificantlylessinnumberthanwerefarmers108(54%).
Frequency of predisposing ocular conditions in bacterial
keratitis
Contact lens (CL) wear was the most common risk factor. This
wasencounteredin127eyes(50.8%).SoftCLwasnotedin41.7%
7576
V.Nithya & Anusha Bhaskar et al./Int J Biol Med Res.14(2):7575- 7581
3. of cases, extended wear contact lens in 44.8%, and disposable
contact lens wear in 13.5% of cases were found in CL cases (Table
1).
In 59 cases of ocular surface diseases, 45.8% cases of
aphakic bullous keratopathy, 16.9% cases of epidermal dysplasia
and 37.3% cases of chronic bullous keratopathy were found. In 37
cases of ocular trauma, 59.1% of workplace-related trauma,
13.9% of home-related trauma and 27% motor vehicle-related
trauma were found in corneal patients. In 11 cases of corneal
related surgery, 45.5% of Penetrating Keratoplasty (PK), 36.4% of
Anterior Lamellar Keratoplasty (LK) and 18.1% Endothelial
Keratoplasty (EK) were found. In 16 cases of corneal sutures,
18.8% of facial nerve plasy (leprosy), 31.2% of eyelid trauma,
12.5% of Exposure keratopathy, 12.5% of Secondary to cataract
extraction and 25% of Thyroid Exophthalmopathy were found in
cornealulcersareshowninTable2.
Microbiologicaldiagnosis
Cultures were positive and fulfilled the criteria established for
the presence of infection in 110 (55%) of the 200 corneal ulcers
(Table 3). Pure bacterial growth was present in 70 (28%) of the
250 cultures performed and pure fungal growth in 67 (26.8%).
Mixed microbial growth was present in the cultures of 16 (6.4%)
of the 250 patients. A total of 354 bacterial organisms were
cultured from160 corneal ulcers (Table 4). Of the 70 isolates, 228
(64.5%) were Gram positive and 126 (35.5%) were Gram-
negative bacteria. Staphylococcus sp. was the most commonly
isolatedbacterialorganismrepresenting99(66.3%)ofallpositive
bacterial cultures. The next most commonly isolated Gram-
positive organism was propionibacterium acnes with 31
(39.24%)positivecultures.Ofthese31cultures,19werepure,and
12 were mixed with fungi. Pseudomonas aeruginosa was isolated
from40cultures(34.48%)andfollowedbyserratiasp.38cultures
were the most frequently occurring Gram-negative organism. A
total of 116 fungal organisms were cultured from equal number of
corneal ulcers (Table 6) of which 38 (32.75%) were Fusarium sp.,
24 (20.68%) were Aspergillus sp., 17 (14.65%) were Candida sp.,
9 (7.75%) were Byssochlamys nivea, 10 (8.62%) were Sebipora
aquosa and 18 (5.51%) were A. fumigates were found in fungal
species. Of the 116 fungal ulcers 39 were positive for fungal
elementsonKOHexamination.
In CL wearers group, 65.3% of the corneal scrapings were
positive mostly Staphylococcus sp. (34.7) of isolated bacteria and
Gram negative, mostly Pseudomonas aeruginosa. Contact lens
and/or storage cases cultures were performed in 127 cases. In
ocular disease group, 67% of the corneal scrapings were positive
whereas 37% of isolated bacteria were found to be Gram negative.
In ocular trauma group, 58.2% of the corneal scrapings were
positive whereas 41.8% of isolated bacteria were Gram negative
was found. In corneal surgery group, 65.3% of the corneal
scrapings were positive whereas 34.7% of isolated bacteria were
Gram negativewas found.In cornealsuturesdiseasegroup, 72.7%
ofthecornealscrapingswere
AntibioticResistanceincausativeBacteria
In vitro resistance to at least one antibiotic was found in 55
(33%) bacterial isolates (Table 8). Resistance to multiple
antibioticswasfoundin250isolates,S.aureushadthehighestrate
of resistance, whereas no isolates of P. aeruginosa were resistant
to tested antibiotics. Scrapings from cases related to prior ocular
surgery were more likely to culture resistant bacteria than
scrapingsrelatedtootherkeratitisriskfactors.
Antifungalsensitivitytest
Antifungal sensitivity test as were performed and Fusarium
spp. were found to be more resistance, while Candida spp. is most
sensitive among all isolates. In these tests the Azoles drugs were
mostsensitivityorganismswhencomparedwithPolyenes
Molecularidentification
Inordertodeveloptheproposedmethodology,theprimersfor
conserved regions of the rDNA for the bacterial and fungal
pathogens were synthesized. The DNA isolated from the bacteria
and fungi were amplified and sequenced and the sequences were
used for the strain identification. A BLAST was carried out to
confirm the organism from the database. The sequences were
submitted to the GenBank. A unique primer was designed for
bacteria and fungi and multiplex PCR was carried out and the
results obtained are shown in Fig 3. This could be positively
developed into an unique diagnostic tool for identification of the
causativeorganismofkeratitis.
Table 1: Frequency of predisposing ocular conditions in
bacterialkeratitis
Table 2: Ocular Factors Predisposing to Culture-Proven
BacterialKeratitis
7577
V.Nithya & Anusha Bhaskar et al./Int J Biol Med Res.14(2):7575- 7581
4. 7578
Table 3: Microbial growth pattern of cultures from corneal
ulcers (n=250) from the Eye clinic of the General Hospital in
Thiruchirappalli,TamilNadu,India.
Table 5: Organisms isolated in bacterial corneal ulcers.
Polybacterialinfectionwasnotedinthiscases
Table 4: Bacterial isolates from corneal ulcers
V.Nithya & Anusha Bhaskar et al./Int J Biol Med Res.14(2):7575- 7581
5. 7579
Table 10: Drug sensitivity of fungi isolated from cases of
keratitis
GM: Gentamycin; AM: Ampicillin; TE: Tetracycline; SXT: Co-
trimaxazole; CRO: Ceftriaxon; CF: Cephalotin; CTX:
Cefotaxime;TOB:Ceftriaxon;K:Kanamycin;–:notused
V.Nithya & Anusha Bhaskar et al./Int J Biol Med Res.14(2):7575- 7581
6. 7580
Fig 1 Age of patients with corneal ulceration (n=250) in the
Eye clinic of the General Hospital in Thiruchirappalli, Tamil
Nadu,India.
DISCUSSION
This study evaluated demographic, clinical, microbiological,
and treatment information taken from medical records of all
patients that had corneal scrapings over a 3 year period at a public
hospital in Tiruchirappalli India. In this population, we found
there average age of patients was 51 years, and 60% of patients
were men. Contact lens wear and ocular trauma were more
common risk factors in young patients, whereas prior ocular
surface disease and previous ocular surgery were more common
riskfactorsinolderpatients.
Thispredominanceofmenpresentingwithtraumatickeratitis
is reflected in another recent study, which found that 90% of
traumatickeratitisoccurredinmen.
TherehasbeeninthepastseveralyearsasteadyincreaseinCL
wearers. Subsequently, CL wear is now the major predisposing
factor for corneal infection in the United States and Western
Europe [16] and a matter of public health concern. Soft CL have
greatly increased the risk of bacterial keratitis, which is estimated
tobe10–20timeshigherwiththeuseofextendedweardisposable
CL.8 hypoxia and hypercapnia of the cornea induced effects were
alreadyreportedincaseofcontactlensewearers[17].
Multiple organisms have been reported from the microbial
keratitis seen in association with CL wear. We found a moderate
shift towards higher prevalence of Gram negative rods compared
with that prevalence in the absence of CL wear and we confirmed
the results of previous studies. These results confirm that Gram
negative bacteria are more often associated with soft CL wear.
However, conversely to what might be expected, the percentage of
Gram negative organisms is lower than those of Gram positive
organisms. Approximately two thirds of CL related bacterial
keratitis are associated with Gram positive cocci, such as
staphylococci and streptococci and one third is associated with
Gram negative rods, especially Pseudomonas aeruginosa and
Serratiamarcescens,
This result suggests that contact lenses and cases may tend to
preferentially harbour Gram negative organisms, but that the
causative organism may well not match that organism, and indeed
Gram positive organisms are important in contact lens related
keratitis.
Of 57 fungal isolates cultured from equal number of corneal
ulcers 29.82% were Fusarium spp., 21.05% were Aspergillus spp.
and there were no onfilamentous fungus because these are
specifically overcomes the Fusarium sp, is similar reported from
South Florida and from Ghana by previous researchers. The
numerous species occurred in india are like Fusarium spp. as the
predominantfungalspeciesarereportedbyBharathietal[12].
More than three quarters of cases were treated initially on an
outpatient basis. All the patients have been traeted with good
traditional antibiotics. Thus most have undergoes the
combination of antibiotics, whereas broad spectrum antibiotic
therapy. There was no evidence of proven fungal keratitis above
the age of 61 yr. in north India reported in 2005 [18], but in this
study we have a broad spectrum of infection up to the age of 71 yrs
[19]. Status of antibiotic resistance also a huge interruption in the
line of treatment of keratitis with some routine antibiotics like
cyclohexamide,amphoterin,incaseoffungalspectrum[20,21].
This case audit has defined the common risk factors, causative
organisms, antibiotic resistance, patient demographics, clinical
presentations, of patients with keratitis presenting to a major
hospitalinregionofsouthIndia.easabestmodeofdiagnosis.
Fig 2 Occupation of patients with corneal ulceration (n=250)
in the Eye clinic of the General Hospital in Thiruchirappalli,
TamilNadu,India.
*Anindividualeithermaleorfemale,whodoesheavymanual
labor, lifting loading & carrying of material usually balanced
onhead;
† Middle class workers such as mechanics, stonemasons,
electricians, carpenters, pipe fitters, welders, and fishermen.
This category also includes profession as teachers, police,
officeworkers,factoryworkers,driversandmerchants
Fig:3 Electrophoresis of multiplex PCR products from
organismscausingkeratitis
L1-DNA100bpstepladder(Promega)
Agarose Gel (1%) M – 1Kb DNA Ladder,AG – Aspergillus
fumigatus,CA–Candidaalbicans
FM – Fusarium sp., KA – Klebsiella pneumoniae, PM –
Pseudomonasaeruginosa
SA – Staphylococcus aureus,MPL – Multiplex PCR (for all
organisms
V.Nithya & Anusha Bhaskar et al./Int J Biol Med Res.14(2):7575- 7581
7. 7581
We, also investigated the possibility of using PCR amplification
and 16S rDNA and 18S rDNA of bacteria and fungi respectively to
identify the microorganisms in the setting of the disease. The PCR
based technique has a certain advantage over the standard culture
methods because the results are obtained with shorter time than
the conventional methods. We have sequenced the conserved
regions of the rDNA and they have been submitted to the Genbank.
Onthebasisoftheresultsobtainedwehavedesignedaprimerone
common for bacteria and one for fungi and the multiplex PCR
carriedoutgaveverygoodandreproducibleresults.
Conclusion
The clinical histories, an agricultural status for most patients,
recollection of a preceding trauma from vegetable matter and soil
andthepatternsofcausativefungiandbacteria,viz.,P.aeruginosa,
S. penumoniae, among the bacteria and Aspergillus sp. Fusarium
oxysporum Penicillium sp. Paecilomyces farinosus among the
fungi. The filamentous fungi showed sensitivity in decreasing
order to flucytosine, nystatin, amphotericin B, and econazole. In
conclusion, this study demonstrated that culture-proven bacterial
keratitis mainly affected middle-aged males who sustained ocular
trauma. Gram-negative bacteria in general and Pseudomonas
aeruginosa in particular were the most common microorganisms
isolated. In this study we have carried out the molecular
identification of the organisms causing keratitis and we also
recommendmultiplexPCRtechniqu
All rights reserved.
Copyright 2019 BioMedSciDirect Publications IJBMR - ISSN: 0976:6685.
c
1. Wilhelmus KR. Bacterial keratitis, p. 970–1031. In JS. Pepose, GN. Holland,
and KR. Wilhelmus editors. Ocular infection and immunity. Mosby Year Book
Inc.,St.Louis,Mo.1996
2. Whitcher JP, Srinivasan M, Upadhyay MP. Corneal blindness A Global
perspective.BullWHO;2001;79;214-21
3. Chin GN, Hundiuk RA, Kwasny GP, Schultz GP. Keratomycosis in Wisconsin.
AmJOphthalmol1975;79:121-25.
4. Cunningham, ET, Short GA, Irvine AR. AIDS-associated herpes simplex virus
retinitis: clinical description and use of a polymerase chain reaction-based
assayasadiagnostictool.Arch.Ophthalmol.1996;114:834–840.
5. Dean D, Millman K. Molecular and mutation trends analysis ofomp1 alleles
forserovarEofChlamydiatrachomatis.J.Clin.Invest.1997;99:475–483.
6. Dean D, Schachter JS, Dawson CR, Stephens RS.. Comparison of the major
outer membrane protein variant sequence regions of B/Ba isolates: a
molecular epidemiological approach to Chlamydia trachomatis infections. J.
Infect.Dis.1992;166:383–392.
7. Hykin PG, Tobal K, McIntyre G, Matheson MM, Towler HMA, Lightman SL.
The diagnosis of delayed post-operative endophthalmitis by polymerase
chain reaction of bacterial DNA in vitreous samples. J. Med. Microbiol. 1994;
40:408–415.
8. Knox CM, Margolis TP, Chandler D, Short GA. PCR based assays for the
diagnosis of viral retinitis: use in diagnostic dilemmas. Ophthalmology
1998;105:37–45.
9. McCann JD, Margolis TP, Wong MG. A sensitive and specific polymerase chain
reaction-based assay for the diagnosis of cytomegalovirus retinitis. Am. J.
Ophthalmol.1995;120:219–226.
10. Short GA, Margolis TP, Kuppermann BD. A polymerase chain reaction-based
assay for the diagnosis of varicella-zoster virus retinitis in patients with
AIDS.Am.J.Ophthalmol.1997;123:157–164.
11. Relman DA.Theidentificationofunculturedmicrobialpathogens.
J.Infect.Dis.1993;168:1–8.
12. Bharathi MJ, Ramakrishnan R, Vasu S; Meenakshi, Palaniappan R.
Aetiological diagnosis of microbial keratitis in South India - a study of 1618
cases.IndianJMedMicrobiol.2002;20(1):19-24.
13. Duru ME, Cakir A, Kordali S, Zengin H, Harmandar M, Izumi S, Hirata T.
Chemical composition and antifungal properties of essential oils of three
Pistaciaspecies.Fitoterapia2003;74,170–176.
14. Casal JB. Tablet sensitivity testing on pathogenic fungi. J Clin Pathol, 1979;
32,719–722.
15. Joseph J, Sharma S, Somasheila IM, Krishna PV, Garg P, Nutheti R, Kenneth J,
Balasubramanian D. Microsporidial keratitis in India: 16S rRNA gene-based
PCR assay for diagnosis and species identification of microspordia in clinical
samples.InvestOphthalandVisualScience2006;47:4468–4473.
16. Jones DB. Pathogenesis of bacterial and fungal keratitis. Trans Ophthalmol
SocUK1978;98:367-71.
17. Wood TO, Williford W. Treatment of keratitis with amphotericin B 0.15%.
AmJOphthalmol1976;75:847-49
18. Chowdhary A , Singh K. Spectrum of Fungal Keratitis in North India, Cornea:
2005–24;8-15
19. Kaufman E, Wood RM. Mycotic keratitis. Am J Ophthalmol 1965; 59: 993-
1000.
20. Flynn JR. Fusarium keratitis treated with cycloheximide. Am J Ophtbalmol
1964;58:637-41.
21. Iwata K. Drug resistance in human pathogenic fungi. Eur J Epidemiol 1992;
8:407-21
References
V.Nithya & Anusha Bhaskar et al./Int J Biol Med Res.14(2):7575- 7581