Incidence of fluconazole resistance in candida spp. among immunocompromised patients

Objective: To determine fluconazole resistance in Candida spp. Among immunocompromised patients. Methodology: This study included of total 100 Bronchoalveolar lavage fluid samples of cancer patients, which comprised 37 lung cancer, 19 lymphoma, 18 leukemia, 4 oral cancer, 7 GI cancer, 6 bone cancer and 9 breast cancer proved by histology reports. Samples were collected from Jinnah Hospital, Lahore, Pakistan. Sabouraud’s dextrose agar was used for the isolation of Candida spp. and further confirmed by Gram staining and germ tube test. Disk diffusion method was used to check the susceptibility against fluconazole. Results: Out of patients, 65 were male and 35 females. Out of 100 samples, 40 were positive for lung cancer, 30 for leukemia and 30 for other malignancies proved by histopathology reports. From the samples, 28 (28%) were positive for Candida spp. and within these positive samples, 12 (42.71%) strains were C. albicans and 16 (57.14%) were non albicans species. Out of these 28 positive strains, 4 (14.85%) were resistant, 2 (7.14%) susceptible dose dependent and 22 (78.57%) sensitive to fluconazole. Conclusion: Isolation of candida spp. other than the Candida albicans and presence of resistance to most common drug that was used for the treatment is making the situation alarming. It needs more research to find out new antifungal drugs and mechanisms of drug resistance among Candida sp

Circulating microRNAs in oncogenic viral infections: potential diagnostic biomarkers

Cancer is a leading cause of high death rate worldwide. One strategy to control the disease is the early diagnosis by novel biomarkers that express during early stage of the disease. The recent diagnostic strategies in cancer don’t have enough specificity to promote the detection of cancer at its beginning. Many biomarkers like protein biomarkers and metabolites are being used for diagnosis of various cancer types but miRNAs are excellent among them, because they have distinctive biochemical characteristics. Moreover, to raise the precision and capability of miRNA to diagnose cancer, the analyzing of both miRNAs and as well as selective mRNA will help in creating a more complete categorizer. Virus constitutes the cause of 20% of entire human cancer cases and both RNA and DNA viruses are linked with tumors in both animal and man. Even though many viruses can cause different tumors in animals, only some of them are linked with human cancers and are presently regarded as oncogenic viruses. These viruses include Human Papillomavirus (HPV), Hepatitis B (HBV) and Hepatitis C Virus (HCV), Epstein Barr Virus (EBV), Human Herpes Virus 8 (HHV8), Human T cell Leukemia Virus (HTLV) and Merkel Cell Polyomavirus (MCPyV). Expression data of miRNA in several cancers reveal that miRNA profile is different in cancer cells as compared to normal cells. So, miRNA could be useful biomarker for the detection of cancer. The present study strengthens a foundation and gives a logic to investigate the ability of miRNAs as circulating biomarkers in various cancers

Carbapenem resistance gene crisis in A. baumannii: A computational analysis

Acinetobacter baumannii (A. baumannii) is one of the members of ESKAPE bacteria which is considered multidrug resistant globally. The objective of this study is to determine the protein docking of different ARGs in A. baumannii. In silico analysis of antibiotic resistance genes against carbapenem are the blaOXA-51, blaOXA-23, blaOXA-58, blaOXA-24, blaOXA-143, NMD-1 and IMP-1 in A. baumannii. The doripenem, imipenem and meropenem were docked to blaOXA-51 and blaOXA-23 using PyRx. The top docking energy was − 5.5 Kcal/mol by imipenem and doripenem and meropenem showed a binding score of -5. 2Kcal/mol each and blaOXA-23 energy was − 4.3 Kcal/mol by imipenem and meropenem showed a binding score of -2.3 Kcal/mol, while doripenem showed the binding score of -3.4 Kcal/mol. Similarly, doripenem imipenem and meropenem were docked to blaOXA-58, IMP-1, Rec A and blaOXA-143, with docking energy was − 8.8Kcal/mol by doripenem and meropenem each while imipenem showed a binding score of -4.2Kcal/mol and with IMP-1 demonstrated their binding energies. was − 5.7 Kcal/mol by meropenem and doripenem showed a binding score of -5.3Kcal/mol, while imipenem showed a binding score of -4.5 Kcal/mol. And docking energy was − 4.9Kcal/mol by imipenem and meropenem showed binding energy of -3.6Kcal/mol each while doripenem showed a binding score of -3.9Kcal/mol in RecA and with blaOXA-143 docking energy was − 3.0 Kcal/mol by imipenem and meropenem showed a binding score of -1.9Kcal/mol, while doripenem showed the binding score of -2.5 Kcal/mol respectively. Doripenem, imipenem, and meropenem docking findings with blaOXA-24 confirmed their binding energies. Doripenem had the highest docking energy of -5.5 Kcal/mol, meropenem had a binding score of -4.0 Kcal/mol, and imipenem had a binding score of -3.9 Kcal/mol. PyRx was used to dock the doripenem, imipenem, and meropenem to NMD-1. Docking energies for doripenem were all – 4.0Kcal/mol, whereas meropenem had docking energy of -3.3 Kcal/mol and imipenem was − 1.50Kcal/mol

Evaluation of 16S rRNA methyltransferase gene in aminoglycosides resistant isolates of cancer patients

Aminoglycosides are used in empiric treatment of critically ill patients. Efficacy of aminoglycoside has been reduced due to dissemination of resistance. The aim of this study was to evaluate aminoglycoside resistance in cancer patients with pneumoniae. A total of 150 Bronchoalveolar lavage and Bronchial washing samples were collected from cancer patients. The samples were identified with standard microbiological procedures. Phenotypic susceptibility pattern of the isolates was determined against various groups of antibiotics such as Penicillins, Cephalosporins, Carbapenems, Monobactams, Aminoglycosides, Tetracyclins, Glycopeptides and Sulphonamides. The isolates with phenotypic resistant to aminoglycosides were further evaluated for the presence of armA gene. The strains of E. coli (12.5%), S. aureus (15.6%), Streptococcus (15.6%), Pseudomonas (18.7%) and K. pneumoniae (37.5%) were isolated. The phenotypic resistance profile showed highest resistance against aminoglycosides (Tobramycin, 53.1% Gentamicin and 50% Amikacin) followed by cephalosporins and sulfonamides group. The armA gene was detected in aminoglycoside resistant isolates. The overall genotypic resistance was evaluated as 21.8%. The armA gene was found in K.pneumoniae 23.5%, Pseudomonas 11.8% (4/24) and E. coli 5.9%. High level resistance to aminoglycosides raises therapeutic concern to health care professionals. These findings highlight the importance of effective monitoring and surveillance to the use of broad-spectrum antibiotics

Evaluation of Antibiotic Resistance and Virulence Genes among Clinical Isolates of Pseudomonas aeruginosa from Cancer Patients

Objectives: The objectives of this study were to evaluate P. Aeruginosa isolates from cancer patients for the phenotypic pattern of antibiotic resistance and to detect the gene responsible for virulence as well as antibiotic resistance. Methods: A total of 227 P. aeruginosa isolates were studied and 11 antibiotics were applied for susceptibility testing. PCR detection of the genes BIC, TEM, IMP, SPM, AIM, KPC, NDM, GIM, VIM, OXA, toxA and oprI was done. Finally, the carbapenem resistant isolates were tested for phenotypic identification of carbapenemase enzyme by Modified Hodge test. Results: The results showed that the isolates were resistant to imipenem (95%), cefipime (93%), meropenem (90%), polymixin B (71%), gentamicin (65%), ciprofloxacin (48%), ceftazidime (40%), levofloxacin (39%), amikacin (32%), tobramycin (28%) and tazobactum (24%). The PCR detection of the carbapenem resistant genes showed 51% isolates were positive for IMP, GIM and VIM, 38% for AIM and SPM, 30% for BIC, 20% for TEM and NDM, 17% for KPC and 15% for OXA. However, toxA and oprI genes were not detected. 154 carbapenem resistant isolates were found positive phenotypically for carbapenemase enzyme identification by Modified Hodge test. Conclusion: The co-existence of multiple drug-resistant bodies and virulent genes has important implications for the treatment of patients. This study provides information about treating drug-resistant P. Aeruginosa and the relationship of virulent genes with phenotypic resistance patterns

Biological and physical characterization of bacteriophage JHA against multidrug-resistant Acinetobacter baumannii

Due to the emergence of antibiotic resistance, bacteriophage therapy appears to be an ideal weapon to utilize against pathogenic bacteria. This study aimed to isolate, identify and characterize the lytic bacteriophage effective against the multidrug-resistant Acinetobacter baumannii clinical isolates. The isolated bacteriophage caused lysis by applying the double-layer agar technique on A. baumannii up to 99% in 18 hours of incubation at 37ºC. The bacterial growth reduction assay exhibited that JHA phage had high adsorption rates and could rapidly inhibit bacterial growth. The pH and thermal stability testing showed that JHA phage was stable in vast ranges of pH from 5 to 9 but its activity was highest at pH7 (1860000±1000 pfu/mL). It was stable in broad ranges of temperatures from 25ºC to 60ºC but the highest activity was found at 37ºC (1300000±30000 pfu/mL). One-step growth test results showed that it has a short latent period, strong lytic ability, high burst size, and adsorption rates and was host specific. Scanning electron microscopy (SEM) of JHA phage demonstrated icosahedral heads and tailless particles. Transmission electron microscopy (TEM) revealed JHA phage belongs to Tectiviridae family. All the characteristics of JHA phage possess lytic activity against A. baumannii strains and exhibit novel candidates to use as an alternative competitor to antibiotics in controlling such infections

Potential impact of microbial consortia in biomining and bioleaching of commercial metals

Biomining is the use of microorganisms for the commercial extraction of lavish metals from ores and mines with least effect on environment. Microbes play vital role in bioleaching procedures in commercial mining. The bacterial cells are used to detoxify/replace waste cyanide, marginal biomass and activated carbon. These methods are preferred over conventional techniques due to energy efficient, low cost, environment friendly and production of useful by-products. At industrial scale, different microbial strains (Acidophilic, Sulphobacillus, Rhodococcus, Ferrimicrobium &chemolithotrophic) are deployed to boost the processes of copper and uranium bioleaching. About 20% of the world’s copper is extracted by using this technique. These extraction procedures involve oxidation of insoluble metal sulphides to soluble sulphates. The isolation of thermophilic microbes for mineral biooxidation increase the commercial extraction of minerals at industrial scale. The conventional pyrometallurgical techniques have environmental concerns as they result in depletion of high grade ores and release harmful gaseous. The microbe-assisted gold mining is expected to double the yield of gold and needs to be fully explored using diverse array of microbes. Bioleaching is simple and low cost method for the developing countries with large ore deposits. About 30 strains of microbes have been discovered so for with potential impact on bioleaching. With advances in molecular genetics, physiology and microbial genomics, more promising strains with increased bioactivities are possible. Further efforts are underway to culture diverse range of archaea and improving its genetic potential to be used as industrial tool for commercial bioleaching. The currents review enlightens the recent trends in biomining/bioleaching and implementation of modern biological approaches to engineer target microbes for commercial use

MRSA Clinical Isolates Harboring mecC Gene Imply Zoonotic Transmission to Humans and Colonization by Biofilm Formation

This study was conducted for the molecular detection of the mecA, mecC, and nuc gene among MRSA and to investigate biofilm formation among the methicillin-resistant Staphylococcus aureus (MRSA) clinical isolates. A total of 208 different samples were collected and processed for phenotypic and genotypic identification of MRSA. All MRSA isolates were subjected to antibiotics sensitivity, cefoxitin disk diffusion test, and vancomycin minimum inhibitory concentration (MIC) E-test. The MRSA isolates were detected for the presence of mecA, mecC, and nuc genes. Congo red agar (CRA) method was used to assess the ability of isolates to form biofilms. The results of the study showed that the prevalence of MRSA was 48%. The MRSA isolates were highly resistant (100%) to penicillin, β lactamase inhibitors, cephalosporins, and macrolides. All the MRSA isolates were susceptible to vancomycin antibiotic drugs. Cefoxitin (30 µg) disk diffusion test showed 100% sensitivity and specificity for the identification of MRSA phenotypically. A total of 100 MRSA clinical isolates were positive for the mecA and nuc gene. Only 3 MRSA isolates were positive for the mecC gene. Congo red agar method showed that 20 (20%) isolates formed moderate biofilm while 80 (80%) isolates were non-biofilm forming. Multi drugs resistant and mecC gene-positive MRSA isolates are rapidly emerging in Pakistan. Therefore, the mecC gene should be detected along with the mecA gene for the identification of MRSA clinical isolates. It also requires early identification of biofilm formation and necessary interventions for its effective treatment and contro

Molecular Characterization of Mercury Resistant Bacteria Isolated from Tannery Wastewater

Mercury resistant (HgR) bacteria were isolated from heavy metal polluted wastewater and soil, collected from the proximity of some tanneries from Kasur, Pakistan. Three out of 30 bacterial strains were screened out on the basis of resistance level against various concentrations of HgCl2. Bacterial isolates AZ-1, AZ-2 and AZ-3 showed resistance up to 40 μg/mL of HgCl2 and mercury sensitive (HgS) isolate ZA-15 was taken as a negative control. 16S rDNA ribotyping and phylogenetic analysis were performed for the characterization of isolates as Bacillus sp. AZ-1 (KT270477), B. cereus AZ-2 (KT270478), B. cereus AZ-3 (KT270479) and Enterobacter cloacae ZA-15 (KJ728671). Phylogenetic relationship on the basis of merA nucleotide sequence confirmed 51-100% homology with the corresponding region of the merA gene of already reported mercury resistant Gram-positive bacteria. Restriction fragment length polymorphism (RFLP) analysis was applied to the amplification products of 16S rRNA and merA genes and a specific restriction patterns was successfully obtained after treatment with different endonucleases. A small-scale reservoir of Luria Bertani (LB) medium supplemented with 30 μg/mL of HgCl2 was designed to check the detoxification ability of the selected strains. The results demonstrated 83% detoxification of mercury by both B. cereus AZ-2 and B. cereus AZ-3, and 76% detoxification by Bacillus sp. AZ-1 (p<0.05)

Study of Gc-ms And Hplc Characterized Metabolic Compounds in Guava (Psidium Guajava L.) Leaves

Psidium guajava leaves are rich source of nutrients, antioxidants, phytoconstituents and biological active compounds. The study was designed to elucidate secondary metabolites like alkaloids, saponins, flavonoids, tannins and glycosides in extracts of guava leaves through Gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) by qualitative as well as quantitative procedures. These metabolites were further tested for their antimicrobial potential against two-gram positive (Bacillus subtilis and Staphylococcus aureus) and two-gram negative (Escherichia coli and Pasteurella multocida) bacteria and three pathogenic fungal strains (Asprrgillus niger, Fusarium solani and Aspergillus flavus). GC-MS analysis revealed the presence of major constituents like Ca- Caryophyllene (22.70%), α cubebene (11.2%) and alpha Humulene (5.91%). The ethyl-acetate, methanol, n-hexane and chloroform extracts were tested for antibacterial and antifungal activities against above mentioned microbes. Among all the tested solvent extracts, Chloroform and ethyl acetate extracts of P. guajava demonstrated more sensitivity towards the growth of B. subtilis and P. multocida with MIC of 230±3.02, 316. ±6.2 and 237±5.09 and 288±1.55 μg/ml, respectively. Methanolic extracts showed higher MIC against S. aureus (233±5.51 μg/ml) and E. coli (192±2.05 μg/ml), respectively. The findings of this current study would provide the way to use guava as a potential therapeutic agent to combat antimicrobial and antifungal resistance