Multidrug Resistance Properties Research Articles

In vitro lytic efficacy of bacteriophages against multidrug-resistant pathogenic bacterial species isolated from pyogenic skin infections

Bacterial multidrug resistance has worsened the situation by adding to economic burden but also poses a greater risk of patient death. The aim of the study was to characterize the multidrug resistance (MDR) properties of the bacterial isolates from skin infections and then to isolate and evaluate lytic efficacy of bacteriophages against the pathogenic bacteria. Antimicrobial susceptibilities of the isolates (n=84) from pyogenic skin infections against 14 antibiotics was studied using CLSI guidelines. Phylogenetic analyses of the MDR strains from each species was performed. Lytic efficacy of the sewage-derived phages was assessed by spot test. was the most predominant (57, 68%) of the total of 84 isolates. The number of Gram-negative isolates that were resistant to all antibiotics (except amikacin) were significantly higher (P<0.05). On the other hand, significant number of strains were susceptible only to clindamycin and erythromycin (P<0.05). Phylogenetic analysis based on the 16S rRNA gene revealed close relatedness of the strains with MDR strains previously reported. analysis of select MDR strains (n=20) showed that the bacteriophages ΦDMSA-2, ΦDMEC-1 and ΦDMPA-1 against , , and , respectively, showed lytic efficacy against 4 of 5 MDR strains tested from each species. These preliminary, but still important results emphasize the potential of phages as an effective alternative therapy against MDR bacteria. Further, the lytic efficacy of phages underscores the importance of developing need-based and locally isolated bacteriophages as potential antimicrobial therapy alternative to antibiotics.

Antibacterial Activity of Composite Mixture of Senna siamea Leaves and Tamarind Pulp Extracts on Multidrug Resistant Salmonella typhi

Background: For many years, antimicrobial chemotherapeutic approach have been challenged by drugs resistant bacteria such as Salmonella associated with gastrointestinal infections. To overcome these effects, several attempts by herbalists and traditional healers were in progress, using some plants parts to treat Typhoid fever in Nigeria.
 Aim: The aim of this work was to determine the antibacterial activity of Senna siamea leaves and Tamarindus indica fruit pulp against multidrug resistant Salmonella typhi.
 Methods: A total of 168 samples of stool and blood from patients with suspected cases of typhoid fever were collected and only three (3) Salmonella typhi isolates were recovered (from the 168 samples) by conventional microbiological techniques. The isolates were screened for multidrug resistant properties according to Kirby -Bauer disc diffusion method. Water and ethanol were used to extract phytochemical components from powdered leaves of Senna siamea and Tamarindus indica fruit pulp via percolation method. The extracts were tested for the antibacterial activity against the clinical isolates of Salmonella typhi.
 Results: All the isolates (100%) were resistant to Ampicillin, Amoxicillin, while two (66.7%) to Erythromycin and Tetracycline and sensitive (100%) to Streptomycin, Gentamycin, Nalidixic acid Ciprofloxacin, Cotrimoxazole, Augmentin and Chloramphenicol, The phytochemical screening o of both plants extracts showed the presence of alkaloids, terpenoids, flavonoids, saponins, tannins, carbohydrates and cardiac glycosides. The result shows that aqueous and ethanolic extracts of combined Tamarind and Senna siamea were more active (2.50µg/ml) against Salmonella typhi when compared to individual extracts.
 Conclusion: This study shows that aqueous and ethanol extracts of both plants exhibited activity on S. Typhi, hence, possess antimicrobial potentials that it can be used in treatment of typhoid.

Deciphering the Role of S-adenosyl Homocysteine Nucleosidase in Quorum Sensing Mediated Biofilm Formation.

S-adenosylhomocysteine nucleosidase (MTAN) is a protein that plays a crucial role in several pathways of bacteria that are essential for its survival and pathogenesis. In addition to the role of MTAN in methyl-transfer reactions, methionine biosynthesis, and polyamine synthesis, MTAN is also involved in bacterial quorum sensing (QS). In QS, chemical signaling autoinducer (AI) secreted by bacteria assists cell to cell communication and is regulated in a cell density-dependent manner. They play a significant role in the formation of bacterial biofilm. MTAN plays a major role in the synthesis of these autoinducers. Signaling molecules secreted by bacteria, i.e., AI-1 are recognized as acylated homoserine lactones (AHL) that function as signaling molecules within bacteria. QS enables bacteria to establish physical interactions leading to biofilm formation. The formation of biofilm is a primary reason for the development of multidrug-resistant properties in pathogenic bacteria like Enterococcus faecalis (E. faecalis). In this regard, inhibition of E. faecalis MTAN (EfMTAN) will block the QS and alter the bacterial biofilm formation. In addition to this, it will also block methionine biosynthesis and many other critical metabolic processes. It should also be noted that inhibition of EfMTAN will not have any effect on human beings as this enzyme is not present in humans. This review provides a comprehensive overview of the structural-functional relationship of MTAN. We have also highlighted the current status, enigmas that warrant further studies, and the prospects for identifying potential inhibitors of EfMTAN for the treatment of E. faecalis infections. In addition to this, we have also reported structural studies of EfMTAN using homology modeling and highlighted the putative binding sites of the protein.

Role of Bioaerosols on the Short-Distance Transmission of Multidrug-Resistant Methicillin-Resistant Staphylococcus aureus (MRSA) in a Chicken Farm Environment.

Methicillin-resistant Staphylococcus aureus (MRSA) is a dynamic and tenacious pathogenic bacterium which is prevalent in livestock farming environments. This study investigated the possibility of MRSA spread via bioaerosol transmission from an indoor chicken farm environment to outdoors downwind (up to 50 m). The concentration of total airborne bacteria colony formation units (CFUs) was decreased with increasing sampling distance ranging from 9.18 × 101 to 3.67 × 103 per air volume (m3). Among the 21 MRSA isolates, 15 were isolated from indoor chicken sheds and exposure square areas, whereas 6 were isolated from downwind bioaerosol samples. Molecular characterization revealed that all of them carried the staphylococcal cassette chromosome mec (SCCmec) VIII, and they were remarkably linked with the hospital-associated MRSA group. Spa typing analysis determined that all MRSA isolates belonged to spa type t002. Virulence analysis showed that 100% of total isolates possessed exfoliative toxin A (eta), whereas 38.09% and 23.80% strains carried exfoliative toxin B (etb) and enterotoxin A (entA). Additionally, all of these MRSA isolates carried multidrug resistance properties and showed their resistance against chloramphenicol, ciprofloxacin, clindamycin, tetracycline, and erythromycin. In addition, chi-squared statistical analysis displayed a significant distributional relationship of gene phenotypes between MRSA isolates from chicken farm indoor and downwind bioaerosol samples. The results of this study revealed that chicken farm indoor air might act as a hotspot of MRSA local community-level outbreak, wherein the short-distance dispersal of MRSA could be supported by bioaerosols.

Nimbolide-encapsulated PLGA nanoparticles induces Mesenchymal-to-Epithelial Transition by dual inhibition of AKT and mTOR in pancreatic cancer stem cells

Pancreatic ductal adenocarcinoma (PDAC) is associated with poor prognosis and remains highly aggressive despite current advancements in therapies. Chemoresistance and high metastatic nature of PDAC is attributed to a small subset of stem-like cells within the tumor known as Cancer Stem Cells (CSCs). Here, we developed a strategy for targeting pancreatic CSCs through forceful induction of mesenchymal-to-epithelial transition driven by encapsulating a phytochemical Nimbolide in nanoparticles. Binding of Nimbolide with the key regulator proteins of CSCs were studied through molecular docking and molecular dynamic simulation studies, which revealed that it binds to AKT and mTOR with high affinity. Further, in vitro studies revealed that Nim NPs are capable of inducing forceful mesenchymal-to-epithelial transition of pancreatospheres that leads to loss of multidrug resistance and self-renewal properties of pancreatospheres. Our study gives a proof of concept that encapsulation of Nim in PLGA nanoparticles increases its therapeutic effect on pancreatospheres. Further, binding of Nim to AKT and mTOR negatively regulates their activity that ultimately leads to mesenchymal-to-epithelial transition of pancreatic CSCs.

Targeting Omp-A Protein of Acinetobacter Baumannii with the Bio-Active Compounds from Azadirachta Indica - an in-silico Approach

Background: Acinetobacter baumannii is a gram negative bacterium which is typically short, round, coccobacillus and was named after the bacteriologist Paul Baumann. It is an emerging dental pathogen since it acquires drug resistance and expression of several virulence genes. It is an opportunistic pathogen in humans, affecting people with compromised immune systems. Acinetobacter baumannii is an arising nosocomial microorganism causing serious complications because of the propensity of its multi-drug resistant property.
 Aim: The aim of the present study was to target omp-A protein of Acinetobacter baumannii with the bio active compounds from Azadirachta indica an in-silico approach.
 Materials and Methods: The crystal structure of ompA protein was obtained from the PDB protein data bank. The structures of the bio-active derivatives of A. indica were obtained from the chemsketch software. The generated 3D structures were then optimised. Auto Dock instrument was utilized for docking investigation to interpret the affinity between bio-compounds of A. indica against ompA protein of A. baumannii.
 Results: The 3D crystal structure of OmpA-like domain from A.baumannii was retrieved from PDB database and its PDB ID was 3TD3 – A chain. 3D Structure of OmpA visualization using Biovia-Discovery studio visualizer. The 2D structure of compounds from Azadirachta indica was drawn using ACD chemsketch and saved in MDL-mol format and converted to PDB format using open babel converter. The final docked structures for the drug ligand interactions were assessed for their binding energies and hydrogen bonds.
 Conclusion: The present study had achieved the anti-biofilm inhibitory effect of imidazole-2-carboxylic acid from A. indica exhibiting a great interaction between activity with ompA utilizing computational investigation.

A Markerless CRISPR-Mediated System for Genome Editing in Candida auris Reveals a Conserved Role for Cas5 in the Caspofungin Response.

ABSTRACTCandida auris is a multidrug-resistant human fungal pathogen that has recently emerged worldwide. It can cause life-threatening disseminated infections in humans, with mortality rates upwards of 50%. The molecular mechanisms underlying its multidrug resistance and pathogenic properties are largely unknown. Few methods exist for genome editing in C. auris, all of which rely on selectable markers that limit the number of modifications that can be made. Here, we present a markerless CRISPR/Cas9-mediated genome editing system in C. auris. Using this system, we successfully deleted genes of interest and subsequently reconstituted them at their native loci in isolates across all five C. auris clades. This system also enabled us to introduce precision genome edits to create translational fusions and single point mutations. Using Cas5 as a test case for this system, we discovered a conserved role for Cas5 in the caspofungin response between Candida albicans and C. auris. Overall, the development of a system for precise and facile genome editing in C. auris that can allow edits to be made in a high-throughput manner is a major step forward in improving our understanding of this important human fungal pathogen.IMPORTANCE Candida auris is a recently emerged multidrug-resistant fungal pathogen capable of causing life-threatening systemic infections in humans. Few tools are available for genome editing in C. auris. Here, we present a markerless genome editing system for C. auris that relies on CRISPR/Cas9 technology and works to modify the genomes of all known C. auris clades. Using this system, we discovered a conserved role for Cas5 in the caspofungin response between C. albicans and C. auris. Overall, the development of a system for facile genome editing in C. auris is a major step forward in improving our understanding of this important human fungal pathogen.

Cytotoxic Potential of Ficus palmata Extracts on Lung Cancer Cell Lines (A549)

Cancer accounts for 10 million deaths globally whereas the treatments available either have several side effects or non-effective due to multi-drug resistant property of cancer cells for long term use. Therefore, it is utmost important to find safe therapeutic drugs that have cytotoxic potentials against cancer cells. Medicinal plants are believed to have lesser side effects with huge therapeutic efficacy. One such species is Ficus palmata which is known to have several medicinal properties and great antioxidant potentials. Earlier it was reported that methanolic extract of leaves (FPLM) and aqueous extract stembark (FPBA) of this plant have decent antioxidant properties. Therefore, the anticancer potentials of these plant extracts have been evaluated in addition of hydroxyl radical scavenging property. The results shown that FPBA represents higher antioxidant (IC50 value= 242.46±11.26 µg/ml) and anticancer effects against lung cancer (A549) cell lines (IC50 value= 69.74±2.12) whereas this plant extract have lower toxicity (IC50 value= 249.61±7.31) on normal cell lines (3T3-L1) which indicate that FPBA can be a potential therapeutic option for cancer and oxidative stress. The study concluded that the stem bark aqueous extract having therapeutic properties against lung cancer. It is to be recommended that the bioactive compound responsible for its therapeutic properties need to explore against cancer cell by evaluating various other parameters such as apoptosis, cell cycle arrest, and toxic effect on cell morphology.

LeuO, a LysR-Type Transcriptional Regulator, Is Involved in Biofilm Formation and Virulence of Acinetobacter baumannii.

Acinetobacter baumannii is an important nosocomial pathogen that can survive in different environmental conditions and poses a severe threat to public health due to its multidrug resistance properties. Research on transcriptional regulators, which play an essential role in adjusting to new environments, could provide new insights into A. baumannii pathogenesis. LysR-type transcriptional regulators (LTTRs) are structurally conserved among bacterial species and regulate virulence in many pathogens. We identified a novel LTTR, designated as LeuO encoded in the A. baumannii genome. After construction of LeuO mutant strain, transcriptome analysis showed that LeuO regulates the expression of 194 upregulated genes and 108 downregulated genes responsible for various functions and our qPCR validation of several differentially expressed genes support transcriptome data. Our results demonstrated that disruption of LeuO led to increased biofilm formation and increased pathogenicity in an animal model. However, the adherence and surface motility of the LeuO mutant were reduced compared with those of the wild-type strain. We observed some mutations on amino acids sequence of LeuO in clinical isolates. These mutations in the A. baumannii biofilm regulator LeuO may cause hyper-biofilm in the tested clinical isolates. This study is the first to demonstrate the association between the LTTR member LeuO and virulence traits of A. baumannii.

A Comparative Analysis of Virulence Factors in Klebsiella Species

Klebsiella is a genus of bacteria frequently associated with nosocomial infections due to their multi-drug resistant properties. Klebsiella pneumoniae accounts for a large majority of hospital-acquired pneumonia, urinary tract infections, and septicemia. The pathogenicity of K. pneumoniae clinical isolates has been greatly established to better understand and treat these infections. Klebsiella oxytoca is also primarily healthcare associated and is considered to be an emerging nosocomial pathogen, as it causes the second most frequent infections from the Klebsiella genus. Although it shares many characteristics with K. pneumoniae, its virulence factors have not been well studied due to its lesser degree of incidence. As an emerging pathogen, more information must be gathered in order to learn how to best treat this infectious, antibiotic-resistant bacteria. This study aims to first determine if the pathogenic factors found in clinical Klebsiella pneumoniae isolates are also present in Klebsiella oxytoca and then compare the production of these virulence factors under normal and inducing conditions. This work compares the expression of pathogenic factors in five isolates of K. oxytoca, three isolates of K. pneumoniae, and one strain of Escherichia coli. The pathogenic factors chosen for this study include antibiotics resistance, siderophore production, biofilm formation abilities, and reactive oxygen species (ROS) detoxification. To determine antibiotic resistance, the minimum inhibitory concentration of nine different antibiotics was calculated for each isolate. Siderophore production was measured using a Chrome Azurol Sulfonate colorimetric test in both the absence and presence of 2,2’-dipyridyl, an iron chelator. Biofilm formation was detected with Crystal Violet dye in both the absence and presence of acetaminophen and hydrochlorothiazide, both of which are commonly used therapeutic pharmaceuticals in nosocomial settings, as well as 2,2’-dipyridyl. ROS detoxification via superoxide dismutase enzyme activity was measured through native gel activity assays. Our findings indicate that K. pneumoniae has resistance to more antibiotics than K. oxytoca, but the two species have comparable siderophore production, biofilm formation capabilities, and ROS detoxification capabilities. In conclusion, Klebsiella pneumoniae and Klebsiella oxytoca share several mechanisms of pathogenicity under normal and inducing conditions and drug resistance is the most notable difference among the strains tested in this work.

Isolation and molecular detection of Streptococcus agalactiae from popped eye disease of cultured Tilapia and Vietnamese koi fishes in Bangladesh.

ObjectivePresent research aims to isolate, identify, and determine the virulence of the Streptococcus agalactiae (group B Streptococcus; GBS), isolated from popped eye disease affected Tilapia and Vietnamese Koi (V. Koi) fishes.Materials and MethodsA total of 330 fish samples were collected, of which Tilapia (n = 180) and V. Koi (n = 150), were collected from 35 affected ponds of four selected districts of Bangladesh. Isolation of the bacterium was done using different culture media (Nutrient broth, Plate count agar, Tryptic Soy Agar, and Blood agar), and identification by using various biochemical tests (conventional and using API 20 Strep kit) and polymerase chain reaction (PCR) using primers against 16S rRNA gene of S. agalactiae. Antibiotic susceptibility of the bacteria was performed using seven different antibiotics disc (Tetracycline, Oxytetracycline, Chlortetracycline, Streptomycin, Ciprofloxacin, Gentamicin, and Neomycin). Virulence of the isolated S. agalactiae was determined by infecting healthy Tilapia and V. Koi fishes through experimental infection.ResultsIsolated bacteria were found Gram-positive paired and chained cocci, β-hemolytic and non-motile. Findings of biochemical and serological tests indicate that the isolated bacterium belongs to Group B Streptococcus of Lancefield classification. PCR result also confirmed that the bacteria were S. agalactiae. The bacterial isolates possessed resistance property against all the seven antibiotics used in this study. The isolated GBS was found highly virulent and showed 80%–90% mortality for Tilapia and V. Koi fishes in experimental infection within 1–6 days of post-infection.ConclusionFrom the findings of this study, it may be concluded that isolated GBS from the Tilapia and V. Koi fishes were highly virulent and possessed multidrug-resistance properties.

Isolation, Molecular Detection and Antibiogram of Multi-drug Resistant Salmonella Typhimurium DT104 from Selected Dairy Farms in Mymensingh, Bangladesh

The Salmonella typhimurium DT104, an emerging cause of human illness has received an increasing attention due to its multidrug resistant properties. Since it has been isolated from human and other sources including food-producing animals around the world; it has become a worldwide public health concern. Therefore, the present study was designed to isolate, identify and study the antibiogram profile of multidrug resistant S. typhimurium DT104 from several dairy farms in Mymensingh district, Bangladesh. A total of 135 fecal samples from diarrhoeic cattle were collected aseptically and subjected for bacterial isolation, molecular detection using PCR followed by antibiogram study. Salmonella spp. could be isolated from a total of 39 (28.88%) samples based on cultural and staining methods which were further confirmed by PCR using invA gene specific primers. However, out of 39 Salmonella spp., 6 isolates were confirmed as S. typhimurium DT104 strain. Results of the antibiotic resistance patterns demonstrated that 100% (39/39) isolates were resistant to erythromycin followed by tetracycline (73.68%), colistin (89.47%), ampicillin (47.36%), gentamicin (21.05%), ciprofloxacin (31.57%), streptomycin (42.10%), enrofloxacin (10.52%) and chlormphenicol (31.57%). Moreover, about 23.07% isolates were resistant to more than 5 antibiotics. However, all the isolates were found to be sensitive to amikacin. These results suggest that antibiotic resistant S. typhimurium DT104 strain has been circulating in dairy cattle in Bangladesh which is alarming and may impose threat to livestock and public health due to lack of proper hygienic management. This study will be helpful for the selection of proper antibiotics against salmonellosis in cattle.

Candida auris: Epidemiology, biology, antifungal resistance, and virulence.

First described in 2009 in Japan, the emerging multidrug-resistant fungal pathogen Candida auris is becoming a worldwide public health threat that has been attracting considerable attention due to its rapid and widespread emergence over the past decade. The reasons behind the recent emergence of this fungus remain a mystery to date. Genetic analyses indicate that this fungal pathogen emerged simultaneously in several different continents, where 5 genetically distinct clades of C. auris were isolated from distinct geographical locations. Although C. auris belongs to the CTG clade (its constituent species translate the CTG codon as serine instead of leucine, as in the standard code), C. auris is a haploid fungal species that is more closely related to the haploid and often multidrug-resistant species Candida haemulonii and Candida lusitaniae and is distantly related to the diploid and clinically common fungal pathogens Candida albicans and Candida tropicalis. Infections and outbreaks caused by C. auris in hospitals settings have been rising over the past several years. Difficulty in its identification, multidrug resistance properties, evolution of virulence factors, associated high mortality rates in patients, and long-term survival on surfaces in the environment make C. auris particularly problematic in clinical settings. Here, we review progress made over the past decade on the biological and clinical aspects of C. auris. Future efforts should be directed toward understanding the mechanistic details of its biology, epidemiology, antifungal resistance, and pathogenesis with a goal of developing novel tools and methods for the prevention, diagnosis, and treatment of C. auris infections.

Profile of Multidrug Resistance (MDR) and Methicillin Resistant Staphylococcus aureus (MRSA) on Dairy Farms in East Java Province, Indonesia

Staphylococcus aureus is one of the causes in the case of Foodborne Disease, with the resulting toxintriggering the occurrence of Foodborne Intoxication. The purpose of this study was to identify multidrugresistant patterns and detect Methicillin Resistance Staphylococcus aureus (MRSA) in cow’s milk in EastJava, Indonesia. A total of 170 cow’s milk samples were taken from dairy farms on 3 Districs, then thebacteria were cultured and purified using Mannitol Salt Agar media. Resistance tests were carried out with5 antibiotics on Mueller Hinton Agar media and Methicillin Resistant Staphylococcus aureus (MRSA) byOxacillin Resistance Screen Agar Base (ORSAB). The results showed there were 76 (44.7%) S. aureus, thenantibiotic testing showed 13 (17.1%) S. aureus isolates had multidrug resistant (MDR) properties on theantibiotics tested and 84.6% isolates with MDR confirmed MRSA properties by ORSAB. It was concludedthat the raw milk can be a potential reservoir for MDR and MRSA strains to threat public health.

SATP‑binding cassette subfamilyG member2 enhances the multidrug resistance properties of human nasal natural killer/Tcell lymphoma side population cells.

Extranodal natural killer (NK)/T cell lymphoma, nasal type (ENKL) is a rare type of non-Hodgkin's lymphoma that is associated with limited effective treatment options and unfavorable survival rate, which is partly the result of multidrug resistance (MDR). The presence of side population (SP) cells-SNK-6/ADM-SP (SSP) cells has been previously used to explore mechanisms of drug resistance. ATP-binding cassette subfamily G member 2 (ABCG2) is a gene involved in MDR and is closely associated with SPs. However, the function of ABCG2 in SSP cells is unclear. The present study verified the high expression of ABCG2 in SSP cells. The IC50 values of doxorubicin, cytarabine, cisplatin, gemcitabine and l-asparaginase were tested to evaluate drug sensitivity in SSP cells with different levels of ABCG2 expression. ABCG2 was identified as a gene promoting in MDR. ABCG2 upregulated cell proliferation, increased clonogenicity, increased invasive ability and decreased apoptosis, in vivo and in vitro, when cells were treated with gemcitabine. To conclude, ABCG2 enhanced MDR and increased the typical biological characteristics associated with cancer cells in SP cells. With further investigation of the ABCG2 gene could have the potential to reverse MDR in ENKL.

Inhibition of Wnt/β-catenin pathway reverses multi-drug resistance and EMT in Oct4+/Nanog+ NSCLC cells

Cancer drug resistance and epithelial-mesenchymal transition (EMT), a critical process of cancer invasion and metastasis, have recently been associated with the existence of cancer stem cells (CSCs). However, there are no appropriate CSC-markers of non-small cell lung cancer (NSCLC)-associated drug resistance and EMT. It is unknown if and how the drug-resistant and EMT phenotypes in NSCLC cells link to specific stemness-related pathways. Here, we found a significant elevated expression of both Oct4 and Nanog in gefitinib-resistant NSCLC cells, which displayed multi-drug resistance (MDR) properties and exhibited EMT phenotype. Ectopic co-expression of Oct4/Nanog empowered NSCLC cells with cancer stem cell properties, including self-renewal, drug resistance, EMT and high tumorigenic capacity. Following molecular mechanism investigation indicated Oct4/Nanog-regulated drug resistance and EMT change through Wnt/β-catenin signaling activation. Moreover, silencing β-catenin abrogated Oct4/Nanog-mediated MDR and EMT process in NSCLC cells. Our findings propose Wnt/β-catenin pathway as a promising therapeutic target for the treatment of progression and metastasis of NSCLC with CSC-like signatures and epithelial-mesenchymal transition phenotype.

Inhibition of multidrug resistance property of Candida albicans by natural compounds of parthenium hysterophorus L. An In-Silico approach

Objectives: In this study, we targeted enzymes (Erg11, Erg5, Erg3), transporters (CDR1, CDR2), and cytochrome 450 (CaALK8) involved in MDR of Candida albicans, which caused fungal disease. ATP-binding cassette (ABC) and some other major facilitator superfamilies (MFS) of transporters are responsible for MDR in Candida Albicans. Material and methods: The compounds present in Parthenium hysterophorus L. were docked against the proteins involved in MDR of Candida Albicans. PyRx-Python prescription 0.8. was used to identify binding affinities of compounds against the proteins. Result and Discussion: Erg11, Erg5, Erg3, CDR1, CDR2 and CaALK8 proteins docked with β-Sitosterol (-10.6, -9.6, -9.6, -9.6, -9.6, and -8.5) ç-Sitosterol (-9.9, -9.2, -9.3, -9.4, -9.6, and -8.5). Piperine (-10.0, -8.3, -9.3, -8.4, -8.5, and -8.4) Kcal/mol respectively and found to show good hydrophobic interactions. Conclusion: In this study, we may conclude that compounds isolated from parthenium hysterophorus might be effective against the fungal disease caused by Candida Albicans.

Antimicrobial resistance situation in animal health of Bangladesh.

Antimicrobial resistance (AMR) is a crucial multifactorial and complex global problem and Bangladesh poses a regional and global threat with a high degree of antibiotic resistance. Although the routine application of antimicrobials in the livestock industry has largely contributed to the health and productivity, it correspondingly plays a significant role in the evolution of different pathogenic bacterial strains having multidrug resistance (MDR) properties. Bangladesh is implementing the National Action Plan (NAP) for containing AMR in human, animal, and environment sectors through “One Health” approach where the Department of Livestock Services (DLS) is the mandated body to implement NAP strategies in the animal health sector of the country. This review presents a “snapshot” of the predisposing factors, and current situations of AMR along with the weakness and strength of DLS to contain the problem in animal farming practices in Bangladesh. In the present review, resistance monitoring data and risk assessment identified several direct and/or indirect predisposing factors to be potentially associated with AMR development in the animal health sector of Bangladesh. The predisposing factors are inadequate veterinary healthcare, monitoring and regulatory services, intervention of excessive informal animal health service providers, and farmers’ knowledge gap on drugs, and AMR which have resulted in the misuse and overuse of antibiotics, ultimate in the evolution of antibiotic-resistant bacteria and genes in all types of animal farming settings of Bangladesh. MDR bacteria with extreme resistance against antibiotics recommended to use in both animals and humans have been reported and been being a potential public health hazard in Bangladesh. Execution of extensive AMR surveillance in veterinary practices and awareness-building programs for stakeholders along with the strengthening of the capacity of DLS are recommended for effective containment of AMR emergence and dissemination in the animal health sector of Bangladesh.

Alteration of Multi-drug Resistance Activities by Ethanolic Extracts of Nigella sativa Against Urinary Pathogens

Abstract: Background and Objective: Most of the urinary pathogens cause urinary tract infection (UTI) have developed multidrug-antibiotics resistance (MDR) and had forewarned the interest in researching natural products to increase the usage of medicinal plants as alternative therapies for infectious diseases. The study was aimed to investigate the alteration of multidrug-resistant properties of ethanolic extract from Nigella sativa against clinically isolated MDR urinary pathogens. Materials and Methods: Antimicrobial activity of ethanolic extract from Nigella sativa was investigated against clinical isolates of Proteus mirabilis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli and Acinetobacter baumannii using agar disc diffusion technique, minimal bactericidal concentration (MBC) and minimal inhibitory concentration (MIC) assays. The data was analyzed using Microsoft Excel. Results: Ethanol extract of Nigella sativa possesses strong microbial growth inhibitory effects in a dose-dependent manner. The minimum MIC values were observed for Proteus mirabilis (1.5 mg mL–1), followed by Pseudomonas aeruginosa (2 mg mL–1), extended-spectrum ?-lactamase (ESBL) producing Klebsiella pneumoniae (2.5 mg mL–1), ESBL producing Escherichia coli (2.75 mg mL–1) and Acinetobacter baumannii (2.5 mg mL–1). The MBC inhibitory effects of ethanolic extracts were greater than that of corresponding MIC results for all clinical isolated urinary pathogens. The bactericidal concentration of ethanol extracts was slightly increased than the corresponding MIC. Conclusion: This study augmented the effective alteration of MDR properties by the ethanolic extract of Nigella sativa seeds. Thus, it might be used as an effective natural source of a safe antimicrobial agent against emerging MDR clinical isolates in the future.

Four LysR-type transcriptional regulator family proteins (LTTRs) involved in antibiotic resistance in Aeromonas hydrophila.

Aeromonas hydrophila is a Gram-negative bacterium that causes serious infections in aquaculture and exhibits significant multidrug resistance. The LysR-type transcriptional regulator (LTTR) family proteins are a well-known group of transcriptional regulators involved in diverse physiological functions. However, the role of LTTRs in the regulation of bacterial resistance to antibiotics is still largely unknown. In this study, to further investigate the role of four putative LTTR family proteins (A0KIU1, A0KJ82, A0KPK0, and A0KQ63) in antibiotic resistance in A. hydrophila, their genes were cloned and overexpressed in engineered Escherichia coli. After the optimization of experimental conditions including incubation time, temperature, and IPTG concentration, these proteins were successfully purified, and their specific antibodies against mice were obtained. Using western blot analysis, we found that these LTTR family proteins were downregulated in A. hydrophila following antibiotic treatment, indicating that they may be involved in the regulation of antibiotic resistance. Additionally, minimum inhibitory concentration (MIC) assays of chloramphenicol (CM), chlortetracycline (CTC), ciprofloxacin (CF), furazolidone (FZ), and balofloxacin (BF) in E. coli showed that overexpression of these LTTRs led to increased sensitivity to several antibiotics. To further validate their functional role in antibiotic resistance, we demonstrated that bacteria with loss of A0KQ63 (ΔAHA_3980) exhibited multi-drug resistance properties. Our results indicate that these LTTR family proteins may play an important role in the antibiotic resistance of A. hydrophila, and the that underlying mechanisms controlling antibiotic resistance should be further investigated.