MAR Index Research Articles

Karadeniz'in deniz sedimentlerinden izole edilen Gram-negatif heterotrofik bakterilerin antibiyotik direnci-CLSI ve EUCAST'ın çevresel izolatlar üzerinde karşılaştırılması

Marine sediments are the important reservoirs of antibiotics and resistant genes. Antibiotic resistance becomes a global concern today and bacteria from marine sediments show resistance to multiple microbial agents. This study investigates antibiotic resistance of Gram-negative heterotrophic bacteria from the marine sediment of the Black Sea. The samples were gathered from various sampling locations on the marine environments of the Black Sea between May 2019 and February 2020. The VITEK 2 system was utilized to detect the MIC values against different antibiotics automatically. The MIC values were ranged from 0.12 µg/ml to 80 µg/ml. The highest resistance ratios among all antibiotics were detected as 90% in CLSI and 81% in EUCAST against cefazolin. In general, the resistance ratios were determined as 13.4% in CLSI and 26.1% in EUCAST. All isolates were detected as susceptible to meropenem, cefepime, ceftazidime, colistin, ertapenem, and piperacillin/tazobactam. MAR indexes were ranged between 0 and 0.45. MAR index were detected above or equal the value of 0.2 as 27% in CLSI and 81% in EUCAST among all isolates. This study shows that EUCAST may be better than CLSI in terms of showing the antibiotic exposure among natural isolates and the MAR indexes may be a useful tool for defining the environmental status. The results of this study indicate that antibiotic resistance may be a serious concern in sediments of the Black Sea.

Molecular characterisation and antibiotic resistance profile of Escherichia coli isolated from seafood

The present study made an attempt to investigate the occurrence of pathogenic Escherichia coli in seafood, its antimicrobial susceptibility patterns and the biofilm forming capacity. Total of 43 seafood samples collected from ten commercial markets of Kochi was screened for the presence of pathogenic E. coli. The PCR results revealed that out of 21 isolates screened, only one isolate from fish sample showed positive for rfb E gene of Enterohaemorrhagic E. coli (EHEC). Antibiotic susceptibility pattern of the isolates revealed that 90% of them were multidrug resistant to more than 5 antibiotics of four structurally different classes. The MAR index of all the isolates was found between 0.2 and 0.5. Out of the 15 moderate biofilm forming E. coli obtained, 2 of them exhibited maximum multidrug resistance to 8 antibiotics of six different classes whereas remaining 13 isolates showed multidrug resistance to 5 antibiotics. Hence, this study revealed the emergence of antibiotic resistance in biofilm forming E. coli in seafood from Kochi markets, which may pose future threat to develop control strategies in the fish processing industry and also pose significant treatment challenge to physicians. Keywords: Biofilm, Multidrug resistance, Pathogenic E. coli, Seafood

Isolation and study of MRSA-resistant Staphylococcus aureus from clinical samples in Al-Diwaniyah Hospitals

This study included the collection of 554 samples from different clinical sources and cases for patients of different ages, visit the different Diwaniyah hospitals for the period from November 2021 to March 2022. Staphylococcus aureus was investigated. The results showed the yield of 100 isolates of S aureus bacteria, which included 11 isolates from burns, 11 from wounds, 24 from sputum, and 55 from urine. MRSA was investigated using the method of spreading on the agar, and it was 80%, 50% urine, 25% sputum, 13% burns, and 11%wound. An antibiotic assay was done towards MRSA isolates, and the percentages were as follows: Penicillin 100%, Cefoxitin 100%, Moxifloxacin (MFX) 76.2%, Erythromycin 70%, azithromycin 63.7%, levofloxacin 51.5, CLARITHROMYCIN Amikacin 41, 38% 12, Clindamycin 287%, Ofloxacin 41.2%, Tetracycline 36.2%, Doxycycline 31.2%, Rifampin 23.7%, Norfloxacin 18.7%,Trimethoprim 18.7%, Gentamicin 13,3% Trimethoprim-Sulfamethoxazole 12 and Ciprofloxacin and 12% and Nitrofurantoin 10% and Chloramphenicol 6.1%, while the values ​​of the MAR index ranged from 0.75 to 0.05

PREVALENCE AND ANTIBIOGRAM PROFILE OF LISTERIA MONOCYTOGENES ASSOCIATED WITH CATTLE FARMS IN LAFIA, NASARAWA STATE

Listeriosis, mostly caused by ubiquitous Listeria spp., notably L. monocytogenes is a foodborne disease of great public health concern with high mortality and severe symptoms. However, it is greatly underreported and diagnosed. Using standard microbiological procedures (EN ISO 11290-12017) and Kirby-Bauer antimicrobial disk diffusion method, the prevalence and antibiogram profile of listerial contaminants from four cattle farms were assessed. A total of 160 samples, 40 samples per farm comprising ten samples each of cattle dung, feed, milk and water were collected. Of the total studied samples, 20.63% (33/160) were positive for listerial contaminants. Farm A (6.88%; 11/160) and cattle feed samples (33%; 13/40) accounted for the highest prevalence among farm and sample type, respectively. Seven different species of Listeria were isolated with L. monocytogenes being the most prevalent (42.42%; 14/33) and L. welshimeri and L. grayi the least prevalent (6.06%; 2/33). 28.57% (4/14) of L. monocytogenes isolates harbored the hlyA virulence gene. A high antimicrobial-resistant rate and MAR index range of 0.07-0.73 to tested antibiotics, including to last-resort drugs, was observed in this study. L. monocytogenes isolates were 100% susceptible to Clarithromycin, with high resistance observed to nalidixic acid (100%), streptomycin (64.29%) and ofloxacin (42.86%). This study findings revealed a significant level of listerial contamination in cattle farm samples. The presence of Listeria spp., especially antimicrobial-resistant L. monocytogenes is a cause for alarm. Appropriate regulatory and monitoring authorities are encouraged to sensitize cattle-breeders and consumers appropriately and step-up checks in the sales of cattle products in Lafia.

Detection of carbapenemases blaOXA48-blaKPC-blaNDM-blaVIM and extended-spectrum-β-lactamase blaOXA1-blaSHV-blaTEM genes in Gram-negative bacterial isolates from ICU burns patients

Background and objectivesBurn patients are highly susceptible to invasion by multidrug-resistant Gram-negative bacteria (MDR-GNB) through post-burn damage. The prevalence of MDR-GNB isolated from burns patients has increased dramatically in the last decade, representing a serious risk to patients admitted to burns units worldwide. The challenges of managing infected burns patients are exacerbated in poor resource settings. This study was designed to develop a pathway for the rapid diagnosis of multidrug-resistant (MDR) Gram-negative infections and identify the bacterial genes including blaOXA1, blaTEM, and blaSHV encoding ESBLs and blaOXA48, blaKPC, blaNDM, and blaVIM encoding carbapenemases from the patient of post burns infection. MethodsClinical isolates were collected (August 2017 to August 2018) from Intensive care unit (ICU) of Burn Centre. Antibiotic susceptibility testing and phenotypic detection of ESBLs and carbapenemases was performed by disk diffusion, double disk synergy test (DDST), combination disk test (CDT), and Imipenem + EDTA combined disk test (IMP + EDTA CDT). Polymerase chain reaction (PCR) detection was performed for ESBLs blaOXA1-blaSHV-blaTEM and carbapenemases genes blaOXA48-blaKPC-blaNDM-blaVIMResultsIn total, of 170 Gram-negative isolates, 104 (61.2%) were confirmed as multidrug-resistant (MDR); Pseudomonas aeruginosa was found to be the most prevalent 43/104 (41.4%), followed by Klebsiella pneumoniae 17/104 (16.4%), Acinetobacter baumannii12/104 (11.5%), and 6/104 Proteus mirabilis (5.8%). All isolates (100%) were resistant to cefotaxime and ceftazidime, while the meropenem resistance was 58.7%. ESBL and carbapenemase genotypes were found to be associated with higher MAR index (0.65–0.88) and MIC (> 32 µg/ml) values P. aeruginosa was the major ESBL and carbapenemase producer as determined by phenotypic testing and PCR. blaTEM positive isolates among ESBLs producers were predominant 81.8% (27/33), followed by 27.3% blaOXA1 and blaSHV, respectively. blaVIM positive isolates among carbapenemase producers were predominant 47.7% (21/44), followed by 27.3% blaKPC, 20.5% blaOXA48, and 11.4% blaNDM positive isolates.ConclusionsThe predominant organism causing burn infections was ESBL and carbapenemase-producing Pseudomonas aeruginosa. There are only limited effective antibiotics against such strains. blaVIM and blaTEM individually and in co-existence with blaKPC, blaOXA48, blaSHV, and blaOXA1 confer antimicrobial resistance in burns patients. Rapid detection of ESBL and carbapenemase genes will inform treatment strategies improving the outcome for post-burn patients in ICU.

Genetic Diversity, Biofilm Formation, and Antibiotic Resistance of Pseudomonas aeruginosa Isolated from Cow, Camel, and Mare with Clinical Endometritis.

Pseudomonas aeruginosa is a ubiquitous opportunistic bacterium that causes diseases in animals and humans. This study aimed to investigate the genetic diversity, antimicrobial resistance, biofilm formation, and virulence and antibiotic resistance genes of P. aeruginosa isolated from the uterus of cow, camel, and mare with clinical endometritis and their drinking water. Among the 180 uterine swabs and 90 drinking water samples analysed, 54 (20%) P. aeruginosa isolates were recovered. Isolates were identified biochemically to the genus level by the automated Vitek 2 system and genetically by the amplification of the gyrB gene and the sequencing of the 16S rRNA gene. Multilocus sequence typing identified ten different sequence types for the P. aeruginosa isolates. The identification of ST2012 was significantly (p ≤ 0.05) higher than that of ST296, ST308, ST111, and ST241. The isolates exhibited significantly (p ≤ 0.05) increased resistance to piperacillin (77.8%), ciprofloxacin (59.3%), gentamicin (50%), and ceftazidime (38.9%). Eight (14.8%) isolates showed resistance to imipenem; however, none of the isolates showed resistance to colistin. Multidrug resistance (MDR) was observed in 24 isolates (44.4%) with a multiple antibiotic resistance index ranging from 0.44 to 0.77. MDR was identified in 30 (33.3%) isolates. Furthermore, 38.8% and 9.2% of the isolates exhibited a positive extended-spectrum-β-lactamase (ESBL) and metallo-β-lactamase (MBL) phenotype, respectively. The most prevalent β-lactamase encoding genes were blaTEM and blaCTX-M, however, the blaIPM gene was not detected in any of the isolates. Biofilm formation was observed in 49 (90.7%) isolates classified as: 11.1% weak biofilm producers; 38.9% moderate biofilm producers; 40.7% strong biofilm producers. A positive correlation was observed between the MAR index and biofilm formation. In conclusion, the results highlighted that farm animals with clinical endometritis could act as a reservoir for MDR and virulent P. aeruginosa. The emergence of ESBLs and MBLs producing P. aeruginosa in different farm animals is a public health concern. Therefore, surveillance programs to monitor and control MDR P. aeruginosa in animals are required.

Multidrug-resistant epi-endophytic bacterial community in Posidonia oceanica of Mahdia coast as biomonitoring factor for antibiotic contamination.

Faced with the significant disturbances, mainly of anthropogenic origin, which affect the Mediterranean coastal ecosystem, Posidonia oceanica (L.) Delile has often been used to assess the state of health of this environment. The present study aims to determine the multidrug resistance patterns among isolated and identified epi-endophytic bacterial strains in P. oceanica seagrass collected from Mahdia coastal seawater (Tunisia). To investigate the bacterial community structure and diversity from coastal seawater samples from Mahdia, total DNA extraction and 16S rRNA gene amplification were performed and analyzed by denaturing gradient gel electrophoresis (DGGE). The DGGE profiles showed that some bands were specific to a given site, while other bands were found to be common to more than one sample. In the other hand, bacterial strains were isolated from 1mL of leaves and epiphytes suspension of P. oceanica seagrass in marine agar. Forty-three isolates were obtained, seven of them were selected and identified on the basis of 16S rRNA gene sequence analysis. These isolates belonged to the genus Bacillus, exhibiting 98-100% of identity with known sequences. Susceptibility patterns of these strains were studied toward commonly used antibiotics in Tunisia. All identified isolates were resistant to Aztreonam (72.1%), Ceftazidime (60.5%), Amoxicillin (56%) and Rifampicin (51.2%). S5-L13 strain had presented the highest multidrug resistance with a MAR index of 0.67.

Antimicrobial Resistance, Biofilm Formation, and Virulence Genes in Enterococcus Species from Small Backyard Chicken Flocks.

Backyard birds are small flocks that are more common in developing countries. They are used for poultry meat and egg production. However, they are also implicated in the maintenance and transmission of several zoonotic diseases, including multidrug-resistant bacteria. Enterococci are one of the most common zoonotic bacteria. They colonize numerous body sites and cause a wide range of serious nosocomial infections in humans. Therefore, the objective of the present study was to investigate the diversity in Enterococcus spp. in healthy birds and to determine the occurrence of multidrug resistance (MDR), multi-locus sequence types, and virulence genes and biofilm formation. From March 2019 to December 2020, cloacal swabs were collected from 15 healthy backyard broiler flocks. A total of 90 enterococci strains were recovered and classified according to the 16S rRNA sequence into Enterococcus faecalis (50%); Enterococcus faecium (33.33%), Enterococcus hirae (13.33%), and Enterococcus avium (3.33%). The isolates exhibited high resistance to tetracycline (55.6%), erythromycin (31.1%), and ampicillin (30%). However, all of the isolates were susceptible to linezolid. Multidrug resistance (MDR) was identified in 30 (33.3%) isolates. The enterococci AMR-associated genes ermB, ermA, tetM, tetL, vanA, cat, and pbp5 were identified in 24 (26.6%), 11 (12.2%), 39 (43.3%), 34 (37.7%), 1 (1.1%), 4 (4.4%), and 23 (25.5%) isolates, respectively. Of the 90 enterococci, 21 (23.3%), 27 (30%), and 36 (40%) isolates showed the presence of cylA, gelE, and agg virulence-associated genes, respectively. Seventy-three (81.1%) isolates exhibited biofilm formation. A statistically significant correlation was obtained for biofilm formation versus the MAR index and MDR. Multi-locus sequence typing (MLST) identified eleven and eight different STs for E. faecalis and E. faecium, respectively. Seven different rep-family plasmid genes (rep1–2, rep3, rep5–6, rep9, and rep11) were detected in the MDR enterococci. Two-thirds (20/30; 66.6%) of the enterococci were positive for one or two rep-families. In conclusion, the results show that healthy backyard chickens could act as a reservoir for MDR and virulent Enterococcus spp. Thus, an effective antimicrobial stewardship program and further studies using a One Health approach are required to investigate the role of backyard chickens as vectors for AMR transmission to humans.

Molecular identification and antibiotic resistance patterns of diverse bacteria associated with shrimp PL nurseries of Bangladesh: suspecting Acinetobacter venetianus as future threat.

Shrimp aquaculture has been accomplished with breeding and nursing of shrimp in an artificial environment to fulfill the increasing demand of shrimp consumption worldwide. However, the microbial diseases appear as a serious problem in this industry. The study was designed to identify the diverse bacteria from shrimp PL (post-larvae) nurseries and to profile antibiotic resistance patterns. The rearing water (raw seawater, treated and outlet water) and shrimp PL were collected from eight nurseries of south-west Bangladesh. Using selective agar plates, thirty representative isolates were selected for 16S rRNA gene sequencing, antibiotic susceptibility test and MAR index calculation. Representative isolates were identified as Aeromonas caviae, Pseudomonas monteilii, Shewanella algae, Vibrio alginolyticus, V. brasiliensis, V. natriegens, V. parahaemolyticus, V. shilonii, V. xuii, Zobellella denitrificans which are Gram-negative, and Bacillus licheniformis and B. pumilus which are Gram-positive. Notably, six strains identified as Acinetobacter venetianus might be a concern of risk for shrimp industry. The antibiotic resistance pattern reveals that the strain YWO8-97 (identified as P. monteilii) was resistant to all twelve antibiotics. Ceftazidime was the most powerful antibiotic since most of the studied strains were sensitive against it. The six strains of A. venetianus showed multiple antibiotic resistance patterns. MAR index were ranged from 0.08 to 1.0, and values of 26 isolates were more than 0.2 which means prior high exposure to the antibiotics. From the present study, it can be concluded that shrimp PL nurseries in southern part of Bangladesh are getting contaminated with antibiotic resistant pathogenic bacteria.

Antibiotic Profile of Pathogenic Bacteria Isolated from Postsurgical Site Infections in Public Hospitals in Northern Jordan.

BackgroundSurgical site infections are hospital-acquired, vary from one hospital to another, and can cause significant postoperative morbidity, mortality, and prolonged hospital stay.ObjectiveThe main aim of the study was to identify the bacterial pathogens associated with surgical site infections and their antibiotic susceptibility in a public hospital in northern Jordan.MethodsPostsurgical wound samples were collected and processed in a microbiology laboratory using standard microbiological techniques. Antibiotic susceptibility tests were performed using 13 antibiotics covering the gram-positive and gram-negative bacteria using the disc diffusion test.ResultsThe bacterial species that were identified in this study include Escherichia coli 8 (29%), Pseudomonas aeruginosa 3 (11%), Proteus mirabilis 1 (3.5%), Klebsiella pneumoniae 4 (14%), Salmonella enterica 2 (7%), Staphylococcus aureus 8 (29%), Staphylococcus epidermidis 1 (3.5%), and Streptococcus pyogenes 1 (4%). The antibiotic profiles of these bacteria showed high resistance. The MAR indices showed that 17 of 28 bacteria isolated were above 0.2 indicating high resistance.ConclusionResistant bacteria are becoming more dominant in wound infections with a high prevalence of multidrug resistant isolates. Hospital disinfection and treatment protocols regarding the use of antibiotics should be practiced vigorously and monitored regularly to combat resistance.

English

Multiple antibiotic resistance is a major cause of clinical infections worldwide. This study determined the pattern of multidrug bacterial resistance in hospital sewage samples from the Marathwada region of India. Forty-eight isolates of bacteria were obtained from 6 locations of Aurangabad.  An antibiotic sensitivity test was carried out using the disc diffusion method. Among all the antibiotics tested, the highest level of resistance was observed in the beta lactam class (85%), followed by Tetracycline (58%), Cephalosporin (58%), quinolones (52%) and gentamycin (45%). Escherichia coli and Klebsiella pneumoniae are the most prevalent bacteria, showing antibiotic resistance to all tested antibiotics with a MAR index of 1. It is concluded that hospital sewage water could be a reservoir of antibiotic resistant bacteria, which may further contaminate drinking water bodies, potentially presenting a public health risk to the general populace.   Key words: Antibiotics, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas species, Staphylococcus aureus, Streptococcus pyogenes.

Comparison Distribution of Vibrio Species in Stocking to Harvesting Process of Shrimp at Commercialize Shrimp Farm

Vibrio species is one of the pathogenic bacteria infecting shrimps in aquaculture farm has caused severe loss to the aquaculture farm. The aim of this research is to isolate and compare the presence of Vibrio spp. in commercialize shrimp farm during the stocking process and the final harvest phase of shrimp productions. Quantification and isolation were conducted and measured using Log CFU (colony forming units) counts and morphological investigation gram staining and conventional biochemical test identification. A total of sixty-four (n=64) sediment and water samples were collected from the inlet and outlet of the two selected shrimp ponds (Pond A and Pond B) in Kampung Telaga Air, Kuching, Sarawak. As a result, the bacteria count from water samples during stocking period at both shrimps ponnds were 6.6 Log CFU/ml and 5.6 Log CFU/ml while during harvesting period were 5.3 Log CFU/ml and 5.8 Log CFU/ml. Meanwhile, bacteria count sediments samples during stocking period at both shrimp ponds were 5.5 Log CFU/g and 5.7 Log CFU/g whereas harvesting period at showed both shrimps pond were 5.2 Log CFU/g and 4.8 Log CFU/g. The finding showed the Vibrio sp. identified in sediment samples were V. parahemolyticus with 27%, followed by V. alginolyticus (22%), V. cholerae (16%). V. fluvialis (14%) and 7% of V. fulmisi and V. vulnificus and V. mimicus. Meanwhile, the Vibrio sp. identified in water samples were V. fluvialis with 29%, followed by V. alginolyticus (22%), V. parahemolyticus (19%) and 10 % were V. fulmisi , V. mimicus and V. vulnificus. The study has been carried out further by investigated the Antibiotic Susceptibility Test (AST) using erythromycin (15 ug/ml), ampicillin (10 ug/ml), and streptomycin (25 ug/ml). The positive cultures from sediment samples showed Ampicillin resistant profile was > 0.2 MAR index while fifty percent (50%) of the isolates from water samples were resistant to ampicillin. Therefore, this study has profound implications for further monitoring process from the stocking process towards the harvesting process in order to prevent any Vibrio sp. infection which caused shrimp’s disease or food safety issue related to public consumption or of shrimps in future.

A peek into mass mortality caused by antimicrobial resistant Edwardsiella tarda in goldfish, Carassius auratus in Kerala

Edwardsiella tarda is one of the serious threats affecting the worldwide aquaculture. In the present study, four isolates were recovered from diseased goldfish, showing hemorrhages, reported with 60% mass mortality in an ornamental fish farm, Ernakulam, Kerala. Based on the phenotypic and genotypic analysis, the bacteria were identified as Edwardsiella tarda, Citrobacter freundii, Acinetobacter junii and Comammonas testosteronii. Experimental challenge studies using healthy goldfish revealed that among the four isolates, E. tarda alone leads to 100% mortality of experimental fish within 175 degree days and the pathogen could be successfully re-isolated from the moribund fish. The LD50 value of E. tarda was calculated as 9.9 × 105 CFU/fish. The histopathology of the infected tissues of goldfish had shown the typical features of E .tarda infection. The pathogen was found positive for the virulence genes viz., hly, etfA, etfD and eseD as detected using PCR. Thus E. tarda was confirmed as the real causative agent of the disease outbreak. Multiple antimicrobial resistance (AMR) exhibited by the pathogen towards 19 tested antibiotics with the MAR index of 0.46 highlighted the exposure of antibiotics to the fish in the farm. The existence of antibiotic resistant genes within the plasmid as revealed through plasmid curing studies pointed out the possibility of rapid dissemination of AMR in aquaculture. Hence proper surveillance and appropriate diagnostic methods need to be implemented at regular intervals to mitigate the menace.

Transmission of Antibiotic-Resistant Escherichia coli from Chicken Litter to Agricultural Soil

A growing concern regarding the use of animal manure as fertilizer is the contamination of soil, plants, and the environment with a variety of antibiotic-resistant and pathogenic bacteria. This study quantified and characterized the antibiotic resistance profiles of Escherichia coli in soil before and after chicken litter application to determine the impact of manure on the soil resistome. Litter and soil samples were collected from a sugarcane field before and after litter application. E. coli was isolated and quantified using the Colilert®-18/Quanti-tray® 2000 and 10 randomly selected isolates from the positive wells of each Quanti-tray were putatively identified on eosin methylene blue agar. Real-time PCR was used to confirm the isolates by targeting the uidA gene. Antibiotic susceptibility test against 18 antibiotics was conducted using the disk diffusion method, and the multiple antibiotic resistance index was calculated. Soil amendment with chicken litter significantly increased the number of antibiotic-resistant E. coli in the soil. Among the 126 E. coli isolates purified from all the samples, 76% showed resistance to at least one antibiotic, of which 54.2% were multidrug-resistant (MDR). The highest percentage resistance was to tetracycline (78.1%), with the least percentage resistance (3.1%) to imipenem, tigecycline, and gentamicin. The isolates also showed resistance to chloramphenicol (63.5%), ampicillin (58.3%), trimethoprim-sulfamethoxazole (39.6%), cefotaxime (30.2%), ceftriaxone (26.0%), cephalexin (20.8%), cefepime (11.5%), amoxicillin-clavulanic acid (11.5%), cefoxitin (10.4%), Nalidixic acid (9.4%), amikacin (6.3%), and ciprofloxacin (4.2%). Of the 54.2% (52/96) MDR, the highest number was isolated from the litter-amended soil (61.5%) and the least isolates from soil samples collected before litter application (1.9%). The relatively higher mean MAR index of the litter-amended soil (0.14), compared to the soil before the amendment (0.04), suggests soil pollution with antibiotic-resistant E. coli from sources of high antibiotic use. E. coli could only be detected in the soil up to 42 days following manure application, making it a suitable short-term indicator of antibiotic resistance contamination. Notwithstanding its relatively short detectability/survival, the application of chicken litter appeared to transfer antibiotic-resistant E. coli to the soil, enhancing the soil resistome and highlighting the consequences of such agricultural practices on public health.

Molecular Confirmation of Vancomycin-Resistant Staphylococcus aureus with vanA Gene from a Hospital in Kathmandu.

Staphylococcus aureus, a commensal on the skin and in the nasal cavity of humans, is one of the most serious cases of nosocomial infections. Moreover, methicillin-resistant S. aureus (MRSA) is a leading cause of morbidity and mortality worldwide. For the treatment of MRSA infections, vancomycin is considered as a drug of choice. However, the emergence of vancomycin resistance among MRSA isolates has been perceived as a formidable threat in therapeutic management. To estimate the rate of vancomycin-resistant S. aureus (VRSA) and to detect the vancomycin-resistant genes, namely, vanA and vanB, among the isolates, a hospital-based cross-sectional study was conducted from July to December 2018 in Annapurna Neurological Institute and Allied Science, Kathmandu, Nepal. S. aureus was isolated and identified from different clinical samples and processed for antibiotic susceptibility testing by the modified Kirby–Bauer disc diffusion method. The screening of MRSA was performed as per Clinical and Laboratory Standard Institute (CLSI) guidelines. VRSA was confirmed by the minimum inhibitory concentration (MIC) method by employing E-test strips. All the phenotypically confirmed VRSA were further processed to detect the vanA and vanB gene by using the conventional polymerase chain reaction (PCR) method. A total of 74 (20.3%) S. aureus were isolated, and the highest percentage of S. aureus was from the wound samples (36.5%). Of 74 S. aureus isolates, the highest number (89.2%) was resistant to penicillin, and on the other hand, linezolid was found to be an effective drug. Likewise, 45 (60.81%) were found to be MRSA, five (11.11%) were VRSA, and 93.2% of S. aureus isolates showed an MAR index greater than 0.2. Two VRSA isolates (40%) were positive for the vanA gene. The higher prevalence of MRSA and significant rate of VRSA in this study recommend routine surveillance for the MRSA and VRSA in hospital settings before empirical therapy.

Multi-Drug Resistance Bacteria: A Case Study in Western Peninsular Malaysia Freshwaters

The emergence of antibiotic resistant bacteria in aquatic environment has led to an increase in waterborne health risks to an alarming extent. This study attempts to investigate the population of certain antibiotic resistant strains in Peninsular Malaysia. From the samples of 14 rivers from 7 different states in Peninsular Malaysia, a total of 203 isolates were successfully isolated. These are from rivers in Negeri Sembilan, Melaka, Selangor, Kuala Lumpur, Kedah, Kelantan and Terengganu with 35, 15, 37, 39, 24, 26 and 27 isolates, respectively. The isolates were tested for their susceptibility towards 6 broad spectrum of antibiotics which are gentamicin, ampicillin, rifampicin, chloramphenicol, tetracycline, and ciprofloxacin. A total of 117 isolates were identified to have more than 20% MAR index value with 47 of the isolates possess minimum MAR index value of 50%. Based on MAR index value, 59% of isolates are high risk threats indicating a serious emergence of antibiotic resistant bacteria in the rivers in Peninsular Malaysia. Isolates with MAR index value of more than 50% were selected for 16S rRNA gene sequencing for further identification.Based on 16s rRNA gene sequencing, the isolates are a mixture of pathogenic and commensals bacteria. This also implies that the environment especially rivers can be a reservoir for genetic jugglery

The Effectiveness of the Transaction Systems on the Dax Index

The purpose of the article/hypothesis: The aim of this article is to examine the effectiveness of trading systems built on the basis of technical analysis tools in 2015–2020 on the DAX stock exchange index. Efficiency is understood as generating positive rates of return, taking into account the risk incurred by the investor, as well as achieving better results than passive strategies. Presenting empirical evidence implying the value of technical analysis is a difficult task not only because of a huge number of instruments used on a daily basis, but also due to their almost unlimited possibility to modify parameters and often subjective evaluation.
 Methodology: The effectiveness of technical analysis tools was tested using selected investment strategies based on oscillators and indicators following the trend. All transactions were carried out on the Meta Trader 4 platform. The analyzed strategies were comprehensively assessed using the portfolio management quality measures, such as the Sharpe measure or the MAR ratio (Managed Account Ratio).
 Results of the research: The test results confirmed that the application of described investment strategies contributes to the achievement of effective results and, above all, protects the portfolio against a significant loss in the period of strong turmoil on the stock exchange. During the research period, only two strategies (Ichimoku and ETF- Exchange traded fund) would produce negative returns at the worst possible end of the investment. At the best moment, however, the „passive” investment achieved the lowest result. Looking at the final balance at the end of 2019, as many as four systems based on technical analysis were more effective than the „buy and hold” strategy, and at the end of the first quarter of 2020 – all of them. When analyzing the management quality measures, it turned out that taking into account the 21 quarters, the passive strategy had the lowest MAR index. The Sharpe’s measure is also relatively weak compared to the four leading strategies.

Plasmid profiling of multiple antibiotic-resistant Pseudomonas aeruginosa isolated from soil of the industrial area in Chittagong, Bangladesh

BackgroundMultiple antibiotic-resistant (MAR) Pseudomonas aeruginosa (P. aeruginosa) plays a significant role in triggering nosocomial infection in clinical settings. While P. aeruginosa isolated from the environment is often regarded as non-pathogenic, the progressive development of antibiotic resistance necessitates exploring the MAR patterns and transposable genetic elements like plasmid in the isolates.ResultsUsing ecfX gene-based PCR, 32 P. aeruginosa isolates among 48 soil samples collected from the industrial region have been confirmed. The antibiotic susceptibility pattern of those isolates revealed that 5 (15.63%) of them were resistant to a range of antibiotics, and they were categorized as MAR isolates. Nevertheless, all MAR isolates were found resistant to piperacillin and gentamicin, but none of them to ceftazidime, aztreonam, and ciprofloxacin. Moreover, the isolates were also showed resistance to amikacin (60%), tobramycin (80%), netilmicin (80%), imipenem (60%), doripenem (40%), meropenem (60%), and cefixime (40%). Furthermore, 60% of MAR isolates possessed double plasmids of 1000–2000 bp sizes which indicates the distribution of antibiotic resistance genes in MAR P. aeruginosa might be correlated with the presence of those plasmids. The MAR index’s high threshold values (> 0.20) implied that the isolates were from high-risk environmental sites where the presence of numerous antibiotic residues happened.ConclusionsThese findings highlighted the presence of multiple antibiotic resistance in P. aeruginosa of the industrial soil and a considerable prospect of transferring antibiotic resistance genes in the microbial community by plasmids. We recommend taking immediate stringent measures to prohibit the unnecessary and overuse of antibiotics in agricultural, industrial, or other purposes.

Occurrence and antimicrobial resistance of Campylobacter isolates from broiler chicken and slaughter house environment in India

Campylobacteriosis is among the most frequently reported foodborne zoonoses. A total of 848 samples were screened for Campylobacter spp. and occurrence was found to be 8.7%, 2.3% and 1.65% in broiler cecum samples, chicken meat samples and slaughter house environment swabs, respectively. High level of antimicrobial resistance was found against tetracycline (64.1%), doxycycline (54.4%), ampicillin (46.6%), nalidixic acid (42.7%), kanamycin (35.9%), and ciprofloxacin (33.33%). Resistance to co-amoxiclav (19.4%) and erythromycin (21.4%) was less common. The MAR index of the isolates was in the range of 0.11–0.78. Multi-drug resistance was observed in 54.4% of the isolates, with 53.2% C. jejuni and 55.3% C. coli isolates found resistant against three or more classes of antimicrobials. Presence of mutations in gyrA and 23S rRNA genes was investigated, which revealed that all the fluoroquinolone resistant isolates possessed Thr-86-Ile point mutation, whereas only 68% of erythromycin resistant isolates had A2075G mutation. The tetO gene was present in 91.7% tetracycline resistant isolates and bla OXA-61 gene was detected in 97.9% of the ampicillin resistant isolates. The occurrence of antimicrobial resistant Campylobacter spp. in broiler chicken samples and slaughter house settings is a public health risk and calls for judicial use of antimicrobials.

Assessment of Antibiotics Susceptibility Pattern of Vancomycin Resistant Enterococci Isolated from Clinical Specimens in Port Harcourt

Aim: The aim of this study was to assess the prevalence of vancomycin resistance among Enterococcal species isolated from clinical specimens of patients attending two hospitals in Port Harcourt, Rivers State.
 Study Design: The study employs statistical analysis of the data and interpretation.
 Place and Duration of Study: Two hospitals which are Meridian hospital Port Harcourt and University of Port Harcourt Teaching Hospital, located in the city of Port-Harcourt, Rivers State were used for this study. Specimen collection lasted for 3 weeks and the analysis was carried out daily and it lasted for six months.
 Methodology: A total of one hundred and eighteen (118) urine and stool specimens (60 urine and 58 stool specimens) were collected from Fifty nine (59) patients for a period of three months from Meridian hospital and University of Port Harcourt Teaching Hospital, Port-Harcourt, Rivers State. The specimens collected were grouped inpatients and outpatients and were subjected to standard microbiological procedures which include standard plate counts, identification, and sensitivity testing using Kirby-Bauer disk diffusion method, Minimum inhibitory concentration and molecular identification of the isolates.
 Results: A total of 48 enterococcal isolates were isolated from the different specimens (hospitalized and non-hospitalized patients) of urine and stool specimens. All Enterococcal isolates showed high level of resistance to Ceftazidime and Cefuroxime (100%) followed by cloxacillin (95.8%), augumentin (85.4%) and Ceftriaxone (75.0%). The isolates showed higher sensitive rates to Ofloxacin (95.5%), followed by Gentamicin (77.1%) and Vancomycin (39.6%). All Enterococal isolates from this study had a MAR index > 0.2. A total of the 48 Enterococci were isolated, the 23 (47.9%) isolates were identified as vancomyin resistant during this study were subjected to MIC (Minimum Inhibitory Concentration) for vancomycin as a confirmatory test. Of the 23 isolates, 12 isolates were vancomycin resistant with 11 isolates showing vancomycin MIC values of 8-16μg /ml (vancomycin intermediate).
 Conclusion: Conclusively, this study revealed varying Antibiotic susceptibility pattern of the isolated bacteria. Treatment guidelines for use of antibiotics should be based on the hospital formulary and the sensitivity patterns is advocated. This should be reviewed occasionally to ensure rational use of antibiotics