Isolation of Antibiotic Resistant Plasmid Deoxyribonucleic Acid from MDR Diarrheal Pathogens

Cholera is an acute infectious disease in countries with poor sanitation. The main clinical symptom of cholera is gastroenteritis and a symptom of the disease includes mild to moderate dehydration, vomiting, fever and abdominal pain. Antibiotic drug resistance in V. cholera has become a serious problem mostly in developing countries and muti-drug resistance to different antibiotics as well as Tetracycline, Ampicillin, Kanamycin, Trimethoprim, Sulphonamides Streptomycin and Gentamicin. Multi-drug resistant V. cholera showed resistance against three or more three contrasting classes of antibiotics for recent decades. Fifty diarrheal samples were collected from the different hospitals in and around the Tirupur district. 25 positive V. cholerae were isolated. The isolates were characterized morphologically and biochemically. The confirmed strains were taken to decide susceptibility patterns by the disc diffusion method. Vibrio cholerae isolated in this study was subjected to an antibiogram against 10 commonly used antibiotics. All the tested isolates showed maximum resistance to Erythromycin (97%) and minimum resistance was noted in Trimethoprime (50%) respectively. Electrophoretic examination of plasmid DNA was carried out by Agarose gel Electrophoresis on 0.7%. More than 90% of resists showed were taken for plasmid isolation. The molecular weights of the fragments were evaluated by using a 10,000 bp DNA ladder and it was determined to be 1500 bp respectively. It is crucial to know the sensitivity and resistance pattern of any bacterial species for intercession an effective drug of choice in future

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Isolation, Molecular Identification and Multidrug Resistance Profiling of Bacteria Causing Clinical Mastitis in Cows

Agricultural Science Digest - A Research Journal ◽

10.18805/ag.d-5220 ◽

2020 ◽

Author(s):

D.J. Vatalia ◽

B.B. Bhanderi ◽

V.R. Nimavat ◽

M.K. Jhala

Keyword(s):

Drug Resistance ◽

Antibiotic Resistance ◽

Multiple Drug Resistance ◽

Bacterial Species ◽

Research Work ◽

Resistance Index ◽

Diffusion Method ◽

Multiple Drug ◽

Milk Samples ◽

Sensitivity Pattern

Background: Mastitis, the inflammation of parenchyma of mammary gland is frequently considered to be costliest and complex disease prevalent in India. Mastitis is caused by pathogens like Staphylococcus spp., Streptococcus spp., Mycoplasma bovis, E. coli, Klebsiella spp., Citrobacter spp., Enterobacter spp. and Entercoccus. The treatment of mastitis in animals is carried out using antibiotics. Treatment failure in mastitis is due to increased antibiotic resistance of mastitis pathogens and also due to indiscriminate use of antibiotics without testing in vitro antibiotic sensitivity test against causal organisms. In comparison to cultural method, PCR assays takes less time for detection of bacteria from the mastitis milk samples. Present research work was carried out regarding isolation, identification and multiple drug resistance profile of clinical bovine mastitis associated pathogens using conventional as well as molecular approach. Methods: In the present study, 73 mastitis milk samples were collected from Anand and Panchmahal district of Gujarat. The milk samples were subjected for cultural isolation and DNA extraction for identification of bacteria by cultural and PCR method. Antimicrobial sensitivity pattern of the isolates were carried by disc diffusion method and isolates were categorized in multiple drug resistant. Result: In the present study, Out of 73 mastitis milk samples collected from cows 48 (65.75%) cows were positive for bacterial isolation and S. aureus was the most predominant bacterial species. PCR from the mastitis milk additionally detected bacteria in culturally negative milk samples. Most sensitive drug was gentamicin and most of the isolates (90.19%) showed the multiple drug resistance for the two to nine drugs with 0.1 to 0.6 multiple antibiotic resistance index.

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Antibiotic Resistance in the Veterinary Perspective

Global Applications of One Health Practice and Care - Advances in Medical Diagnosis, Treatment, and Care ◽

10.4018/978-1-5225-6304-4.ch006 ◽

2019 ◽

pp. 142-157

Author(s):

Sophia Inbaraj ◽

Vamshi Krishna Sriram ◽

Prasad Thomas ◽

Abhishek Verma ◽

Pallab Chaudhuri

Keyword(s):

Drug Resistance ◽

Antibiotic Resistance ◽

Food Chain ◽

Bacterial Species ◽

Animal Husbandry ◽

Drug Resistant ◽

Antimicrobial Drug Resistance ◽

Antibiotic Drug ◽

Antibiotics Use ◽

Genetic Mechanisms

Antibiotic resistance is an emerging threat to achieving one health all over the globe. The phenomenon leads to the emergence of drug-resistant microbes previously susceptible to an antibiotic. Drug-resistant microbes are the major reasons for medical complications like patient mortality and treatment failure. Unregulated use of antibiotics in animal husbandry is one of the major reasons for the emergence of antibiotic resistance. The resistance enters the human population mainly through the food chain. The genetic markers associated with drug resistance spread among different bacterial species by horizontal gene transfer mechanisms. Therefore, regulation of antibiotics use in animal husbandry and proper safety measures at farm level are necessary to check drug-resistant microbes entering the food chain. This chapter discusses the antibiotics, antibiotic resistance, genetic mechanisms involved, the spread of resistance, and also the available strategies to combat antimicrobial drug resistance.

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Detection of Pan drug resistance OXA-48 producing Providencia in an ICU patient for the first time in Nepal

Antimicrobial Resistance and Infection Control ◽

10.1186/s13756-019-0608-1 ◽

2019 ◽

Vol 8 (1) ◽

Cited By ~ 3

Author(s):

Ranjit Sah ◽

Shusila Khadka ◽

Gentle Sunder Shrestha ◽

Subhash Acharya ◽

Diptesh Aryal ◽

...

Keyword(s):

Drug Resistance ◽

Pathogenic Bacteria ◽

Antimicrobial Agents ◽

Vital Signs ◽

Carbapenem Resistance ◽

Diffusion Method ◽

Resistance Pattern ◽

Common Mechanism ◽

Disk Diffusion Method ◽

Drug Resistance Pattern

Abstracts Background Resistance to antimicrobial agents of pathogenic bacteria has become a major problem in routine medical practices. Carbapenem resistance has long been increasing. The production of carbapenem- hydrolysing β-lactamases (carbapenamases), which include NDM, KPC, OXA-48, IMP-1 and VIM is the most common mechanism. Case presentation A 56 years old male presented with fever and mental changes with progressively decreasing sensorium for the last 3 days. He was admitted to Intensive care unit (ICU) with a diagnosis of meningoencephalitis. On day seven, he developed ventilator associated pneumonia due Klebsiella pnemoniae and Acinetobacter baumannii. He was on meropenem, but the isolates were susceptible to colistin, tigecyclin and amikacin solely. Hence, amikacin was started with addition of intravenous and nebulized colistin. Subsequently, vital signs improved with resolution of fever. However, on day 18, he developed fever once again with a drop in blood pressure. Inotropic support was maintained, and echinocandins and tigecycline were added to the regimen. Repeat blood and urine culture grew Providencia species, which were resistant to most of the drugs on phenotypic Kirby-Bauer disk diffusion method and are intrinsically resistant to colistin and tigecycline. Phenotypic detection of ESBL (combined disk method), MBL, KPCs, AmpC and co-producer were tested according to updated CLSI guideline and all were negative. But the Modified Hodges test was found to be positive. Consequenty, OXA-48 drug resistance pattern was brought into action by blank disc method according to A Tsakris et al., which revealed indentation of growth toward both EDTA and EDTA/PBA disk indicating production of OXA-48 carbapenamase. To confirm the resistance pattern we processed the isolated colonies for Xpert Carba-R (Cepheid) assay, which detected blaOXA-48 gene and confirmed the OXA-48 drug resistance pattern. Hence, the infecting organism was not susceptible to any of the antibiotics. The patient was kept under isolation and on 31th day of admission, he died of septic shock. Conclusions Carbapenamase production along with intrinsic colistin resistance in infecting bacterial pathogens can cause fatal outcomes in the resource limited countries like Nepal where new antibiotic combinations ceftazidime+ Avibactam, or aztreonam +avibactam are not available. Drug resistance patterns including OXA 48 producer should be characterized in all cases by standard phenotypic methods or by Xpert Carba-R assay and larger studies are required to know the exact burden of OXA 48 producer in Nepal.

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Antibiotic Resistance, Plasmids and Integron Profile of Salmonella Species Isolated from Poultry Farm and Patients

Dhaka University Journal of Pharmaceutical Sciences ◽

10.3329/dujps.v15i2.30939 ◽

2017 ◽

Vol 15 (2) ◽

pp. 209-214 ◽

Cited By ~ 3

Author(s):

Kohinur Begum ◽

Sultana Juhara Mannan ◽

Aliza Ahmed

Keyword(s):

Drug Resistance ◽

Nalidixic Acid ◽

Plasmid Profile ◽

Class I ◽

Diffusion Method ◽

Resistance Pattern ◽

Poultry Farm ◽

Serological Tests ◽

Clinical Strains ◽

Drug Resistance Pattern

A total of ten selected strains of Salmonella species, five from the environment of poultry farm and five from patients were included in this study. All strains were isolated and identified by using selective media, standard biochemical and serological tests. Antimicrobial susceptibility tests were performed by disc diffusion method using twelve commercial antibiotic discs of aztreonam, ceftriaxone, kanamycin, ciprofloxacin, gentamycin, chloramphenicol, ampicillin, erythromycin, cephalexin, tetracycline, cotrimoxazole and nalidixic acid. Plasmid profile and integron gene detection were conducted by Kado-Liu method and PCR, respectively. Only aztreonam, ceftriaxone, kanamycin and gentamycin were shown to be inhibitory to all strains. However, ciprofloxacin, chloramphenicol, amoxicilin, erythromycin, cephalexin, tetracycline, cotrimoxazole and nalidixic acid revealed different degrees of resistance pattern against environmental and clinical strains. Analysis of plasmid demonstrated that three environmental strains contained both plasmids of 140 Mda and 62 Mda. On the other hand, another three strains, one environmental and two clinical isolate only contained 140 Mda plasmid. All the plasmid containing strains (140 Mda and 62 Mda) exhibited same type of drug resistance pattern, whereas strains containing the 140 Mda plasmid only did not show similar type of resistance pattern. Therefore, no correlation was found between plasmid containing strains and drug resistance. Four environmental strains were also found to be positive for the class I integron and one clinical isolated was positive for class I integron gene conferring resistance to common antibiotics. However, none of the strains were found to contain class II integron. Therefore, the present study demonstrated that both environmental and clinical strains contain both large to middle size plasmids and integron I but not integron II. The plasmid and integron I containing strains experienced resistance to different antibiotics, used in the experiments.Dhaka Univ. J. Pharm. Sci. 15(2): 209-214, 2016 (December)

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Antibiotic Resistance ofSalmonellaspp. Isolated from Shrimp Farming Freshwater Environment in Northeast Region of Brazil

Journal of Pathogens ◽

10.1155/2013/685193 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5 ◽

Cited By ~ 11

Author(s):

Fátima C. T. Carvalho ◽

Oscarina V. Sousa ◽

Edirsana M. R. Carvalho ◽

Ernesto Hofer ◽

Regine H. S. F. Vieira

Keyword(s):

Drug Resistance ◽

Antibiotic Resistance ◽

Shrimp Farming ◽

Diffusion Method ◽

Resistance Pattern ◽

Plasmid Curing ◽

Freshwater Environment ◽

Disk Diffusion Method ◽

Northeast Region ◽

Drug Resistance Pattern

This study investigated the presence and antibiotic resistance ofSalmonellaspp. in a shrimp farming environment in Northeast Region of Brazil. Samples of water and sediments from two farms rearing freshwater-acclimatedLitopenaeus vannameiwere examined for the presence ofSalmonella. Afterwards,Salmonellaisolates were serotyped, the antimicrobial resistance was determined by a disk diffusion method, and the plasmid curing was performed for resistant isolates. A total of 30 (16.12%) of the 186 isolates were confirmed to beSalmonellaspp., belonging to five serovars:S. serovar Saintpaul,S. serovar Infantis,S. serovar Panama,S. serovar Madelia, andS. serovar Braenderup, along with 2 subspecies:S. entericaserovar houtenae andS. entericaserovar enterica. About twenty-three percent of the isolates were resistant to at least one antibiotic, and twenty percent were resistant to at least two antibiotics. Three strains isolated from water samples (pond and inlet canal) exhibited multiresistance to ampicillin, tetracycline, oxytetracycline, and nitrofurantoin. One of them had a plasmid with genes conferring resistance to nitrofurantoin and ampicillin. The incidence of bacteria pathogenic to humans in a shrimp farming environment, as well as their drug-resistance pattern revealed in this study, emphasizes the need for a more rigorous attention to this area.

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Bacterial Tonsillitis and Antimicrobial Resistance Profiles Among Children Within Five Years of Age At Hargeisa Group of Hospital, Somaliland: A Cross-Sectional Study

10.21203/rs.3.rs-586545/v1 ◽

2021 ◽

Author(s):

Hamda Hussein Darod ◽

Addisu Melese ◽

Mulugeta Kibret ◽

Wondemagegn Mulu

Keyword(s):

Drug Resistance ◽

Antimicrobial Resistance ◽

Antimicrobial Susceptibility ◽

School Attendance ◽

Bacterial Species ◽

Cross Sectional Study ◽

Diffusion Method ◽

Sectional Study ◽

Cross Sectional ◽

History Of

Abstract Background Tonsillitis is the third most frequently diagnosed infection in the pediatrics age group around the world. It causes significant morbidity and loss of school attendance in children. The emergence of drug resistance in bacterial tonsillitis is getting higher every year. However, data on the drug resistance profiles of bacterial causes of tonsillitis among children within five years of age is not available in Somaliland.Therefore; this study determined the bacterial causes of tonsillitis and their antimicrobial resistance profiles among children within five years of age at Hargeisa Group of Hospital, Somaliland. Methods A cross-sectional study was conducted from March to July 2020. A total of 374 children within five years of age were included using convenient sampling method. Throat swabs were collected from children, processed and bacterial species were identified using standard bacteriological procedures. Antimicrobial susceptibility was done using disc diffusion method. Data on demographic variables and clinical profiles were collected using structured questionnaires. Logistic regression analysis was computed to identify factors associated with bacterial tonsillitis. P-values < 0.05 were taken as statistically significant. Results The median age of children included in the study was 4 years. Overall, 120(32.1%)(95% CI 27.4–36.8%) of children had culture confirmed bacterial tonsillitis. Of them,23(19.2%) had mixed infections.The most frequent bacterial isolates were Streptococcus pyogenes 78(55%), Staphylococcus aureus 42 (29%) and Streptococcus pneumoniae 10(7%). Isolates revealed 83.3–100 % rate of resistance to ampicillin. S. aureus was resistant to clarithromycin (38%) while 60% of S. pneumoniae isolates were resistant to gentamicin. The overall multidrug resistance (MDR) was 50.4% and 52.6% of S.pyogenes and 60% of S.pneumoniae were MDR. History of tonsillitis(AOR = 0.12; 95% CI = 0.06–0.21), difficulty of swallowing(AOR = 6.99; 95% CI = 3.56–13.73), weight loss (AOR = 0.33; 95% CI = 0.186–0.597) and attending school (AOR = 2.98; 95% CI = 1.64–5.42) were found to be associated with tonsillitis among children within five years of age. Conclusions Bacterial tonsillitis with high degree of ampicillin resistance, mixed infections and MDR isolates are major concerns in children within five years of age at Hargeisa, Somaliland. Therefore, treatment of cases should be guided by regular culture and antimicrobial susceptibility testing to prevent disease consequences and critical drug resistance.

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Evaluation of antimicrobial resistance pattern of uropathogens in a tertiary care hospital in Dhaka city, Bangladesh

South East Asia Journal of Public Health ◽

10.3329/seajph.v7i2.38851 ◽

2018 ◽

Vol 7 (2) ◽

pp. 12-18 ◽

Cited By ~ 1

Author(s):

Azizun Nahar ◽

Shahed Hasnat ◽

Hasina Akhter ◽

Nilufar Begum

Keyword(s):

Urinary Tract ◽

Bacterial Infections ◽

Bacterial Species ◽

Tertiary Care ◽

Urine Samples ◽

Diffusion Method ◽

Resistance Pattern ◽

National Committee ◽

Care Hospital ◽

Dhaka City

Urinary tract infection (UTI) is one of the most common bacterial infections seen in developing countries like Bangladesh. This study is, therefore, designed to determine the bacterial uropathogens and their antibiotic resistance pattern among patients with complaints of UTIs in Dhaka city. This study was carried out in the laboratory of the Department of Microbiology, Bangladesh Medical College, Dhaka, Bangladesh from January to June 2015. A total of 2541 urine samples were collected in sterile containers from suspected urinary tract infected cases. A specimen was considered positive for UTI if an organism was cultured at a concentration of ≥105CFU/ml or when an organism was cultured at a concentration of 104CFU/ml and >5 pus cells per high power field. Antimicrobial susceptibility testing of the isolated bacterial species was performed by disc diffusion method following the National Committee for Clinical laboratory Standards (NCCLS) guidelines. A total of 303 (11.92%) bacterial uropathogens were isolated from 2541 urine samples. Among the 303 isolates, majority of the isolates 197 (65.02%) were from females. Both the age groups up to 18 years and above 18 years the highest prevalence was found in females 34(11.22%) and 163 (53.79%) respectively. Most predominant organism was Escherichia coli 262(86.46%)followed by Pseudomonas 12(3.96%), Enterococci 12(3.96%), Klebsiella11(3.63%). Escherichiacoli showed very high resistance to amoxycillin 95.41%, cefradin 90.45%, nalidixic acid and Klebsiella to amoxycillin 90.90%, nitrofurantoin 90.90%. Again Pseudomonas was highly resistant to cefuroxime 100%, cefexime 100%and ceftriaxone 83.33%. Enterococci were found highly resistance to cefexime 91.66%, cloxacillin 83.33%, and erythromycin 83.33%.Due to wide scale resistance of the drugs used to treat UTI, choice of drugs in the treatment of UTI is quite narrow. In country like ours awareness for prevention of UTI should be encouraged among the community level as it affects all age groups.South East Asia Journal of Public Health Vol.7(2) 2017: 12-18

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Molecular Detection of Carbapenem Resistance in Acinetobacter Baumannii Isolated From Patients in Khorramabad City, Iran

Infectious Disorders - Drug Targets ◽

10.2174/1871526519666190517124314 ◽

2020 ◽

Vol 20 (4) ◽

pp. 543-549

Author(s):

Zeinab Babaie ◽

Somayeh Delfani ◽

Faranak Rezaei ◽

Fatemeh Norolahi ◽

Somayeh Mahdian ◽

...

Keyword(s):

Drug Resistance ◽

Acinetobacter Baumannii ◽

Resistance Genes ◽

High Resistance ◽

Diffusion Method ◽

Resistance Pattern ◽

Biochemical Tests ◽

Transfer Resistance ◽

Drug Resistance Pattern ◽

Wide Range

Background: Acinetobacter baumannii is an opportunistic pathogen, which causes a wide range of infections in hospitals, especially in intensive care units. Nowadays, due to the high resistance of Acinetobacter bumanni to antibiotics, this study, in addition to the phenotypic and genotypic investigations of drug resistance, focused on determining the molecular types of Acinetobacter baumannii isolated from patients in Khorramabad city by the pulsed-field gel electrophoresis (PFGE) method. Materials and Methods: In this cross-sectional study, 50 samples of Acinetobacter baumannii were collected from educational hospitals in Khorramabad city, Iran, from January to August 2015. They were identified in the laboratory using biochemical tests and culture methods. After determining the drug resistance pattern by the disc diffusion method and percentage of resistance genes to carbapenems, Acinetobacter baumannii isolates were analyzed using the PFGE method using the Apa1 enzyme. Results: The highest antibiotic resistance observed for Acinetobacter baumannii strains was against ampicillin-sulbactam (100%) and aztreonam (98%). The highest sensitivity was to polymixin B (100%) and colistin (94%), and also to the OXA-51-like gene present in all samples. The OXA-23-like gene was positive in 44 (88%) samples. PFGE results showed that Acinetobacterbaumannii strains had 33 different pulsotype patterns, of which 27 patterns had more than one strain and 23 had only one strain. Conclusion: Due to the high resistance of Acinetobacter baumannii and its ease of spread and its ability to transfer resistance genes, resistance control methods should be used in the disinfection of hospital areas. Hospital staff should observe hygiene standards and there should also be a reduction in antibiotic use.

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Prevalence and Antibiotic Resistance Pattern of Acinetobacter Isolated from Patients Admitted in ICUs in Mazandaran, Northern Iran

Global Journal of Health Science ◽

10.5539/gjhs.v8n11p112 ◽

2016 ◽

Vol 8 (11) ◽

pp. 112 ◽

Cited By ~ 1

Author(s):

Roya Ghasemian ◽

Mohammad Ahanjan ◽

Ebrahim Fatehi ◽

Mehran Shokri

Keyword(s):

Antibiotic Resistance ◽

Resistance Rate ◽

Test Method ◽

Diffusion Method ◽

Resistance Pattern ◽

Disc Diffusion ◽

Disc Diffusion Method ◽

Northern Iran ◽

Cross Sectional ◽

Antibiotic Resistant

BACKGROUND & PURPOSE: Antibiotic resistance rate is increasing in Acinetobacter species, especially in Acinetobacter baumannii, as the most important pathogen of hospital and ICU . This research aimed to evaluate antibiotic resistant rate of Acinetobacter spp. isolated from patients admitted to ICUs in educational hospitals affiliated with Mazandaran University of Medical Sciences.METHODS: In this cross-sectional descriptive study, 50 Acinetobacter isolates were collected during 2013- 2014. After confirming Acinetobacter species, antibacterial sensitivity test was done using disc diffusion method and minimal inhibitor concentration (MIC) was evaluated by E-test in all isolates.RESULTS: Disc diffusion method revealed that 100% of isolates were resistant to Amikacin and Cefepim and 96% were resistant to both Meropenem and Ciprofloxacin antibiotics, 6% were sensitive, 18% were intermediate and 76% were resistant to imipenem. Also, 84% of isolates were sensitive and 16% were resistant to colistin. In E-test method, 92% of isolates were sensitive and 8% were resistant to colistin. Moreover, an isolate was sensitive, one was intermediate and the remaining isolates were resistant to ciprofloxacin, and 100% of isolates were resistant to other antibiotics in E-test. Over 96% of Acinetobacter isolates were resistant to the antibiotics frequently used in ICU (ciprofloxacin, meropenem, amikacin, and cefepim). Colistin was found as the only appropriate antibiotic that could be used for patients in ICU.CONCLUSION: We hope these results could change the attitude of physicians toward using antibiotics in ICUs and encourage them to follow antibiotic stewardship as the only effective strategy to somewhat control antibiotic resistances.

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Purification, crystallization and preliminary X-ray diffraction analysis of theN-acetyltransferase SAV0826 fromStaphylococcus aureus

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x13034493 ◽

2014 ◽

Vol 70 (2) ◽

pp. 211-214

Author(s):

Parul Srivastava ◽

Yogesh B. Khandokar ◽

Jade K. Forwood

Keyword(s):

Diffraction Analysis ◽

Single Molecule ◽

Protein Function ◽

Bacterial Species ◽

Diffusion Method ◽

X Ray Diffraction ◽

X Ray ◽

Cell Parameters ◽

Wide Range ◽

Antibiotic Drug

Staphylococcus aureusis a prevalent microorganism that is capable of causing a wide range of infections and diseases. Several strains of this bacterial species have developed antibiotic resistance to methicillin and vancomycin, and higher death rates are still being reported each year owing to multidrug-resistant strains. Certain GCN5-relatedN-acetyltransferases (GNATs) exhibit a broad substrate range, including aminoglycosides, histones, other proteins and serotonin, and have been implicated in antibiotic drug resistance. Here, the expression, purification, crystallization and preliminary X-ray diffraction analysis of a GNAT fromS. aureus(SaNAT) are reported. SaNAT was recombinantly expressed and crystallized by the hanging-drop vapour-diffusion method at 296 K, and the crystals diffracted to 1.7 Å resolution on the MX2 beamline at the Australian Synchrotron. The crystals belonged to space groupP43212, with unit-cell parametersa=b= 84.86,c= 49.06 Å, α = β = γ = 90°. A single molecule is likely to be present in the asymmetric unit. A full structural and functional analysis is currently being undertaken to provide novel insights into the protein function, which in turn may provide a basis for drug design.

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