Antibacterial effects of Octenicept, and benzalkonium chloride on Acinetobacter baumannii strains isolated from clinical samples and determination of genetic diversity of isolates by RAPD-PCR method

Author(s):  
Azar Dokht Khosravi ◽  
Effat Abbasi Montazeri ◽  
Seyyedeh Roya Maki
2021 ◽  
Author(s):  
Azar Dokht Khosravi ◽  
Effat Abasi Montazeri ◽  
Seyedeh Roya Maki

Abstract Due to the emergence of antibiotic resistance in Acinetobacter baumannii which is one of the important causes of nosocomial infections, many problems have been raised in the successful treatment of patients with the subsequent mortality. So, the present study was performed to evaluate the antibacterial effect of Actinidine dehydrochloride, Actinisept, and Benzalkanium chloride against Acinetobacter baumannii strains isolated from clinical samples and to determine the genetic diversity of strains by RAPD-PCR. A total of 119 non-duplicate, suspected Acinetobacter baumannii isolates were collected and confirmed by conventional culture and biochemical tests and PCR technique. Susceptibility of the isolates to antibiotics was evaluated by standard Antibiotic susceptibility testing (AST). For antiseptics Octenidine dihydrochloride (OCT), Actinisept, and Benzalkonium chloride (BZK), Minimal inhibitory concentration (MIC) was assessed. The prevalence of Qac E and Qac delta E genes related to antiseptics was estimated by PCR. Finally, genetic diversity of strains was determined by RAPD-PCR. All 119 suspected isolates were confirmed as Acinetobacter baumannii using conventional tests and PCR. The isolates were mostly originated from blood samples. In AST, the lowest resistance was seen for ciprofloxacin and gentamicin. The MIC values were reported as OCT (15.26 µg) and BZK (640 µg). The antiseptic genes of qacE and qac ΔE1 were found to be present in 56 (47.05%) and 59 (49.57%) of isolates respectively. RAPD typing method revealed great diversity among A. baumannii isolates, with 37 clusters in isolates from ICU, of which 32 isolates were single and 5 were multiple. In conclusion, considering the increase of resistance to antiseptics, it is of importance to monitor the susceptibility of A. baumannii to antiseptics and to promote antiseptic stewardship in hospitals. Furthermore, in this study great diversity among A. baumannii was observed making it difficult to properly carry out infection control policies. analysis of RAPD-PCR typing results, and we found 37 clusters, among them 32 isolates were single and 5 were multiple. So, the method generated 37 RAPD type which shows great diversity among 57 out of 62 A. baumannii isolates at 80% cutoff.


Author(s):  
Riva Hafidah ◽  
Ayi Yustiati ◽  
Yuniar Mulyani ◽  
Ibnu Bangkit Bioshina Suryadi

This research aims to determine genetic diversity of four strains guppy, respectively are japan blue double sword (JBD), japan blue tiger double sword (JBTD), blue moscow (BM), and panda guppy (PG) with RAPD-PCR method. The obtained genetic diversity data is used as guide reference for hybridization between four strains. The research was conducted in September 2020 to April 2021 with explorative methods and in qualitative and quantitative descriptive analysis. The research was carried out in biotechnology Laboratory, Fishery and Marine Sciences Faculty and Central Laboratory,Padjadjaran University, Indonesia.Strains of JBD, JBTD, BM obtained fromCilengkrangSubdistrict, Bandung and PG strain obtained from Parung market, Bogor. Primary OPA-03 (AGTCAGCCAC) is used for standard parameters to interpret genetic diversity among four strains of guppy. Based on results, amplification with OPA-03 primary visualize 25 bands that include five polymorphic bands and 20 monomorphic bands. The phylogenetic tree result show that there are two relationship groups. The first group are JBD, JBTD, and BM with similarity index in the range of 80-89%The first group consist two sub groups of relationship. The first sub group are JBD and JBTD with similarity index of 89%. The second sub group is BM with similarity index of 80%. The second group isPG with similarity index of 65.5%.


2017 ◽  
Vol 7 (1) ◽  
pp. 34
Author(s):  
Alireza Khakzad ◽  
Fatemeh Keshavarzi

Salmonella species are gram negative bacteria and members of Enterobacteriaceae family. It has a rod-shaped appearance; it is catalase positive, oxidase negative, non-spore. Salmonella classified into two species, Salmonella Enterica and Salmonella Bangori. Salmonella is now one of the main reasons of diarrhea and vomiting in humans in many countries and especially in industrialized. In a study in Japan 164 Salmonella digestions were collected during 2006 to 2008 which 81 digestions were Salmonella Infantis. Salmonella-specific characteristics are studied in the two phenotype and genotype methods. In this research, with using genotype methods based on PCR, genetic diversity was evaluated; this PCR includes rep-PCR based on repetitive sequence elements (method was done by the use of three primers ERIC, REP and BOX). Studied showed most isolated strains were relevant to Salmonella Enteritidis and dendorogram study showed that the bacteria were grouped in one cluster in dendrogram that all 37 strains were put in a large cluster of Salmonella’s type which is divided into two clusters: Salmonella Enterica and Bongori. The results also in this experiment reflect the efficiency of rep-PCR method by using three ERIC, REP and BOX primers.


2011 ◽  
Vol 8 (1) ◽  
pp. 1-7
Author(s):  
Purnama Hidayat ◽  
Noor Aidawati ◽  
Sri Hendrastuti Hidayat ◽  
Dewi Sartiami

Indicator Plant and PCR-RAPD for Biotype Determination of Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae).B. tabaci has been known world wide as a major pest and virus vector of horticulture. In Indonesia the presence of B.tabaci was reported since 1980 and its role as virus vector in tomato and chilli pepper has becoming more importantrecently. Genetic diversity of B. tabaci has been well recognized, but very little information available for diversity of B.tabaci in Indonesia. This research was conducted in Bogor, West Java from May 2004 to June 2005. The aim of thisresearch was to initiate basic information regarding genetic diversity of B. tabaci in Indonesia, particularly in Java Island.Whiteflies population collected from different crops, i.e. tomato, broccoli, chill pepper, eggplant, cucumber, soybean, andedamame, was evaluated using silverleaf-induction test, and RAPD-PCR. It was evidenced that only B. tabaci populationfrom broccoli was able to induce silverleaf. Two genetic types of B. tabaci, i.e. biotype B and non B, were identified basedon polymorphism character of DNA. Population from broccoli was belong to biotype B, whereas other populations fromtomato, chill pepper, eggplant, cucumber, soybean, and edamame were belong to biotype non B.


Author(s):  
Merih Şimşek ◽  
Cengiz Demir

Objective: Acinetobacter baumannii (A. baumannii) can develop resistance to various antimicrobial agents via different mechanisms. Hence, the aim of this study was to investigate, by using different methods, the resistance profiles of A. baumannii strains isolated from different clinical specimens; from colistin and tigecycline antibiotics, and also the distribution of this resistance according to the clinical samples. Material and Methods: For this study, 1,265 clinical samples (a samples from each patient) were obtained from various clinics, between; January 2015/December 2018. Identification was conducted by VITEK® 2 compact (bioMerieux, USA) and conventional biochemical tests. Antibiotic susceptibility tests were performed by VITEK 2, and the results of colistin and tigecycline were confirmed by E test and the broth microdilution method. Results: A. baumannii strains (1,265) were most frequently isolated from tracheal aspirate, sputum and blood samples. At the same time, strains were obtained from intensive care units (70.4%) as well as other clinics (29.6%). The rates of colistin and tigecycline-resistant strains were determined using VITEK 2, E test and the broth microdilution methods as: 3.0%, 5.7%, 9.0% and 21.7%, 24.5%, 33.0%, respectively. Conclusion: The determination of appropriate antibioticis are important for empirical treatment. Colistin and tigecycline have become prominent as an important, alternative agent in the treatment of A. baumannii-related infections. The results of this study show that colistin and tigecycline resistance rates in intensive care units have been increasing gradually over the years. Monitoring of resistance patterns of nonfermentative bacteria, isolated from intensive care units, is important for the immediate initiation of appropriate empirical treatment. In-vitro studies with A. baumannii strains should also be supported by clinical trials.


2017 ◽  
Vol 62 (4) ◽  
pp. 301 ◽  
Author(s):  
I. MASTRANESTASIS (Ι. ΜΑΣΤΡΑΝΕΣΤΑΣΗΣ) ◽  
Ch. LIGDA (Χρ. ΛΙΓΔΑ) ◽  
K. THEODOROU (Κ. ΘΕΟΔΩΡΟΥ) ◽  
L. V. EKATERINIADOU (Λ.Β. ΑΙΚΑΤΕΡΙΝΙΑΔΟΥ)

Genetic structure and diversity of 120 animals from three Greek local breeds were investigated by Random Amplified Polymorphic DNA (RAPD) - PCR method. Sheep samples originated from the Lesvos, Chios and Karagouniko breeds were treated with 11 random primers to estimate their genetic diversity and phylogenetic relationships. Our analysis comprised two levels of the breeds' genetic structure: i) the genetic differentiation among the three breeds and ii) the genetic differentiation among the flocks within each breed. This combined approach gave two main findings: i) the study of genetic distances and identity revealed that the Karagouniko sheep breed is genetically distinct from Chios (GD=0.1979) and Lesvos (GD=0.1691) breeds, while ii) the Chios and Lesvos breeds are genetically similar (GI=0.9631); half of the flocks of Lesvos have a relatively closer relationship with those of Chios than with the other Lesvos flocks. This is the first study that reports the close genetic relationship between the Chios and Lesvos breeds and gives strong evidence to hypotheses about their related origin. Furthermore, the study of polymorphic loci revealed particular indicators located in Karagouniko breed, as definitional datum of genetic identity or as a fingerprint of breed. Therefore, RAPD-DNA methods can be an efficient tool for the determination of phylogenetic relationships and genetic identity among the bloodstock breed of sheep.


2017 ◽  
Vol 11 (07) ◽  
pp. 543-548
Author(s):  
Sibel Aydogan ◽  
Ziya Cibal Acikgoz ◽  
Aysegul Gozalan ◽  
Fisun Kirca ◽  
Tuba Muderris ◽  
...  

Introduction: The purpose of this study was to evaluate and compare two manual isolation and real-time (RT) polymerase chain reaction (PCR) kits (RTA RT-PCR with RTA isolation kit and Artus RG RT-PCR with QIAamp isolation kit) for molecular diagnosis of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Methodology: The study was conducted on 121 and 54 clinical samples for the detection of HBV DNA and HCV RNA, respectively, with an additional 8 HCV RNA external quality control samples. Results: Though a high correlation was observed between the two kits for the HBV DNA (r = 0.955, p = 0.001) and HCV RNA quantifications (r = 0.828, p = 0.001), discordant results were found in nine of the HBV DNA and in six of the HCV RNA samples. The mean difference between the two systems was found to be 0.4 log IU/mL in Quality Control for Molecular Diagnostics (QCMD) HCV RNA samples by Bland-Altman analysis. Conclusions: Although there was a high correlation between HBV DNA and HCV RNA tests according to the results of the study, the RTA system requires improvement for the determination of HCV RNA.


2020 ◽  
Vol 14 (3) ◽  
pp. 1723-1736
Author(s):  
Mahyar Porbaran ◽  
Reza Habibipour

Bacteria are capable of evolving high doses of the drug in various infections by forming biofilms. Perhaps, biofilm regulator genes have different frequencies in β-lactam producing non-fermentative Gram-negative Bacilli (NFGNB). In this study, we investigated the role of biofilm operons of Pseudomonas aeruginosa and Acinetobacter baumannii on the prevalence of different β-lactamase enzymes. One-hundred twenty (120) nosocomial NFGNB isolates were collected from different clinical samples of patients. PCR method was used for the amplification of resistance genes. Isolates were collected, including 50 isolates (41.66%) of P. aeruginosa and 70 isolates (58.33%) of A. baumannii. The distribution of ESBL, AmpC, KPC, and MBL β-lactamase enzymes in P. aeruginosa and A. baumannii isolates were 64%, 58%, 38%,44%, and 57.14%, 60%, 32.85%, 34.28%, respectively. The frequency of csuABC, pgaABC operon in A. baumannii were as follows: pgaA (45.71%), pgaB (32.85%), pgaC (42.85%), csuA (34.28%), csuB (32.85%), csuC (41.42%), and ompA (38.57%). Further, the prevalence of pslABC and pelABC operons in P. aeruginosa isolates were as follows: pslA (58%), pslB (58%), pslD (60%), pelA (64%), pelB (38%), pelC (44%), and algD (68%). This study revealed that the abundance of biofilm regulator genes in NFGNB strains is affected by different β-lactamase enzymes.


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