scholarly journals Antibiotic Resistance Profiling and Molecular Phylogeny of Biofilm Forming Bacteria From Clinical and Non-clinical Environment in Southern Part of Bangladesh

2019 ◽  
Vol 7 (2) ◽  
pp. 37-43 ◽  
Author(s):  
Zulkar Nain ◽  
Md. Ariful Islam ◽  
Mohammad Minnatul Karim

Background: Biofilm is a surface adhered extracellular polymer matrix produced by bacteria. The establishment of biofilms is considered as an important pathogenic trait in many chronic infections and antibiotic resistance. Objective: The present study was intended to evaluate biofilm forming potency and antibiotic resistance (AR) pattern in clinical and non-clinical bacterial isolates, and their phylogenetic characterization. Materials and Methods: A total of 82 bacterial isolates were obtained from clinical settings and animal farms from southern (Kushtia-Jhenaidah) region of Bangladesh. Biofilm forming potentials and AR profile were evaluated by standard biofilm assay and Kirby-Bauer disk diffusion method, respectively. Further, antibiotic exposure was assessed by multiple antibiotic resistance (MAR) value indexing. Furthermore, statistical methods were applied to estimate the relationship between AR and biofilm formation. Finally, selected isolates were characterized by morphological and biochemical tests, as well as 16S rRNA gene sequencing. Results: Clinical isolates showed higher biofilm formation (OD595=1.17±0.03) than non-clinical isolates (OD595=0.68±0.03). Among all, Pseudomonas isolates produced the highest amount of biofilms (OD595=2.08±0.02). The AR profiles fell within 46.67-86.67% and MAR index ranged from 0.47 to 0.87. Moreover, a significant positive correlation (P<0.05) was found between biofilm formation and AR. Eventually, heavy biofilm producers with ≥60% resistance profile were characterized and identified as Escherichia coli, Cronobacter sakazakii, Pseudomonas aeruginosa, Staphylococcus sciuri, and Staphylococcus aureus. Conclusion: In general, biofilm formation and MAR were highly correlated regardless of the source, type, and environment of the isolates. Therefore, a rigorous evaluation of both biofilm formation and AR is demanded to minimize AR and associated problems.

Author(s):  
Pakhshan A. Hassan ◽  
Adel K. Khider

Acinetobacter baumannii is an opportunistic pathogen that is reported as a major cause of nosocomial infections. The aim of this study was to investigate the biofilm formation by A. baumannii clinical and soil isolates, to display their susceptibility to 11 antibiotics and to study a possible relationship between formation of biofilm and multidrug resistance. During 8 months period, from June 2016 to January 2017, a total of 52 clinical and 22 soil isolates of A. baumannii were collected and identified through conventional phenotypic, chromo agar, biochemical tests, API 20E system, and confirmed genotypically by PCR for blaOXA-51-like gene. Antibiotic susceptibility of isolates was determined by standard disk diffusion method according to Clinical and Laboratory Standard Institute. The biofilm formation was studied using Congo red agar, test tube, and microtiter plate methods. The clinical isolates were 100% resistance to ciprofloxacin, ceftazidime, piperacillin, 96.15% to gentamicin, 96.15% to imipenem, 92.31% to meropenem, and 78.85% to amikacin. The soil A. baumannii isolates were 100% sensitive to imipenem, meropenem, and gentamicin, and 90.1% to ciprofloxacin. All A. baumannii isolates (clinical and soil) were susceptible to polymyxin B. The percentage of biofilm formation in Congo red agar, test tube, and microtiter plate assays was 10.81%, 63.51%, and 86.48%, respectively. More robust biofilm former population was mainly among non-MDR isolates. Isolates with a higher level of resistance tended to form weaker biofilms. The soil isolates exhibited less resistance to antibiotics than clinical isolates. However, the soil isolates produce stronger biofilms than clinical isolates.


2020 ◽  
Vol 8 (3) ◽  
pp. 101-106
Author(s):  
Hosein Akhlaghi ◽  
Seyed Hesamodin Emadi Chashmi ◽  
Ashkan Jebelli Javan

Background: Helicobacter pullorum predominantly colonizes the gut of apparently healthy chickens and the livers and intestinal contents of hens with enteritis and vibrionic hepatitis. Objective: The aim of this study was to assess the prevalence and antibiotic resistance of Helicobacter pullorum in broiler chickens, laying hens, and turkeys in Semnan province. Materials and Methods: A total of 300 samples were collected from 60 poultry farms in Semnan province, including 240 cecal samples from 48 broiler farms, 30 fecal samples from 6 laying hen farms, and 30 cecal samples from 6 turkey farms. Each sample was analyzed by conventional culture method and biochemical tests. The suspected colonies were subjected to polymerase chain reaction (PCR) using 16S rRNA gene. Antibiotic resistance of the confirmed colonies was determined using disk diffusion method. Results: Of 300 samples, 85 (28.3%) samples obtained from 36 (60%) poultry farms were positive for H. pullorum. Of these samples, 72 (30%) were from 30 (62.5%) broiler farms, 4 (13.3%) were from 2 (33.3%) laying hen farms, and 9 (30%) were from 4 (66.7%) turkey farms. Moreover, resistance to ciprofloxacin was observed in all of the H. pullorum isolates. Conclusion: This study demonstrated the moderate prevalence of H. pullorum in poultry in Semnan province for the first time, while the prevalence of this pathogen in laying hen and turkey has not been determined in Iran. In addition, this study could reveal the antibiotic resistance profile of H. pullorum as the first report in Iran. Therefore, more studies are needed to focus on the prevalence and antibiotic resistance of H. pullorum in poultry in other regions of Iran.


2020 ◽  
Vol 10 (1) ◽  
pp. 1-4
Author(s):  
Omor Ahmed Chowdhury ◽  
Md Raihan Ahmed ◽  
Md Raihan Dipu ◽  
Md Aftab Uddin

The use of earphones has increased in recent times throughout the world especially among the different level of students such as school, college or university who have a higher tendency of sharing these among them. Unlike airline headsets, headphones and stethoscope ear-pieces, ear phones are often shared by multiple users and can be a potential medium for transmission of pathogens, which can give rise to various ear related infections. The objective of this study was to detect the pathogenic bacteria from the ear-phones used by the students of Stamford University Bangladesh. A total of 16 ear-phone swabs were collected by sterile cotton swabs. The swabs were inoculated onto blood agar and incubated aerobically overnight at 37oC. Microscopic observation and standard biochemical tests were performed to confirm the identification of all the bacterial isolates. Six presumptively identified Staphylococcus spp. (38%) were tested against six different types of antibiotics following Kirby-Bauer disk diffusion method. Isolates were found to be 84% resistant against Cotrimoxazole and demonstrated 100% sensitivity to Vancomycin and Ciprorofloxacin. The findings of this study suggest the users to disinfect their respective ear phones and not to exchange them as they may act as a potential source to transfer pathogenic and antibiotic resistant bacteria among the ear phone users. Stamford Journal of Microbiology, Vol.10 (1) 2020: 1-4


2021 ◽  
Vol 14 (8) ◽  
Author(s):  
Seyed Ali Bazghandi ◽  
Mohsen Arzanlou ◽  
Hadi Peeridogaheh ◽  
Hamid Vaez ◽  
Amirhossein Sahebkar ◽  
...  

Background: Drug resistance and virulence genes are two key factors for the colonization of Pseudomonas aeruginosa in settings with high antibiotic pressure, such as hospitals, and the development of hospital-acquired infections. Objectives: The objective of this study was to investigate the prevalence of drug resistance and virulence gene profiles in clinical isolates of P. aeruginosa in Ardabil, Iran. Methods: A total of 84 P. aeruginosa isolates were collected from clinical specimens of Ardabil hospitals and confirmed using laboratory standard tests. The disk diffusion method was used for antibiotic susceptibility testing and polymerase chain reaction (PCR) for the identification of P. aeruginosa virulence genes. Results: The highest and the lowest antibiotic resistance rates of P. aeruginosa strains were against ticarcillin-clavulanate (94%) and doripenem (33.3%), respectively. In addition, the frequency of multidrug-resistant (MDR) P. aeruginosa was 55.9%. The prevalence of virulence factor genes was as follows: algD 84.5%, lasB 86.9%, plcH 86.9%, plcN 86.9%, exoU 56%, exoS 51.2%, toxA 81%, nan1 13.1%, and pilB 33.3%. A significant association was observed between resistance to some antibiotics and the prevalence of virulence genes in P. aeruginosa. Conclusions: Our results revealed a high prevalence of antibiotic resistance, especially MDR, and virulence-associated genes in clinical isolates of P. aeruginosa in Ardabil hospitals. Owing to the low resistance rates against doripenem, gentamicin, and tobramycin, these antibiotics are recommended for the treatment of infections caused by highly resistant and virulent P. aeruginosa strains.


2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Kiana Karimi ◽  
Omid Zarei ◽  
Parinaz Sedighi ◽  
Mohammad Taheri ◽  
Amin Doosti-Irani ◽  
...  

Aim. Klebsiella pneumoniae (K. pneumoniae) is an encapsulated Gram-negative bacterium that can lead to 14–20% of nosocomial infections. The ability of biofilm formation in this bacterium decreases the host immune response and antibiotic efficacy. This may impose a huge impact on patients and healthcare settings. This study aimed to evaluate the antibiotic resistance pattern and biofilm formation in K. pneumoniae strains isolated from two major Hamadan hospitals, west of Iran. Methods. A total of 83 K. pneumoniae strains were isolated from clinical samples of patients in different wards of Hamadan hospitals from September 2018 to March 2019. Determination of antimicrobial susceptibility was performed using the disk diffusion method. Biofilm formation was evaluated by the crystal violet method. Data were analyzed by the SPSS software and chi-square test. Results. The results showed that clinical samples included 18 urinary tract samples (22%), 6 wound samples (7%), 6 blood samples (7%), 17 tracheal tube aspiration samples (20%), 32 throat cultures (38%), 2 sputum samples (2.5%), and 2 abscess drain cultures (2.5%). High-level resistance to cefotaxime was detected in 92%, and all of isolates were susceptible to colistin. Biofilm formation was seen in 62 (75%) isolates. Strong biofilm formation was observed in 17 (20%) strains. A significant correlation was seen between biofilm formation and antibiotic resistance ( P value <0.05). Conclusion. Our findings emphasize the need for proper diagnosis, control, and treatment of infections caused by K. pneumoniae especially in respiratory tract infections due to the strong biofilm formation and high antibiotic resistance in these strains.


Diseases ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 15
Author(s):  
Ram Shankar Prasad Sah ◽  
Binod Dhungel ◽  
Binod Kumar Yadav ◽  
Nabaraj Adhikari ◽  
Upendra Thapa Shrestha ◽  
...  

Background: Antimicrobial resistance (AMR) among Gram-negative pathogens, predominantly ESBL-producing clinical isolates, are increasing worldwide. The main aim of this study was to determine the prevalence of ESBL-producing clinical isolates, their antibiogram, and the frequency of ESBL genes (blaTEM and blaCTX-M) in the clinical samples from patients. Methods: A total of 1065 clinical specimens from patients suspected of heart infections were collected between February and August 2019. Bacterial isolates were identified on colony morphology and biochemical properties. Thus, obtained clinical isolates were screened for antimicrobial susceptibility testing (AST) using modified Kirby–Bauer disk diffusion method, while ESBL producers were identified by using a combination disk diffusion method. ESBL positive isolates were further assessed using conventional polymerase chain reaction (PCR) to detect the ESBL genes blaTEM and blaCTX-M. Results: Out of 1065 clinical specimens, 17.8% (190/1065) showed bacterial growth. Among 190 bacterial isolates, 57.4% (109/190) were Gram-negative bacteria. Among 109 Gram-negative bacteria, 40.3% (44/109) were E. coli, and 30.2% (33/109) were K. pneumoniae. In AST, 57.7% (n = 63) Gram-negative bacterial isolates were resistant to ampicillin and 47.7% (n = 52) were resistant to nalidixic acid. Over half of the isolates (51.3%; 56/109) were multidrug resistant (MDR). Of 44 E. coli, 27.3% (12/44) were ESBL producers. Among ESBL producer E. coli isolates, 58.4% (7/12) tested positive for the blaCTX-M gene and 41.6% (5/12) tested positive for the blaTEM gene. Conclusion: Half of the Gram-negative bacteria in our study were MDR. Routine identification of an infectious agent followed by AST is critical to optimize the treatment and prevent antimicrobial resistance.


Author(s):  
H. F. Massawe ◽  
R. H. Mdegela ◽  
L. R. Kurwijila

Aim: The study determined and evaluated the prevalence and antibiotic resistance of Staphylococcus aureus isolated from milk collected along the milk value chain from farm herds, milk collection center, and milk shops in Mbeya rural and Mbozi districts, Tanzania. Materials and Methods: A total of 150 milk samples were collected; 96 from farmers' herds, 18 from milk collection centers, and 36 from milk shops. The samples were cultured in Mannitol salt agar for pathogen isolation and biochemical tests performed for confirmation of S. aureus. Kirby-Bauer disk diffusion method was employed for antibiotic resistance testing. Results: One hundred and forty samples yielded Staphylococcus species; these were from farmer's herd (92), milk collection center (18), and milk shops (30), respectively. Biochemical tests showed that 21 (15%) were positive for S. aureus. The corresponding prevalence rates from the value chain nodes were 14.1%, 16.7%, and 16.7%, respectively. Resistance to penicillin was frequently observed (57.1%) and vancomycin was sensitive to all S. aureus isolates tested. Resistance along the sampling points showed a significant positive correlation (r=0.82, p<0.0001; r=0.65, p<0.003; and r=0.61, p<0.01) between farmers, milk collection points, and milk shops, respectively. More than half (57.1%) of the isolates exhibited resistance to three or more of the antibiotics used in this study. S. aureus isolates were shown to have a multiple antimicrobial resistance patterns, particularly with respect to penicillin, ampicillin, erythromycin, and tetracycline. Conclusion: The level of staphylococcal isolates and the antibiotic resistance of S. aureus found in this study is an indication of subclinical mastitis, poor hygiene, and inappropriate use of antibiotics; therefore, education of farmers on subclinical mastitis control and proper use of antibiotics would be of benefits in these areas.


2019 ◽  
Vol 34 (2) ◽  
pp. 61-66
Author(s):  
Sunjukta Ahsan ◽  
Mayen Uddin ◽  
Juthika Mandal ◽  
Marufa Zerin Akhter

Antibiotic resistant E. coli are prevalent in Bangladesh. The indiscriminate use of antimicrobials and ready availability of over the counter drugs are responsible for this. This study was conducted to investigate the susceptibility of clinical Escherichia coli to the antibiotics Imipenem, Ceftriaxone, Ceftazidime and Azithromycin. Kirby-Bauer disk diffusion method was used to determine sensitivity to antimicrobials. Agar based assay was employed for the detection of efflux pumps. PCR was used amplify antibiotic resistance genes.All isolates were resistant to Ceftriaxone whereas most were sensitive to Imipenem. The MICs of Ceftazidime and Azithromycin ranged between 128 μg/ml and 256 μg/ml. The prevalence of ²-lactamase producers was 57.89 % with 36.84 % of the isolates exhibiting ESBL activity. No specific correlation could be found between plasmid sizes and antibiotic resistance patterns. Efflux pump was found to be involved in Azithromycin resistance in 63.15% of the isolates. The gene for phosphotransferase, mph(A) was the most common among the macrolide modifying genes, being present in 73.68% (14/19) of the isolates followed by both erm(A) anderm(C) esterases each present in 10.53% (2/19) isolates. This study concluded that clinical isolates of E. coli in Bangladesh could be resistant to multiple classes of antibiotics through different mechanisms of resistance. Bangladesh J Microbiol, Volume 34 Number 2 December 2017, pp 61-66


Author(s):  
Salwa Nurhasanah ◽  
Edy Fachrial ◽  
Nyoman Ehrich Lister

Aims: This study aims to isolate and identify the indigenous bacteria of almonds fermentation. Methods: Characterization of the indigeneous bacteria are using gram staining, biochemical tests, 16SrRNA gene sequencing, and the antimicrobial activity against Escherichia coli bacteria. Results: Approximately 28 x 106 CFU / mL bacteria were obtained from almonds fermentations with 14 isolates from enrichment results. Three randomly selected isolates were gram-positive rod-shaped with a negative catalase and positive fermentation test. However, one isolate showed positive results on the motility test. The antimicrobial test results from the three randomly selected isolates using the disk diffusion method obtained inhibition zones of 7 mm, 6.7 mm, and 7 mm, respectively. Therefore, by using 16S rRNA gene sequencing, three different microorganisms were found, namely Bacillus subtilis strain IAM 12118, Bacillus Piscis strain 16MFT21, and Bacillus licheniformis strain BaDB27. Conclusion: It was found that Bacillus subtilis strain IAM 12118, Bacillus Piscis strain 16MFT21, and Bacillus licheniformis strain BaDB27 in almonds fermentation and also can be used as probiotic bacteria.


2020 ◽  
Author(s):  
Shadi Shadkam ◽  
Hamid Reza Goli ◽  
Bahman Mirzaei ◽  
Mehrdad Gholami ◽  
Mohammad Ahanjan

Abstract BackgroundKlebsiella pneumoniae (K. pneumoniae) is a common cause of nosocomial infections. Antibiotic resistance and ability to form biofilm, as two key virulence factors of K. pneumoniae, involved in persistent of the infections. The purpose of this study is to investigate the correlation between antimicrobial resistance and biofilm formation capability among K. pneumoniae strains isolated from hospitalized patients in Iran.MethodsOver a 10-month period, a total of 100 non-duplicate K. pneumoniae strains were collected. Antibiotic susceptibility test was determined by Kirby-Bauer disk diffusion method according to CLSI. Biofilm formation was assessed by tissue culture plate method. Finally, polymerase chain reaction was conducted to detect four families of carbapenemase: blaIMP, blaVIM, blaNDM, blaOXA-48, biofilm formation associated genes; treC, wza, luxS and K. pneumoniae confirming gene; rpoB.ResultsMost of the isolates were resistant to co-trimoxazole (52%), cefotaxime (51%), cefepime (43%), and ceftriaxone (43%). Among all the 100 isolates, 67 were multidrug-resistant (MDR), and 11 were extensively drug-resistant (XDR). The prevalence of the blaVIM, blaIMP, blaNDM, and blaOXA-48 genes were 7%, 11%, 5%, and 28%, respectively. Among these isolates, 25% formed fully established biofilms, 19% were categorized as moderately biofilm-producing, 31% formed weak biofilms, and 25% were non-biofilm-producers. Molecular distribution of biofilm formation genes revealed that 98%, 96%, and 34% of the isolates carried luxS, treC, and wza genes, respectively. ConclusionThe rise of antibiotic resistance among biofilm-producer strains, demonstrating a serious alarm about limited treatment options in hospital setting. Also, fundamental actions and introduction of novel strategies for controlling of K. pneumoniae biofilm-related infections is essential.


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