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Proteus strains isolated from tap water are assessed for their twitching motility and biofilm formation on urethral catheter strips of two selected brands, Protos and Huaian, since these strains are the common cause of catheter associated urinary tract infection (CAUTI). Three types of movement identified in the genus Proteus are swimming, swarming and twitching.
An important virulence factor in Proteus is biofilm formation: an attached structure with microbial cells and populations embedded in a polysaccharide layer.
A biofilm is an aggregate of microorganisms immersed in a polysaccharide matrix adherent to each other and to the airway mucosa. The adhesion of bacteria to the catheter depends on many factors, for example surface charge, hydrophobicity or hydrophilicity of the catheter and bacterial cell, and specific genes for adhesion [32].
Although bacteria of the genus Proteus can move by means of twitching motility, and this virulence factor plays an important role in biofilm formation in Pseudomonas [37], it has not previously been explored in detail in Proteus. AcknowledgementsThe authors wish to thank the Department of Pharmacy, University of Rajshahi, Bangladesh for providing laboratory facilities to carry out the entire experiments. Authors’ Contribution AC designed and guided the study, and drafted the manuscript; AW worked out the experimental part.
Hello readers, in this article you can get information about Antimicrobial Resistance Patterns Of Proteus Isolates From . Prevalence of antimicrobial resistance among urinary tract pathogens isolated in canada from 2007 to 2009: results from the canward surveillance study. Intjcurrmicrobiolappsci (2013) 2(8): 330337 334 result for citrate utilization, vogesproskauer and urease tests (figure 2) klebsiella grown on chromagar. Download Antimicrobial Resistance Patterns Of Proteus Isolates From 19 levofloxacin for the treatment review of urosepsis introduction urinary tract infections (utis) can manifest themselves in a wide range of clinical forms,.
Download Antimicrobial Resistance Patterns Of Proteus Isolates From Resistance antibiotics + §. Cefobid ® (sterile cefoperazone) for intravenous or intramuscular use to reduce the development of drug-resistant bacteria and maintain the effectiveness of. Tables table 1 resistance of proteus species to selected antibiotics (3708 p mirabilis isolates and p vulgaris isolates were included from references 1, 2, 14, 21. Background the antimicrobials misuse in clinical medicine has led to an increase of the microbial resistance and the consequent spread of bacterial. Review article mechanisms of disease franklin h epstein, md, editor new mechanisms of bacterial resistance to antimicrobial agents george a jacoby, md, and. Since their discovery, antimicrobial drugs have proved remarkably effective for the control of bacterial infections however, it was soon evident that bacterial.
Research from jama — antibiotic resistance among gramnegative bacilli in us intensive care units — implications for fluoroquinolone use — contextprevious. Above you can read article and ebook that discuss about Antimicrobial Resistance Patterns Of Proteus Isolates From . Photos by user 'listentoreason', with default photo number, size, and sorting options (can change). Better way to display more photos from your contacts - you must be logged in for this to work.
Flickr requires that I inform you that Flickr Hive Mind uses the Flickr API, but is neither endorsed nor certified by them.
If you find your login does not work, perhaps your browser has disabled the cookies that I use to track your session. Any revenue generated through advertisement on this site is used only to cover the cost of keeping the site online, beyond that Fiveprime donates to charitable organizations (also, there are no ads on User pages). Urinary Tract Infection (UTI) defines a condition in which the urinary tract is infected with a pathogen causing inflammation which is a common, distressing and occasionally life threatening condition.
RESULTS AND DISCUSSIONA total of 44 mid stream urine samples from diabetic patients were collected between the age range of 30-80 years which consists of 30 male (68%) and 14 female (32%) and was found that 68% of the patients were having asymptomatic bacteriuria.When the data were classified into different age and gender groups, it appeared that the cases of Urinary Tract Infections (UTIs) were more in men than women and a total of 38 strains were isolated. It was observed that oxytetracycline has failed to control the growth of all isolates throughout the study. Neither the strains of high swarming motility nor of high antimicrobial resistance enabled to twitch over the catheter strips.
Urease test and carbohydrate fermentation, A: Urea hydrolysis by Proteus isolates turned the color of Christensen’s broth medium from reddish to pink. The biofilm enables better adaptation of bacterial cells to the conditions of the external environment and facilitates survival at adverse external environment [11]. Biofilm-forming bacteria are protected from phagocytosis, antimicrobial agents, and the ciliary action of the airway epithelial cells.
Preparation of Catheter StripBased on the market price two catheter bags from two different companies, Protos, USA (Highest price) and Huaian, China (lowest price), were purchased from local market and were aseptically cut into strips (1 ? 8 cm2).
Test for Twitching Motility over Catheter StripThe catheter strips (1 ? 8 cm2) were cut aseptically and placed on the LB agar plate across the diameter line. Assay of Biofilm FormationBiofilm formation was assayed qualitatively based on the ability of bacterial cells to adhere on the surface of polyvinylchloride plastic (PVC), a material used to make catheter lines (Urine drainage bag and tubing) [24]. Kinetics of Swarming MotilityEach isolate was tested for its swarming growth which is connected with the cell surface components of bacteria and is an important virulence factor in cases of UTIs especially for the genus Proteus. Twitching Motility over Catheter StripNext we aimed to investigate whether the isolates of enhanced swarming motility are able to twitch over the catheter bridge since the ascending CAUTIs are very common in patients undergoing catheterization. Biofilm Formation and QuantificationUrine drainage bag from two different brand chosen on the basis of their market price were used for biofilm study. Therefore, the risk of infection depends on the duration of catheterization and catheter management.
Cephalosporin Resistance and Amoxicillin SusceptibilityAntibiotics kill the pathogenic bacteria in a way of their different mode of actions [33] and are accordingly classified in different groups. DiscussionProteus are dimorphic bacteria, which in liquid media are motile, peritrichously flagellated short rods (1-2 µm in length with 6-10 flagella).
This virulence factor was more expressed in CAUTI isolates [38], but none of the isolates moved over the whole plate [15]. Therefore, the high incidence of multi-drug resistant might be due to indiscriminate use of antibiotics, which may eventually supersede the drug resistant microorganisms from antibiotic saturated environment. ConclusionThe rate of swarming growth of Proteus on LB-agar plate was decreasing with increasing incubation time. To also remove yourself from searches for specific user names, you will need to set your Flickr profile to be hidden from searches. Flickr Hive Mind is currently consuming about 13.6 terabytes of network bandwidth per year (not including the photos themselves)!
These samples were processed to monitor urinary tract infection and antibiotic sensitivity patterns of pathogenic bacteria. By using Bergey's Manual of Determinative Bacteriology, the isolates were biochemically characterized and identified. The other antibiotics erythromycin, rifampicin and streptomycin have exhibited moderate effect on the growth of isolates. All of high antibiotic resistant strains were found to form biofilm on PVC strip, few strains on Huaian catheter strip and one strain (Pv) on both catheter strips.

Catheter tubes filled with BHI medium were incubated at 37°C with (11, 911, 912) and without (C) inoculums.
Biofilm formed on PVC strip (gradient filled bar) by isolates 11(Pv), 911(Pm), 912(Pm), 661(Pp), 662(Ph) and 667(Pp) were significantly high compared to that on Protos strip (open bar) and Huaian strip (filled bar).
Six to ten flagella are present on the bacterial surface required for their swimming motility. Furthermore, microorganisms gathered in a biofilm significantly develop different genetic properties compared to planktonic species. Bacterial Strain, Culture Medium, Medical DeviceEleven Proteus strains belonging four species, Proteus mirabilis (Pm), P. The cells were Gram-negative, rod in shape, singly arranged non-diploid or clump predicting the family Enterobacteriaceae. The strip was submerged with 10ml of BHI medium in a test tube and used for biofilm formation and quantification assay.
After placement the strip was gently pressed to make the contiguity of horizontal plane and uniform adherence of the strip onto the surface of agar medium. The catheter tube filled with BHI medium was inoculated with Proteus isolates followed by the overnight incubation at room temperature with mild shaking.
Isolation and Identification of Proteus SpeciesTotal 100 samples of municipal supplied water were subjected to a systematic analysis for the isolation and identification of Proteus species. In practice, 10–50% of patients undergoing short-term catheterization (up to 1 week) develop infection [29]. A qualitative biofilm formation assay was carried out using catheter tube with highly resistant isolates of Pm (911, 912) and Pv (11) (Figure 6) since antibiotic resistance was found more often in strong biofilm formers than in weak biofilm formers. Biofilm forming abilities of tested strains on catheter strips from two brands, Protos and Huaian, were compared to that on PVC strip (Figure 7). Twelve antibiotics belonging four modes of actions (Protein synthesis inhibitor, Folate synthesis inhibitor, DNA synthesis inhibitor, Cell wall synthesis inhibitor) were tested to evaluate their current status of susceptibility to Proteus isolates. In our study no Proteus isolates from tap water were able to twitch over the catheter bridge, it is probably due to their less pathogenic potentialities than CAUTI isolates. Urine drainage bag from Protos brand was relatively safer than that of Huaian brand to prevent CAUTI.
But in this post i will explain Since their discovery, antimicrobial drugs have proved remarkably effective for the control of bacterial infections.
To ensure appropriate therapy, current knowledge of the organisms that cause UTI and their antibiotic susceptibility is mandatory.
All the bacterial isolates were preserved on nutrient agar slants at 4°C and subcultured periodically. It was found that nalidixic acid, chloramphenicol and kanamycin had played a significant role in controlling the growth of urinary isolates. Based on biofilm quantification study, the urethral catheter from Protos brand was relatively safer than that of Huaian brand to prevent CAUTI.
A- 11 (Pv), B- 911 (Pm), C- 912 (Pm), D- 661 (Pp), E- 662 (Ph), F- 663 (Pp), G- 664 (Pp), H- 665 (Pp), I- 666 (Pp), J- 667 (Pp), K- 668 (Pp). Swarming motility requires an increased number of flagella and this type of motility is used for movement on solid surfaces. Antibiotic resistance is found more often in strong biofilm formers than in weak biofilm formers. Donlan 2001 isolated Proteus mirabilis from indwelling medical device urinary catheter, one of the biofilm-associated microorganisms among others [20]. By sequential biochemical analyses the bacterial isolates were identified as Proteus species [21]. The medium was discarded and the tube was repeatedly rinsed with water, PBS pH 7.2 and water. Briefly, the black centered bacterial colonies formed on sample inoculated KIA culture plate at 37°C were picked up and analyzed for the presence of the genus Proteus. Here the isolates' competitive swarming motilities up to 44 hours on LB agar medium were kinetically assessed. Patients undergoing long-term catheterization (> 1 month), even with high standards of care, will inevitably develop bacteriuria. Based on the spectrophotometric data (OD595nm) isolates 11(Pv), 911(Pm), 912(Pm), 661(Pp), 662(Ph) and 667(Pp) were found to form biofilm on PVC strip and isolates 912(Pm), 662(Ph) and 667(Pp) positively formed biofilm on Huaian strip since the OD595nm values were greater than 0.24 (Figure 7). Strong antimicrobial resistance was found in folate synthesis inhibitor, cotrimoxazole, and in cell wall synthesis inhibitor, cephalosporins (Table 2).
However, when transferred onto solid media these short rods change into elongated (20–80 µm in length), hyperflagellated, multinucleated, nonseptated swarmer cells. Now-a-days, the extensive use of cephalosporins in Bangladesh offered the pathogens to be resistant to them but the drugs of protein and DNA synthesis inhibitors (Table 2) were effective enough for the infection control and management. This study focused on the frequency of uropathogens and their antibiotic susceptibility in different gender of diabetic patients. All urine specimens were obtained aseptically in well labeled, screw capped universal containers and transported promptly to the laboratory.
All the isolates have failed to grow in the presence of trimethoprim and thus proved which can be used to treat urinary tract infections effectively. Moreover, Proteus isolates were strong resistant to cotrimoxazole (folate synthesis inhibitor) and cephalosporins (cell wall synthesis inhibitor).
It causes urine stone due to presence of urease and also causes wound infection, septicemia, and pneumonia mostly in hospitalized patients. Swarming motility is the typical mode of movement of Proteus grown on solid agar, the Rauss phenomenon. Five basic sugars- glucose, lactose, mannitol, maltose and sucrose were used in carbohydrate fermentation test for the genus and species identification. After drying under laminar air flow the adherent cells on the inner surface of the tube was stained with 1% crystal violate (CV) solution that stained the cells but not the PVC. The solution was then discarded and the strips were rinsed repeatedly with water, PBS, pH 7.2 and water. Eleven Proteus isolates of Pm, Pv, Ph and Pp designated as 11(Pv), 661(Pp), 662(Ph), 663(Pp), 664(Pp), 665(Pp), 666(Pp), 667(Pp), 668(Pp), 911(Pm), and 912(Pm) were identified. Four isolates, 911(Pm), 912(Pm), 662(Ph) and 665(Pp), showed enhanced swarming motility than others (Figure 4). Swarming motility has correlated very well with the ability to swarm over the various catheters [15].
The intensity of 1% CV staining revealed that the bacterial cells of these tested strains were attached on the inner surface of the catheter tube compared to that of negative control (Figure 6). Among the ?-lactam antibiotics underlying the cell wall synthesis inhibitor, cephalosporin resistance were strong whereas amoxicillin susceptibility underlying the same group was pronounced in all tested isolates. The multinucleated cells migrate out from the inoculation site as long as the population of swarmer cells is reduced on solid surfaces [34].
Apparently biofilm quantification was directly proportional to the spectrum of resistance and swarming motility of isolates.

The amine group of amoxicillin provides increased polarity to expand its spectrum to include Gram negative bacteria. Antibiotics underlying the protein and DNA synthesis inhibitors were satisfactorily effective and amoxicillin was the best of choice for the inhibition of Proteus isolates. Biochemical characterization of uropathogens revealed the prevalence of gram negative organisms and E. Prevalence, Microbiologic Profile of Urinary Tract Infection and its Treatment with Trimethoprim in Diabetic Patients. In any case when it was not possible to reach the laboratory in time, the specimens were kept in refrigerator not more than 6 h. It is stated that UTI is predominantly a disease of the females due to a short urethra and proximity to the anal opening and our findings revealed that there was a male preponderance for this infection. The antibiotic sensitivity pattern showed that most isolates were sensitive to trimethoprim while they are resistant to oxytetracycline which indicates that increased consumption of a particular antibiotic can be a pathway to its resistance.CONCLUSIONThe results of antibiotic susceptibility pattern on the casual agents in this study revealed that most of the isolates were resistant to oxytetracycline which is relatively cheaper and more frequently prescribed.
Underlying the ?-lactam group, strong cephalosporin resistance and amoxicillin susceptibility in isolates indicated that the isolates were non ?-lactamase producer.
About 50% patients requiring an indwelling urethral catheter for 5 days or longer will develop bacteriuria or candiduria [Care of the urinary catheter: Urinary tract infections, detection, prevention and management, 5th Ed.
2012 showed that this trend in Proteus bacteria isolated from urinary catheter is generally strongest for co-trimoxazole and ciprofloxacin resistance [15]. The tubes were visualized to check the intensity of violet color and the rough scores were noted compared to that of control.
After drying, staining for adherent biofilm was performed using 1% CV which was then discarded and the strip with adherent cells were rinsed three times with PBS, pH 7.2 to remove weakly adhered cells before drying for 24 h at room temperature. Two representative biochemical tests for the identification of Proteus species are shown in Figure 2 and Figure 3. To date, Pm is thought to be a strong swarmer cell among Proteus species and here two isolates, 911 and 912, of Pm exhibited faster swarming (Figure 4B,C) than other species resembling with other reports [28].
Therefore, the four isolates were selected based on their high swarming motility (912, 665) and high antimicrobial resistance (11, 911) to test for their abilities to twitch over the catheter strip (Figure 5). The result indicated that the tested strains enabled to form biofilm on the inner surface of catherter tube. However, the urine drainage bag from Protos brand was relatively safer than that of Huaian brand.
The catheter bag from Protos brand showed better properties than that of Huaian brand regarding the prevention of biofilm formation of Proteus isolates. In addition, the phenolic acid present in amoxicillin is an ionizable group, which prevents zwitterions formation. The resistance rates among common uropathogens continue to evolve and appear to be increasing too many commonly used antimicrobial agents such as rifampicin, erythromycin and streptomycin.
Tested antibiotics underlying the protein and DNA synthesis inhibitors were satisfactorily effective for the inhibition of Proteus isolates. Therefore, the purpose of the present study was to assess (1) the biofilm forming abilities of Proteus isolates in relation to their drug resistances, (2) the kinetic study of catheter-associated swarming and twitching motilities, and (3) the cause of amoxicillin potentiality in multi-drug resistant Proteus inhibition. All chemicals were purchased from Sigma-Aldrich, Difco, Thermo Scientific, Oxoid unless it is mentioned otherwise. All isolates turned the red color of McConkey agar medium to colorless (Figure 2-I) and were urease positive in urea hydrolyzing reaction (Figure 3A). However, in this study we found an isolate of Ph (662) which showed strong swarming motility comparable to Pm (Figure 6 B,C,E).
We then attempted to quantify the biofilm formed on catheter strip using spectrophotometric method at OD595nm. Currently our in vitro data make evidence that the antibiotics underlying the protein and DNA synthesis inhibitors are most effective drugs for the infection caused by Proteus. Flagellin, a surfactant secreted by the flagellum acts as lubricant to facilitate this movement. Here obtained results suggest that low cost catheter bags are relatively unsafe to prevent CAUTI in ascending route. Among the antibiotics tested, trimethoprim was found to be effective for empirical treatment of UTI and has covered the majority of urinary pathogens followed by nalidixic acid, chloramphenicol and kanamycin. Continued surveillance of resistance rates among uropathogens is needed to ensure appropriate recommendations for the treatment of the infections.
Brain-Heart infusion (BHI) medium was purchased from Hi-media, India and MacConkey agar medium from Accumedia, Spectrum Chemicals and Laboratory Products, USA. The intensity of coloration was determined at 595 nm using UV-spectrophotometer (UVmini-1240, Shimadzu, Japan).
The kinetics of swarming motility was studied for these top scorer isolates (Table 1) where we found that the rate of swarming was decreasing with increasing incubation time. After 24 h incubation at 37?C, the ability of isolates to twitch over the tested catheter strip was assessed. Although both ampicillin and amoxicillin are the broad spectrum semi-synthetic antibiotics of group penicillin, ampicillin was less effective than amoxicillin. Only streptomycin and amoxicillin were most effective antibiotics among tested others to inhibit the Proteus strains.
Most of the isolates were resistant to oxytetracycline which is more frequently prescribed and indicates that increased consumption of a particular antibiotic can be a pathway to its resistance by the uropathogens. The obtained result should connect with their chemical structures since chemically amoxicillin is more polar compound than ampicillin. In this study, the kinetic analysis of swarming motility revealed that the rate of swarming is decreasing with increasing incubation time.
Conclusively, resistance rates among common uropathogens continue to evolve and appear to be increasing too many commonly used antimicrobial agents and a continued surveillance of resistance rates among uropathogens is needed to ensure appropriate recommendations for the treatment of the urinary tract infections. Ltd, Houston, Texus, USA, (2) National Urine Drainage Bag 2000 ml, Huaian Angel Medical Instruments Co. Acid production by the isolates was assessed with the color change from red to yellow in carbohydrate fermentation tests (Figure 3B, C).
The isolates were found to swarm on the inoculated half of the agar plates but did not cross the catheter bridge (Figure 5). The radical attached to ?-lactam ring may contain a polar group that broadens the spectrum of activity by allowing more drugs to enter Gram negative bacterial cell wall via porins. Recently O’Loughlin et al 2013 showed that the human pathogen Pseudomonas aeruginosa uses quorum sensing to control virulence and biofilm formation [36]. We, therefore, speculated that after a certain period of migration the isolates use quorum sensing to express new pathogenic factors like biofilm. 2012 found the clinical Proteus isolates which enabled to swarm over the catheter bridge but in this study Proteus isolates from tap water were unable to form a bridge over the catheter strips.

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