Blood Agar Medium Research Articles

Halophilic Bacteria Producing Protease from Salted Fish in Ponrang, Luwu Regency, South Sulawesi

The need for protease enzymes for medical and industrial purposes. The need for proteases in the world reaches 65% of the total sales of enzymes, and in Indonesia can reach 2.500 tons every year, and 99% percent of the enzyme needs are still imported from abroad. Salted fish is one of the foods that contain a lot of protein, which is about 42% in 100 g of salted fish. It allows the presence of proteolytic bacteria that have halophilic properties in salted fish. This study aims to explore the presence of proteolytic bacteria with halophilic properties in salted fish. Proteolytic isolates were isolated using SMA media from salted fish from Ponrang District, Luwu Regency, South Sulawesi. A qualitative test was carried out by measuring the clear zone formed in Skim Milk Agar (SMA). From the isolation process, 51 isolates were obtained. However, after the screening, only 25 pure isolates were found that formed a clear zone, followed by a quantitative test to see which isolates had stable activity at incubation times of 24, 48, and 72 hours using Tryptic Soy Broth (TSB) media. The results obtained four superior isolates, P1A1K, P2B2PS, P3C6PS, and P3C6P, then continued to quantify halophilic bacteria properties by administering NaCl with 15% and 20% concentration into TSB media for 24 and 48 hours incubation. Two isolates with the highest protease activity were P3C6P isolates with the activity of 43.23 ± 7.11 U.mL-1 at 15% salt concentration and 42.83 ± 3.04 U.mL-1 at 20% salt concentration and P2B2PS isolates of 38.05 ± 4.05 U.mL-1 at 15% salt concentration and 38.15 ± 1.47 U.mL-1 at 20% salt concentration. The two isolates were then tested for pathogenicity on blood agar media. It was found that only P3C6P isolates did not have pathogenic properties, so P3C6P isolates were continued with catalase, oxidase, and gram staining tests, which showed negative catalase and oxidase results and were gram-positive, followed by identification based on sequences. 16S rDNA and phylogenetic tree construction where isolate P3C6P was identified as Bacillus cereus with a 100% similarity level to the WHX1 strain. Keywords: Enzyme, Protease, Proteolytic, Salted Fish, 16s-rDNA.

Correlation of extracellular enzymes activity of Candida glabrata clinical isolates with in vivo pathogenicity in Galleria mellonella larvae

Objective: To explore the relationship between extracellular enzymes activity and virulence of Candida glabrata clinical isolates based on the infection model of Galleria mellonella larvae. Methods: Using experimental research methods, 71 strains of non-repetitive Candida glabrata were collected from Qinghai Provincial People's Hospital from June 2021 to January 2022. Bovine serum protein agar medium, egg yolk agar medium, sheep blood agar medium, Tween-80 agar medium and triglyceride agar medium were used to detect the aspartyl protease activity, phospholipase activity, hemolysis activity, esterase activity and lipase activity of Candida glabrata. Median lethal concentration (LC50) was calculated by using 1.25×108 CFU/ml,2.50×108 CFU/ml,3.75×108 CFU/ml,5.00×108 CFU/ml suspension of Candida glabrata ATCC2001 to infect Galleria mellonella larvae. Histopathological and etiological analysis was performed to determine whether the infection model was successfully established. The clinical isolates of Candida glabrata were configured to infect Galleria mellonella larvae with LC50 concentration to detect the pathogenicity of Galleria mellonella larvae.Spearman test or Pearson test were used to analyze the correlation between the extracellular enzyme activity of Candida glabrata clinical isolates and the pathogenicity of Galleria mellonella larvae. Results: 71 strains of Candida glabrata isolated clinically were detected to have low hemolytic activity after 2 days of culture. Aspartyl protease was detected after 4 days of culture, among which 7 strains (9.86%), 19 strains (26.76%) and 45 strains (63.38%) showed low, medium and high aspartyl protease activity. After 7 days of culture, 71 strains did not detect phospholipase, esterase and lipase activities. Candida glabrata on Galleria mellonella larvae of LC50=2.5×108 CFU/ml Fungal spore were found in the intestinal tissue pathological section of Galleria mellonella larvae in the experimental group, and Candida glabrata was identified by the microbial Mass Spectrometry after culture, while no fungi were found in the pathological section and culture of the control group. Spearman test shows that, there was a linear positive correlation between aspartyl protease activity and the survival rate of Galleria mellonella larvae (r = 0.73, P<0.01), the difference was statistically significant.Pearson test shows that, there was no significant linear relationship between hemolytic activity and survival rate of Galleria mellonella larvae (r = 0.16, P = 0.34), the difference was not statistically significant. Conclusion: The clinical isolates of Candida glabrata in this study had aspartyl protease activity and low hemolytic activity, but no phospholipase, esterase and lipase activity. The activity of aspartyl aspartyl protease of Candida glabrata was positively correlated with the pathogenicity of Galleria mellonella larvae.

Optimization and Chemical Characterization of Biosurfactant Produced from a Novel Pseudomonas guguanensis Strain Iraqi ZG.K.M.

Microbial surfactants are widely used in medical, pharmaceutical, agricultural, industrial, food, and cosmetics applications. In the present study, 85 indigenous bacteria were isolated from petroleum-contaminated soils of the Al Dourah refinery, electric power station, and electric generators in Baghdad, Iraq. Twenty nine isolates gave positive results in both blood agar and blue agar medium and were secondarily screened. One isolate was selected as a potent biosurfactant producer and molecularly identified and recorded in the NCBI GenBank nucleotide sequence database as Pseudomonas guguanensis strain Iraqi ZG.K.M. In optimized conditions, this strain can produce about 3.01 g/l of biosurfactant. The product could reduce the surface tension from 72 to 38 ± 0.33 mN/m and have E24% of 52 ± 0.33%. This biosurfactant was preliminarily specified to be a glycolipid and characterized as a rhamnolipid with anionic nature, usually to be a monorhamnolipid as evident from TLC, FTIR, and GC-MS analyses.

Species diversity of microorganisms previously identified as nontuberculous mycobacteria by DNA hybridization.

Over the past 10 years, the clinical importance of opportunistic bacteria of the order Actinomycetales has increased significantly. While many problems for the Mycobacterium tuberculosis complex have been solved, for nontuberculous mycobacteria, some questions remain open. These pathogens have a number of structural features that allow them to persist in the external environment for a long time. The main inclusion criteria were cultural characteristics in assessing the growth of microorganisms on solid egg media. If nontuberculous mycobacteria (NTM) growth was detected, identification signs were carried out using the DNA hybridization method. Subsequently, these cultures were identified using the matrix-assisted laser desorption/ionization-time-of-flight (MALDI-ToF) mass spectrometry method. In case of obtaining unacceptable results of identification from primary inoculations, re-identification to obtain pure cultures was carried out after transferring the material from primary media to agar media: 5% blood agar and universal chromogenic medium. When re-identifying isolated cultures using MALDI-ToF mass spectrometry, all isolated cultures were analyzed, regardless of whether they belonged to the NTM group or not. DNA hybridization, which accounted for 59.5% of the total number of cultures included in the study, performed species identification of 188 strains. Using MALDI-ToF mass spectrometry, 345 strains were identified. The use of methods based on DNA hybridization makes it possible to identify quite accurately some of the most common NTM species. MALDI-ToF mass spectrometry is an important technique to allow species identification of most Actinomycetales. However, algorithms to standardize methods for their isolation from clinical material are needed.

Potentially Virulent Multi-Drug Resistant Escherichia fergusonii Isolated from Inanimate Surface in a Medical University: Omphisa fuscidentalis as an Alternative for Bacterial Virulence Determination.

Multi-drug resistant (MDR) bacteria are becoming a worldwide problem due to limited options for treatment. Moreover, patients infected by MDR with highly virulent accessories are worsening the symptoms, even to the point of causing death. In this study, we isolated bacteria from 14 inanimate surfaces that could potentially be reservoirs for the spread of bacterial infections in the medical university. Blood agar media was used for bacterial isolation. The bacterial colony that showed hemolytic activities on each surface was tested for antimicrobial susceptibility against eight different antibiotics. We found that MDR bacterium, namely TB1, which was isolated from a toilet bowl, was non-susceptible to ampicillin, imipenem, chloramphenicol, amoxicillin-clavulanic acid, gentamicin, and tetracycline. Another MDR bacterium isolated from the mobile phone screen of security officers, namely HSO, was resistant to chloramphenicol, gentamicin, tetracycline, and cefixime. An in vivo virulence test of bacterial isolates used Omphisa fuscidentalis larvae as an alternative to Galleria mellonella larvae for the infection model. A virulence test of TB1 in O. fuscidentalis larvae revealed 20% survival in the bacterial density of 104 and 105 CFU/larvae; and 0% survival in the bacterial density of 106 CFU/larvae at 24 h after injection. Bacterial identification was performed for TB1 as a potential virulent isolate. Bacterial identification using partial 16s rRNA gene showed that TB1 exhibited 99.84% identity to Escherichia fergusonii 2611. This study concludes that TB1 is a potentially virulent MDR E. fergusonii isolated from toilet bowls at a medical university.

Comparative Evaluation of Antimicrobial Efficacy of Simvastatin Paste and Aloe Vera Paste as a Pulpotomy MedicamentAn In-vitro Study

Introduction: Pulpotomy is the complete removal of the coronal part of the dental pulp, followed by the application of an appropriate dressing or medicament to promote healing and maintain the viability of the teeth in the dental arch. Aloe vera has anti-inflammatory, anti-bacterial properties which can be used as a pulpotomy medicament. Simvastatin, a lipid-lowering medication has recently emerged as a potential medicament in regenerative dentistry. It has an anti-inflammatory and anti-oxidant properties. Hence, it can be used as a pulpotomy medicament. Aim: To evaluate and compare the antimicrobial efficacy of Aloe Vera paste and Simvastatin paste as pulpotomy medicament against Pseudomonas aeruginosa and E. faecalis. Materials and Methods: An in-vitro study will be conducted in Department of Paediatric and Preventive Dentistry at Sharad Pawar Dental College, Wardha, Maharashtra, India, in which Aloe Vera paste and Simvastatin paste will be prepared in an aseptic environment. The antimicrobial activity of simvastatin paste and Aloe Vera paste against E.faecalis and pseudomonas will be assessed using the agar diffusion method. It will be cultured overnight in brain heart infusion broth. On blood agar media, pseudomonas aeruginosa will be inoculated. 4 mm wells will be punched out on plates with a puncher and will be instantly loaded with freshly customised test materials. The agar plates shall then be incubated in an incubator at 37°C for 24 hours and 72 hours before being analysed. Statistical analysis will be done using Chi-square test, ANOVA test and multiple comparision 2k test. Expected Results: Aloe Vera and Simvastatin in the paste form shall be able to show its antimicrobial efficacy as a pulpotomy medicament. Conclusion: Aloe Vera paste and Simvastatin paste shall provide antimicrobial efficacy against E. faecalis and pseudomonas and prove to be a better pulpotomy medicament for primary teeth.

First report of Aliarcobacter cryaerophilus in ready-to-cook chicken meat samples from super shops in Bangladesh.

This study aimed to isolate Aliarcobacter cryaerophilus in ready-to-cook poultry meat in Bangladesh. Thirty drumstick samples were collected from super shops in Dhaka city (n = 10), Mymensingh city (n = 10), and Patuakhali town (n = 10). After sample processing, they were cultured in Blood agar media with Campylobacter base using a microfilter (0.42 nm). Suspected colonies were subjected to DNA extraction and PCR assay targeting 16SrRNA genes. Then, sequencing was performed for confirmation. Of 30 samples, 3 (10%) were positive for A. cryaerophilus. Phylogenetic analysis shows that our isolate has strong similarities with one of the isolates from China. The presence of this organism in ready-to-cook poultry meat is a significant concern for consumers as it bears zoonotic importance.

Hemolysins profile of clinical isolates of S. aureus from tertiary care hospitals in India

Background: Staphylococcus aureus is an important pathogenic bacterium causing both hospital and community acquired infections. Hemolysins are important virulence factors of S. aureus which help in the pathogenesis of diseases. Screening of hemolysins is important in infection control and prevention. Methods: Aim of the study was to characterize hemolysins of Staphylococcus aureus (n=152) isolated from tertiary hospitals in India, by phenotypic and genotypic methods. Phenotypic characterization of hemolysins of Staphylococcus aureus was done on either sheep or rabbit blood agar plates. Genes encoding hemolysin were amplified with specific primers in a multiplex Polymerase Chain Reaction (PCR) and analyzed by agarose gel electrophoresis. Results: Genes of all four hemolysins namely hla, hlb, hlg and hld were screened by multiplex PCR. Genotyping revealed that the isolates contained patterns of four, three or two hemolysin genes in multiple combinations. The most common profile comprised of all four hemolysin genes, namely hla, hlb, hlg, hld in a majority of isolates (47.3%, 72/152). Sixty one isolates (40.1%, 61/152) carried three hemolysin genes in three different patterns: 56 (37%) isolates contained hla, hlb, hld genes, second pattern combined 3 genes namely (1.9%) hla, hlg, hld and the third pattern was found in 2 (1.3%) isolates with hlb, hlg, hld. Five (3.28%) isolates in this study had only two hemolysin genes in three different combinations: 2 (1.3%) isolates with hlb:hld, 2 (1.3%) isolates with hla:hld pattern and 1 (0.7%) isolate with hlg:hld pattern. 14 (9.2%) isolates carried only hld gene. Phenotyping showed that all the isolates had α, β and δ activities and high activity of α, β and δ hemolysins were seen in isolates which were obtained from pus, blood and other body fluids. All these sites suffer enormous tissue damage in severe S. aureus infection. The genotype data correlated well with the phenotype analysis. Conclusions: Four hemolysin genes were detected in all clinical isolates of S. aureus. We could identify the activity of α, β and δ hemolysins on blood agar media. We found out that the isolates displayed varying combination of the four hemolysin genes. These differences in genotype of four genes and the hemolysin activity may indicate a preference mandated by the host tissue or site during the pathogenesis of the disease since each hemolysin has different activity profile. Body fluids in general, pus, blood, sputum in particular carried S. aureus with higher hemolysin activities compared to isolates of specimen from other sites. The results of this study reinforce the importance of hemolysins in S. aureus diseases and suggest that the tissue damages and symptoms are related to the hemolysins activity during pathogenesis.

A study on the microbial spectrum of otitis externa in a tertiary care teaching hospital

Background: Otitis externa is frequently seen illness by ENT practitioners predominantly in children and old age people. The severity sometimes varies from it ranges in severity from a trivial infection to dangerous malignant otitis externa. Furthermore, due to absurd consumption of antibiotics, though there is a fall in development of complications, there is a rise in resistance to various infectious organisms. Mostly, we start treating patients with empirical therapy where sometimes, it leads to occurrence of various antibiotic-resistant infections. For that reason, recent evidence about the organism which is mostly involved in infection and also most importantly the pattern of their response to various pharmacological agents is crucial for the sensible use of the medications for the management. Aim and Objectives: Our study was planned to evaluate and find out the causative organism which is most commonly seen in otitis externa, particularly focusing on drug sensitivity for those organisms, so that efficient management of otitis externa can be done. Materials and Methods: It is a retrospective study conducted in acute otitis externa patients from Novemver 2019 to January 2020. Patients who all presented with ear discharge within 3 months of period, samples were collected with clinically diagnosed acute otitis externa and sent for culture in blood agar, chocolate agar, and Mcconkey medium in vitro and drug sensitivity pattern was identified for individual organisms. Inclusion criteria: Patients more than 18 years having ear discharge within 3 months were included in the study. Exclusion criteria: Chronically discharging ear &gt;3 months were excluded from the study. Results: Pseudomonas organism was frequently encountered pathogen in our study, trailed by Staphylococcus aureus. The organisms identified were very much sensitive to drugs like fluoroquinolones particularly Ciprofloxacin, and Aminoglycosides like Amikacin and almost all were responding to treatment with Piperacillin-Tazobactam, these drugs have an cidal activity over pathogens and are suitable for any age and safety-wise better too. Conclusion: Intermittent assessment and awareness of the bacteriological summary and their drug response profile in a particular geographical area will only help in rational drug prescription. Furthermore, rational usage of drugs will help in reducing any morbidity due to infection which may sometimes develop when treated inappropriately, further decreasing development of dangerous drug resistance.

Multidrug resistant and extended spectrum β-lactamase producing gram negative bacterial uropathogens among females in a tertiary hospital, Pokhara

Introduction: Extended spectrum β-lactamase producing gram negative bacilli has emerged and spread worldwide as an important cause of urinary tract infections and indicates a major threat. The study aimed to determine the prevalence of multidrug resistant and extended spectrum beta-lactamase producing gram negative bacterial uropathogens among females in a tertiary level hospital. Methods: A hospital-based cross-sectional study was conducted in Pokhara Academy of Health Sciences, Pokhara from February to November, 2019. A total number of 301 midstream urine samples were collected and examined using MacConkey agar and blood agar medium. Antimicrobial susceptibility testing was done by Kirby Bauer disk diffusion method on Mueller Hinton agar using Clinical and Laboratory Standards Institute guidelines. Results: Out of 301 mid-stream urine samples, 99(33%) sample showed significant bacterial growth. Among them, 78(79%) were gram negative bacteria. Escherichia coli were the predominant organism. Multidrug resistant gram negative isolates were 65.4%. Among 78 Gram negative isolates, 31(39.7%) were extended spectrum β-lactamase producers. Among extended spectrum β-lactamase producers, 27(87.1%) were MDR. Highest frequency of extended spectrum β-lactamases production was seen in E. coli, 23(74.2%). Majority of gram negative bacteria showed susceptibility toward colistin and nitrofurantoin. Ampicillin was found to be highly resistant towards gram negative uropathogen. Conclusions: This study found that higher proportion of multi-drug resistants were among gram negative isolates and further more among extended spectrum β-lactamase producing gram negative isolates. Thus, there is urgent need to address the issue of antimicrobial resistant and promote rational use of the antibiotics in our region.

The Comparative Efficacy of Imipenem and Meropenem On Different Bacterial Strains Obtained from Clinical Samples

The most serious threat to patient's and the public's health is the resistance of clinically relevant microorganisms to antimicrobials. Objective: This study's primary objective was to compare the susceptibility profiles of imipenem and meropenem on various bacterial strains. Methods: 101 distinct patients' positive samples of blood and pus were collected and sent to a pathology lab in Lahore (Mughal Diagnostic and Research laboratory Lahore). On Macconkey, CLED, and Blood Agar media, five bacterial strains; E. coli, P. aeruginosa, Enterococcus species, Klebsiella species and S. typhi were isolated and resurrected. Following microscopical (gram staining) and biochemical tests to identify these bacterial strains, the antibiotic sensitivity of these bacterial strains was assessed. Results: The findings of this investigation clearly demonstrated that imipenem is more sensitive than meropenem. Imipenem demonstrated improved sensitivity to all of the bacterial strains included in the study, especially E. coli, P. aeruginosa, various Enterococcus, Klebsiella, and S. typhi, imipenem showed a sensitivity of 84.15%, while meropenem showed a sensitivity of 40.59%. Conclusions: The current investigation came to the conclusion that antibiotics (imipenem and meropenem) are becoming more resistant to bacteria as a result of their growing and frequent use. Physicians started to worry about the developing antibiotic resistance due to the indiscriminate use of these treatments after more than a decade of increased antibiotic consumption in the era

Effect of Culture Conditions on Fatty Acid Profiles of Bacteria and Lipopolysaccharides of the Genus Pseudomonas-GC-MS Analysis on Ionic Liquid-Based Column.

The profiling of bacterial fatty acids is a well-established technique in identifying and classifying bacteria. Cultivation conditions may affect the biosynthesis, thereby, changing the fatty acid profile in bacteria. The effect of the culture conditions on the fatty acid components of Pseudomonas aeruginosa PAO1, Pseudomonas aeruginosa ATCC 27853, Pseudomonas aeruginosa polyresistant and Pseudomonas putida all are aligned to the genus Pseudomonas. The fatty acids in the lipopolysaccharides of Pseudomonas aeruginosa PAO1 were also examined. The effects of the cultivation conditions were followed by using agar and blood agar media at the characteristic temperatures, 25 °C, 37 °C and 42 °C, respectively, and an analysis was made during the 1st, 3rd and 5th day following inoculation. In addition to quantitative differences, we also experienced qualitative differences in the fatty acid profiles which detect newly appearing fatty acids, due to changes in environmental factors. The application of ionic liquid-based column unveils new possibilities for the analyses of fatty acids in GC-MS experiments for bacterial fatty acid profiling. The validation results (response linearity, limit of detection, limit of quantification, system suitability, intraday and interday repeatability and accuracy) show the high separation efficiency of the ionic liquid-based column in the analyses.

High serum AMH inhibits pathological growth of the low biomass endometrial microbiome.

Although host microbiome play a role in both hormonal status and fertility, this issue has not yet been clarified. Since the endometrium is a sterile tissue, it is accepted that microbiota does not grow under normal conditions. The aim of the study was to reveal the characteristics of endometrial microbiota according to serum AMH levels in women with implantation failure. Forty-five women aged 20-30 years with two or more implantation failures were included in the study. They were divided into 3 groups according to their serum AMH values: Group 1 -AMH <1.3 ng/ml; Group 2 - AMH between 1.3-2.6 ng/ml; Group 3 - AMH >2.6 ng/ml. Twenty-two healthy fertile women who were the same age as the infertile group and applied for cervical smear screening were accepted as the control group. Following the embryo transfer, the tip of the catheter was inserted into the transport medium under sterile conditions. Sowing was carried out by touching the tips of the catheter to the blood agar medium. After the evaluation of the petri dishes at the end of 48 hours of incubation, colonies were stained with Gram stain. Microorganisms in the colonies were identified with the Vitek-2 device according to their gram-staining characteristics and their antibiograms were made. A negative correlation was detected between low AMH values and the microbiome detection rates in endometrial cultures. In patients with low serum AMH levels, the chance of endometrial microbiota growth was higher in the endometrial culture medium. The most common bacteria were found to be MSSA, MRKNS and lactobacillus. Clinical pregnancy rates were found to be significantly higher in the group with high AMH levels. As AMH levels increased, positive flora detection rates decreased, while clinical pregnancy rates increased. Low serum AMH level increases the rate of positive endometrial microbiome in culture and decreases clinical pregnancy rates.

Phenotypic and Molecular Investigation of Streptococcus pneumoniae Hemolysin

The current study aimed to detect S.pneumoniae, which is one of the gram-positive bacteria that gives partial α-hemolysis on blood agar media, as well as has the ability to complete blood hemolysis (beta)-hemolysis under anaerobic conditions. Fifty sputum specimens were collected from patients in Ibn sena and Al Salam hospitals in Mosul/ Iraq from august 2021 to March 2022. Five isolates were diagnosed as S.pneumoniae which exhibited an ability to produce hemolysin when phenotypicallyinvestigated. Polymerase chain reaction (PCR) technique was used to detect hlys gene Electrophoresis results showed four bands (80% of isolates) with a molecular size of 296 bp the PCR product were then sequenced.

Development of respirator design for children using bamboo-based activated carbon filter and bipolar ionization

This paper proposes a respirator for children integrated with bamboo-based activated carbon filters and bipolar ionization as the sterilizer to increase protection from Covid-19 and other pathogens. The respirator, named as Bion-Kids, was designed based on the children's head-and-face anthropometry from direct measurement. The duration of sterilization process is controlled based on the activities, which are classified using the Recurrent Neural Network (RNN) model with dataset acquired from the accelerometer. Manufacturing process of the main body prototype comprises heat bending and 3D printing. The bamboo-based activated carbon filter was synthesized by applying heat and pressure at the strewed activated carbon particles on the N95 filter. Observation using scanning electron microscope showed the ability of the bamboo-based activated carbon filter to adsorb and trap the particles. Result of the qualitative fit test and questionnaire survey indicated that Bion-Kids is suitable and comfortable for the children's face. The result of filtration and bipolar ionization sterilization system analysis have also met the ISO14698 standard by not showing any infiltration of microorganisms on the blood agar media. This device may become an early-stage personal protective equipment against the transmission of Covid-19 and other pathogens.

&lt;strong&gt;Antibacterial activity of cocoa bean husk extract on the growth of Streptococcus alpha&lt;/strong&gt;

ABSTRACTIntroduction: Streptococcus alpha is the most common bacteria found in dental plaques of children­ and roled as pioneer bacteria in plaque formation. One of the natural agents which has antibacterial activity is cocoa bean husk (Theobroma cacao, L.). This study aimed to analyze antibacterial activity of cocoa bean husks extract concentration 12.5%, 25%, 37.5% and 50% on the growth of S. alpha. Methods: This study was a laboratory experiment conducted in March, 2021 at Microbiology Laboratory, Faculty of Medicine Universitas Negeri Surakarta. The bacteria was isolated from the buccal tooth surface plaque of caries-free-9-years-old girl, then incubated on the blood agar medium to grow the S. alpha. The extract of cocoa bean husk concentration 12.5%, 25%, 37.5%, 50% and 0,2% Chlorhexidine Gluconate(CHX) as positive control were used to identify the antibacterial activity by measure the diameter ofinhibition zone growth of S. alpha in the blood agar medium. The data was analyzed using one-way Anova. Results: Significant differences were found among the treatment groups (p&lt;0.05). The highest mean of the inhibitory zone was the 50% concentration of cocoa bean husk extract group (17.07 mm ± 1.01), but lower than CHX. The lowest inhibitory zone was the 12.5% (10.12 ± 0.05 mm). Conclusion: The cocoa bean husk extract has antibacterial activity on the growth of Streptococcus alpha, but it is lowerthan Chlorhexidine Gluconate.Keywords: cocoa bean husk extract; antibacterial activity; S. alpha’s growth; child’s dental plaque

Evaluation of microbiological contamination in the clinics of the faculty of dentistry of the university of Cuenca

Objective: To determine the bacterial load in the dental units of the undergraduate clinics and the radiology area of the Faculty of Dentistry of the University of Cuenca after the disinfection protocol used by the cleaning staff. Introduction: Dentistry is considered a high-risk profession for that reason people who are part of the work team and the patients are exposed to pathogenic microorganisms. For this reason, it is too important and necessary to establish asepsis, antisepsis and biosafety protocols in order to reduce and prevent contamination and proliferation. Materials and methods: A total number of 13 different surfaces were evaluated for the collection of the sample, the sterile Stuart transport medium was used to subsequently seed it in the blood agar media. Results: At 48 hours there was no growth of colonies, however at 96 hours the following colonies were obtained: Adult clinic (Bacillus spp: 4000 CFU/ML), child clinic (Bacillus spp: 2000 CFU/ML, Coagulase-negative Staphylococcus: 1000 UFC/ML) and radiology (Bacillus spp: 1000 CFU/ML). Conclusions: The cleaning and disinfection protocols used in the clinics and radiology service of the Faculty of Dentistry of the University of Cuenca are not enough to kill microorganisms completely. Although the CFUs were relatively lower compared to other studies, it is still a predisposing factor for infections in patients, operators, teachers and even cleaning staff.

Molecular Detection of Pseudomonas Aeruginosa Isolated from Different Clinical Cases and Test Antibiotics Sensitivity on the Bacterial Growth

150 samples were collected from different clinical causes of hospitals in Al-Diwaniyah city. All samples were cultured on blood agar and Maconkey agar medium and incubated for 24 hat 37°C. The results showed 112 (74.6%) bacterial growth. Initially, identification of the bacterial isolates were carried out viabiochemical tests, 25 isolates Pseudomonasaeruginosa were detected. Identity was confirmed by polymerase chain reaction (PCR) targeting the areE genewith size 498pb. This bacterium was isolated from burn infection 30% and wound infection 20%, while it was isolated from Respiratorytract 16.6%. The antibiotics susceptibilitytest for 16 bacterial isolates have been tested. It has been found that the bacterial isolates were resistance 100% for Ampicillin, Amoxicillin, Piperacillin, Cefotaxime and Cefepim and 18.75% forMeropenem. While they were sensitive 100% for Amikacin. Keywords: antibiotics, areE gene, Pseudomonas aeruginosa, PCR.

Molecular detention of efflux pumps in Pseudomonas aeruginosa obtained from different clinical cases

150 samples were collected from different clinical sources from Al-Diwaniyah city hospitals from September 2021 to January 2022. The samples were grown on Maconkey agar and blood agar medium and incubated at 37°C for 24 hours. Bacterial isolates were diagnosed through culture characteristics, microscopic examination and biochemical tests, as 25 isolates belonging to p. aeruginosa. The phenotypic detection of these isolates was conducted and it was found that 16 isolates contain efflux pumps, after which the genetic detection was done by PCR technique. The isolates were subjected to the mexAB-OprM efflux pump detection using PCR technique, and after the replication product was migrated in agarose gel, it was observed that 93.75 %) of the isolates possess the mexA gene encoding the above efflux pump, and (43.75%) the oprM gene encoding the above efflux pump also.

Comparison of the frequency of bacteriuria in diabetic and non-diabetic patients without symptoms of urinary tract infection

Introduction: Urinary tract infections (UTIs) are the most common infections among either community or hospital-acquired infections. UTIs are common in diabetes mellitus, with glycosuria and neurogenic bladder being the predisposing factors. Methods: This was a retrospective observational study. The study population consisted of 100 diabetic and 40 non-diabetic patients. The rates of bacteriuria and pyuria were investigated in diabetic and non-diabetic patients who had been hospitalized and who did not have any complaints of UTI. Urine samples of patients were collected and each sample was tested through microscopy and culture. Furthermore, the urine samples were inoculated on blood agar and eosin methylene blue agar medium and incubated for 18–24 h at 37°C. Diabetic patients were evaluated to obtain data on age, gender, duration and type of diabetes, body mass index, retinopathy, nephropathy, glycated hemoglobin (HbA1c) level, and fasting plasma glucose (FPG) level. Results: A higher rate of bacteriuria was detected in diabetic patients (21%) than in non-diabetic patients (5%) (p = 0.02). Infections were detected more frequently in patients with high HbA1c level (p = 0.001), those in the advanced age group (p = 0.014), and those with nephropathy (p &lt; 0.01). Conclusion: Asymptomatic bacteriuria was more prevalent in diabetic than in non-diabetic patients, with poor glycemic control, age, and nephropathy being significant risk factors. Escherichia coli is the most common organism that causes bacteriuria in diabetic patients.