Optimization of Quantitative Analysis of Biofilm Cell from Pipe Materials

The quantitative analysis of biofilm can be used not only to assess the microbiological stability of tap water but also on its basis can assess: the degree of colonization of materials by bacterial cells, the rate of biofilm formation on the surfaces of pipes and determine their composition and number. The article presents the results of research on the development of an effective method of biofilm detachment from the surface of the galvanized steel. The number of biofilm cells was determined by methods: (1) luminometric ATP determination, (2) flow cytometry and (3) heterotrophic plate counts (HPC). The presence of the biofilm was confirmed by SEM and fractal analysis. The analysis of the obtained results showed that the most effective method of detaching the biofilm cells from the galvanized steel surface was the mechanical separation with a sterile cotton swab. The variant with the use of a sterile swab enables rapid collection of the biofilm from the surface of the ducts forming internal installations or water supply networks. Due to the simplicity and speed of obtaining results, the luminometric ATP measurement has been established as the best method for the quantification of biofilm cells. The results of this study were intended to provide reliable and useful data on the quantification of biofilm cells.

Evaluation of Candidal Carriage Among Smokers and Non-Smokers

Objectives: To determine the Candidal carriage among smokers and non-smokers and with different intra-oral sites including examination of various biotypes of Candida. Study design and setting: Cross-sectional based study conducted at Dr. Ishrat ul Ebad Khan Institute of Oral Health Sciences and Dow International Dental College, Karachi, from May 2017 till April 2018. Methodology: Comprised 100 patients (50 smokers and 50 nonsmokers) between 20 and 60 years of age. The collection was performed through sterile cotton swab to evaluate oral Candidal carriage and the colonizing Candida species using Sabouraud Dextrose Agar (SDA) and API20C AUX (BIOMERIEUX). Data was analyzed Spss version 20. Results: A total of 100 participants (50 smokers and 50 non-smokers) were evaluated for candidal carriage. The common age group was 20-30 years in both the groups, without significant difference (p-value 0.79). Frequency of candidal carriage was comparable among smokers 14 (28.0%) to non-smokers 10 (20.0%), with a statistically insignificant p-value 0.35. Based on various biotypes among smokers and non-smokers, Candida albicans was 9(18%) and 7(14%), Candida glabrata was 4(8%) and 2(4%); and Candida tropicallis was 1(2%) each for both smokers and non smokers. Dorsum of tongue harbored all prevalent biotypes i.e. Candida albicans, Candida glabrata and Candida tropicalis as statistically significant among smokers (p-value 0.04). Conclusion: Candidal carriage was comparable among smokers and non-smokers. Candida albicans and Candida glabrata were the common biotypes predominantly among smokers

Assessment of conjunctival flora in eyes with lacrimal passage obstruction before and after successful dacryoendoscopic recanalisation

BackgroundTo investigate conjunctival bacterial flora in eyes with lacrimal passage obstruction before and after dacryoendoscopic recanalisation with lacrimal tube intubation.MethodsOne-hundred fifty eyes with lacrimal passage obstruction that were successfully treated by dacryoendoscopic recanalisation were enrolled. Conjunctival sampling was done for each eye before and 4 months after surgery. The lower fornix was rubbed by a sterile cotton swab, and the collected samples were cultured with several agar plates. Colonies were differentiated and enumerated by standard bacteriological laboratory techniques.ResultsPositive bacterial growth was detected in 42.0% of all the samples before surgery, and the positivity rate significantly decreased to 26.0% after surgery (p=0.0051). The number of strains detected also decreased from 20 before surgery to 9 after surgery, especially pathogenic microorganisms decreased. In addition, drug-resistant bacteria such as penicillin-resistant Streptococcus pneumoniae and methicillin-resistant Staphylococcus aureus were detected in nine eyes before surgery but were detected only in one eye after surgery. Coagulase-negative Staphylococci and Corynebacterium spp., which are common in normal conjunctival flora, accounted for 46.5% of all the isolates before surgery and 80.9% after surgery, showing a significant increase in the rate after surgery (p<0.0001).ConclusionsThis study showed that physiological recanalisation of lacrimal passage after dacryoendoscopic surgery significantly decreased the culture positivity rate of conjunctival sac and the number of microorganism strains detected. It also decreased the number of potentially pathogenic and drug-resistant bacteria and increased the percentages of indigenous bacteria, causing the normalisation of conjunctival flora.

Potency of stored DNA derived from buccal mucosal epithelial cells for genotype analysis of interleukin 4 -590 C/T

The genotype variation of the interleukin 4 (IL-4) gene on spot -590 C/T is known to correlate with IL-4 protein expression in periodontitis, but the results found in the population are inconsistent. Sequencial genotype analysis of multiple genes often used stored DNA samples obtained from buccal mucosal epithelial cells. This study aims to determine the use of stored DNA samples obtained from buccal mucosal epithelial cells for genotype analysis specifically on -590 C/T IL-4 genes. This is a descriptive analytical study. The samples were 58 DNA derived from the buccal mucosal epithelial cells taken by sterile cotton swab isolated with PrestoTM kit (GeneAid). The DNA has been stored for 2 years at -20 °C. The genotype analysis was conducted using PCR RFLP technique with BSmFI enzyme and the data were presented descriptively. The results show that 51.72% of the stored DNA is still in good condition and 37.9% can be used for genotype analysis. It was concluded that DNA derived from buccal mucosal epithelial cells stored at -20 °C for 2 years can still be used for genotype analysis, but the quality of DNA is affected by storage time.

Geographic Tongue - Does Candida Play a Role in its Pathogenesis

Summary Background/Aim: Geographic tongue (GT), a benign self-limiting condition (inflammation) is commonly seen in practice. Although quite easy for clinical diagnosis, in clinical practice it is commonly misdiagnosed and treated as Candida infection. The main aim of this study was to compare the prevalence of Candida spp. on tongue in patients with GT and subjects with clinically healthy oral mucosa. Additional aims were to evaluate subjective symptoms, uncontrolled usage of prescribed or non-prescribed antifungal topical medications and the presence of the cancerophobia or other health concerns in patients with GT. Material and Methods: A total of 70 subjects were divided into two groups: Group B- patients with diagnosed Geographic tongue and group A - aged and gender matched controls with clinically health oral mucosa. Anamnestic charts designed for this study included information about symptoms (measured by Visual Analogue Scale), previous knowledge or fear about presence of GT and received therapy for this condition. Detection of Candida spp. from tongue was done using sterile cotton swab and Sabouraud dextrose agar. Results: At the time of this study, 18 (52.94%) of subjects with GT were aware of having this condition and even 12 of them (66.7%) used topical antifungal drugs prescribed by physician. Also, 66.7% of them experienced fear about the presence of GT at some moment. Detection of Candida spp. was similarly distributed in both groups (22.2% in group A and 17.6% in group B). Conclusions: Geographic tongue has frequently been treated by topical antifungal drugs. In this study, GT was not associated with presence of Candida spp.

Stethoscope as potential occupational hazards for intra hospital carries of pathogenic microorganisms

Background: The stethoscope is a tool that doctors use daily in the examination of patients and it can take part in the transmission of health care-associated infections. In a single day it may come in direct contact with multiple patients and the intra hospital environment may be contaminated by various type of bacteria and possibly transmit to others. Objective:- The study was to know the attitude and knowledge about the stethoscope hygiene behavior among physicians and to determine the types of bacterial agents that can contaminate stethoscopes. Type of the study: The study was a cross-sectional study Methods:- It was conducted from 1st of July to end of October 2014 at AL-Emmamain Alkadhomain Medical City in which a convenient sample of 150 physicians were included. A semi constructed questionnaire was used to collect demographic data and hygiene practice among participants. Specimen was collected using moisten sterile cotton swab and then cultured following standard microbiological techniques. The results:- showed a total of 121 (80.6%) of the stethoscopes had bacterial contamination only 29(19.40%) had negative cultures. There is statistically significant association between stethoscope contamination and frequency of cleaning it. Of the studied group only 26 subjects (17.3%) received education regarding stethoscope cleaning. Conclusion:-There was evidence that bacteria can transfer from the skin of the patient to the stethoscope and from the stethoscope to the skin and there was poor education and assessing cleaning practices of stethoscopes.

Cytological diagnosis of infectious conjunctivitis in cats

Cytological examination of conjunctival smears obtained from cats with conjunctivitis can reveal the severity and type of inflammation and diagnose conjunctivitis caused by Chlamydophila felis. The diagnosis of chlamydia was made by the study of material taken from the conjunctival sac with a sterile cotton swab by PCR. Material for cytological examination was taken from these cats. Cytological examination of inclusions interpreted as chlamydial, was detected in all cytologic smears from sick cats, and has not been detected in none of healthy cats. Thus, cytological examination is a convenient, non-invasive, easily feasible method, which is suitable for rapid diagnosis of chlamydial conjunctivitis in cats.

Cytological and bacteriological sampling from filters used for embryo recovery to evaluate the uterine status of donor mares

While testing for uterine bacterial infection is usually performed prior to artificial insemination (AI), samples taken during or after embryo flushing are generally not assessed either in subfertile and old mares or in fertile mares, even though knowledge of the status of the uterine environment in which the embryo is to develop would help to predict the outcome of embryo transfer programmes. The presence of bacteria and inflammatory cells in the liquid retained in the filter after uterine flushing in donors was determined at the moment of embryo recovery. Primarily, a group of mares (n = 8) displaying evident clinical signs of endometritis was selected to evaluate the cytological and bacteriological findings in filters after uterine flushing and in uterine cotton swabs. Two uterine samples (for cytological and bacterial evaluation) were taken with cotton swabs and, subsequently, the uterus was flushed and the efflux was also subjected to bacteriological and cytological analysis. Later, a group of donors (n = 20) was also involved to evaluate the presence of bacteria and polymorphonuclear leukocytes (PMN). After embryo flushing and collection, the efflux retained in the filter was evaluated by cytology and bacteriology. A sterile cotton swab was then scrubbed on the filter mesh, and a bacterial culture was performed. The embryo recovery rate was 30% (n = 6); Escherichia coli was isolated in one efflux sample collected from embryo-productive flushings, while the other five samples were negative by culture. Bacterial growth (not considered as contamination) was observed in a total of three samples, although no inflammatory cells were detected. Bacteria were isolated in endometrial samples collected after embryo flushing in donor mares, although inflammatory cells were never present in the uterus of mares from which embryos were recovered. In the absence of clinical signs, cytological and/or bacteriological samplings are not very useful for estimating the success of embryo recovery in donor mares, but evaluation of the filter and efflux after uterine flushing in donors may provide valuable information regarding uterine status at embryo collection.

Correction of vaginal dysbiosis in mice caused by a film-forming strain Staphylococcus aureus, using bacteriophages and probiotics

The complex use of bacteriophages and probiotics is a promising trend in improving prevention and treatment of gynecological lesions. Our study of their influence on the microflora was performed on the model of vaginal dysbiosis of white laboratory mice induced by introduction of a filmforming strain of Staphylococcus aureus. For correction of dysbiosis, staphylococcal bacteriophage liquid, piobacteriophage polyvalent, intesti-bacteriophage liquid and probiotic "Vahilak" were used. For the identification of the microflora of the reproductive tract, samples of biological material from the vagina were obtained by sterile cotton swab and plated on nutrient media to determine the nature and extent of growth of the cultures. The maximal effect was found to occur with the correctional complex "bacteriophage staphylococcal liquid – vahilak" that led to decrease of total microbial number to 4.77 × 104 CFU/ml and to the restoration of the ratio of aerobic to anaerobic bacteria 1 : 52 when indicators of the norm were 4,69 × 104 CFU/ml and 1 : 52. In this case, 24 hours after the last injection of the preparations the amount of microaerophilic and anaerobic lactobacilli had increased by 20.8 and 2.1 times respectively. The frequency of isolation of microaerophilic lactobacilli increased to 100%, and anaerobic – up to 70%. Also the number of staphylococci, streptococci, enterococci, bacilli and enterobacteria decreased by 30.1, 1.1, 1.5, 2.2 and 11.8 times respectively. Also, there was a decrease in the detection rate of enterococci, micrococci and enterobacteria by 10% and bacilli by 20% compared to the control dysbiosis. The number of anaerobic bacteria also underwent significant changes. Thus, the number of fusobacteria decreased by 33.2 times, peptococci – 2.3, peptostreptococci – 6.6 and Bacteroides – 7.9 times, which is almost consistent with indicators of the norm. In addition, the frequency of detection of peptostreptococci decreased by 10%. Therefore, it can be concluded that medical bacteriophages are active against lesions caused by able to film-forming staphylococci, in vivo, so they are appropriate to use in medical practice both independently and in combination with other agents.

ПОРІВНЯЛЬНА ОЦІНКА ДІЇ ДЕЗІНФІКУЮЧИХ ЗАСОБІВ З МИЮЧИМ ЕФЕКТОМ ПРИ ЗНЕЗАРАЖЕННІ E. СOLI ТА S. АUREUS

A comparative analysis of the use detergents disinfectants with different active ingredients: heotsyd (polihexametylenhuanidin hydrochloride, benzalkonium chloride) virosan (alkildymetylbenzylamoniya chloride, glutaraldehyde) neohlor (sodium hypochlorite) orhasept (benzalkonium chloride, lactic acid) and determination of their minimum effective concentration and exposure required for removal of E. coli and S. aureus on test sites: stainless steel, tile, concrete, brick was were conducted. At sterilized in an autoclave test objects applied to 1 cm3 culture E. coli and S. aureus at a concentration of 2 billion. mikr.til per cm3. Contaminated test objects dried and placed into cuvettes horizontally and vertically and spray disinfectants applied explored solutions, while noting their exposure concentration and amount spent. Objects whish were exposure by the same scheme, but with using sterile water were used as control. After a certain period of time by sterile cotton swab from experimental and control test objects were taken probes. Then 1 cm3 initial suspension of each of these tubes were placed in the appropriate environment for 24 hours. at thermostat at 37 °C. By the changes in defined media presence or absence of these microorganisms were determined.Stated that disinfectants orhasept and virosan in 0.1% concentration and heotsyd neohlor in 0.5% concentration completely disinfected from E. coli and S. aureus on surfaces of the test objects for 10 min, confirms their detergent disinfectant effect. Effective concentrations of disinfectants regarding test organisms consistent with regulations for their use.