Microbiological Diagnostic Methods Research Articles

INTERDISCIPLINARY INTEGRATION OF MICROBIOLOGY, VIROLOGY, AND IMMUNOLOGY IN THERAPEUTIC STOMATOLOGY

Introduction: This article examines the potential for integrating Therapeutic Stomatology with Microbiology, Virology, and Immunology. It highlights the significance of studying key concepts in these disciplines, which underpin various methods of microbiological diagnostics utilized in dentistry. Practical skills and theoretical knowledge essential for the study of Therapeutic Stomatology are exemplified. The aim of this paper is to underscore the significance of interdisciplinary integration between Microbiology, Virology, and Immunology with Therapeutic Stomatology in the education and training of future dental practitioners.
 Results and Discussion: A solid grasp of microbiology is crucial for fostering the clinical acumen of future dental professionals. The integration of disciplines should be systematically woven into all phases of the educational curriculum. By merging fundamental and clinical disciplines, a multidimensional approach to problem-solving is facilitated, highlighting the interconnectedness between fields. This integration enhances students' intellectual, cognitive, and communicative abilities, broadening their educational horizons and enabling the application of foundational knowledge in novel contexts. Consequently, this approach nurtures the development of clinical reasoning skills among aspiring dental practitioners.
 Conclusions: The interdisciplinary integration of Microbiology, Virology, and Immunology with Therapeutic Stomatology in professional training ensures educational coherence, enhances the organization and refinement of knowledge and skills, and lays the groundwork for cultivating professionals characterized by versatile thinking and comprehensive expertise in their field.

Pathogenesis and clinical forms of human listeriosis

Listeriosis is a saprozoonotic infection caused in humans mainly by the species Listeria monocytogenes. Its source can be objects of the external environment (soil and water of open reservoirs with a high content of organic compounds, plants) and more than 90 species of animals. The high metabolic activity of listeria and resistance to adverse factors create the possibility of an easy transition from a saprophytic to a parasitic type of metabolism when ingested by the host. A variety of transmission routes and "entrance gates" cause the emergence of a variety of primary localized forms of infection, the most common of which is gastrointestinal. Invasive forms of listeriosis develop mainly in people with immunological insufficiency. "Risk groups" are pregnant women, newborns, people with comorbid conditions, over 60 years old, drug addicts, etc. They are characterized by polymorphism of symptoms, the absence of specific manifestations. Pathogenicity factors of L. are important in the pathogenesis of infection. monocytogenes, providing their incomplete phagocytosis, intracellular parasitism, high rate of colonization of neighboring healthy cells, the formation of antibiotic resistance. The pathological expression of changes caused by listeria in cells is the formation of small granulomas — listeriomas. The "diversity" of listeriosis, the high mortality rate (up to 50% or more) and the low alertness of doctors regarding this infection create difficulties in making a diagnosis, and therefore the improvement of microbiological diagnostic methods is of particular importance.

Evaluation of analyte-specific reagents for the direct detection of Pneumocystis jirovecii.

Our study is unique compared to other previously published studies on the DiaSorin analyte-specific reagent (ASR) because we focused on microbiological diagnostic methods commonly used (immunofluorescent assay) as opposed to pathology findings or reference PCR. In addition, in our materials and methods, we describe the protocol for the use of the DiaSorin ASR as a singleplex assay, which will allow other users to evaluate the ASR for clinical use in their lab.

The Challenges of The Diagnostic and Therapeutic Approach of Patients with Infectious Pathology in Emergency Medicine.

A retrospective observational study was performed on 657 patients, from whom clinical samples (urine, purulent secretions, blood cultures, etc.) were collected for microbiological diagnosis in the first 3 days after presentation in the ED. The identification of pathogens and the antimicrobial susceptibility testing with minimum inhibitory concentration determination were carried out according to the laboratory protocols. From the 767 biological samples analyzed, 903 microbial isolates were identified. E. coli was most frequently isolated (24.25%), followed by Klebsiella spp., S. aureus (SA), and non-fermentative Gram-negative bacilli. E. coli strains maintained their natural susceptibility to most antibiotics tested. In the case of Pseudomonas spp. and Acinetobacter spp., increased rates of AMR were identified. Also, 32.3% of SA strains were community-acquired MRSA. The introduction of rapid microbiological diagnostic methods in emergency medicine is imperative in order to timely identify AMR strains and improve therapeutic protocols.

Rapid identification of canine uropathogens by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and the clinical factors that correlated bacterial species and antimicrobial resistance.

Using the matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) method for bacterial diagnosis, rapid urine sample preparation can reduce time relapsing of diagnosis and improve discriminatory power in coinfection cases. We aimed to evaluate rapid urine preparation procedures before MALDI-TOF MS application using dog clinical urine samples in comparison with standard microbiological diagnostic methods by agreement analysis. We determined the frequency and distribution of bacteria and bacterial resistance and their correlations to clinical history. Three experimental procedures comprising direct centrifugation, 10% sodium dodecyl sulfate digestion, and ultrasonic preparation were performed for method validation and sensitivity. Sterile urine containing Escherichia coli and/or Staphylococcus aureus were used as simulated samples. By ultrasonic preparation, the microorganisms could be detected 1.46-1.51 × 105 CFU, which was considered the most suitable technique. This preparation was significantly consistent with the routine method based on data from Hospital Information Systems for 50 urine samples from canine cystitis. By standard protocol, Enterobacteriaceae and Staphylococcus pseudintermedius were found in most of the 155 urine samples with cystitis. Extended-spectrum beta-lactamase-producing Enterobacteriaceae was found in 25-30% of the samples. Imipenem resistance was found in 70% of Acinetobacter baumannii cases; almost all were resistant to second-generation fluoroquinolones and tetracyclines. The most efficient antibiotic for treating bacterial urinary tract infection was amoxicillin plus clavulanic acid. A. baumannii and Pseudomonas aeruginosa were susceptible to pradofloxacin. Prolonged urine catheterization was linked to lower urinary tract infections by Enterobacter spp., which also correlated with chronic kidney disease.

МОНИТОРИНГ И ОЦЕНКА МИКРОБИОЛОГИЧЕСКОЙ ДИАГНОСТИКИ ТУБЕРКУЛЁЗА: РЕСУРСЫ И ДЕЯТЕЛЬНОСТЬ МИКРОБИОЛОГИЧЕСКИХ ЛАБОРАТОРИЙ

Significance. The system of microbiological diagnostics is of key importance for organizing microbiological diagnostics in accordance with the modern recommendations, and fulfilling political commitments of the Russian Federation within the framework of the End TB Strategy Purpose: to assess the availability, use and results of the modern methods of microbiological diagnostics of tuberculosis in Russia in 2021. Material and methods. The study has analyzed forms #7-TB, ВР-4БЛ, ВР-1Ф of official statistical observation, results of external quality assessment of laboratories (Interlaboratory comparison system “Federal System of External Quality Assessment of Clinical Laboratory Testing (FSEQA)”). Results. The share of patients with pulmonary tuberculosis with a smear positive Ziehl-Nelsen, detected in medical organizations providing primary health care (PHC) decreased to 12.8%, while the share of new smear positive pulmonary TB patients increased to 37.4%; the share of newly diagnosed patients with pulmonary tuberculosis with a positive result of sputum bacterioscopy among patients with cavities increased to 60.6%.The share of laboratories with satisfactory results of the FSEQA participation regarding smear microscopy increased to 92.8%; the share of laboratories with the confirmed quality increased to 14.3%. The culture coverage of new pulmonary TB patients equaled to 98.0%. Culture-based confirmation of the diagnosis was obtained in 55.3% of the patients; the share of smear positive new pulmonary TB patients with the negative culture result decreased to 3.8%. The share of laboratories participating in FSEQA increased to 44.5%; the share of laboratories that showed satisfactory results - up to 90.8%, the share of laboratories with the confirmed quality - up to 40.4%. The coverage with molecular genetic testing added up to 90.8%. The drug susceptibility testing (DST) coverage equaled to 97.7%. The patient coverage with rifampicin resistance DST to fluoroquinolones reached 93.2%; coverage with pre-XDR-TB DST to linezolid - 35.0%, bedaquiline - 12.4%. Conclusion. Microbiological diagnostics of tuberculosis in PHC by smear microscopy is ineffective in modern conditions; there are prospects for its use in new Russian regions. The COVID-19 pandemic failed to produce any significant negative impact on the patient coverage with microbiological examination methods. In 2020–2021, the quality of bacterioscopic and culture testing for tuberculosis increased. DST coverage for bedaquiline and linezolid is poor; this does not allow to meet the current clinical guideline requirements. The cost of consumables per patient increased 3.1 times between 2015 and 2021.In the structure of financial expenses on TB microbiological diagnostics, consumables that require import substitution prevail. Scope of application. The study results can be used by heads of health authorities, chief physicians of TB facilities, heads of microbiological laboratories in order to increase effectiveness of TB care, timely detection of people excreting bacteria and MBT drug susceptibility testing.

Rapid diagnosis of TB using Aptamer-based assays for Mycobacterium tuberculosis antigens in children and adolescents.

Despite advances establishing microbiological evidence of tuberculosis (TB) is still a concern in children due to the limitation of availability of sample and predominance of extrapulmonary TB, there is unmet need for diagnostic tests which are low cost, rapid and sensitive and specific. This study evaluated the utility of aptamer-based assay for detecting mycobacterium tuberculosis antigens HspX and MPT 64 in rapid diagnosis of TB in children up to 18 years of age in a tertiary medical college. A total of 100 children were sequentially enrolled with presumptive pulmonary (n = 52 and extrapulmonary n = 48) TB based on clinico-radiological characteristics. The samples were evaluated with ALISA technique for TB antigens and compared with the results of ZN microscopy, GeneXpert and mycobacterial culture MGIT. The enrolled children had mean age (11.7 + 4.4 years) with both pulmonary (n = 52) and extrapulmonary TB (n = 48). Our study results concluded poor results of smears (11% positivity, sensitivity: 17.7%, NPV: 42.7%) and better of GeneXpert (positivity: 42%, sensitivity of 67.4%, NPV: 65.5%) and culture (positivity 57%, sensitivity 91.9%, NPV 88.3%). HspX antigen by ALISA had comparable results (positivity: 49%, sensitivity: 62.9%; NPV: 54.9%). MPT 64 antigen by ALISA also had similar results (positivity: 45%, sensitivity: 58% and NPV 52, 3%). Sensitivity and specificity were higher in pulmonary TB compared to EPTB for both antigens. HspX antigen assay by ALISA and MPT 64 ALISA over existing microbiological diagnostic methods had additional of 13%. ALISA technique for mycobacterium antigens HspX and MPT 64 was rapid, low-cost test (1-3$/test) high sensitivity and specificity and comparable to currently available methods.

About the use of antibiotics as a prevention of inflammatory complications in patients during dental implantation

Dentists often resort to systemic antibiotics to minimize the risk of inflammatory complications when performing dental implant surgery Literature data are ambiguous: a number of studies prove the need for systemic prescription of antibiotics for dental implantation, others say that antibiotic prophylaxis is an excessive measure that does not affect the outcome of treatment when other aseptic and antiseptic rules are followed.
 Materials and methods. The search was carried out on the online platforms Scopus, Web of Science, PubMed, Cochrane, ResearchGate, Online library, Elibrary. For the review, a search was performed on articles, mainly from the last 10 years, using randomization of patients, with a minimum follow-up period of 1 year. We compared 3 strategies for patient management: without antibiotic prophylaxis, using a prophylactic dose of the antibiotic amoxicillin, using a prolonged dose of amoxicillin. Clinical signs of wound healing, general well-being of the patient were assessed.
 Results. It has been shown in clinical practice that prescribing antibiotics at the time of dental implant placement is recommended, as this approach increases the likelihood of success and makes treatment more effective, but attention should be paid to rational antibiotic prescribing, with an adequate number of days of drug administration, to reduce the likelihood of the formation of antibiotic-resistant strains of microorganisms and minimize side effects. Most scientific studies indicate that the appointment of an antibiotic in the postoperative period does not increase the effectiveness of treatment in a contingent of patients without a burdened anamnesis.
 Conclusion. The results of the research allow to substantiate the advantages of perioperative antibiotic prophylaxis carried out before the dental implantation. A prophylactic dose of the antibiotic amoxicillin is the dominant approach for the prevention of early inflammatory complications of dental implantation. The wider use of microbiological diagnostic methods (including PCR diagnostics) to identify the carriage of representatives of periodontopathogenic bacteria in preparing a patient for dental implant surgery allows optimizing the indications. In this case, perioperative prophylaxis should be recognized as necessary.

Research on the Use of Thermal Imaging as a Method for Detecting Fungal Growth in Apples

This study assesses whether thermal imaging can be utilized for detecting and monitoring the level of plant product contamination with apples used as an example. The growth of Penicillium expansum, Botrytis cinerea, and Rhizopus stolonifer on apples contaminated with these fungi was investigated by measuring temperature changes using a thermovision camera. The results showed a significant relationship between the temperature of apples and the growth of microorganisms, as well as that the temperature changes occurred in two stages (temperature of the contaminated apples increased on the first day but then decreased). Significant differences were found between the temperature of the apples showing microbial growth and the noncontaminated control sample, which indicates that the thermal imaging technique has a potential application in microbial quality control. Automation of the production process and attempts on the intensification of production capacity have resulted in the need to improve individual stages of product quality control. Thermovision-based methods have a high potential in this field, as they represent an innovative and noninvasive alternative to conventional microbiological diagnostic methods.

Etiological Characteristics in Causative Agents of Infectious Complications in Cancer Patients

Background . Infectious complications are the most frequent and severe among all complications in cancer patients. The development of fungal-bacterial infections is particularly difficult. The aim was to develop an etiological description for the causative agents of infectious complications in oncological patients. Materials and methods . The study was conducted from January, 2020 to December (inclusive both dates), 2021. 3662 hospitalized patients (men and women) aged from 1 to 85 years with clinical manifestations of an infection were examined. The traditional microbiological method was used to clarify the etiology of the infectious process and to verify the pathogen. Phenotypic and real-time PCR methods were used for detection of carbapenemase and ESBL production. Results . The organs of the respiratory system, skin, soft tissues, and blood are most susceptible to infectious complications. In the etiology of infectious complications, the leading place is occupied by gram-negative bacteria. The conducted microbiological study made it possible to establish that resistance to cephalosporins and carbapenems in enterobacteria is realized through various resistance mechanisms. Extended-spectrum β-lactamases and carbapenemases are particularly relevant and concerning. Monitoring revealed the growth of carbapenemase-producing isolates of Klebsiella pneumoniae (34.9%), the spread of carbapenemase among Escherichia coli strains (5.1%), the increase in the spread of Acinetobacter baumannii strains with the production of NDM groups (32.6%). Conclusion . Prescription of antibacterial drugs should be based only on the results of microbiological diagnostics. The current situation requires modern methods of microbiological diagnostics: the use of only up-to-date (periodically updated) criteria for determining sensitivity to antimicrobial drugs, as well as the introduction of phenotypic tests that provide reliable information without molecular genetic methods.

The Evaluation of the Diagnostic Performance of the BioFire FilmArray Meningitis/Encephalitis Panel in Children: A Retrospective Multicenter Study

Abstract Objective Acute bacterial meningitis (ABM) declined after implementing conjugate Haemophilus influenzae type B and the pneumococcal vaccines worldwide. However, it still contributes to significant morbidity and mortality. The Biofire FilmArray Meningitis Encephalitis (FAME) panel can rapidly diagnose common bacterial and viral pathogens. Several studies suggested that the use of FAME may accelerate diagnosis and decrease the time to pathogen-specific therapy. However, the clinical utility is still controversial due to scarce data and relatively high costs. Therefore, we aimed to evaluate the diagnostic performance of FAME in children. Methods A retrospective multicenter cross-sectional study was conducted to evaluate FAME in diagnosing ABM in children with a suspected central nervous system infection between January 2017 and May 2021. Results This study consisted of 179 children diagnosed with central nervous system infection who had parallel testing done using FAME and traditional microbiological diagnostic methods. Twenty-two FAME results were positive; 8 (36.3%) were bacterial pathogens and 14 (53.7%) were viral pathogens. The most common viral pathogen was human herpesvirus 6 (n = 6; 27.2%), followed by herpes simplex virus 1 (n = 4; 18.1%), Enterovirus spp. (n = 2; 9%), Parechovirus (n = 2; 9%), and Cytomegalovirus (n = 1; 4.5%). Bacterial pathogens included S. pneumoniae (n = 3; 13.6%), H. influenzae (n = 3; 13.6%), Neisseria meningitidis (n = 1; 4.5%), and Streptococcus agalactiae (n = 1; 4.5%). Bacterial culture confirmed S. pneumoniae infection in only 1 of 8 (12.5%) patients, while 7 of 8 bacterial meningitis were only detected by FAME. Conclusion FAME may also help with diagnosis and pathogen identification in patients who have already had antibiotics before cerebrospinal fluid collection. The use of FAME to detect infections quickly may minimize the improper use of medications, treatment duration, and the cost of hospitalization.

Detection of Streptococcus suis using the optimized real-time polymerase chain reaction protocol

The article presents the results of studies on the detection of Streptococcus suis by real-time polymerase chain reaction. Isolation and species identification of the studied isolates of streptococci was carried out according to morphological, cultural, biochemical and biological properties by conventional methods. The study of cultural characteristics of growth was carried out using conventional bacteriological methods on the brain heart infusion broth (BHI) and BHI agar with the addition of 5% sheep blood (blood BHI agar). To confirm biochemical properties as a confirmatory method, API 20 STREP test kit (bioMerieux, France) was used. In addition, to differentiate S. suis from the non-pathogenic species of streptococci, the hemolysis test was used. As a result of the studies, it was found that the use of the real-time PCR (polymerase chain reaction) method makes it possible to detect S. suis in an amount of 1 x 104 genome copies in the sample. All described validation parameters for the qualitative detection of S. suis DNA by real-time PCR meet international requirements, which guarantees accurate and reliable results. In Ukraine only a diagnostic test kit for convential PCR has been developed for the detection of swine streptococcosis. This approach is more time consuming and complex in comparison with the real-time PCR approach. We recommend that diagnostic laboratories implement this method in their practice. This will increase the number of effective diagnostic tools available to veterinarians on pig farms when they order laboratory tests. The high analytical sensitivity limit of a test is an essential parameter when screening is the focus, and obtaining false negative results causes a risk of the development of infection process among pig populations within infected herds. Our study showed that microbiological diagnostic methods to determine morphological and cultural properties can identify S. suis at the genus level. Determination of biochemical properties using the API 20 STREP test kit can be used to identify S. suis 1 and 2 serotypes. The conventional method and real-time PCR have 100% specificity and can be used to identify S. suis of different serotypes. Real-time PCR is a 2 to 4 times more sensitive limit than conventional PCR depending on the serotype being studied, and can be used to more accurately identify streptococcal DNA. It was found that the use of the real-time PCR method makes it possible to detect S. suis in an amount of 1 x 104 copies of the genome in the sample. Additionally, it was found that all the studied validation parameters of the qualitative method for determining S. suis DNA by real-time PCR meet international requirements, which guarantees accurate and reliable results.

Microbiological Laboratory Diagnosis of Human Brucellosis: An Overview.

Brucella spp. are Gram-negative, non-motile, non-spore-forming, slow-growing, facultative intracellular bacteria causing brucellosis. Brucellosis is an endemic of specific geographic areas and, although underreported, represents the most common zoonotic infection, with an annual global incidence of 500,000 cases among humans. Humans represent an occasional host where the infection is mainly caused by B. melitensis, which is the most virulent; B. abortus; B. suis; and B. canis. A microbiological analysis is crucial to identifying human cases because clinical symptoms of human brucellosis are variable and aspecific. The laboratory diagnosis is based on three different microbiological approaches: (i) direct diagnosis by culture, (ii) indirect diagnosis by serological tests, and (iii) direct rapid diagnosis by molecular PCR-based methods. Despite the established experience with serological tests and highly sensitive nucleic acid amplification tests (NAATs), a culture is still considered the “gold standard” in the laboratory diagnosis of brucellosis due to its clinical and epidemiological relevance. Moreover, the automated BC systems now available have increased the sensitivity of BCs and shortened the time to detection of Brucella species. The main limitations of serological tests are the lack of common interpretative criteria, the suboptimal specificity due to interspecies cross-reactivity, and the low sensitivity during the early stage of disease. Despite that, serological tests remain the main diagnostic tool, especially in endemic areas because they are inexpensive, user friendly, and have high negative predictive value. Promising serological tests based on new synthetic antigens have been recently developed together with novel point-of-care tests without the need for dedicated equipment and expertise. NAATs are rapid tests that can help diagnose brucellosis in a few hours with high sensitivity and specificity. Nevertheless, the interpretation of NAAT-positive results requires attention because it may not necessarily indicate an active infection but rather a low bacterial inoculum, DNA from dead bacteria, or a patient that has recovered. Refined NAATs should be developed, and their performances should be compared with those of commercial and home-made molecular tests before being commercialized for the diagnosis of brucellosis. Here, we review and report the most common and updated microbiological diagnostic methods currently available for the laboratory diagnosis of brucellosis.

Stroke in a Patient with Recurrent Meningeal Tuberculosis and HIV Coinfection

Recurrent tuberculous meningitis can have a non-specific clinical presentation when it presents as a co-infection with HIV and can resemble other causes of subacute and chronic meningitis, leading to unwanted outcomes; for this reason, timely diagnoses are required. The most widely used microbiological diagnostic methods can be of low sensitivity or have delayed results. The aim of this article is to present a clinical case of recurrent tuberculous meningitis with a clinical presentation of stroke due to a basal cistern ischaemic lesion in a patient co-infected with HIV, who had a microbiological confirmation and a drug sensitivity study with phenotypic and molecular tests.

Molecular Detection of Bacterial Agents of Atypical Pneumonia: Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophila in Suez Canal region

Background: Atypical bacterial infections played an important role in community-acquired pneumonia (CAP). It is difficult to detect atypical pathogens by conventional microbiological diagnostic methods. Atypical bacteria do not respond to beta-lactam antibiotics. The use of multiplex polymerase chain reaction (PCR) methods enables rapid and simultaneous detection of many pathogens in a single analysis. Aim: detecting the prevalence of atypical bacterial pathogens as etiologic agents of atypical pneumonia, in the Suez Canal region. Materials and Methods: This cross-sectional descriptive study was conducted throughout 18 months, from October 2018 to April 2020. It included 84 Egyptians suffered from CA atypical pneumonic patients of all age groups from Suez-Canal region, Egypt. Sputum samples were collected for identification of Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella pneumophilia by using multiplex PCR. Results: Among the 84 atypical pneumonia patients, L. pneumophila were detected in 12 (14%) patients. M. pneumoniae and C. pneumoniae were not detected in our samples. Compared with L. pneumophila -negative cases, L. pneumophila -positive cases were more prevalent in middle aged males, smokers, COPD, diabetic and asthmatic patients (P values = 0.048). Persistent cough, elevated levels of C-reactive protein (C-RP), bilateral pulmonary infiltration are significant clues for predicting L. pneumophila pneumonia. Conclusions: L. pneumophila incidence is not low in our geographical region in atypical pneumonia patients. Clinicians should consider atypical bacterial pathogens while prescribing antimicrobial management plan. Keywords: Atypical pneumonia, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophilia, multiplex-PCR.

Clinical metagenomics assessments improve diagnosis and outcomes in community-acquired pneumonia

BackgroundIdentifying the causes of community-acquired pneumonia (CAP) is challenging due to the disease’s complex etiology and the limitations of traditional microbiological diagnostic methods. Recent advances in next generation sequencing (NGS)-based metagenomics allow pan-pathogen detection in a single assay, and may have significant advantages over culture-based techniques.ResultsWe conducted a cohort study of 159 CAP patients to assess the diagnostic performance of a clinical metagenomics assay and its impact on clinical management and patient outcomes. When compared to other techniques, clinical metagenomics detected more pathogens in more CAP cases, and identified a substantial number of polymicrobial infections. Moreover, metagenomics results led to changes in or confirmation of clinical management in 35 of 59 cases; these 35 cases also had significantly improved patient outcomes.ConclusionsClinical metagenomics could be a valuable tool for the diagnosis and treatment of CAP.Trial registrationTrial registration number with the Chinese Clinical Trial Registry: ChiCTR2100043628.

COVID-19 Pandemisinde Mikrobiyolojik Tanı Yöntemleri

COVID-19 pandemic that caused the death of more than one million people worldwide, has clearly demonstrated the critical importance of microbiological diagnostic methods in preventing infectious diseases and controlling outbreaks. The most effective way for combating SARS-CoV-2, is reducing the spread of the virus as much as possible. To decrease the spread of the virus is only possible with diagnostic methods that enable the detection of all symptomatic or asymptomatic patients infected with the virus accurately and as early as possible. Various microbiological methods (serological tests, nucleic acid amplification methods) recommended by international health authorities are widely used in the diagnosis and follow-up of COVID-19 disease. The challenges experienced in application of diagnostic methods, including real-time polymerase chain reaction (RT-PCR) method, which is specified as the gold standard method in the diagnosis of COVID-19 by the World Health Organization, were once again acknowledged during the pandemic. To achieve higher success rates in combating infections, it is necessary to better determine the application and evaluation standards in current microbiological diagnostic methods, to develop new cost-effective methods that are easily applicable and yielding results within a short time.

Role of medicinal plants in the treatment of eumycetoma: A review

Mycetoma is a resultant infection of either fungi (eumycetoma) or bacteria (actinomycetoma). It is categorized under neglected tropical diseases, as it typically affects the economically backward communities of the endemic regions. The infection occurs with the invasion of etiological agents through the open wounds of the body. The cutaneous and subcutaneous tissue is characterized by the sinus formation and discharge. Primarily affecting the foot, it further spreads to other parts of the body. Eumycetoma has a worldwide distribution yet considered endemic, as it is restricted only to few countries. Diagnosis and treatment of the disease has been burdensome since poor response and unavailability of medical support in endemic regions. Microbiological and serological diagnostic methods have been unreliable except molecular diagnostics. Treatment with antifungal agents has given a mixed response, with only itraconazole being effective. These techniques are relatively expensive and cannot be afforded by people with weak economic background. In the context, it becomes essential to introduce phytotherapy to relieve the health and economic burden. Herein, we discuss few medicinal plants with profound antifungal activity towards eumycetoma. We highlight the possible course of actions that needs to be put forward to deliver the plant-based drugs to cure eumycetoma.

Effects of Ciprofloxacin on the Dynamics of Biofilm Formation by Staphylococcus Epidermidis Strains Isolated from Implant-Associated Infection

Introduction. The formation of a microbial biofilm in implant-associated infection after arthroplasty of large joints reduces the informative value of traditional microbiological diagnostic methods and limits the range of effective antimicrobial drugs. When prescribing etiotropic therapy for periprosthetic joint infection, it is necessary to take into account not only the antibacterial effect of the drug, but also its effect on biofilm formation. Ciprofloxacin therapy may be a risk factor for the development of biofilm periprosthetic infection caused by multidrug-resistant staphylococcal strains.The aim of research was to study the effect of sub-inhibiting and therapeutic doses of ciprofloxacin on biofilm formation by Staphylococcus epidermidis strains isolated from implant-associated infection.Materials and methods. The authors studied the effect of various concentrations of ciprofloxacin on 15 strains of St. epidermidis isolated from 83 patients with deep periprosthetic joint infection after primary knee arthroplasty, treated at NIITON SSMU in 2018-2019. The effect of the calculated concentrations of ciprofloxacin on the planktonic culture, forming and preformed biofilms was investigated. Biofilm simulation was performed according to the method described by G.D. Christensen under in vitro conditions with determination of the optical density of alcohol eluates of gentian violet in polystyrene microplates.Results. It was demonstrated that ciprofloxacin in a dose 0.01 g/ml inhibits the growth of planktonic forms by 50% and statistically significantly (p = 0.001) stimulates formation of microbial biofilms as compared to the control without antibiotic addition. Concentration of ciprofloxacin equal 0.03 g/ml inhibits the growth of planktonic forms by 90%, statistically significantly (p = 0.001) stimulates formation of biofilms and activates further increase in the mass of pre-formed microbial biofilms. An increase in the concentration of ciprofloxacin to 0.05 g/ml completely inhibits the growth of planktonic forms and statistically significantly stimulates further growth of preformed biofilms.The use of ciprofloxacin at concentrations equal 1-3 g/ml statistically significantly (p = 0.001) inhibits the formation of microbial biofilms, but does not affect the preformed biofilm.Conclusions. Thus, there has been found a dose-dependent effect of ciprofloxacin towards clinical strains of St. epidermidis: subinhibitory and therapeutic concentrations of the drug have a stimulating effect on the formation and further increase in the mass of the preformed microbial biofilms. This fact must be taken into account when prescribing etiotropic therapy for implant-associated complications following large joint replacements.

Vasculitis due to Candidatus Neoehrlichia mikurensis: A Cohort Study of 40 Swedish Patients.

Candidatus (Ca.) Neoehrlichia (N.) mikurensis is an emerging tick-borne pathogen of humans that is closely related to Ehrlichia and Anaplasma species. This strict intracellular bacterium escapes detection by routine microbiologic diagnostic methods such as blood culture, leading to considerable under-diagnosis of the infectious disease it causes, neoehrlichiosis. Here, we describe the vascular and thromboembolic events afflicting a series of 40 patients diagnosed with neoehrlichiosis in Sweden during a 10-year period (2009-2019). The majority of the patients (60%) developed vascular events ranging from repeated thrombophlebitis, deep vein thrombosis, pulmonary embolism, transitory ischemic attacks, to arteritis. Younger age was a risk factor for vascular complications. In contrast, there was no difference in the incidence of vascular events between immunosuppressed and immunocompetent patients. However, there were qualitative differences, such that deep vein thrombosis exclusively afflicted the immunosuppressed patients, whereas arteritis was restricted to the immunocompetent persons. We also present the case histories of two patients who developed vasculitis mimicking polyarteritis nodosa and giant cell arteritis. Both were cured by doxycycline treatment. Ca. N. mikurensis infection should be considered in patients living in tick-endemic areas of Europe and northern Asia who present with atypical vascular and/or thromboembolic events. Early diagnosis and antibiotics targeting this emerging infectious agent can eradicate the infection and prevent the development of new vascular events.