Resident Microflora Research Articles

Impact of Bacterial Etiology on Procalcitonin, C-reactive Protein and Hematological Parameters: Evaluating Mean Platelet Volume for Differentiating Gram-Negative and Gram-Positive Bacteria in Odontogenic Versus Non-odontogenic Head and Neck Abscesses.

Introduction Head and neck abscesses, which can originate from odontogenic or non-odontogenic sources, pose significant diagnostic challenges due to their diverse bacterial etiologies. This study aims to investigate the impact of bacterial etiology on procalcitonin (PCT), C-reactive protein (CRP), and various hematological parameters, and to assess the diagnostic performance of mean platelet volume (MPV) in differentiating between Gram-negativebacteria (GNB) and Gram-positive bacteria (GPB) in adults with odontogenic and non-odontogenic head and neck abscesses. Materials and methods Our retrospective analysis of a prospective study comprised 80 patients: 50 individuals (56% men, average age 41.6±18.18 years) with odontogenic and 30 patients (66.7% men, average age 44.53±15.49 years) with non-odontogenic head and neck abscesses during the period from July 2021 to June 2022. White blood cell count (WBC); neutrophil (Neu) and lymphocyte (Ly) count; MPV, and platelet count (PLT) were derived from the results of complete blood count. MPV/PLT (MPI) was calculated by dividing MPV by PLT. CRP levels (mg/l) were quantified via immunoturbidimetric analysis utilizing latex-enhanced particlesand PCT levels (ng/ml) by latex-enhanced immunoturbidimetric assay. Results In 25 (31.3%) of all 80 patients, no microorganisms were isolated (sterile cultures); in 28 (35%) resident microflora were isolated; in seven (8.8%) GNB were isolated; and in 17 (21.3%) GPB were isolated. CRP and Neu were significantly higher in patients with odontogenic abscesses compared to non-odontogenic ones. PLT and PCT were lower in patients with odontogenic abscesses vs those with non-odontogenic abscesses. Additionally, according to bacterial type, MPV, MPI and PCT were significantly higher in GPB compared to GNB. WBC, Neu and PLT were higher in patients with GNB vs GPB. Significant correlations were found between MPV andLy, and between MPV and Neu, regardless of the abscess origin or etiological factor. MPI exhibited an area under the curve of the receiver operating characteristic (AUC-ROC)=0.776, MPV of 0.541, and PCT of 0.568in distinguishing patients with GPB from GNB. A cut-off value of 0.029 was derived for MPI (70.6% sensitivity and 80% specificity). Conclusions This study highlights the impact of bacterial etiology on inflammatory and hematological markers in head and neck abscesses. Odontogenic abscesses showed higher CRP and Neu, indicating a stronger inflammatory response, while non-odontogenic abscesses had higher PLT, Ly, and PCT. MPI proved to be a more effective diagnostic marker (cut-off value of 0.029) than MPV or PCT for distinguishing between GPB and GNB, suggesting its valuable role in clinical practice for accurate and timely diagnosis.

Surgical antisepsis in Russia: historical parallels

The origins of surgery and the improvement of surgical practices did not always guarantee a favorable surgical outcome. Even a masterfully performed operation could lead to a purulent-inflammatory process. Even at the stage of empirical development of medicine, before the great microbiological discoveries, an understanding was formed about a certain “substrate” that penetrates the surgical wound from the external environment and causes its suppuration. Supporters of the contagionist theory called it contagium, and supporters of the miasmatic theory considered the external (air) environment as such a source. The ideas of the supporters of the contagionist theory of the occurrence of purulent-septic complications in surgery in the middle of the 18th century are reflected in the works of D.S. Samoilovich and I.F. Semmelweis; the latter proved the transmission of the infectious principle by the surgeon’s hands, which became the starting point for the development of preventive antiseptics. In historical retrospect, the technology and means used for surgical hand antisepsis in Russia were analyzed from the mid-18th to the early 20th centuries. Moreover, literary sources for the period from 1783 to 2023 were analyzed, containing information about the antiseptic treatment of surgeons’ hands. Historical, descriptive, evaluative, analytical, and bacteriological research methods were used. As an experiment, the original antiseptic recipe described by D.S. Samoilovich (1783) was restored, and its effectiveness was confirmed. During surgical practices in Russia, starting from the mid-18th century, facts about the use of drugs with antiseptic activity for hand treatment are described. It is shown how, in historical retrospect, views on the final result of surgical antisepsis changed: from partial to complete destruction of microorganisms, preventing their entry from the deep layers of the skin, to the destruction of transient microflora and reduction of the abundance of resident microflora. Despite the influence of the English (J. Lister) and German (P. Fürbringer, J.F. Ahlfeld) surgical schools on the development of antiseptics in Russia, it retained its originality, determined by the needs of society and the possibility of using certain chemical antiseptics and their compositions for its implementation. The contribution of domestic surgeons to the improvement of antiseptic technologies and drugs in military field conditions is shown (N.I. Pirogov, A.M. Zabludovsky, D.I. Tatarinov et al.).

The influence of different methods of decontamination of microbial biofilms formed on eggshells

According to "food legislation" requirements, all eggs entering the production of egg products must be disinfected. Therefore, developing technologies for decontaminating chicken eggs before use for food purposes is a promising work direction in chicken egg production and storage. This research aimed to identify the microbiota of chicken eggs with varying degrees of shell contamination and determine the influence of different methods of decontaminating microbial biofilms formed on eggshells. It was set up that the quantitative content of microorganisms on the surface of chicken eggs ranged from 103 CFU to 106 CFU/ml of washing and depended on the contamination of the shell with droppings. Lactobacillus spp., Bacillus spp., Corynebacterium, Staphylococcus were among the genera of bacteria that prevailed on the clean chicken shell, which were isolated in 30-50% of cases, and gram-negative microbiota was practically absent. The constant release of gram-positive bacteria is noted on the contaminated eggshell, and the frequency of identification of gram-negative microbiota of the Enterobacteriaceae genus and non-fermenting genera Pseudomonas and Psychrobacter increases. That is, the microbial scape of the microbiota of the chicken shell depends on its cleanliness, and the presence of a dirty surface increases the frequency of allocation of the resident microflora of the gastrointestinal tract. It was found that the working solution of the disinfectant Vircon S destroyed planktonic bacteria applied to the eggshell in an average of 2 minutes of exposure, stabilised water ozone for 1 minute, gaseous ozone for 3 minutes, and the action of ultraviolet rays with a length of 253.7 nm for 25-30 min. At the same time, using these disinfection methods on bacteria formed in a biofilm on the eggshell did not cause a bactericidal action during this time. To significantly reduce bacteria in the biofilm using these methods, it is necessary to increase the exposure time of the biocide by 2-3 times. Therefore, the complex structure of the eggshell and the multi-layered matrix of biofilms provide better protection for bacteria against the influence of the investigated disinfection methods.

Isolation, Characterisation and Identification of Bacteria from Three Different Unpasteurized Fruit Juices Sold at a Local Market in Guyana

Fruit juices are widely consumed for their nutritional value. However, fruit juices that are unpasteurised may be unsafe to drink because they are not subject to heat processing and much of their human handling results in contamination. The fruits used to make these juices have resident microflora, transient bacteria from handling and equipment, and bacteria from water used to wash fruits and in some cases make the juice. The objective of this research was to determine the bacteria present in unpasteurised passionfruit, cherry, and tamarind juices respectively. Through the use of a survey, and standard methods for isolation, identification and characterisation of bacteria, the results showed that temperature and pH did not significantly affect the diversity of bacteria. Cherry juice had the highest number of Colony Forming Units while passion fruit juice had the lowest. In total seven (7) species of bacteria were found including E.coli, S. aureus, Lactobacillus sp, Acetobacter sp, Bacillus subtilis, Bacillus cereus, and Micrococcus. The results for the Colony Forming Units were found to be above maximum acceptable levels for fruit juices. The results of this study demonstrate that unpasteurised juices may be a threat to public health and hazardous to humans if not processed in a sanitary manner.

The enhanced effect of key microorganisms in chromium contaminated soil in Cr(VI) reduction

The escalating threat of Cr(VI) pollution to the environment and human health can be effectively controlled through microbial methods, which are promising, safe, and ecofriendly. To enhance Cr(VI) removal efficiency, scholars have been optimizing strains. However, synergies between in-situ soil particles and crucial microorganisms in soil have rarely been investigated. In this study, Cr(VI) was removed by collaborating with in-situ soil particles and key microorganisms in the soil. The results indicated that within 48 hours, the removal rate of Cr(VI) reached over 99% in the soils+microflora system, which was 45% higher than that of the microflora system alone. Factors such as Cr(VI) concentration, soil dosage, pH level, oxygen availability, and electron donors influenced the removal efficiency of Cr(VI) in the soils+microflora system. The cyclic experiments showed that soil particles effectively prevented chromium invasion on microflora, promoting the growth of crucial microorganisms. The addition of microflora can effectively regulate the composition of soil flora and enhance the efficiency of chromium reduction. Moreover, two strains each of Ochrobactrum sp. and Paenarthrobacter sp., exhibiting remarkable tolerance to Cr(VI), were successfully isolated from these soils, significantly enhancing the reduction capacity of the indigenous microflora towards Cr(VI). Additionally, 16S rRNA-PCR sequence analysis revealed that in-situ soil particles not only synergistically collaborated with the resident microflora for efficient removal of Cr(VI), but also facilitated the proliferation of key microbiota such as Ochrobactrum sp. and Paenarthrobacter sp. Remarkably, when exposed to an initial concentration of 50 mg/L Cr(VI), complete removal was achieved by Paenarthrobacter-2 within a time frame as short as 60 hours. This research found four novel highly efficient strains for reducing Cr(VI) and provides an innovative method for the synergistic interaction between indigenous soil microflora and soil particles to remove heavy metal ions from wastewater.

Antibiotic resistance among Aerobic Gram-Negative Bacilli isolated from patients with oral inflammatory dysbiotic conditions—a retrospective study

IntroductionAerobic Gram-Negative Bacilli (AGNB) are not part of the resident oral microflora but are occasionally found in high abundance under inflammatory dysbiotic conditions at various oral niches. The aim of the present study was to investigate the identity and antibiotic susceptibility of AGNB isolated from patients in Sweden with mucosal lesions, periodontitis, and peri-implantitis, with special attention to antibiotic resistance and on the presence of phenotypic Extended Spectrum Beta-Lactamase (ESBL) isolates.Materials and methodsMicrobiolgical samples were harvested from 211 patients in total, experiencing mucosal lesions (N = 113), periodontitis (N = 62), or peri-implantitis (N = 36). The growth of AGNB's was semiquantified by selective and non-selective culture and the strains were isolated, identified, and tested for antibiotic susceptibility. A total of 251 AGNB strains, occurring in moderate to heavy growth (>100 CFU/ml sample), indicating a dysbiotic microbiota, were identified. The disc diffusion method was used for screening of the antibiotic susceptibility of the isolates. Phenotypic identification of ESBL isolates was based on resistance to ceftazidime and/or cefotaxime.ResultsThe most commonly detected AGNB isolates in oral inflammatory dysbiotic conditions were fermentative species belonging to Enterobacteriaceae e.g. Citrobacter spp., Enterobacter spp., Escherichia coli, Klebsiella spp, and the non-fermentative environmental Burkholderia cepacia, Pseudomonas spp., and Stenotrophomonas maltophilia. No clear trends were seen in frequency of the various species in samples from mucosal lesions, severe periodontitis, and peri-implantitis cases. The 138 Enterobacteriaceae isolates and 113 environmental AGNB isolated showed a high antibiotic resistance in general against antibiotics commonly used in dentistry (Amoxicillin, Amoxicillin + Clavulanic acid, Ampicillin, Clindamycin, Doxycycline, Erythromycin, Oxacillin, PenicillinV, and Tetracycline). The majority of these isolates were susceptible to ciprofloxacin. Ten isolates (4.1%) were phenotypically classified as ESBL positive. The ESBL isolates were predominantly found among isolates of S. maltophilia, while only one ESBL positive isolate was found among Enterobacteriaceae.ConclusionsPhenotypically identified ESBL isolates can occasionally be present among oral AGNB strains isolated in abundance from the dysbiotic microbiota occurring in cases with oral mucosal lesions, severe periodontitis, or peri-implantitis.

The place and role of oral microflora in the pathogenesis of oral mucositis in malignant neoplasms (literature review)

Background. Oral mucositis (OM) is one of the most frequent complications of systemic drug therapy of oncological diseases. Clinically, the disease manifests itself in the form of erythema and ulceration of the non-corneating mucous membrane. Although this condition is self-healing, it can affect the conduct of comprehensive antitumor treatment. Currently, the pathophysiological model of the development of OM is supplemented with new data obtained during studies of the oral microflora. Antitumor treatment can lead to changes in the composition of the resident oral microflora and it is quite possible that the changes that occur may affect the development of damage to the oral mucosa. The expansion of knowledge in this area allows researchers to look for new ways to include oral microbiota in OM management strategies.
 Aim. To summarize and update the accumulated data regarding the role of the oral microbiota in the pathogenesis of OM in patients with malignant neoplasms.
 Materials and methods. Literature search was performed in Medline, Cochrane Library, Elibrary and Pubmed, including publications demonstrating the current ability to assess the impact of the oral microbiome on OM, as well as developments in this area relating to OM management.
 Results. In the review the current views on the position of the oral microbiome in the pathogenesis of OM were highlighted and the promising directions for the use of agents influencing the state of the oral microbiota in various strategies to control this disease were emphasised.
 Conclusion. A better understanding of the pathogenesis of OM and the inclusion of new factors, such as the oral microbiome, into the picture of pathogenesis is likely to enable the formation of more effective management strategies for this disease in the future. This direction seems very promising, and developments in this area show promising results.

Nanobiomaterials in the Management of Oral Biofilms

The oral cavity inhabits diverse microorganisms with members of all age groups including children being associated with oral infections. Bacteria being the principle component of this resident microflora along with the diverse group of other species reflect the wide range of viscerously derived nutrients, the heterogenous habitats for colonization contributing to the survival of the biofilm. Biofilms constitute surface- adherent communities of microorganisms like Streptococcus mutans along with other bacterial species embedded in an extracellular matrix material. The commensal relationship between the host and the microflora can be disrupted in numerous ways, resulting in compromised oral health causing conditions like dental caries and endodontic infections, gingival and periodontal conditions, oral candidiasis and peri-implantitis. Biofilm-related infections prove to be a major threat from both an economical and health perspective. Innovative treatment options remain limited despite of advancements in understanding the mechanisms of biofilm formation. The concept of nanotechnology is based on controlling atoms on an individual basis and creating structures less than 100 nm (10_7 m) in size thereby greatly influencing the properties. Incorporation of metals and other nanoparticles with polymers and coating of surfaces provide antimicrobial and anti-adhesive benefits within the oral cavity. Therefore, this paper seeks to throw light into the current advancements in nanoparticle mediated treatment approaches of oral biofilms and its future implications in pediatric dentistry. Keywords: Biofilm, Oral Cavity, Metal Oxides, Nanoparticles, Reactive Oxygen Species, Antibiofilm Efficacy

DEMOGRAPHIC DISTRIBUTION, ETIOLOGICAL BACTERIAL SPECTRUM AND ANTIBACTERIAL TREATMENT OF PATIENTS WITH ODONTOGENIC ABSCESSES AND PHLEGMONS OF THE HEAD AND NECK

Background: Odontogenic abscesses are the most common pathology in maxillofacial surgery. Their frequency is significantly higher in people with poor social status. Materials and methods: The study retrospectively included 81 patients with odontogenic abscesses and phlegmons of the head and neck, hospitalized and operated on as an emergency case. Incision wound secretion for microbiological examination and preparation of an antibiogram were taken from each of them as a standard routine procedure. Results and discussion: The most affected are young patients (till 44 years), followed by patients in middle age (45-59 years). The reason for this is that with age, teeth gradually fall out of the dentition of mature individuals, and with this, the possibility of the occurrence of odontogenic infections decreases. The lower jaw is more affected. A mixed resident microflora comprising more than one bacterial species is isolated in 62% of all patients. Gram-positive bacteria isolated in the samples of the studied patients were coagulase-negative staphylococci (Staphylococcus coagulase negative, CNS), Staphylococcus aureus, Streptococcus anginosus and Streptococcus viridans. Gram-negative bacteria were Escherichia coli, Enterobacter cloacae and Stenotrophomonas maltophilia. Obligate anaerobes were found in only one sample. The isolated fungi were Candida albicans and Candidanonalbicans. Conclusion: In the studied group of patients with odontogenic abscesses, Gram-positive bacteria In the studied group of patients with odontogenic abscesses, Gram-positive bacteria were 2.5 times more than Gram-negative. Single antibacterial therapy is preferable for patients with small-area odontogenic abscesses, while triple antibacterial therapy is recommended for large-area odontogenic abscesses and their phlegmonous forms.

Demographic Distribution and Etiological Bacterial Spectrum of Patients with Non-Odontogenic Abscesses and Phlegmons of the Head and Neck

The aims of the present study are to prove that in non-odontogenic abscesses and phlegmons in the head and neck region, the main microbiological agents are representatives of the oral resident microflora, as in odontogenic purulent inflammatory diseases in the same area, and to determine and compare the demo¬graphic distribution between the two analyzed etiological groups. It includes and studies 57 patients with non-odontogenic abscesses and phlegmons of the head and neck. In all of them, as a routine procedure, mate¬rial was taken from the surgical wound for microbiological examination and the identification of the isolated bacteria was confirmed by a microbiological analyzer “VITEK” (“BioMérieux”, France). After analyzing the obtained results, the goal was confirmed - the largest part is of the resident microflora (59.5%, n=34) of which 50.75% (n=29) mixed resident microflora and 8.75% (n=5) mono infections, followed by represen¬tatives of Gram-positive bacteria (15.75 %, n=9), Gram-negative ones (5.25%, n=3) and obligate anaerobes (1.75%, n=1). In the studied group of patients with non-odontogenic abscesses, Gram-positive bacteria are 3 times more than Gram-negative. The causative agents of phlegmons of non-odontogenic origin in the maxil¬lofacial region are Gram-negative bacteria and obligate anaerobes in a 1:1 ratio. Unlike patients with odonto¬genic purulent diseases, in which the young population under the age of 44 is most affected, non-odontogenic abscesses and phlegmons most often affect mature individuals between the ages of 45 and 59.

METABOLIC CHANGES IN BURNS DURING ORAL ADMINISTRATION OF AEROCOCCUS VIRIDANS IN UNCOMFORTABLE TEMPERATURE CONDITIONS

The organism of farm animals is in an inextricable connection with the environment, with which there is a constant exchange of substances, energy and information, that is, it is an open metabolic system that depends on the conditions of existence. Therefore, it is very important to create the most favorable technologies for exploiting the physiological capabilities of animals in conditions of physiological well-being and epidemiological well-being. This is the foundation for obtaining high-quality and bio-safe animal husbandry products. One of the common negative factors of violations of the physiological needs of the animal body is the effect of low-intensity permanent stress of multivalent etiology. During the development of negative pathophysiological disorders, the quantitative and qualitative composition of the microbiota of the large intestine changes, as a dynamic microbial community of transient and resident populations that develop according to the chemo-static principle of functioning. Restoration and correction of the microbiota of the large intestine and transformation of the disturbed equilibrium of the interpopulation ratio is an effective means of eliminating the negative effects of stress on the animal body. The resident probiotic microflora is represented by a wide variety of prokaryotic microbiota and many reference strains are used for therapeutic and prophylactic purposes. Aerococcus viridans is very promising and effective, which lives in various biotopes of the macro-organism and is an indicator of the physiological well-being of the organism, because it is sensitive to toxic compounds and antimicrobial substances, but with permanent use it exhibits pronounced antagonistic properties against transient microflora with potentially pathogenic properties due to inhibition of their colonization abilities and exhibits detoxification potentials. Ants were used to simulate low-intensity permanent stress. as heat-loving animals sensitive to uncomfortable temperatures of the external environment and placed them in a household refrigerator for ten days at a temperature of 6-8 oC. Probiotic culture of production strain BI-07 Aerococcus viridans in a dose of 2×109 CFU/cm3 was administered orally as an anti-stress factor. A similar experiment was conducted in comfortable conditions, that is, at a temperature of 20-22 oC. Metabolic changes in the body of ants were recorded during a laboratory blood test. Probiotic culture does not affect the normergic processes of the macro-organism and does not change indicators beyond the physiological norm, but when used in conditions of low-intensity temperature stress, it has a positive effect on the adaptation-compensatory mechanisms of the macro-organism and shows protective potential in relation to non-resident microbiota.

Structure of the probiotic market in Russia

The article provides an overview of the Russian market of probiotic products, taking into account the registration of products of domestic and foreign production in the segments of medicines for veterinary use and animal feed additives by conducting a content analysis of the state information system. The increase in consumer preferences in favor of natural products is the main factor influencing the growth of the market. The growing demands of consumers for the maintenance and breeding of animals using biologically active feed additives, including complex functional probiotic products, stimulate the expansion of the market. The demand for veterinary probiotics has also increased due to the global trend of combating antibiotic resistance and the rational use of antibacterial drugs, pesticides and agrochemicals. Rational use of available antimicrobial veterinary drugs is becoming increasingly necessary, taking into account the spectrum of their activity and the profile of antibiotic resistance of the main pathogens, as well as the development of comprehensive animal treatment programs using alternative products. In many countries, scientists recommend the use of probiotics for the preservation and correction of intestinal biocenosis starting from the first hours of animal life. The ways of using probiotics, their component composition, and the types of target animals have expanded. Probiotics are used as preparations for veterinary use, feed additives to correct intestinal dysbiosis in animals, restore the resident microflora of animals, as well as to improve various production indicators.

The Etiological Bacterial Spectrum of Neck Abscesses of Lymph Node Origin - Gram-Positive and Gram-Negative Bacteria.

According to our medical practice as maxillofacial and oral surgeons, operated patients with purulent infections of the cervical lymph nodes are not many in number. On the other hand, the presence of a purulent infection requires not only the surgical evacuation of the pusbut also the application of antimicrobial preparations. This necessitatesgood knowledge of the spectrum of the bacterial causative agents of the disease, the determination and analysis of which is the purpose of this original article. The bacteria studied in 181 patients with a mean age of 26.25 years, ranging between 29 days and 82 years, who underwent surgery for suppurating cervical lymph nodes, were retrospectively analyzed over a period of eight years. No bacteria were found in 69 of them. In 83 (74.11%) of the remaining 112 studied patients, the isolated microorganisms were of the gram-positive spectrum - Staphylococcus aureus (n=34), gram-positive resident microflora represented by more than one bacterial species (n=21), Staphylococcus hemolyticus (n=10), Staphylococcus epidermidis (n=9) and beta-hemolytic streptococci (n=9). Gram-negative bacteria were 25.89% (n=29) - Klebsiella pneumoniae (n=8), Bartonella henselae (n=7), Klebsiella oxytoca (n=6), Enterobacter cloacae (n=5) and Flavimonas oryzihabitans (n=3). No anaerobic and fungal microorganisms were isolated. Therefore, antimicrobial therapy in these patients should be directed against both gram-positive and gram-negative bacteria, which in our study were represented in a ratio of approximately three to onein favor of gram-positive microorganisms. Otherwise, we create a prerequisite for the formation of phlegmon on the neck, which hides real chances for the lives of patients.

Etiological Bacterial Spectrum of Purulent Inflammation of Cervical Lymph Nodes in Children With Unknown Causes of Lymphadenopathy.

In the management of the non-specific purulent inflammation of the cervical lymph nodes in children, not only surgical treatment to evacuate the available pusbut also antibacterial therapy is mandatory. Knowledge of the spectrum of the bacterial causative agents of this disease is of fundamental importance for it. This retrospective study included 66 patients with purulent inflammation of the cervical lymph nodes with a mean age of 5.79 years, ranging from 29 days to 17 years, who were hospitalized for the period 2015-2022 in the three pediatric clinics at the St. Marina University Multispecialty Hospital for Active Treatment in Varna, Bulgaria, and operated by an oral or maxillofacial surgeon, in which for microbiological examination material from the suppurated lymph node was taken and analyzed first by direct microscopy for gram-positivity and then on a biochemical machine identifier VITEK(bioMérieux, Marcy-l'Étoile, France). In all of them, the patients and their parents did not provide anamnestic data explaining the etiology of their infectious diseases of the cervical lymph nodes. There was no clear cause that would explain the occurrence of cervical lymphadenopathy - skin infections, dental diseases, diseases of the ears and upper and lower respiratory tracts, onco-hematological diseases, and others. During the medical examination by pediatricians and surgeons, no entrances to the infection were found, but only local signs of inflammation (redness, swelling, and pain), increased levels of blood markers of inflammation (leukocytes, neutrophils, erythrocyte sedimentation rate, and C-reactive protein) and there was imaging evidence (ultrasound, magnetic resonance imaging, and computed tomography) of a purulent collection in the lymph nodes and/or in the soft tissues surrounding them. Exudate without microbial growth was found in 22 of the cases. The main causative agents of purulent lymphadenopathy are gram-positive bacteria (n=32, 72,73%) - Staphylococcus aureus (n=16), Staphylococcus haemolyticus (n=8), Beta-hemolytic streptococcus(n=2), and gram-positive mixed resident microflora (n=6). However, in 27.27% (n=12) of all 44 patients described in this article with isolated pathogens, cultures were gram-negative. These are Bartonella henselae (n=4), Klebsiella pneumoniae (n=4), Klebsiella oxytoca (n=2), and Flavimonas oryzihabitans (n=2).

ANALYSIS OF MICROBIOCENOSIS OF ORAL FLUID IN PATIENTS WITH DEFECTS OF DENTITION, ZAKARPATTIA REGION RESIDENTS

Dental diseases are among the most frequent diseases of the human body. The microflora of the oral cavity is specific, unlike the microflora of other cavities both in composition, quantity, and functions. The resident microflora will play an important role both in maintaining the normal physiological state of the oral cavity and in the development of dental diseases.
 The aim of the study is to establish insemination with clinically significant microorganisms of oral fluid in patients with defects of dentition, Zakarpattia region residents.
 The species and quantitative composition of microbiocenosis of the oral fluid of 282 (94 men and 188 women) patients with dentition defects, residents of the Zakarpattia region, confirms the formation and progression of inflammatory and inflammatory-dystrophic changes in the periodontal tissues.
 In patients with bilateral end defects according to Kennedy, the predominance of anaerobic microflora was determined in the form of Klebsiella pneumonia (108), Prevotella spp. (7×105), Enterobacter spp. (102), Porphyromonas spp. (5×105), Staphylococcus epidermidis (104) and Staphylococcus aureus (1,2×103), Streptococcus pneumonia (108), Streptococcus viridans (103), Enterobacter spp. (102) and opportunistic microorganisms Candida albicans (104).
 Among patients with unilateral end defects according to Kennedy, Enterococcus faecalis was defined. (103), Staphylococcus epidermidis (102–103) and Staphylococcus aureus (105), Streptococcus pneumonia (105–107), Streptococcus viridans (102–5×105), Stahylococcus haemolyticus (102), Candida albicans (5×102) and Escherichia coli coli (109).
 In patients with included defects in the lateral areas according to Kennedy, there were cultured Klebsiella pneumonia (101–3.5×106), Prevotella spp. (4-5×101), Enterococcus faecalis (107), Peptostreptococcus micros (3.5×103–5×103), Staphylococcus haemolyticus (101–105) and Staphylococcus aureus (101–105), Streptococcus pneumonia (106–108 ), Streptococcus pyogenes (105–109), Candida albicans (102), Escherichia coli (107), Sarcina spp. (103) and Citrobacter spp. (104).
 Microbiocenosis of the oral fluid of the fourth clinical group with included defects in the frontal areas according to Kennedy is presented Lactobacillus acidophilus (108-1011), Streptococcu mitis (108-1011), Bifidobacterium (108-1010), Streptococcus salivarius (1081010), Streptococcus mutans (107-109), Streptococcus pneumonia (104 –108), Streptococcus sanguis (105–107).
 Therefore, in patients with dentition defects, the microflora of the oral fluid confirms the formation of inflammatory and inflammatory-dystrophic processes in the periodontal tissue due to the prevalence of anaerobic flora, while in the presence of inclusions in the frontal area, the prevalence of normal obligate microflora is diagnosed.

Shift in fungal communities associated with Pinus sylvestris stands affected by root rot

Introduction
 Scots pine (Pinus sylvestris) covers large areas in European regions with significant economic importance to Ukrainian forest industry. Root rot caused by the wood-decay fungus Heterobasidion annosum damages both below- and above-ground parts of Scots pines. The disease progress is likely to be affected by reshaping in the forest such as soil properties, vegetation composition, and tree age. These changes are apparently followed up by paralleled shifts in fungal community composition on forest soil with potential feedback on ecosystem functioning.
 The objective of the study was to screen fungal groups associated with the root system of P. sylvestris in stands affected by H.annosum s.s. to better understand the pathogenesis and development of root rot infection, as well as to recognize whether root size and disease severity affect diversity of fungi of the root system in the forest-steppe conditions of Ukraine. The additional object was to study other resident microflora of P. sylvestris root infested by H.annosum s.s. to find out whether the H. annosum s.s. impacts the overall diversity of other fungi.
 Materials and Methods
 The field study was carried out in 2018–2020. Field study sites were pure pine forest stands located in Kharkiv region (compartment 126, subcompartment 7, tract Bugri, Kharkiv Forest Research Station). Wood core and root samples from P. sylvestris were collected from the five infected (50–100 m apart from each other) and five non-infected trees (up to 500 m apart from the infested area and 50–100 m apart from each other). Wood and root samples were used for fungal culturing and direct sequencing using ITS1F and ITS4 primers.
 Results
 In the present study, we tried to evaluate fungal communities across diseased Pinus sylvestris stands and investigated correlations between taxonomic composition and forest health. Not surprisingly, root rot infestation had a significant effect on root-associated fungal abundance and diversity. During disease development, the root-associated fungal community shifted in composition from dominance by saprotrophic fungi to ectomycorrhizal and pathogenic fungal species. Our results suggested that maintenance of functional diversity in the root-associated fungal community may sustain long-term forest health or even root rot resistance to some extent by retaining a capacity for symbiosis-driven recycling of organic nutrients; however, this hypothesis is necessary to carefully examine and prove further.
 Conclusions
 Fungal culturing from 10 surface-sterilized wood cores resulted in 21 fungal cultures, 2.1 per wood segment. Direct sequencing from 40 surface-sterilized segments of lateral roots resulted in 247 fungal sequences or 6.2 per root segment on average. The most dominant fungi from the infested trees of Pinus sylvestris were Dactylonectria macrodidyma (4.98%), Acremonium sp (4.52%), Cladosporium cladosporioides (4.07%) from Ascomycota and Heterobasidion annosum s.s. (4.07%) from Basidiomycota, while for non-infested group Unidentified Ascomycota175244 (13.19%), Penicillium spinulosum (9.89%), Acremonium sp. (8.79%), Bionectriaceae sp. (8.79%) were the most common.
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Effect of different concentrations of garlic extract on dental plaque isolates obtained from patients attending Federal Medical Center Makurdi

Instead of the traditional microbiological pathogens, the microbes that make up the mouth's resident micro-flora are what cause oral diseases like dental caries/plaque and periodontal disorders. The aim of this study was determine the effect of different concentrations of garlic extract on dental plaque isolates obtained from patients attending Federal Medical Center Makurdi. Swap samples were collected from teeth of patients and were transported to the laboratory, analysis was done on different culture media and identification of isolates was done using the Bergy’s manual of determinative bacteriology, biochemical characterization was carried following standard microbiological methods. The antibacterial activity was done using agar well diffusion method in which the garlic extract was introduced in concentrate. Concentrations of 500 mg/mL, 250 mg/mL, 125 mg/mL, 65.3 mg/mL and 35.5mg/mL were prepared and used for the susceptibility. The Isolates from results were Streptococcus oralis, Streptococcus mutans, Enterococcus feacalis and Staphylococcus aureus. The effect observed was concentration dependent as all the graphs obtained shows a linear series, the lowest inhibitory concentration shows that 250mg/ml was the lowest concentration that inhibits each of the isolates in ethanolic garlic extract while 500 mg/mL was the minimum inhibitory concentration for the aqueous garlic extract. Oral hygiene was recommended in order to help reduce the accumulation of dental plaque which usually harbours primary microbes that colonize the teeth surfaces resulting to caries and periodontal diseases. The use of mouth wash containing garlic and or toothpaste incorporated with alicin was recommended as this can help reduce the accumulation of dental plaque biofilm thereby minimising the incidence of dental caries and periodontal diseases.

ETIOLOGICAL BACTERIAL SPECTRUM OF PATIENTS WITH ODONTOGENIC AND NON-ODONTOGENIC ABSCESSES AND PHLEGMONS IN THE MAXILLOFACIAL AREA

Background: Clarifying the bacterial causes of inflammatory diseases has a major role in the treatment of this type of pathology. Materials and methods: In a retrospective study of 138 adult patients with abscesses and phlegmons of the head and neck, hospitalized and operated on urgently in the Clinic for Maxillofacial Surgery at University Multispecialty Hospital for Active Treatment "Sveta Marina" EAD – city of Varna, Bulgaria. Depending on their origin, the patients were divided into two groups - with odontogenic and non-odontogenic abscesses. Results: The study group consists of 73 men (52.9%) and 65 women (47.1%) with a mean age of 43 (18-84) years. The mixed resident microflora, including more than one bacterial species, is isolated in the largest number of cultures – 64.2%. The total number of cultures with Gram-positive bacteria isolated in the samples of the studied 92 patients with isolates is 20,33%. Representatives of Gram-negative bacteria are isolated in 7.49% of all. Obligate anaerobes are found in 2.14% of the isolates. Microorganism fungi are encountered in 3.21% of the crops. In patients with phlegmons of odontogenic origin the ratio between Gram-positive and Gram-negative bacteria is 3:1. In phlegmons of non-odontogenic origin, they are entirely of the Gram-negative spectrum. Conclusion: Microorganisms are not isolated in a third of the examined patients, which are most likely long errors in taking material for microbiological examination or in its improper storage and transportation. Of the other crops, the share of resident microorganisms as opportunistic pathogens is the largest. Of the conditionally pathogenic microorganisms, Gram-positive ones predominate.

Nonbacterial Microflora in Wastewater Treatment Plants: an Underappreciated Potential Source of Pathogens.

Wastewater treatment plants (WWTPs) receive and treat large volumes of domestic, industrial, and urban wastewater containing pathogenic and nonpathogenic microorganisms, chemical compounds, heavy metals, and other potentially hazardous substances. WWTPs play an essential role in preserving human, animal, and environmental health by removing many of these toxic and infectious agents, particularly biological hazards. Wastewater contains complex consortiums of bacterial, viral, archaeal, and eukaryotic species, and while bacteria in WWTP have been extensively studied, the temporal and spatial distribution of nonbacterial microflora (viruses, archaea, and eukaryotes) is less understood. In this study, we analyzed the viral, archaeal, and eukaryotic microflora in wastewater throughout a treatment plant (raw influent, effluent, oxidation pond water, and oxidation pond sediment) in Aotearoa (New Zealand) using Illumina shotgun metagenomic sequencing. Our results suggest a similar trend across many taxa, with an increase in relative abundance in oxidation pond samples compared to influent and effluent samples, except for archaea, which had the opposite trend. Additionally, some microbial families, such as Podoviridae bacteriophages and Apicomplexa alveolates, appeared largely unaffected by the treatment process, with their relative abundance remaining stable throughout. Several groups encompassing pathogenic species, such as Leishmania, Plasmodium, Toxoplasma, Apicomplexa, Cryptococcus, Botrytis, and Ustilago, were identified. If present, these potentially pathogenic species could be a threat to human and animal health and agricultural productivity; therefore, further investigation is warranted. These nonbacterial pathogens should be considered when assessing the potential for vector transmission, distribution of biosolids to land, and discharge of treated wastewater to waterways or land. IMPORTANCE Nonbacterial microflora in wastewater remain understudied compared to their bacterial counterparts despite their importance in the wastewater treatment process. In this study, we report the temporal and spatial distributions of DNA viruses, archaea, protozoa, and fungi in raw wastewater influent, effluent, oxidation pond water, and oxidation pond sediments by using shotgun metagenomic sequencing. Our study indicated the presence of groups of nonbacterial taxa which encompass pathogenic species that may have potential to cause disease in humans, animals, and agricultural crops. We also observed higher alpha diversity in viruses, archaea, and fungi in effluent samples than in influent samples. This suggests that the resident microflora in the wastewater treatment plant may be making a greater contribution to the diversity of taxa observed in wastewater effluent than previously thought. This study provides important insights to better understand the potential human, animal, and environmental health impacts of discharged treated wastewater.

GENETIC BARCODING FOR SURVEILLANCE OF VIRULENCE PLASMIDS IN METAGENOMIC SAMPLES OF MICE GUT MICROFLORA

Outbreaks of nosocomial infections strike the public health system around the world. Current surveillance systems for infectious agents either take too long to identify pathogens or lack the necessary sensitivity to distinguish the virulent strains from the benign ones in the resident microflora. To address this problem, we propose a method for improving the sensitivity and specificity of detection of virulent clonal lines of bacteria or individual virulence plasmids in metagenomic samples. In this study, we used previously developed computational tools, Barcode Generator and Barcoding 2.0, to analyse gut microflora of laboratory mice infected with a pathogenic multidrug-resistant S. aureus and treated with cefazolin, an iodine-containing complex CC-195, and by a combinatorial treatment. We searched the metagenomic samples for representatives of pathogenic microflora using diagnostic genetic barcodes, which were designed based on the whole genome sequences of a collection of clinical isolates. Our results demonstrated a practical applicability of these diagnostic barcodes to monitor clonal lines of pathogens and even individual virulence plasmids in the environment. We also found that the novel drug CC-195 promotes a speedy restoration of the normal gut microflora disturbed by infection and administration of the antibiotic cefazolin.