Oligonucleotide primers used in the study.
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More than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\\n\\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\\n\\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\\n\\nAdditionally, each book published by IntechOpen contains original content and research findings.
\\n\\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Simba Information has released its Open Access Book Publishing 2020 - 2024 report and has again identified IntechOpen as the world’s largest Open Access book publisher by title count.
\n\nSimba Information is a leading provider for market intelligence and forecasts in the media and publishing industry. The report, published every year, provides an overview and financial outlook for the global professional e-book publishing market.
\n\nIntechOpen, De Gruyter, and Frontiers are the largest OA book publishers by title count, with IntechOpen coming in at first place with 5,101 OA books published, a good 1,782 titles ahead of the nearest competitor.
\n\nSince the first Open Access Book Publishing report published in 2016, IntechOpen has held the top stop each year.
\n\n\n\nMore than half of the publishers listed alongside IntechOpen (18 out of 30) are Social Science and Humanities publishers. IntechOpen is an exception to this as a leader in not only Open Access content but Open Access content across all scientific disciplines, including Physical Sciences, Engineering and Technology, Health Sciences, Life Science, and Social Sciences and Humanities.
\n\nOur breakdown of titles published demonstrates this with 47% PET, 31% HS, 18% LS, and 4% SSH books published.
\n\n“Even though ItechOpen has shown the potential of sci-tech books using an OA approach,” other publishers “have shown little interest in OA books.”
\n\nAdditionally, each book published by IntechOpen contains original content and research findings.
\n\nWe are honored to be among such prestigious publishers and we hope to continue to spearhead that growth in our quest to promote Open Access as a true pioneer in OA book publishing.
\n\n\n\n
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Urinary tract infection (UTI) is a topical problem of the contemporary pediatrics, pediatric nephrology, and urology, which takes the second or third place in the structure of children’s morbidity [1, 6, 9, 12]. Risk factors of UTI in children include neonatal period, in particular premature birth, family medical history, abnormalities of urinary tract development, disruptions of urodynamics, including vesicoureteric reflux, obstructive uropathy, neurogenic bladder malfunction, urolithiasis, constipation, fecal and perianal colonization, immunocompromising conditions, including AIDS, immunosuppression therapy, frequent bladder catheterization, diabetes, sexual activity in teenagers [1, 14, 15, 18]. Newborns and infants are under high risk of UTI development, among other factors, since their immune system is not yet sufficiently developed [7]. Among the many factors, which affect development and forecast of UTI, biological properties of microorganisms, which inhabit kidney tissue, are of no small importance [10, 22, 35, 43].
Etiological structure of UTI in children has changed recently. Many studies show the growing etiological significance of Enterococcus faecalis [1, 6, 8, 20, 37]. Clinical significance of enterococci, which were earlier considered in different saprophytes, is being reassessed currently [4, 10].
Increase in etiological significance of enterococci is caused by many reasons, including development of antibiotic resistance, in particular, resistance to cephalosporin, widely used for treating UTIs in children [8]. Enterococci can initiate the infectious process due to their genes coding many pathogenicity factors involved in adhesion and invasion processes, development of biofilms, and hystodamaging effect (Esp, Asa1, EfaA, CylA, CylM, GelE, FsrB, etc.) [6, 13, 16, 30, 35, 42]. It is known that virulence of microorganisms depends upon the quantity of these genes [17]. Scientists from Europe, England, America, India, and Japan are doing research in this area [21, 33, 38, 39, 41]. No research related to characteristics of intraspecific genetic variability, including pathogenicity factors, in uropathogenic E. faecalis isolates from children having UTI, has been done in Russia until now. All the above has been determined in the topic of the research, which was undertaken in the present paper.
The aim of this research was to identify genetic variability and phenotypic features of biologic properties in uropathogenic E. faecalis isolates from children having urinary tract infections to assess its virulent potential and clinical significance.
At the first stage, bacterial tests of urine samples (n = 6438) isolated from patients in cases of UTI at the multispecialty regional children’s hospital from 2008 to 2016 were analyzed. Patients were aged from 3 days to 17 years.
At the second stage, biological properties and genetic variability of enterococci were studied. E. faecalis (n = 71) were isolated from urine of children with urinary tract infections and age from 3 days to 17 years in the diagnostic titer from 104 CFU/ml and higher during from 2013 to 2016. E. faecalis NCTC 12697 was used as the typical culture. All strains of uropathogenic E. faecalis were previously investigated using classical microbiological methods [22, 43]. Antimicrobial susceptibility was performed using disk diffusion method on the Muller-Hinton agar according to EUCAST.
Bacterial DNA of E. faecalis was isolated using DNA-express kit (Lytech, Russia). The testing of enterococci (n = 31) pathogenicity genes was made by polymerase chain reaction (PCR), using previously developed primer sets (Table 1) [5, 11, 32, 34], synthesized by Eurogen (Russia), on TProfessional 96 (Biometra, Germany). The amplification products were analyzed in a 1% agarose gel containing 1 μg/ml ethidium bromide in ultraviolet light using BioDocAnalyze (Biometra, Germany).
Gene | Primer sequence | Product size (bp) | Reference |
---|---|---|---|
cylA | TGGATGATAGTGATAGGAAGT TCTACAGTAAATCTTTCGTCA | 517 | [11] |
aggA | AAGAAAAAGTAGACCAAC AACGGCAAGACAAGTAAATA | 1553 | [11] |
efaA | GACAGACCCTCACGAATA AGTTCATCATGCTGCTGTAGTA | 705 | [11] |
eep | GAGCGGGTATTTTAGTTCGT TACTCCAGCATTGGATGCT | 937 | [5] |
esp | TTGCTAATGCTAGTCCACGACC GCGTCAACACTTGCATTGCCGAA | 933 | [32] |
gelE | ACCCCGTATCATTGGTTT ACGCATTGCTTTTCCATC | 419 | [34] |
Oligonucleotide primers used in the study.
The obtained data were processed using the parametric analysis method. From the indicators of descriptive statistics, the relative values (P, %), their errors (mp, %) were calculated. To evaluate the degree of interrelation, the Pearson correlation analysis (R) was performed with calculation of correlation coefficient (r) and reliability of correlation (p). At statistical processing of the received materials, the software package Statistica 10.0 is used in the operating environment Windows 2010.
The research was approved by Interdisciplinary Committee for Ethics of the Federal State Budgetary Educational Institution of Higher Education “Pacific State Medical University” of the Ministry of Healthcare of the Russian Federation (protocol no. 4, 26.12.2016).
The most common uropathogen in children with UTI, who were treated at the multispecialty regional children’s hospital, is Escherichia coli, for which specific gravity equals from 33.92 ± 1.7 to 62.96 ± 1.2%. Most frequently, it was pointed out in outpatients (in 62.96 ± 1.2% cases); less frequently, it was found at departments for newborns (33.92 ± 1.7%). The second significant etiological factor of UTI in children was E. faecalis, for which specific gravity ranged from 16.14 to 32.5% [22]. At the same time, E. faecalis was the most important factor in development of UTIs at departments for newborns: it was found in 57.2% of cases (Figure 1). Frequency of identifying E. faecalis in newborns diagnosed with UTI ranged from 30.8 to 74.5% of cases over 9 years.
The etiological structure of UTI in newborn.
The present study analyzed the features of phenotypic manifestations of biological properties, including antibiotic resistance of E. faecalis (n = 71) isolated from urine of children with UTI. All analyzed uropathogenic E. faecalis had typical properties—morphology (cocci or oval Gram-positive bacteria), biochemical activity against mannitol, methylene blue, its absence in rhamnose fermentation, variability in glucose, lactose, sucrose and 2,3,5-triphenyltetrazolium chloride (TTC), and lack of mobility and catalase (Table 2).
Most E. faecalis isolated from the urine of children with UTI had in vitro enzymatic activity associated with pathogenicity: hemolytic, proteolytic, lipolytic, and lecithinase. A capsule was found in 45.6 ± 6.6% of uropathogenic enterococci. An inverse relationship was established between the presence of a capsule in E. faecalis with α-type hemolysis (r = 0.3, p = 0.0001), fermentation of milk (r = 0.31, p = 0.00), and a positive correlation with the microorganism titer in the urine (r = 0.33, p = 0.0332).
A direct correlation was established among the lecithinase and DNAase activity of E. faecalis (r = 0.31, p = 0.0438), the manifestation of hemolytic activity (α- or β-type) in uropathogenic enterococci, and the presence of the gene gelE (r = 0.49; p˂0.05).
The capsule (р1–2˂0.001; р1–3˂0.01), milk fermentation (р1–2˂0.001; р1–3˂0.01), gelatinous (р1–2˂0.001, р1–3˂0.05), and lipolytic activity with respect to the tween 60 (р1–2˂0.01, р1–3˂0.01) was determined more often in E. faecalis, isolated from the urine of newborn children (group 1) with UTI, than in other age groups.
E. faecalis, isolated from children under 1 year (group 2), have hemolytic activity more often than other age groups (р2–1˂0.05, р2–3˂0.05), but expressed less gelatinous activity (р1–2˂0.001; р2–3˂0.001). In this group, lipolytic activity against Tween 60 was less pronounced, but the differences were significant only between groups 1 and 2 (p˂0.01).
E. faecalis isolated from patients older than 1 year (group 3) showed more lipolytic activity against Tween 80, compared to other age groups (p1–3˂0.001; p2–3˂0.01), gelatin’s fermentation was determined more often than in enterococci of the group 2 (p˂0.001). The hemolytic activity of enterococci in the group 3 differed little compared to the group of newborn children. The other biochemical properties associated with pathogenicity factors of E. faecalis strains of this age group were similar to enterococci isolated from children under 1 year of age (Table 2).
Assessment of antibiotics resistance revealed that all studied cultures of uropathogenic enterococci (n = 71) are sensitive to vancomycin and nitrofurantoin. We found that enterococci are highly resistant to erythromycin (77.1 ± 5.02%), tetracycline (73.2 ± 5.3%), fluoroquinolones of the II and the III generations (ciprofloxacin (55.1 ± 5.9%), norfloxacin (48.6 ± 9.9%), and levofloxacin (46.5 ± 5.9%)). Were identified E. faecalis cultures, which are resistant (20.1 ± 4.9%) and mid-resistant (8.9 ± 3.5%) to the reserve drug linezolid. More than half (59.2 ± 5.8%) of the studied cultures of uropathogenic enterococci were resistant to several antibiotics (four or more antimicrobial agents).
It was found that enterococci, which are sensitive to penicillins, were characterized with lipolytic, lecithinase, and hemolytic (β-type) activity in vitro. Enterococci cultures which are resistant to fluoroquinolones, fermented sucrose, had proteolytic activity, and did not break down lactose. E. faecalis, which are resistant to gentamicin and erythromycin, had a capsule. Furthermore, E. faecalis, which are resistant to linezolid and chloramphenicol, had gelatinase, lecithinase, and lipolytic activities, as compared to other cultures (these data were not published).
In that way, analysis of biological properties of E. faecalis, isolated from urine of children with UTI in Primorsky region, showed that overwhelming majority of cultures had typical properties. The mentioned variability of biochemical and fermentation activities of pathogenicity factors and resistance to antibiotics suggested a certain phenotypic heterogeneity of E. faecalis.
E. faecalis was tested for genes, coding various pathogenicity factors, using PCR method. It was found that clinical strains (n = 30) of enterococci isolated from urine of children with UTI contained two and more studied pathogenicity genes. In this context, 27 out of 30 uropathogenic E. faecalis had four and more of the studied genes (Table 3).
Biological properties | Enzymatic activity of E. faecalis | |||||
---|---|---|---|---|---|---|
Newborns (n = 21) | Children from 29 days to 1 year (n = 18) | Children over 1 year old (n = 21) | ||||
Number of examined cultures, n | Abs. (Р ± mр, %) | Number of examined cultures, n | Abs. (Р ± mр, %) | Number of examined cultures, n | Abs. (Р ± mр, %) | |
Reduction of TTC *■ | 19 | 18 (94.7 ± 5.1) | 14 | 11 (78.6 ± 11.0) | 21 | 20 (95.2 ± 4.7) |
Reduction of methylene blue | 19 | 19 (100) | 14 | 14 (100) | 21 | 21 (100) |
Biochemical activity in relation to: | ||||||
Mannitol | 20 | 20 (100) | 13 | 13 (100) | 19 | 19 (100) |
Glucose ▲ | 20 | 19 (95.0 ± 4.9) | 13 | 12 (92.3 ± 7.4) | 19 | 19 (100) |
Lactose ▲ | 20 | 19 (95.0 ± 4.9) | 13 | 12 (92.3 ± 7.4) | 19 | 16 (84.2 ± 8.4) |
Rhamnose | 21 | 0 | 13 | 0 | 19 | 0 |
Sucrose | 20 | 17 (85.0 ± 8.0) | 13 | 11 (84.6 ± 10.0) | 19 | 16 (84.2 ± 8.4) |
Presence of a capsule *▲ | 21 | 12 (57.1 ± 10.8) | 17 | 6 (35.3 ± 11.6) | 19 | 8 (42.1 ± 11.3) |
Proteolytic activity in relation to: | ||||||
Milk *▲ | 21 | 18 (85.7 ± 7.6) | 18 | 9 (50.0 ± 11.8) | 21 | 10 (47.6 ± 10.9) |
Gelatin *▲■ | 20 | 9 (45.0 ± 11.1) | 17 | 2 (11.8 ± 7.8) | 21 | 7 (33.3 ± 10.3) |
Lecithinase activity | 19 | 5 (26.3 ± 10.1) | 15 | 4 (26.7 ± 11.4) | 21 | 6 (28.6 ± 9.9) |
Lipolytic activity in relation to: | ||||||
Tween 20 | 17 | 14 (82.4 ± 9.2) | 12 | 11 (91.7 ± 7.9) | 18 | 17 (94.4 ± 5.4) |
Tween 60 *▲ | 8 | 4 (50.0 ± 17.2) | 6 | 1 (16.7 ± 15.2) | 11 | 3 (27.3 ± 13.4) |
Tween 80 ▲■ | 21 | 14 (66.7 ± 10.3) | 17 | 12 (70.6 ± 11.0) | 19 | 16 (84.2 ± 8.4) |
Peculiarities of phenotypic manifestations biological properties of E. faecalis depending on the patient’s age.
Note: *, p < 0.05 between 1 and 2 groups; ▲, p < 0.05 between 1 and 3 groups; ■, p < 0.05 between groups 2 and 3; Abs., absolute.
Id | ST | Isolate | Genotype | Total number of genes | |||||
---|---|---|---|---|---|---|---|---|---|
aggA | esp | cylA | efaA | eep | gelE | ||||
1787 | ST116 | PR042 | + | + | + | + | + | + | 6 |
1789 | ST179 | PRV 052 | + | + | + | + | + | + | 6 |
1791 | ST179 | PRV100 | + | + | + | + | + | + | 6 |
1790 | ST179 | PRV105 | + | + | + | + | + | + | 6 |
nd | PRV086 | + | + | + | + | + | + | 6 | |
nd | PR 230 | + | + | + | + | + | + | 6 | |
1788 | ST179 | PRU047 | + | — | + | + | + | + | 5 |
nd | PR 181 | + | + | + | + | + | — | 5 | |
1781 | ST16 | PR050 | + | + | + | + | + | — | 5 |
1786 | ST41 | PRV049 | + | + | — | + | + | + | 5 |
nd | PRL079 | + | + | — | + | + | + | 5 | |
nd | PRV080 | + | + | — | + | + | + | 5 | |
1793 | ST774 | PRAs81 | + | + | — | + | + | + | 5 |
nd | PR 198 | + | — | — | + | + | + | 4 | |
nd | PR 158 | + | — | — | + | + | + | 4 | |
1780 | ST6 | PRV054 | + | — | — | + | + | + | 4 |
1779 | ST6 | PRN030 | + | — | — | + | + | + | 4 |
1795 | ST774 | PRA029 | + | — | — | + | + | + | 4 |
1792 | ST774 | PR51 | + | — | — | + | + | + | 4 |
1784 | ST40 | PR055 | — | + | — | + | + | + | 4 |
1785 | ST40 | PRA038 | — | + | — | + | + | + | 4 |
nd | PR 228 | + | — | + | + | + | — | 4 | |
1782 | ST16 | PRV092 | + | — | + | + | + | — | 4 |
nd | PR 215 | + | + | — | + | + | — | 4 | |
nd | PR 223 | + | + | — | + | + | — | 4 | |
1794 | ST774 | PR040 | + | + | — | + | + | — | 4 |
nd | NCTC 12697 | — | — | + | + | + | + | 4 | |
nd | PR 97 | — | — | + | + | + | + | 4 | |
nd | PR 161 | — | — | — | + | + | + | 3 | |
nd | PR 206 | — | — | — | + | + | + | 3 | |
1783 | ST21 | PRV082 | — | — | — | + | + | — | 2 |
Genotypes of uropathogenic E. faecalis in Primorsky region.
Note: nd, not determined.
Eleven variants of genes combinations, which code pathogenicity factors of E. faecalis, were identified. The most common variants are (aggA-esp-cylA-efaA-eep-gelE) and (aggA, efaA, eep, gelE).
Multilocus sequence typing (MLST) divided 17 E. faecalis strains into eight (ST6, ST16, ST21, ST40, ST41, ST116, ST179, and ST774) sequence types (The results have not been published.) It was noticed that uropathogenic E. faecalis strains attributed to ST6, ST40, ST179, ST774, and ST116 are resistant to four and more groups of antimicrobial agents (Table 4).
Id | Isolate | ST | Phenotype of antibiotic resistance | Genotype |
---|---|---|---|---|
1785 | PRA038 | 40 | ТЕТ-ERT-AmG-CHL-LZD | esp-efaA-eep-gelE |
1784 | PR055 | ТЕТ-ERT-AmG-CHL | ||
1780 | PRV054 | 6 | ТЕТ-ERT-AmG-CHL-FLQ | aggA-efaA-eep-gelE |
1779 | PRN030 | ТЕТ-ERT-AmG-CHL-FLQ | ||
1788 | PRU047 | 179 | ТЕТ-ERT-AmG-LZD | aggA-cylA-efaA-eep-gelE |
1789 | PRV052 | ТЕТ-ERT-AmG | aggA-esp-cylA-efaA-eep-gelE | |
1791 | PRV100 | ТЕТ-ERT-AmG-CHL | ||
1790 | PRV105 | ERT-AmG-CHL-FLQ | ||
1787 | PR042 | 116 | ТЕТ-ERT-AmG-CHL-LZD | |
1781 | PR050 | 16 | ТЕТ-ERT | aggA-esp-cylA-efaA-eep |
1782 | PRV092 | ТЕТ-ERT-AmG | aggA-cylA-efaA-eep | |
1794 | PR040 | 774 | AmG-FX | aggA-esp-efaA-eep |
1793 | PRAs081 | ТЕТ-ERT-PEN-AmG-FLQ | aggA-esp-efaA-eep-gelE | |
1795 | PRA029 | ТЕТ-ERT-PEN-FLQ | aggA-efaA-eep-gelE | |
1792 | PR051 | ТЕТ-ERT-AmG-FLQ | ||
1786 | PRV049 | 41 | ТЕТ | aggA-esp-efaA-eep-gelE |
1783 | PRV082 | 21 | FLQ | efaA-eep |
Antibiotic resistance of uropathogenic E. faecalis depending on the sequence type.
Note: TET, tetracycline; ERT, erythromycin; AmG, aminoglycosides; FLQ, fluoroquinolones; CHL, chloramphenicol; LZD, linezolid; PEN, penicillins.
The results demonstrate broad variability of the range of genes, which code pathogenicity factors and reveal sequence types with multiple resistances to antimicrobial agents among uropathogenic E. faecalis isolated from children with UTI in Primorsky region.
Urinary tract infection is an inflammatory process in the organs of the urinary system without specifying the level of damage or growth of microorganisms in the urinary tract with possible development of local inflammatory changes. UTI refers to the factors that initiate the development of chronic kidney disease and depends on the age of the children [27]. Etiological multifactority is peculiarity of these infections. For a long time, commonly recognized pathogens of uroinfections are Gram-negative enterobacteria, among which Escherichia coli is prevalent [25, 26, 29, 31, 40, 44].
Interest to studying enterococci as participants of infectious diseases has increased in recent years. Our research has shown that E. faecalis are a common pathogen, which causes UTI in children, most often in newborns (from 30.8 up to 75% of cases). Perhaps this is not accidental, as according to some authors, this microorganism is detected in children from the first days of life and its amount exceeds the content of E. coli in the newborn period [19]. The reason for this microbial composition of urine appears to be functional immunodeficiency in this category of patients.
However, until now, the true etiological significance of these microorganisms in the development of the infectious process and unfavorable outcomes remains uncertain due to the ever-changing properties of E. faecalis. It is known that the ability of bacteria to affect the kidneys and urinary tract is determined not by one but by a complex of properties necessary for this process, that at the different stages of the infectious process in the urinary system organs from the microorganism, priority expression of certain pathogenetically significant traits and/or their combinations is required [6].
All analyzed uropathogenic E. faecalis had typical properties—morphology, biochemical activity against mannitol, methylene blue, its absence in rhamnose fermentation, variability in glucose, lactose, sucrose and 2,3,5-TTC, and lack of mobility and catalase.
At the same time, weak and delayed acid formation from lactose was in three cultures (only on the third day), and in 12 isolates during fermentation of sucrose (by day 10). The results obtained with respect to a number of carbohydrates differ from the literature data, as it is known that E. faecalis ferment lactose and sucrose, and in relation to other sugars can be variable (Berdzhi). Possibly, this is associated with the spread of certain E. faecalis biovars in the territory of Primorsky Krai or within a single multispecialty hospital.
According to results of research conducted in recent years, it was determined that enterococci produce many virulence factors, which conduce to development of the infectious process (hemolysin, gelatinase, enterococcal surface protein, aggregation substance, serine protease, capsule, etc.). The greatest number of virulence factors was found in E. faecalis isolated from urine. High proteolytic activity of E. faecalis (hydrolysis of gelatinase, casein, and collagen) causes toxic damage to tissues and conduce to cicatricial changes in kidney [6, 19, 43].
Moreover, the change in the properties of the microorganisms, causing the urinary tract infection, such as the development of resistance factors to antimicrobial drugs and the biofilm formation, makes it difficult to manage patients, especially with chronic persistent and often recurrent infection.
Our research confirmed high virulence properties of E. faecalis isolated from urine of patients with and their manifestations depending on the patient’s age. For example, there is a negative correlation between the proteolytic activity of E. faecalis and the age of the children (r = 0.28, р = 0.002). Most E. faecalis isolated from the urine of children with UTI had in vitro enzymatic activity associated with pathogenicity: hemolytic, proteolytic, lipolytic, and lecithinase.
Currently, lipase is referred to understudied factors of enterococcal persistence, although it is known that lipase may be a potential virulence factor of E. faecalis [12]. Among the studied enterococci isolated from children with UTI, lipolytic activity was determined in 85.0 ± 8.2% of the cultures (more often with respect to Tween 20 and Tween 80). Enterococcus cultures showed heterogeneity in proteolytic and hemolytic activity. A reliable direct correlation between the phenotypic manifestation of β-type hemolytic activity with hydrolysis of gelatin (r = 0.58, p = 0.0001) and lecithinase activity (r = 0.52, p = 0.0004) of this uropathogen has been established. This confirms the combined effect of these pathogenic factors at a certain stage of the inflammatory process. A relationship was established between the phenotypic manifestation of pathogenicity factors and the age of patients.
The most common properties of E. faecalis isolated from urine of newborn children were a capsule, proteolytic, and lipolytic (in relation to Tween 60) activity. Enterococci isolated from 1-year-old children with UTI most frequently were characterized with hemolytic activity. Lipolytic (in relation to Tween 80) activity was most frequently found in cultures isolated from patients older than 1 year. The prevalence of these virulence factors suggests that they are associated with virulence of this species in UTI. Such features of the manifestation of biological properties in vitro indicate the selection of etiologically significant E. faecalis at the level of the macroorganism.
Furthermore, connection between sensitivity to antimicrobial drugs of E. faecalis and its biological properties was identified. It was found that uropathogenic enterococci characterized with proteolytic activity are resistant to antibacterial agents with different action mechanisms. In the work, it was found that E. faecalis, resistant to linezolid and chloramphenicol drugs that suppress protein synthesis at the level of the 50S subunit of the bacterial ribosome, possess a high pathogenic potential. These results require further research in this direction.
Interesting data were obtained with regard to the sensitivity of uropathogenic E. faecalis to the reserve drug linezolid, recommended for treatment of infections caused by strains, which are resistant to vancomycin, aminoglycosides, and betalactams. In Primorsky Krai, E. faecalis cultures were found resistant and intermediate sensitivity to linezolid. However, in Russia in the period from 2005 to 2013, there were isolated single enterococcal strains with reduced sensitivity to linezolid.
This way, the mentioned variability of biochemical and fermentation activity of factors related to pathogenicity demonstrated phenotypic heterogeneity of enterococci and might have a certain diagnostic significance.
Pathogenicity factors of bacteria are genetically determined by properties which are localized in genome of microorganisms in the form of “pathogenicity islands” [17]. These genetic elements can contain various sets of virulence genes, which are important for the development of the enterococcal infectious process, including genes of antibiotics resistance [17, 24]. At the present stage, the association of antibiotic resistance of E. faecalis with pathogenic factors is actively studied. It is known that strains of enterococci resistant to ampicillin, ciprofloxacin, and gentamicin, but sensitive to vancomycin and nitrofurantoin have more pathogenicity factors (hemolysin, gelatinase, hyaluronidase, form biofilms) than vancomycin resistant [2, 28]. E. faecalis having the asa1 gene are more resistant to fluoroquinolones (norfloxacin, ciprofloxacin, and levofloxacin) than isolates lacking this gene. Resistance to ciprofloxacin is significantly higher in E. faecalis having the genes cylL and cylS than in strains with their absence [23, 28]. esp gene-positive E. faecalis are more resistant to doxycycline than esp gene-negative cultures [3]. Among the strains with multidrug resistance, a high prevalence of genes asa1 and esp was observed [23, 36].
The research implemented using PCR method enabled to characterize in greater detail the structure of E. faecalis population isolated from children with UTI from Primorsky region. In our research, significant variability in occurrence frequency of the studied genes was found. Two of them—efaA (coding the surface antigen A (EfaA), which initiates the infectious process) and eep (coding Eep protein, which conduces to formation of a biofilm, making it resistant to various biological stress factors) were found in all studied uropathogenic E. faecalis, which proves their involvement in certain stages of the infectious process.
MLST analysis conducted earlier revealed eight sequence types, five of which were characterized by multidrug resistant.
This way, clinically significant E. faecalis strains have a complex of virulent properties, which allow the bacteria to materialize their pathogenic potential on all stages of the inflammation process in urinary system. This makes further research of the listed factors in clinical E. faecalis necessary to estimate objectively the contribution of these properties of the agent into pathophysiologic mechanism of infectious and inflammatory diseases.
Important role in the etiology of UTI is played not only by Gram-negative bacteria of the Enterobacteriaceae family, but also by gram-positive E. faecalis, which are of paramount importance in the development of UTI in newborns.
E. faecalis, isolated from the urine of children with UTI, have a complex of pathogenicity factors necessary for the development of the inflammatory process and their prolonged persistence in the urinary tract. The relationship between E. faecalis pathogenicity factors and the age of patients was determined.
Uropathogenic E. faecalis possess a polyantibiotic resistance, which is associated with its biological properties and belonging to a particular sequence type.
A set of phenotypic manifestations of the biological properties of E. faecalis (the presence/absence of hemolytic, gelatinase, lecithinase, lipolytic activities) established in the study may determine its clinical significance and serve as an in vitro diagnostic marker of resistance of the studied uropathogen to certain groups of antibacterial drugs.
The research was supported with an internal grant from the university (registration No. AAAA-A18-118031390014-9).
The authors declare that there is no conflict of interests.
If we look at history and especially last century, there were some remarkable incidence happened which leave every lasting impact on our daily life. Moreover, these incidence changes our overall behaviors, as well. Among these incidents was an era of the First and Second World War, then the cold war and 9/11. However, the invention of recent technologies such as mobile phone communication, artificial intelligence, and the internet of things profound effect on our daily life. Besides all wars and inventions, the ongoing pandemic crises of Covid-19 will have permanent and everlasting impacts on our lives, even if we can control it immediately. In terms of economy, due to pandemic crises of Covid-19, IMF predicts that the world economy would contract with 3%; however, the revised estimated figure for the contraction of the global economy is 4.9 to 5.5% in 2020. One of the critical factors, which decline rapidly due to Covid-19, is the tourism sector.
\nAccording to the estimate, the international labor organization tourism sector created approximately 330 million jobs worldwide, and it is 10.3% of the total global employment. The tourism sector creates jobs; promote local economic development and culture. In terms of career, it contributes to direct and indirect jobs for young people and women. Globally, tourism is an essential source of employment. There are some distinctive characteristics of the labor market. In general tourism industry is labor-intensive. Furthermore, approximately 54% of posts are taken by the tourism sector by the woman and young people, which makes the industry inclusive.
\nThe expected loss for 2020 in the tourism sector can be estimated by analyzing the figure of the previous year, as in 2018, approximately 1407 million international tourist arrivals were recorded, which generate total tourist receipts amount 1480 billion dollars. Similarly, the tourism sector also provides a significant amount of indirect employment, such as in construction and infrastructure development, and even a long chain of the supply chain of food and drinks is also associated with this sector. Many staff in the tourism offices, airlines, aircraft, hotels, restaurants, shopping centers, and various tourist attractions often has direct interaction with visitors. Below giveFigure 1 indicate, inbound tourist expenditure recorded in 2018 by top tourist destination in the world.
\nInbound tourism expenditure of 2018 by top tourist destinations. Source: United Nations Conference on Trade and Development.
It is not the first time where the world is experiencing a pandemic outbreak. In the past 20 years, the earth had also experienced two regional pandemic crises, SARS 2004 and Ebola 2014. Both pandemic disasters did not affect the airline and especially the tourism industry. However, due to the Tsunami 2004, the tourism sector of Far East Asian countries. The 9/11 incident did not account for the massive decline in the airline industry. Give below Figure 2 present, these incidences and overall flight operations.
\nImpact of major crises on the global tourism. Source: Gössling et al. [1].
In the European Union, it is expected that 13 billion people might lose their job due to Covid-19, and in terms of revenue, it is approximately 12 billion per year. The key drivers of the tourism industry in Europe are the local festivals, trade fairs, sports events, and concerts. Therefore due to Covid-19 in many tourist places, hotels, restaurants, bars, and theme parks already closed. Similarly, sports events like the Euro 2020 football championship already postpone until 2021. Likewise, on the other hand, at the beginning of the Covid-19 outbreak, many tourists faced difficulty with returning home due to the border controls in massive countries of Europe.
\nThe primarily purpose of this chapter is to explore the nexus between the impact of Covid-19 and tourism. Furthermore, this chapter explains the overall impact of tourism and lockdown on pollution as well.
\nTourism also has an indirect economic effect, in addition to incoming tourism expenditures. Tourism employs workers, ports, and airports as well as a wide variety of intermediate inputs, including financial services, education, food and alcohol, and domestic transport. According to IATA estimation, as compared to April 2019, 80% fewer flights were recorded in 2020 due to the Covid-19 outbreak. In terms of numeric values, the estimated monetary loss recorded in the airline industry is 84.3 billion and 2020. Furthermore, according to IATA estimation, regular international flight schedules will start working until 2023–2024, whereas domestic flights will soon be restored.
\nIn terms of survivor crises of the Covid-19 outbreak, three kinds of potential threats have been indicated by 2020, the United Nations Conference on Trade and Development. These scenarios are listed below in Figure 3.
\nThree kinds of potential threats. Source: United Nations Conference on Trade and Development.
According to the above table, each scenario indicates the annual tourism expenditure is reduced as productivity shock. Similarly, on the other hand, the social effect on the output of the other products and sectors also hit massively in each of the scenarios explained in the table, such as food, drink, logistics, and construction. However, in the case of a moderate scenario, for four-month, the global economy reduced by 1.2 trillion US dollars. Intermediate for eight-month it reduces 2.2 trillion US dollars. In the last, in case of a dramatic scenario, the world would experience a loss of 3.3 trillion US dollars in terms of GDP (2020, United Nations Conference on Trade and Development).
\nSimilarly, Ali and Cobanoglu [2], mention in work once, as initially due to Covid-19 world experience lockdown situation, they predicted that under the current circumstances of international travelers can be shrinking from 1.4 to 1 billion. Furthermore, they also reveal that approximately 50 million jobs could be lost. However, the current statistics are indicating the worse situation. The world has already experienced massive outbreaks, crises, and other natural disasters that directly or indirectly impact the overall tourism sector. In the past, one of the significant crises hit the tourism sector, especially, was the Tsunami of 2004, which ran almost 15 countries of the Indian Ocean, and more than 200,000 people lost their lives. It was a natural disaster, as [3] disasters are unpredictable catastrophic change; usually, it happens all of a sudden and can be responded after the event, furthermore response is possible either by contingency plans or by through reactive responses as well. Although Covid-19 is not a regional issue or any specified geographical disaster, it is pandemic crises spread worldwide, and eventually, every human living in this world is disturbed due to it. It is right to argue that the tourism sector hit and disrupted massively due to Covid-19 due to that sector, every other industry of the world experience a downward trend.
\nIn terms of Covid-19 impact of economy, Barro et al. [4] empirically investigate the economic effect of Covid-19 and reveals that due pandemic crises, on average, 2.1% death rate could cause the decline of averagely 6% of world GDP and 8% decline in private consumption. On the other hand, one more study conducted by Coibion et al. [5] using a survey on the household in the US concluded due to pandemic crises, consumption and employment decreased whereas inflation and economic uncertainty increased.
\nSimilarly, a massive number of empirical studies have been conducted that explain the positive impact of the tourism sector on the economy. Some of these studies which demonstrate a positive relationship between economic growth and tourism are [6] for Turkey, [7] for six Balkan countries, Chen and Chiou-wei [8] for Thailand and Korea [9] for India, and more recently Khan et al. (2019) empirically investigate that relationship between multiple variables for Thailand using ARDL estimation and conclude that the logistics and transportation sector positively impact inbounds tourism. Therefore it is correct to argue that, due to current pandemic crises, the overall economic industry hit massively due to the downfall of the commercial sector. The figure below indicates the global destruction of GDP in terms of the percentage and value of the top 15 tourist destinations in the world (Figure 4).
\nOverall GDP change in top 15 countries due to Covid-19. Source: World Tourism Organization.
The above table indicates that from Asia, Thailand, and Malaysia’s GDP falls with 9 and 3%. In terms of the monetary value of GDP, Indian GDP decreased by 28,120 million US dollars, China bear loss of 104,690 million US dollars, and Korea experienced the loss of 22,092 million US dollars. Jamaican and Dominican republic GDP indicate a downward trend of 11 and 5%. However, European countries’ tourism sector hit massive due to the Covid-19 outbreak. Croatian GDP fall by 8%, Portugal 6%, Greece 4%, Ireland and Spain 3%. In terms of monetary value, France bears losses of 47,289 million US dollars, Germany 46,260 million US dollars, and then Italy lost 34,104 million US dollars. All these losses are due to the lockdown of other sectors, directly or indirectly, with the tourism sector of Europe.
\nSimilarly, the net of loss experience by European countries is due to massively unemployed labor and capital. One of the main reasons for the substantial loss in Europe is the high percentage of Tourism to GDP ratio. According to the estimate, the overall tourism sector is responsible for 30% of the EU’s GDP [10]. Due to this, many people are unemployed or displaced in Europe. However, due to the lockdown situation, all these unemployed people also face difficulty searching for new work.
\nGössling et al. [1] indicate in their research work that various airlines such as Scandinavian Airlines (March 17, 2020), Singapore Airlines (March 27, 2020) and Virgin (March 30, 2020), and German TUI (March 27, 2020) have already requested for state aid due to pandemic crises. According to the Federal Aviation Administration estimate in the USA, approximately 64 million take-offs and landed recorded. However, the Covid-19 pandemic crises became a significant cause of downfall in the airline industry. The below figure indicates the trend of airlines from January to April 2020, and due to lockdown situations, a massive number of airlines top working (Figure 5).
\nFlight operations during for first four mother of 2020. Source: International Energy Agency (2020).
At the beginning of 2020, it was not expected that Covid-19 caused economic down and air restriction. In the middle of February the causalities and death rate due to Covid-19 start increasing especially in the UK, Italy and Spain, most of the European countries lockdown their border, and also put travel bans. Besides that, all the schools, universities, bar-restaurants, and other tourist spots were also closed by authorities in Europe. Given the below figure, indicate travel restriction imposed by countries due to the Covid-19 (Figure 6).
\nTravel restriction on 31 March. Source: Gössling et al. [1].
However, as the number of confirmed cases started increasing, most of the world’s countries put travel restrictions. In the case of tourism, many people also faced issues related to their visa, which was on a visit to other countries as international tourists. However, it is notable that almost every country of the world increased visa validity for an unknown period. Similarly, embassies of the different countries also played an active role during the crises. Many countries arranged chartered flights for their nationals, which were stuck in different countries of the world.
\nBesides the airline industry, exploring and travel by cruise ship is also very much popular in European, American, and Asian countries. However, due to Covid-19 cruise industry also faced a downward trend. The significant number of cruise ships also experienced heavy losses. The cruise industry stops its operation until 2020 September, and also it is expected that it will be until the first quarter of 2021. The top three cruise ship lines which experienced heavy losses are cruise lines Carnival, Norwegian Cruise Line, and Royal Caribbean Cruises. However one of the most significant reasons for imposing bans on cruise ships is indicated by Travel.State.Gov [11], they mention in their work, due to the close environment of the cruise it is a high possibility that the Covid-19 spread among the traveler very quickly and therefore it will also be hard to provide help and assistance immediately.
\nOn the other hand, some of the positive attributes have also been observed due to the Covid-19 impact of the tourism sector—one of the crucial factors found in the rapid downfall of air pollution. Due to the massive lockdown number of airlines grounded their air craft’s, which ultimately reduces the air-pollution. As the airline industry accounts for about 2.5% of global CO2 emission, and it is also predicted that until 2050 the airline industry will averagely increase 1.5 degrees Celsius worldwide [12]. After the lockdown, the clean environment has been observed worldwide, not only in highly industrialized countries, more specifically in European countries due to the lockdown situation, and the concentration of NO2 emission in the air rapidly decreased. It has alone been observed after a long time, in Italy due to lockdown and tourist restriction marine line start running in the canals of Vince. The change in terms of environmental change, which we are experiencing at the moment due to the lockdown situation in terms of clean air and clean water channels will not be long-lasting. It is expected that once in the world normal life cycle starts, we will again experience massive air and other pollutions [13].
\nDue to the deep concentration of tourism in the economy of numerous countries, many tourism business are associated with the global food market due to low cost, which generates employment for the local state. Similarly, on the other hand, there is a high volume of food waste involved in the tourism industry. Due to the Covid-19 outbreak, food waste and supply are declining, but on the other hand, it also becomes the cause of job losses for the people who work in the food industry [14].
\nIt is right to argue that, due to the lockdown situation and downward trend of tourism, the natural beauty around the world is restored. One of the critical factors which are noticed is the reduction of waste and trashes. Covid-19 provides us with a chance to restore our natural beauty. Besides tourism, another significant factor contributing to the decrease in pollutions (water and air) is less transporting activities. Thus due to lockdown, most people working from home, on the other hand, a massive number of industrial units not operational at the moment, which also decreased air pollution.
\nDue to the lockdown activities and the non-operational tourism sector, environmental quality is improved worldwide. Various gases such are CO2, NO2, and SO2 concentration decreased within the air. Furthermore, the critical factor which increases air quality around the world is also due to less operational activities of the airline industry. The figure given below presents the air quality which is observed in some of the cities in Europe (Figure 7).
\nChange in NO2 emission due to lockdown in Europe. Source: Centre for Research on Energy and Clean Air.
On the other hand, in the country, China’s economy is considered one of the world’s top growing economies due to its industrialization. However, due to the Covid-19 outbreak and lockdown in china, its air quality increased as well. Similarly, as we have stated earlier, China also bears massive losses due to the downfall of the tourism sector. One of the critical factor which can be noticed that due to the lockdown situation and decline in tourism sector indicatively impact on overall energy consumption. More especially in China, energy consumption also declined due to the Covid-19 lockdown, which eventually affected its production. Less production of energy increases air quality as less coal consumption is used in the production of energy—figure, present coal consumption in Jingjinji city by five energy-generating plants (Figure 8).
\nCoal consumption by five generating plants in China. Source: Centre for Research on Energy and Clean Air.
However, lockdown and the downward trend of the tourism sector provide an opportunity for the world to move toward renewable sources. It the responsibility of the government at the movement to spread awareness of green economy within their countries and further set upset those on tourist spots, which can efficiently operate with renewable energy sources, ultimately it will also reduce air pollution in the future. Similarly, after once pandemic crises will over, it is also highly recommended that, for local and international tourists, the authorities should arrange those vehicles for traveling purposes which consume renewable energy sources.
\nThis chapter aims to discuss the association between Covid-19’s outbreak and the tourism sector. The debate indicates that Covid-19 indicates an adverse effect on tourism activities. Due to the lockdown, thousands of people lost their jobs. Many airlines, restaurants, bars, and hotels are nearly to become bankrupt. On the other hand, our discussion reveals that global lockdown creates a positive impact on environmental sustainability. Also, air and water pollution have been reduced significantly, which creates a positive impact on fauna and flora.
\nFurther, we predict that in upcoming years, due to the restoration of environmental beauty, the global tourism industry will boom. However, the age of booming may not be very long. Because, once the world engine starts after Covid-19, air and water pollution will occupy the space of greeneries. It is the right time to formulate sustainable strategies and policies to maintain the beauty of the world. The following are the recommendations that will help policymakers and regulatory bodies to develop a master plan and integrate sustainable practices in the tourism industry.
The airlines, and especially tourist operators, should adopt green and eco-friendly practices in their businesses to improve environmental sustainability.
Governmental bodies should formulate strict eco-friendly policies for the tourism sector. Also, regulatory bodies encourage firms to adopt ISO certification
Regulatory bodies may encourage renewable energy and green projects by providing subsidies and tax-exemptions to corporations that adopt eco-friendly practices.
Governmental bodies embossed heavy penalties on the polluting firms, which will not only create pressure on firms to adopt sustainable practices but also motivate the eco-friendly firms.
The regulatory bodies should evaluate enterprises’ environmental performance and publicize the evaluation results, which will create competition between enterprises to be more sustainable.
This work supported by the China Postdoctoral Science Foundation (No. 2019 M660700), the Beijing Key Laboratory of Megaregions Sustainable Development Modeling, Capital University of Economics and Business (No. MCR2019QN09).
\nGeneral requirements for Open Access to Horizon 2020 research project outputs are found within Guidelines on Open Access to Scientific Publication and Research Data in Horizon 2020. The guidelines, in their simplest form, state that if you are a Horizon 2020 recipient, you must ensure open access to your scientific publications by enabling them to be downloaded, printed and read online. Additionally, said publications must be peer reviewed.
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