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Carbapenemase-producing Enterobacteriaceae (CPE) isolates are recognized as one of the most severe threats to public health. However, the population structure and genetic characteristics of CPE isolates among bloodstream infections (BSIs) are largely unknown. To address this knowledge gap, in this study, we included patients with clinically significant BSIs due to Enterobacterales isolates, recruited from 26 sentinel hospitals in China (2014–2015). CPE isolates were microbiologically and genomically characterized, including their susceptibility profiles, molecular typing, phylogenetic features, and genetic context analysis of carbapenemase-encoding genes. Of the 2569 BSI Enterobacterales isolates enrolled, 42 (1.6%) were carbapenemase-positive. Moreover, among the 2242 investigated isolates, 1111 (49.6%) extended-spectrum β-lactamase (ESBL)-producing isolates were identified in Escherichia (E.) coli, Klebsiella (K.) pneumoniae, Proteus (P.) mirabilis, and K. oxytoca. Whole genome sequencing analysis showed the clonal spread of K. pneumoniae carbapenemase (KPC)-2-producing K. pneumoniae sequence type 11 (ST11) and New Delhi metallo-β-lactamase (NDM)-5-producing E. coli ST167 in our collection. Plasmid analysis revealed that carbapenemase-encoding genes were located on multiple plasmids. A high prevalence of biofilm-encoding type 3 fimbriae clusters and yesiniabactin-associated genes was observed in K. pneumoniae isolates. This work demonstrates the high prevalence of ESBLs and the wide dissemination of CPE among BSI isolates in China, which represent real clinical threats. Moreover, our findings first illustrate a more comprehensive genome scenario of CPE isolates among BSIs. The clonal spread of KPC-2-producing K. pneumoniae ST11 and NDM-5-producing E. coli ST167 needs to be closely monitored.

In this study, we report a high incidence of New Delhi metallo-β-lactamase (NDM)-producing and ampicillin-catabolizing bacteria within carbapenem-resistant bacterial populations in the waters of two important rivers, Mahananda and Karala, bisecting two most populous towns, Siliguri and Jalpaiguri, respectively, in the northern West Bengal, India. Isolates producing NDM belonged to four genera, Acinetobacter, Escherichia, Proteus, and Pseudomonas; among which few were phylogenetically determined as putatively novel species. Class 1 integrons with the frequent presence of aadA and aac(6')-Ib gene cassettes in 50% of NDM-bearing isolates are indicative of possible selective pressures generated out of unregulated use of streptomycin, in agriculture practiced by the cultivators and tea planters living in locales drained by these two rivers, in their up- and downstream, and amikacin in the most crowded government-sponsored "sadar" and district hospitals of Siliguri and Jalpaiguri. NDM-delivering bacteria in rivers have genuine consequences for city inhabitants who are dependent on public water and sanitation facilities. Standard reconnaissance of antibiotic resistance, consolidating ecological sampling just as the assessment of clinical isolates, should be set up as a need.

Aims: Investigate the interaction of Salmonella spp. with E. coli and Proteus spp. in biofilm formation as mono and dual-species at different time durations Experimental Design: Salmonella, Proteus, and E. coli were isolated from Broiler chicken meat, and the biofilm-forming ability of these organisms were studied. Place and Duration of Study: The study was conducted at the Laboratory of Livestock Production, Faculty of Agricultural Sciences, Sabaragamuwa University of Sri Lanka, from 2019 December to 2020 May. Methodology: This study investigated the biofilm-forming ability of Salmonella as a mono species and its interaction with E. coli and Proteus in the process of biofilm formation. Microorganisms used for this study were isolated from broiler chicken meat. Biofilm was quantified using a microtitre plate assay. The interaction effects were tested at the temperature of 280C in different time durations (up to 120 hours). Results: Salmonella 1 and Proteus monocultures showed significantly higher biofilm-forming ability than Salmonella 3 isolate at all tested time points. At 120 hr, additionally to the salmonella 1 and Proteus isolates E. coli also formed significantly higher biofilms than Salmonella 3. However, Salmonella 3 was the lowest biofilm former as mono biofilm at all tested time durations. Salmonella 1 interaction with Salmonella 3 isolates formed less biofilms than Salmonella 1 mono biofilm at 48hr and 72hr correspondingly. Salmonella 1 and its interactions with Salmonella 3, Proteus, E. coli showed similar biofilm-forming abilities without significant differences at all other tested time points. Specifically, Salmonella 3 interaction with Salmonella 1 as dual biofilm showed higher biofilm-forming ability than Salmonella 3 mono biofilm at all tested time points. Tested isolates and their interaction achieved the highest biofilm formation at numerous time points. In fact, at 48hr, Salmonella 3 isolates and its interaction of Proteus, E. coli, and Salmonella 1 interaction with Proteus attained their highest biofilm formation abilities. The highest biofilm formation was achieved by Salmonella 1 isolate as mono biofilm and Salmonella 1 interaction with E. coli as dual biofilm at 72hr. Biofilm-forming trend of respective isolates and interactions showed numerous patterns at tested time durations. Specifically, E. coli rapidly enhanced its biofilm-forming ability as monoculture from 24 hr to 120 hr. Proteus, Salmonella 3 as monocultures, Salmonella 3 interaction with Proteus and E. coli as dual cultures showed progressive biofilm development from 24 hr to 48 hr. Salmonella 1 monoculture and its interaction with Salmonella 3, E. coli as dual biofilm improved their biofilm-forming ability from 24 hr to 72 hr. Similar to Salmonella 3 interaction with Proteus, Salmonella 1 interaction with Proteus also increased its biofilm-forming ability from 24 hr to 48 hr. Conclusions: This study concluded that there is a variation among isolates and their combinations in forming the biofilms, where there is an enhancement of biofilm in dual-species over the mono-species in some interaction, and there is a reduction in biofilm formation by dual-species with some combinations. Further, this concluded that Salmonella is interacting with other commonly found bacteria such as Proteus and E. coli in biofilm formation.

Objectives We sought to integrate a VEB-1-encoding gene cassette into the integron of the MDR region of genomic islands (GIs) harboured by Proteus mirabilis strains after antibiotic exposure. Methods An IncP1 plasmid from Achromobacter xylosoxidans carrying the cassette array dfrA14–blaVEB-1–aadB was introduced by conjugation into five strains of P. mirabilis: PmBRI, PmABB, PmSCO and Pm2CHAMA harbouring Salmonella GI 1 and PmESC harbouring Proteus GI 1. Circular intermediates of the cassettes were amplified by PCR. blaVEB-harbouring P. mirabilis were exposed to increasing concentrations of ceftazidime each day. Presence of blaVEB-1 in the GI was assessed by PCR. The complete MDR regions were mapped and sequenced in positive clones. Results Circular intermediates were detected for dfrA14 and blaVEB-1–aadB and dfrA14–blaVEB-1–aadB cassettes arrays in A. xylosoxidans, and for aadA2 in P. mirabilis. Insertion of blaVEB-1 into the GIs occurred under ceftazidime pressure. In all cases, the three cassettes from IncP1 were integrated. They replaced the cassette array of PmBRI, PmABB and PmSCO in which floRc, tet(A)G and blaPSE-1 were conserved, whereas they replaced an integron and the IS26-flanked region in Pm2CHAMA. In PmESC, they only replaced aadB, with aadA2 being conserved. blaVEB-1 integration occurred just after conjugation for Pm2CHAMA but required ceftazidime exposure for the other strains. Conclusion Homologous recombination of gene cassettes conferring resistance to clinically important antibiotics may occur under antibiotic pressure between an integron located on a plasmid and a co-resident GI. This feature participates in the acquisition, maintenance and spread of antibiotic resistance genes.

Context: Proteus species are liable for multitude of infections and associated with resistance to routinely used antibiotics even to reserve drugs such as carbapenems. Aims: The aim of this study was to detect the presence of MBL producers, including blaOXA-833 gene in Proteus spp. along with their antibiotic resistance pattern. Settings and Design: This cross-sectional study was conducted in the Department of Microbiology of a tertiary care hospital of Bangladesh during July 2018 to June 2019. Subjects and Methods: Proteus spp. was isolated from a total of 500 samples. Antibiotic susceptibility was performed by disk-diffusion technique. Minimum inhibitory concentration (MIC) of imipenem was determined by agar dilution method. Carbapenemase producers were phenotypically detected by double disc synergy (DDS) test, combined disc (CD) assay, and modified Hodge test (MHT). Carbapenemase genes (blaKPC, blaVIM, blaIMP, blaNDM-1, blaOXA-23, blaOXA-48-like/blaOXA-833, and blaOXA-58) among imipenem-resistant Proteus spp. were detected by polymerase chain reaction (PCR). Sequencing was performed to differentiate OXA-833 from OXA-48-like gene by capillary method, and the nucleotide sequence of OXA-833 has been deposited to GenBank. Results: Ten (25%) imipenem-resistant isolates were detected during disk-diffusion technique, among them 60%, 70%, 50% carbapenemase producers were detected by DDS test, CD assay, MHT, respectively, and 70% by PCR. A significant increase in MIC was found between 8 and ≥128 μg/ml to imipenem. PCR revealed that 40% imipenem-resistant isolates were positive for blaNDM-1 and blaVIM followed by 20% for blaOXA-48-like/blaOXA-833 and blaOXA-23, respectively. Sequencing of blaOXA-48-like gene established the OXA-833 variant of class D carbapenemase encoding gene. Conclusion: The results of this study showed the presence of high proportion of carbapenemase enzyme-producing Proteus spp. in Bangladesh. blaOXA-833 is emerging in Bangladesh.

Background. Production of numerous biologically active compounds and their metabolites by intestinal microflora, interaction with the immune and other systems is of great importance while studying its changes in various diseases, one of which is thyrotoxicosis. So, the purpose of this study was to determine the severity of intestine microbioma disorder in white rats with experimental thyrotoxicosis (ET). Materials and methods. Studies were carried out on 25 mature male white rats (15 — control group, 10 — research group). ET was simulated by intragastric administration of L-thyroxine for 14 days. Under sterile conditions a laparotomy was performed, a section (2–3 cm) of the large intestine with its contents was taken. Sterile 0.9% NaCl solution was added to the content. Series of ten-fold dilutions with a concentration of the initial mixture of 10–2 to 10–11 was prepared. From each test tube 0.01 ml was seeded on solid nutrient media with subsequent isolation and identification of microbes according to morphological, tinctorial, cultural and biochemical properties. Results. The results of the study demonstrated that in ET animals the main microbioma is represented by bacteria Bifidobacterium, Lactobacillus, Bacteroides, and also opportunistic enterobacteria (Escherichia, Proteus, Klebsiella), peptococcus, staphylococci and clostridia. This is accompanied by the elimination of Peptostreptococcus, Enterococcus from bacterial biotope and the contamination of K. oxytoca and staphylococci. There was a pronounced deficit of bifidobacteria by 42.81 %, lactobacillus by 22.57 %, normal intestinal bacillus by 16.48 %. By the population level, the coefficient of quantitative dominance and the significance factor, the leading place is occupied by bacteroids, role of which is increased by 21.72 %, and lactobacillus role decreases by 39.31 %, bifidobacteria decreases by 51.48 % and E. coli decreases by 57.49 %. In this case, the role of peptococcus 3.37-fold increases, clostridia by 4.53, and by 73.93 % by the number of proteus. Conclusions. Under conditions of ET, there is an elimination of Bifidobacterium, Lactobacillus, Peptostreptococcus, Enterococcus and contamination of the biotope with conditionally pathogenic enterobacteria (Proteus, Klebsiella) and staphylococci. Deficiency of bifidobacteria and lactobacilli leads to changes in taxonomic structure and formation of dysbiosis of II and III stage.

In the years 2006–2011, 617 Proteus spp. strains isolated mostly from urine and wounds or other clinical sources were collected in Łódź, Poland, to determine the offensive O serotypes frequently occurring among patients. P. mirabilis exhibited the most intensive swarming growth and was dominating species (86.9%), followed by P. genomospecies, P. vulgaris, and P. penneri. Ninety four per cent strains were recognized as S (smooth) forms. Serological studies (involving ELISA—enzyme-linked immunosorbent assay and Western blotting using native and adsorbed rabbit antisera) enabled classification of 80% S isolates into respective Proteus O serogroups among the 83 ones, described so far. The remaining strains seemed to be serologically unique. Despite the observed big serological variety of Proteus spp. isolates, we found the O78 serogroup recently described in Poland as dominating and identified other widespread serotypes: O3, O6, O10, O11, O27, O28, and O30 reported earlier as predominating also in other countries; O77 and O79 detected lately in Poland; O16, O18, O20, and O50. No unique structural feature of the prevalent O serotypes has been indicated. However, the prevalence of some O serogroups indicates that particular serotypes may be in some ways beneficial to the strains producing these kinds of O antigen.

Background. The microecosystem of human organism closely cooperates with immune and endocrine systems, and in particular the microbiome of the pre­epithelial biofilm of the large intestine is one of the most sensitive biotopes that is able to change in different diseases. The purpose was to establish the microbiological structure (species composition and population level of microflora) of the pre­epithelial biofilm of the large intestine in albino rats with experimental hypothyroidism. Materials and methods. Experiments were carried out on 25 mature male albino rats weighing 220–240 g, of which 15 animals were included to the control group (intact animals), and 10 rats — to the main group. The experimental thyrotoxicosis was simulated by intragastric administration of mercazolilum for 14 days. Under sterile conditions, laparotomy was performed; a sample (up to 3 cm) of the large intestine with its contents was taken. The washed portion of the intestine was homogenized with a sterile 0.9% NaCl solution. A series of ten­fold dilutions with 10 –2 to 10 –7 concentrations of the initial mixture were prepared. From each tube, 0.01 ml were seeded on solid optimal nutrient media with subsequent isolation and identification of microbes according to morphological, tinctorial, cultural and biochemical properties. Results . Colonization resistance of the pre­epithelial biofilm of the large intestine changed in experimental animals when modeling hypothyroi­dism: the population level of lactobacteria reduced by 51.58 %, bifidobacteria — by 37.35 %, bacteroides — by 22.82 %, peptostreptococci — by 6.5 %. Pre­epithelial biofilm was contaminated by collibacillus and other enterobacteria (Proteus, Klebsiella), P.niger , staphylococci. The increase was detected in the population level of collibacillus by 10.31 % and clostridia by 8.2 %. Dysbiotic violations of I, II and III degrees were determined that requires a correction by means of probiotics. Conclusions. In the experimental research of mercazolilum­induced hypothyroidism on albino rats, the elimination of Bifidobacterium , Lactobacillus was established; contamination and colonization of biotope by opportunistic pathogenic microorganisms of Proteus , Klebsiella , Staphylococcus , Peptococcus were detected on the background of increased population level of collibacillus. There are violations of microbiocenosis in the form of dysbacteriosis degree I–III.

lt;bgt;Background and Objective:lt;/bgt; Ureolytic bacteria are bacteria capable of hydrolyzing urea. In construction, these bacteria are known to help improve soil stability. One of the habitats of ureolytic bacteria is cave ornaments such as gourdam, flowstone, stalagmite and stalactite. This study aims to find isolates and characterization of ureolytic bacteria in cave ornaments.lt;bgt;Materials and Methods:lt;/bgt; Urea-CaCllt;subgt;2lt;/subgt; was used as the isolation medium and urea agar medium was used as a qualitative urease test for cave ornament bacteria isolate. This study applied a survey method and tested for gram staining, spore staining, mannitol test, catalase test and lactose test for characterization.lt;bgt;Results:lt;/bgt; There were 17 isolates positive for urease from 30 isolates from the isolates of cave ornament bacteria. The characteristics of 17 ureolytic bacteria isolates were 2 isolates gram-negative basil with negative lactose test and 1 isolate positive glucose and 1 isolate negative glucose. Total 15 isolates gram-positive basil with spore staining results, 14 isolates spore-positive with 2 isolates positive mannitol and 12 isolates negative mannitol and 1 isolate spore-negative with negative catalase.lt;bgt;Conclusion:lt;/bgt; Total 17 ureolytic bacteria isolates were found from cave ornaments. Biochemical characterization showed 1 isolate oflt;igt;Proteuslt;/igt; spp., 1 isolate oflt;igt;Pseudomonaslt;/igt; spp, 2 isolates suspected of beinglt;igt;Bacillus megateriumlt;/igt; orlt;igt;Bacillus subtilislt;/igt;, 12 isolates oflt;igt;Bacillus cereuslt;/igt; and 1 isolate oflt;igt;Lactobacilluslt;/igt; spp.

Background: T2DM patients are more likely to have UTIs caused by resistant organisms such as ESBLs producing bacteria. Challenging reliable identification and prompt characterization of in-vitro susceptibilities of these bacteria are the first steps of deciding the appropriate antimicrobial therapy for UTIs caused by them. Objectives: To isolate and identify E. coli and K. pneumoniae from urine of T2DM patients with UTIs, to determine antibiotic resistance pattern among isolates, and to identify ESBLs production phenotypically and genotypically. Material and method: All samples were cultured on Cystine-Lactose-Electrolyte-Deficient Agar medium (CLED) by using calibrated loop. Growth of 100 colonies or more, i.e. 105 colony forming units (CFU)/mL urine was considered as signifi- cant bacteriuria. Isolation and identification were done according to standard method. All isolates were tested for antibiotic susceptibility testing by the disc diffusion method according to CLSI guidelines. Phenotypic detection of ESBLs was done by double-disk synergy test. Genotypic detection of blaTEM, blaSHV and blaCTX-M genes by using PCR. Results: Results of this study showed that E. coli and K. pneumoniae were the dominant bacterial isolates, they constituted 103 (91.2%) out of 113 urine isolates. E. coli (58. 4%) K. pneumoniae (32.7%), Enterococcus spp. (4.4%), Proteus spp. (2.7%) and Pseu- domonas spp. (1.8%). About 25 (24.3%) out of 103 E. coli and K. pneumoniae isolates were ESBLs positive by DDST, and 22 (88.0%) out of them had ESBLs encoding genes by conventional PCR. The most common gene detected was blaTEM (59.1%), followed by blaSHV (27.3%). CTX-M had not been detected in any of testes isolates. Conclusion: blaTEM and blaSHV genes were detected in 22 out of 25 ESBLs producing E. coli and K. pneumoniae isolates phenotypically detected by DDST. blaTEM was found to be the predominant gene (59.1%), while blaCTX-Mene was not detected in any of tested isolates. Keywords: Extended Spectrum β-Lactamases; Type 2 diabetes mellitus; Urinary tract infections; Phenotypic; genotypic methods.

Carbapenems, although originally introduced against multidrug-resistant (MDR) Gram negative bacilli (GNB), are now advocated for initial empiric use resulting in increasing carbapenem-resistant (CR) GNB. In this study, we analyzed the frequencies of CR-GNB and compared their resistance patterns against other antibiotics. Overall, 42% (1,014/2,420) of CR-GNB were isolated (range: 29-59%), with similar frequencies among hospitalized and community-acquired infections. However, the CR frequencies in Acinetobacter baumannii were significantly higher in the hospitalized patients (>50%). In addition, the CR-GNB isolates showed significantly higher resistance to the other antibiotics-fluoroquinolones, aminoglycosides, sulfonamides, and ureidopenicillins compared to carbapenem-sensitive isolates, thereby limiting further treatment options. Majority of CR-GNB isolates were extended spectrum β-lactamase producers (38-72%) and MDR (19-61%). Pan-drug resistant (PDR) frequencies among these MDR isolates ranged from 21% (Proteus spp.) to 100% (A. baumannii). Overall, CR-GNB are predominantly MDR or PDR and so warrant continuous antibiotic surveillance to provide better management of the infectious diseases.

Nosocomial infections are enhanced due to a flop in the infection control processes such as disinfection. The aim of this study was to assess long term effectiveness and the stability of disinfectants currently used within healthcare settings in Accra, Ghana against two indicator pathogens. Two locally produced and two imported disinfectants usually used in health care settings were obtained. The efficacy and long-term stability of the disinfectants was done using the in-use method, with identified microbial culture isolates of Pseudomonas aeruginosa and Staphylococcus aureus. Counts of both test organisms (Pseudomonas aeruginosa and Staphylococcus aureus) were over and above the 5 ≥ CFU recommended standard. All the disinfectants failed the test. However, contamination was observed to be higher in local disinfectants as compared to those imported. Furthermore, Enterobacter spp was isolated as contaminant from one imported disinfected and Proteus spp was also isolated from one local disinfectant. Interestingly, one of the imported disinfectants (Disinfectant I) showed more stability and was effective in the long term. All disinfectants did not pass the in-use test. However, disinfectants that are produced locally are more unstable and ineffective. Key words: Infections, disinfectants, drug stability, hospital, pathogens, contamination.

Aim. To evaluate the role of intestinal dysbiosis in the development of urinary tract infections in children admitted to a multidisciplinary hospital.Materials and Methods. We performed a bacteriological analysis of 2,694 urine samples collected from ≤ 3-years-old children who have been admitted to a multidisciplinary hospital (Kemerovo, Russian Federation). Urine specimens were inoculated by the 4-sector technique. Concurrently, we quantified the intestinal microbiota and its antimicrobial resistance in 100 children with urinary tract infections.Results. Titers of pathogenic microbes significantly differed in patients from distinct units (p = 0.02). The highest number of positive samples was detected in the neonatal pathology and neonatal intensive care units (43.8% each) as well as urology unit (37.9%). The most frequent pathogens belonged to Enterobacteriaceae family, in particular Escherichia coli, Klebsiella spp., Enterobacter spp., and Proteus spp. All studied children (100/100) suffered from intestinal dysbiosis. Titers of Bifidobacterium spp. and Lactobacillus spp. were reduced to 6 (5.0; 8.0) and 5 (4.0; 6.0) lg CFU/g, respectively. In 89.2% children, Escherichia coli lac+ levels were elevated to 9-10 lg CFU/g. Of note, 18.9% children had high Escherichia coli lac- titers [8 (6.0; 9.0) lg] and 24.3% had high Escherichia coli hly+ titers [5 (4.5; 6.0) lg] in the intestine. We have also found a high frequency (44.6%) and density [8.1 (7.0; 8.5) lg] of Klebsiella spp., in the intestinal mucosa of such patients. Notably, Enterobacter spp. and Proteus spp. were abundant [7 (5.0; 8.0) CFU/g] in the intestinal microbiota of 10.8% children. The prevalence of resistance strains in the studied setting reached 63%.Conclusions. More than 75% children with urinary tract infections suffer from intestinal dysbiosis. The microbiome of these patients was predominantly composed of Enterobacteriaceae and was characterized by high titers of Escherichia coli lac+, Klebsiella spp., Enterobacte spp., and Proteus spp. Similar profile of antimicrobial resistance in urinary and intestinal isolates of enterobacteria suggests intestinal microbiome as the main source of pathogens causing urinary tract infections in children.

Objective: to study the circulation and prevalence of phage-resistant strains among opportunistic enterobacteria, isolated from children with disorders of intestinal microflora, in relation to domestic preparations of bacteriophages.Materials and methods. A bacteriological study of the fecal microflora of the colon in 970 young children for dysbiosis was carried out. The sensitivity of 720 antibiotic-resistant strains of opportunistic enterobacteriaceae (S. aureus, K. pneumoniae, P. mirabilis and P. vulgaris, P. aeruginosa, E. coli, coagulase-negative staphylococci) isolated from children to domestic mono- and polyvalent drugs was studied (coliprotein, staphylococcal, Pseudomonas aeruginosa, purified Klebsiella pneumonia bacteriophage, “Sextafag”, Intesta bacteriophage). Determination of sensitivity to bacteriophages was carried out by the “sterile spot” method. Antibiotic sensitivity was determined by the disk diffusion method. Statistical processing of the results was carried out using the Microsoft Office Excel 2007 software package.Results. A large percentage of phage-resistant strains with low sensitivity to bacteriophages was revealed among opportunistic bacteria, amounting to 54,2%, with the highest circulation among coagulase-negative staphylococci, Proteus, Klebsiella (more than 50%). A significant spread of intestinal dysbiosis was established in young children (87,5%), characterized by a low content of bifidobacteria and a high level of allocation of opportunistic enterobacteria, including in associations.Conclusion. As a result of the studies, a significant prevalence of phage-resistant strains in children with dysbiosis was revealed, which indicates the need for preliminary determination of their sensitivity to bacteriophages in order to resolve the issue of their possible inclusion in the intestinal microflora correction system.

Surgical site infections (SSIs) are still the most prevalent infections in health care facilities. The magnitude of the problem increased with the development of health care associated infections caused by Gram negative bacilli (GNB), which are resistant to Carbapenem antibiotics. This study aimed to assess the performance of various detection methods of carbapenemase-producing GNB; isolated from healthcare associated SSIs at different surgical units, Mansoura University Hospitals, Al- Dakahliya Governorate, Egypt. A total of 186 wound specimens were collected from patients showing symptoms and signs of SSIs; used for isolation of bacteria and then identification of these bacterial isolates according to colony morphology; microscopic examination and biochemical reactions. About 173 specimens were positive for bacterial pathogens; out of them 83 were GNB isolates. The most commonly isolated bacteria were; Klebsiella spp. 31 (37.3%), followed by Escherichia. coli 22 (26.5%), Pseudomonas. aeruginosa 17 (20.5%), Proteus spp. 10 (12.0%) and Enterobacter spp. 3 (3.6%). The antibacterial sensitivity testing of the total 178 bacterial isolates was assessed using the disc diffusion assay. Bacterial pathogens that were carbapenemase producers were tested using phenotypic, rapid colorimetric (Carba NP test) and genotypic methods. Among these isolated bacteria 31 (83.8%), 26 (70.3%) and 28 (75.7%) were carbapenem resistant; confirmed by MHT, Carba NP test and multiplex Polymerase chain reaction (PCR), respectively. Continuous screening of the bacterial antimicrobial susceptibility at local level and rational use of the antibacterial agents; is essential to decrease the emergence and spread of resistant bacterial pathogens.

The serological classification scheme of the opportunistic Proteus bacilli includes a number of Proteus penneri strains. The tested P. penneri 4034-85 strain turned out to be serologically distinguished in ELISA and Western blotting. The O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of this strain and studied by sugar and methylation analyses and dephosphorylation along with 1H and 13C NMR spectroscopy, including 2D 1H,1H COSY, TOCSY, ROESY, 1H,13C HSQC, HMBC, and HSQC-TOCSY experiments, The O-polysaccharide was found to have a linear repeating unit containing glycerol 1-phosphate and two residues each of Gal and GlcNAc. The following O-polysaccharide structure was established, which, to our knowledge, is unique among known bacterial polysaccharide structures.

Proteus species are common opportunistic bacteria and foodborne pathogens. The proper detection of Proteus can effectively reduce the occurrence of food-borne public health events. Proteus mirabilis and Proteus vulgaris are the two most important pathogens in the Proteus genus. In this study, a dual TaqMan Real-Time PCR method was established to simultaneously detect and distinguish P. mirabilis and P. vulgaris in samples. The method exhibited good specificity, stability, and sensitivity. Specifically, the minimum detection concentrations of P. mirabilis and P. vulgaris in pure bacterial cultures were 6.08 × 102 colony forming units (CFU)/ml and 4.46 × 102 CFU/ml, respectively. Additionally, the minimum detectable number of P. mirabilis and P. vulgaris in meat and milk was 103 CFU/g. In addition, the method can be used to distinguish between strains of P. mirabilis and P. vulgaris within two hours. Overall, it is a sensitive, easy-to-use, and practical test for the identification and classification of Proteus in food.

This work was carried out to determine the incidence of Proteus species infection in patients had diabetic foot wounds admitted to Vascular Surgery Department at Assiut University Hospitals. Proteus isolates were the most isolated organism (37.73%) followed by Klebsiella spp. (22.64%), then Pseudomonas spp. (20.75%) and E. coli (18.87%). Proteus mirabils was the most isolated species represented (78%) followed by Proteus vulgaris (13%) then Proteus penneri (9%). The antimicrobial susceptibility patterns of the isolated Proteus spp. were determined using agar disk diffusion method. The highest sensitivity was to ertapenem 76 isolates (76%). The antibiotic sensitivity then decrease in descending manner to be amikacin (65%) > meropenem (54%) > imipenem (52%) > cefipime (49%), while the highest resistance rates were to amoxicillin–clavulanic acid 75 isolates (75%) > co-trimoxazole (73%) > cefoxitin (63%) > ciprofloxacin (49%). Plasmid DNA profile analysis of 10 MDR Proteus mirabilis that were common resistant to ceftriaxone was studied. Plasmid bands of six resistant Proteus mirabilis isolates were shown at 800bp while the others at 700 bp. Plasmid curing was done by EtBr and SDS. Results of plasmid curing using ethidium bromide sublethal concentration of EtBr 1.25% showed that 7 cured cells become sensitive to ceftriaxone (30 µg), while 3 non cured cell still resistant to ceftriaxone. Plasmid curing using SDS sublethal concentration of SDS 1.2% cured only one of ten Proteus mirabilis which become sensitive to ceftriaxone and lost its band at 800bp, the other Proteus mirabilis not cured by SDS.

NOSOCOMIAL infections are a major challenge, as one third of yearly admissions estimated as deaths. In intensive care units (ICUs), Gram-negative pathogens are responsible for the majority of these deadly infections. Multidrug resistance (MDR) in the seorganisms has been attributed to production of modified enzymes responsible of elevated MICs and MBC/MICs putting them in the transient stages for persistence and tolerance. Bacterial cells those can survive high doses of antibiotics comprise a subpopulation of persistent but not resistant cells. Bacterial isolates (326) were collected from nosocomial infections via microbiology laboratories of police hospitals in Cairo. Frequency-wise, it was found that Escherichia is the most frequent (155), Pseudomonas (96), Staphylococcus (34), Klebsiella (24), Proteus (16) while Citrobacter is the least frequent (1). Pseudomonas, Klebsiella, Proteus and Citrobacter showed the highest MDRs, with the highest MICs in a descending order. Results clearly indicated the high risk of the two Pseudomonas aeruginosa from sputum and urine persisting (inhibited but not killed) against MICs 3 tested antibiotics; CRO, AMC and FEB. It is highly recommended here to run tests for MBC/MICs to choose the antibiotic with least MBC/MIC ratio; less than four, in order to reduce the transient persistent and/ or tolerant stages.

Routine microbiological monitoring of rodent colonies in animal facilities is essential to evaluate the health status of the animals used in research studies. In the present study, animals were examined for the presence of selected microbial infections. In order to determine the contamination rates of mice and rats in Argentina, animals from 102 conventional facilities were monitored from 2012 to 2016. The most frequent bacteria isolated were Pseudomonas aeruginosa and Proteus spp. The common parasites identified were Syphacia spp. and Tritrichomonas spp. Serological assays demonstrated the highest prevalence for Mouse hepatitis virus in mice and Sialodacryoadenitis virus in rats. The results indicate that there is a high incidence of infections, so it is suggested that an efficient management system and effective sanitary barriers should be implemented in conventional facilities in Argentina in order to improve sanitary standards.

Objectives In this study, we aimed at identifying community and hospital-induced uropathogens isolated in urinary tract infection (UTI) determining the regional antibiotic resistance and the antibiotic preferences in empirical treatment in Sanliurfa/Turkey. Methods The urinary culture results of the 842 paediatric patients, who were aged between 0 and 18 years, admitted to Department of Pediatrics, Harran University Medi­cal Faculty Hospital, Sanliurfa, Turkey with UTI complaints, diagnosed with UTI and in whose urine cultures production was detected, were retrospectively evaluated. Age, gender, clinical findings and culture results of the patients were examined in terms of reproducing pathogens, the frequency of their being community and hospital induced, Extended Spectrum Beta Lactamase production of reproduced pathogens, sensitivity and resistance to antibiotics. Results A total of 842 patients, 472 (56.1%) girl were included in the study. According to the results of urine culture, Escherichia coli was detected in (58.9%) of the patients, Klebsiella (17.9%) and Proteus (15.8%). While high resistance to ampicillin (87.3%), cefuroxime (71.6%) and trimethoprim-sulfamethoxazole (60.8%) was found for all microorganisms, the lowest resistance to nitrofurantoin (21.4%), piperacillin/tazobactam (19.1), imipenem (8.6%), meropenem (8.8%), amikacin (6.2%) and cefoperazone/sulbactam (CSL) (4.7%) were determined in descending order. Resistance rates were higher in inpatients with UTI than in outpatients. Conclusions We think that the most appropriate antibiotic to be chosen for the outpatients for empirical treatment in all age groups in our region, is as oral nitrofurantoin and parenteral amikacin. Also the appropriate parenteral antibiotics that should be selected for the empirical treatment of inpatients UTI in all age groups are the CSL, amikacin and carbapenems.

Objectives To report a novel tigecycline resistance gene, tet(X6), and its variants in four bacterial species isolated from chickens and pigs in China. Methods WGS was conducted to identify the suspected resistance genes in the tigecycline-resistant Myroides phaeus 18QD1AZ29W. Functional cloning, homology modelling and molecular docking were performed to compare the function with other Tet(X) variants. Retrospective screening for tet(X6) was conducted for 80 isolates in our WGS data collection, and all genomic environments of tet(X6)-positive isolates were analysed. Results The tigecycline-resistant M. phaeus 18QD1AZ29W isolated from a pig farm in Shandong in 2018 was positive for tet(X2) and a novel tet(X) gene, designated tet(X6). Tet(X6) could increase the MICs of all tested tetracyclines/glycylcyclines for Escherichia coli only 2- to 4-fold, which was possibly due to a lower tetracycline binding capacity of Tet(X6) compared with that of other Tet(X) variants. Retrospective screening showed that seven other isolates (7/80, 8.8%), comprising four Proteus spp. and three Acinetobacter spp. from chickens and pigs in Shandong and Guangdong, were positive for three different variants of tet(X6). The analysis of the genomic environment revealed that two tet(X6)-positive isolates from M. phaeus and Proteus cibarius, respectively, contained ISCR2, which may play a role in tet(X6) transmission. Conclusions This study identified a novel type of tigecycline resistance gene, tet(X6), in Myroides, Acinetobacter and Proteus from chickens and swine. Tet(X6) conferred lower tetracycline/glycylcycline MICs than other Tet(X) variants, and ISCR2 may play a role in the transmission of tet(X6).

Faecal (FM) and colon mucosal associated microbiota (MAM) were studied in a model of colorectal cancer (CRC), the Apc-mutated Pirc rats, and in age-paired wt F344 rats. Principal Coordinates Analysis indicated that samples’ distribution was driven by age, with samples of young rats (1 month old; without tumours) separated from older ones (11-month-old; bearing tumours). Diversity analysis showed significant differences between FM and MAM in older Pirc rats, and between MAM of both Pirc and wt rats and the tumour microbiota, enriched in Enterococcus, Escherichia/Shigella, Proteus and Bifidobacteriaceae. In young animals, Pirc FM was enriched in the genus Delftia, while wt FM was enriched in Lactobacillus and Streptococcus. Some CRC biomarkers and faecal short chain fatty acids (SCFAs) were also measured. Colon proliferation and DClK1 expression, a pro-survival mucosal marker, were higher in Pirc than in wt rats, while the mucin MUC2, was lower in Pirc rats. Branched SCFAs were higher in Pirc than in wt animals. By Spearman analysis CRC biomarkers correlated with FM (in both young and old rats) and with MAM (in young rats), suggesting a specific relationship between the gut microbiota profile and these functional mucosal parameters deserving further investigation.

The prevalence of tet(X) genes threatens the clinical use of last-line tigecycline. The tet(X6) gene has been reported in Proteus strains, but its genetic context is rarely reported. This study aimed to investigate the prevalence and genetic contexts of tet(X6) gene in Proteus spp.A tet(X6) variant-bearing P. terrae subsp. cibarius strain was subjected to susceptibility testing, determination of growth curves, scanning electron microscopy, transmission electron microscopy and whole-genome sequencing (WGS). The genomic contexts of the tet(X6)-positive strain were analysed by sequence comparison and annotation.ZJ19PC, a P. terrae subsp. cibarius strain harbouring the tet(X6) variant, was isolated from 20 cecum samples collected in Zhejiang, China. The chromosome size of ZJ19PC was 3 952 084 bp; the GC content was 38.2%; and hugA, sul2, tet(H), floR, dfra1, aadA1, aac(3)-IV and aph(4)-la were found in addition to the tet(X6) variant. Proteus spp. could be classified into three groups based on the tet(X6) gene contexts. Strain ZJ19PC belongs to group 1 (sra-sul2-ISCR2-floR-ISCR2-floR-ISCR2- tet(X6) variant-tnpA-ISEc59-aph(4)-la-aac(3)-Iva-IS26), and this region of group 1 was inserted between modA and guaA. The common antimicrobial resistance (AMR) genes of the three types of AMR gene islands were sul2, floR, tet(X6) and aac(3). The tet(X6) gene contexts and SNP tree showed that ZJ19PC was homologous to HNCF44W and HNCF43W, which indicated that these strains may be clonally transmitted.This study analysed the genetic contexts of the tet(X6) gene in Proteus spp. and highlighted the significance of monitoring tigecycline-resistant P. terrae subsp. cibarius.

The adherence of Proteus mirabilis to the surface of urinary catheters leads to colonization and eventual blockage of the catheter lumen by unique crystalline biofilms produced by these opportunistic pathogens, making P. mirabilis one of the leading causes of catheter-associated urinary tract infections. The Proteus biofilms reduce efficiency of antibiotic-based treatment, which in turn increases the risk of antibiotic resistance development. Bacteriophages and their enzymes have recently become investigated as alternative treatment options. In this study, a novel Proteus bacteriophage (vB_PmiS_PM-CJR) was isolated from an environmental sample and fully characterized. The phage displayed depolymerase activity and the subsequent genome analysis revealed the presence of a pectate lyase domain in its tail spike protein. The protein was heterologously expressed and purified, the ability of the purified tail spike to degrade Proteus biofilms was tested. We showed that the application of the tail spike protein was able to reduce the adherence of bacterial biofilm to plastic pegs in a MBEC (minimum biofilm eradication concentration) assay and improve the survival of Galleria mellonella larvae infected with Proteus mirabilis. Our study is the first to successfully isolate and characterize a biofilm depolymerase from a Proteus phage, demonstrating the potential of this group of enzymes in treatment of Proteus infections.

Here, we explored the influences of thyme microcapsules on the growth, gene expression, and histamine accumulation by Proteus bacillus isolated from smoked horsemeat sausage. RT-qPCR was employed to evaluate the gene expression level of histidine decarboxylase (HDC) cascade-associated genes. We used HPLC to monitor histamine concentration both in pure culture as well as in the processing of smoked horsemeat sausage. Results showed that histamine accumulation was suppressed by thyme microcapsule inhibitory effect on the histamine-producing bacteria and the reduction in the transcription of hdcA and hdcP genes. Besides, compared with thyme essential oil (EO), thyme microcapsules exhibited higher antibacterial activity and had a higher score for overall acceptance. Therefore, the addition of thyme microcapsules in Xinjiang smoked horsemeat sausage inhibits histamine accumulation.

Resume- La distribution des bacteries, autres que les staphlocoques sur la tige des poils, a la surface cutanee et dans les follicules pileux de 8 chiens est analysee. Sur la tige des poils Micrococcus spp. et les bacteries aerobies gram megatifs sont plus nombreuses avec des numerations variant de 1,12 a 0,84 log10 (colonies formant unites par cm2 ). Des nombres hautement significatifs (p < 0,05) sont egalement trouves. A la surface cutanee Micrococcus spp. des bacteries aerobies gram negatifs et Clostridium sp. sont les plus nombreuses avec des numerations respectives variant de 0,62, 1,12 et 0,84 log10 (colonies formant unites par cm2 ). Des nombres hautement significatifs (p < 0,05) de Micrococcus spp. sont trouves de facon plus importante a l'interieur des follicules pileux qu' a la surface cutanee, les Streptocoques et Bacillus sp. ont ete trouves respectivement sur cinq et quatre chiens. Proteus sp., Pseudomonas sp. Nocardia sp. sont occasion-nellement trouves. [HARVEY, R.G., LLOYD, D.H. The distribution of bacteria (other than Staphylococci and Propionibacterium acnes) on the hair, at the skin surface and within the hair follicles of dogs (Distribution des bacteries autres que Staphylococci et Propionibacterium acnes) sur le poil, a la surface de al peau et dans les follicules pileux). Resumen- Presentamos la localization de bacterias noestafilococicas en el pelo, en la superficie cutanea y dentro del foliculo piloso de ocho perros. En los pelos, Micrococcus spp. abundantes, con contajes medios entre 1.12 a 0.84 Log10 (unidades formadoras de colonias +1) cm-1 , respectivamente. Se encontro a nivel proximal un numero significativamente mayor (p < 0.05) de bacterias aerobicas gram-negativas y Bacillus spp. En la superficie cutanea, Micrococcus spp., las bacterias aerobicas contajes medios de 0.62, 1.12 y 0.84 Log10 (unidades formadoras de colonias +1) cm"2 , respectivamente. Se aislo un numero significativamente mayor (p < 0.05) de Micrococcus spp. dentro de los foliculos pilosos que en la superficie cutanea (p < 0.05). Se aislo Streptococi y Bacillus spp. en cinco y cuatro perros, respectivamente. Proteus spp., Pseudomonas spp. y Nocardia spp. fueron hallados ocasionalmente. [HARVEY, R.G., LLOYD, D.H. The distribution of bacteria (other than Staphylococci and Propionibacterium acnes) on the hair, at the skin surface and within the hair follicles of dogs (Localization de bacterias (exceptuando Staphilococci y Propionibacterium acnes) en el pelo, en la superficie cutanea y dentro de los foliculos pilosos). Abstract- The distribution of bacteria, other than staphylococci, on the hair shaft, at the skin surface and within the hair follicles of eight dogs is reported. On the hair shafts Micrococcus spp. and aerobic Gram-negative bacteria were most numerous, with mean counts ranging from 1.12 to 0.84 Log10 (colony forming units + 1) cm"1 respectively. Significantly higher numbers (p < 0.05) of Gram-negative bacteria and Bacillus sp. were found proximally. At the skin surface Micrococcus spp., aerobic Gram-negative bacteria and Clostridium spp. were the most numerous with mean counts of 0.62, 1.12 and 0.84 Log10 (colony forming units + 1) cm"2 , respectively. Significantly higher numbers (p < 0.05) of Micrococcus spp. were found within the hair follicles than on the skin surface (p < 0.05). Streptococci and Bacillus spp. were found on five and four dogs, respectively. Proteus spp., Pseudomonas spp. and Nocardia spp. were occasionally found.

This work investigated the effect of lipase addition on a Chinese traditional fermented fish product, Suanzhayu. The accumulation of lactic acid and the decrease of pH during the fermentation were mainly caused by the metabolism of Lactobacillus. The addition of lipase had little effect on pH and the bacterial community structure but promoted the growth of Proteus. The addition of lipase promotes the formation of volatile compounds, especially aldehydes and esters. The formation of volatile compounds is mainly divided into three stages, and lipase had accelerated the fermentation process. Lactobacillus, Enterococcus and Proteus played an important role not only in inhibition of the growth of Escherichia-Shigella, but also in the formation of flavor. This study provides a rapid fermentation method for the Suanzhayu process.

Antibiotics are frequently used for treating urinary tract infections (UTI) in dogs and cats. UTI often requires time-consuming and expensive antimicrobial susceptibility testing (AST). Alternatively, clinicians can employ Flexicult Vet, an affordable chromogenic agar with added antibiotics for in-clinic AST. We investigated how well veterinary microbiologists and clinicians, without any prior experience, employ Flexicult Vet for the identification and AST of the most common canine and feline urinary pathogenic bacteria. We prepared 47 monoculture plates containing 10 bacterial species. The test’s mean accuracy was 75.1% for bacteria identification (84.6% and 68.7% for microbiologists and clinicians, respectively) and 79.2% for AST (80.7% and 78.2%). All evaluators employed Flexicult Vet with the accuracies over 90% for the distinctively colored bacteria like Escherichia coli (red), Enterococcus faecalis (turquoise), and Proteus spp. (pale brown). However, the evaluators’ experience proved important in recognizing lightly colored bacteria like Staphylococcus pseudintermedius (accuracies of 82.6% and 40.3%). Misidentifications of E. faecium additionally worsened AST performance since bacterial intrinsic resistance could not be considered. Finally, only 33.3% (3/9) of methicillin-resistant S. pseudintermedius (MRSP) were correctly detected. To conclude, Flexicult Vet proved reliable for certain urinary pathogens. In contrast, light-colored bacteria (e.g., Staphylococcus), often misidentified, require a standard AST.

Introduction. Difficulties in the management and treatment of patients with uncomplicated recurrent lower urinary tract infection (LUTI) are largely because bacterial, viral and other components remain unexplored in the etiological structure of the disease.Purpose of the study. To study the urine bacterial-viral associations in uncomplicated recurrent infections of the lower urinary tract (LUT).Materials and methods. The study included 14 patients with uncomplicated recurrent infections of the LUT, the average age was 33.0 ± 4.7 years. The object of the study was the average portion of morning urine collected prior to the appointment of empirical antibiotic therapy. Urine was divided into 3 aliquots: 1 for general urine analysis, 2 for bacteriological examination, 3 for polymerase chain reaction (PCR).Results. Aerobic-anaerobic associations were found in urine in all cases. Enterobacteriaceae family representatives were discovered in 92.8% of cases; they were represented by E. coli (78.6%), Klebsiella spp. (14.2%), Proteus spp. (7.1%). Moreover, in one patient at the same time (7.1%) E. coli + Klebsiella spp was isolated from urine.The gram-positive microflora pattern consisted of 7 genera and / or species and in various combinations was found in all patients. The taxonomic structure of the gram-positive flora was represented by E. faecalis (64.3%), S. lentus (21.4%), Corynebacterium spp., S. warneri, E. faecium (14.3% each), S. epidermidis + E. coli was recorded in 4 (28.6%) women, E. faecalis + E. faecium + E. coli + Klebsiella spp. - in 1 (7.1%), E. faecalis + E. faecium + E. coli also in 1 (7.1%) patients.Non-clostridial anaerobic bacteria (NAB) were isolated from urine in all cases with Eubacterium spp dominating. (57.1%). The taxonomic structure of NAB was represented by 5 genera: Eubacterium spp., Peptostreptococcus spp., Peptococcus spp., Bifidobacterium spp., Propionibacterium spp.Viruses were detected in urine in 6 (42.9%) patients. Epstein-Barr virus (EBV) was detected in 4 (28.6%) of them, moreover, in 1 case, in association with human papillomavirus (HPV53), in two other cases (7.1% each), cytomegalovirus (CMV) was verified and human herpes simplex virus type 6 (HHV6), respectively.Conclusions. In patients with uncomplicated recurrent LUTI, viruses with a prevalence of EBV were verified in urine in 42.9% of cases (28.6%). Apparently, the treatment of this patients` cohort should be directed not only to the bacterial, but also the viral components of the urinary microbiota.