Investigation of Proteus vulgaris and Elizabethkingia meningoseptica invasion on muscle oxidative stress and autophagy in Chinese soft-shelled turtle (Pelodiscus sinensis).

Muscle is an important structural tissue in aquatic animals and it is susceptible to bacterial and fungal infection, which could affect flesh quality and health. In this study, Chinese soft-shelled turtles were artificially infected with two pathogens, Proteus vulgaris and Elizabethkingia meningoseptica and the effects on muscle nutritional characteristics, oxidative stress and autophagy were assayed. Upon infection, the muscle nutritional composition and muscle fiber structure were notably influenced. Meanwhile, the mRNA expression of Nrf2 was down-regulated and Keap1 up-regulated, thus resulting in a decrease in antioxidant capacity and oxidative stress. However, with N-acetylcysteine treatment, the level of oxidative stress was decreased, accompanied by significant increases in antioxidant enzyme activities and the mRNA levels of SOD, CAT, GSTCD, and GSTO1. Interestingly, there was a significant increase in autophagy in the muscle tissue after the pathogen infection, but this increase could be reduced by N-acetylcysteine treatment. Our findings suggest that muscle nutritional characteristics were dramatically changed after pathogen infection, and oxidative stress and autophagy were induced by pathogen infection. However, N-acetylcysteine treatment could compromise the process perhaps by decreasing the ROS level and regulating Nrf2-antioxidant signaling pathways.

Identification of a novel conjugative plasmid carrying the multiresistance gene cfr in Proteus vulgaris isolated from swine origin in China.

The multiresistance gene cfr has a broad host range encompassing both Gram-positive and Gram-negative bacteria, and can be located on the chromosomes or on plasmids. In this study, a novel conjugative plasmid carrying cfr, designated as pPvSC3, was characterized in a Proteus vulgaris strain isolated from swine in China. Plasmid pPvSC3 is 284,528 bp in size and harbors 10 other antimicrobial resistance genes, making it a novel plasmid that differs from all known plasmids due to its unique backbone and repA gene. BLAST analysis of the plasmid sequence shows no significant homology to any known plasmid backbone, but shows high level homology to Providencia rettgeri strain CCBH11880 Contig_9, a strain isolated from surgical wound in Brazil, 2014. There are two resistance-determining regions in pPvSC3, a cfr-containing region and a multidrug-resistant (MDR) region. The cfr-containing region is flanked by IS26, which could be looped out via IS26-mediated recombination. The MDR region harbors 10 antimicrobial resistance genes carried by various DNA segments that originated from various sources. Plasmid pPvSC3 could be successfully transferred to Escherichia coli by conjugation. In summary, we have characterized a novel conjugative plasmid pPvSC3 carrying the multiresistance gene cfr and 10 other antimicrobial resistance genes, and consider that this novel type of plasmid deserves attention.

Understanding the adhesion mechanism of a mucin binding domain from Lactobacillus fermentum and its role in enteropathogen exclusion.

Lactobacillus species possesses surface exposed Mucin Binding Protein (MucBP) which plays a role in adhesion to gastrointestinal mucin. MucBP contains one or more mucin binding domain (MBD), the functionality of which has yet not been characterized thoroughly. Here, we have characterized a 93-amino acid MBD (MBD93) of MucBP (LAF_0673) from Lactobacillus fermentum. Multiple sequence alignment of L. fermentum MBD93 exhibited ∼60% sequence homology with MBDs from other Lactobacillus species. Further, we cloned, expressed and purified MBD93 from Escherichia coli as N-terminal histidine-tagged protein (6X His-MBD93). The purified MBD93 was able to bind to mucin and showed strong affinity towards the terminally expressed mucin glycans viz. N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), Galactose (Gal), and Sialic acid (N-acetylneuraminic acid; Neu5Ac). In silico experiments further confirmed the interaction between homology modeled MBD93 to mucin glycans through hydrogen-bonding with its surface amino acid residues Ser57, Pro58, Ile60, Tyr63 and Ala65. We also have demonstrated that MBD93 was able to inhibit the adhesion of enteric pathogens, including E. coli, Salmonella Paratyphi A, Shigella sonnei and Proteus vulgaris to mucin. Our results suggested that L. fermentum MBD93 is a functionally sufficient unit to act as an adhesin and to protect from invading enteric pathogens.

Swarming growth and resistance of Proteus penneri and Proteus vulgaris strains to normal human serum.

BACKGROUND: Proteus sp. strains isolated from patients with urinary tract infection (UTI) are often insensitive to the bactericidal action of normal human serum (NHS) which poses a clinical problem. The swarming phenomenon is an especially important factor in cases of UTIs gained through the ascending route. Both these virulence factors are connected with the cell surface components of bacteria, including lipopolysaccharide (LPS). OBJECTIVES: The resistance of Proteus bacilli to the bactericidal activity of NHS and the swarming phenomenon were investigated as well as the possible relationships between these virulence factors and the chemical structure of LPS. MATERIAL AND METHODS: The research was carried out on P. penneri and P. vulgaris species. Two preparations of sera were tested with respect to the bactericidal action of NHS. The ability of bacteria to swarm was checked on broth agar plates. The length of the O-specific part of LPS was estimated after poliacrylamide gel electrophoresis (PAGE) and staining with silver nitrate. RESULTS: Among the 62 tested Proteus strains, over 62% of Proteus vulgaris and 50% of Proteus penneri strains were sensitive to the bactericidal action of NHS. However, the number of resistant strains grew dramatically when serum with blocked complement activation via the alternative pathway was used. From 102 of the Proteus sp. Strains, only few were unable to swarm over the solid surface of the media. The remaining showed diverse ability to translocate. CONCLUSIONS: There was no definite correlation between the chemical structure of the O-specific chains of lipopolysaccharides and sensitivity or resistance of the Proteus sp. strains to NHS. Also, no significant relationships were found between the length or the chemical structure of the O-specific chains of the bacterial LPSs and the swarming phenomenon.

Crystalline bacterial biofilm formation on urinary catheters by urease-producing urinary tract pathogens: a simple method of control.

The problem of catheter encrustation stems from infection by urease-producing bacteria. These organisms generate ammonia from urea, elevate the pH of urine and cause crystals of calcium and magnesium phosphates to form in the urine and the biofilm that develops on the catheter. In this study, a laboratory model was used to compare the ability of 12 urease-positive species of urinary tract pathogens to encrust and block catheters. Proteus mirabilis, Proteus vulgaris and Providencia rettgeri were able to raise the urinary pH above 8.3 and produce catheter-blocking crystalline biofilms within 40 h. Morganella morganii and Staphylococcus aureus elevated the pH of urine to 7.4 and 6.9, respectively, and caused some crystal deposition in the biofilms but did not block catheters in the 96 h experimental period. Isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Serratia marcescens, Pseudomonas aeruginosa and Providencia stuartii were only capable of raising the pH of urine to a maximum of 6.4 and failed to cause crystal deposition in the biofilm. The most effective way to prevent catheter encrustation was shown to be diluting urine and increasing its citrate concentration. This strategy raises the nucleation pH (pH(n)) at which calcium and magnesium phosphates crystallize from urine. Increasing the fluid intake of a healthy volunteer with citrated drinks resulted in urine with a pH(n) of >8.0 in which catheter encrustation was inhibited. It is suggested that this dietary strategy will be an effective means of controlling catheter encrustation, whichever bacterial species is causing the problem.

[Isolation, identification, phylogenetic analysis and related properties of a pathogen in Silurus meridionalis Chen].

In October 2005, a large number of adults of Silurus meridionalis Chen died in the mud fish farming of Sichuan province. Later, three predominate strains of bacteria were isolated from the body of moribund fish. By artificial infection tests, strain TWN3 was confirmed to be the pathogen of the disease. Based on the characteristics of morphology, physiology and biochemistry tests, TWN3 was initially identified as Proteus vulgaris, and its G + C content of DNA is 39.1% . After being amplified, the sequence of its 16S rDNA was analyzed in the database of NCBI and it showed that TWN3 had the highest similarity to P. vulgaris, with 99.52% identity. By constructing the molecular phylogenetic dendrogram with Minimum Evolution method in Mega3.1, it was revealed that TWN3 was in the same branch with P. vulgaris. Based on all the results above, TWN3 is identified as P. vulgaris. However, the result of one biochemistry test, growth in KCN, deviates from the description in Bergey's Manual of Systematic Bacteriology. With reference to the Manual above, Proteus vulgaris is divided into two groups, P. vulgaris BG2 and 3. According to the specific biochemical properties, TWN3 is classified as a member of P. vulgaris BG3. Relevant tests of biological properties were also conducted, which showed that this strain has no haemolysis and is sensitive to four kinds of antibiotic such as gentamicin. Moreover, it can strongly cause diseases to mice. The research on the growth property of strain TWN3 indicated that its growth temperature ranges from 10 degrees C to 43 degrees C , optimum 37 degrees C ; growth pH ranges from 4 to 11, optimum 6. Its optimum salinity varies under different temperatures, and it grows best under 1.5 % salinity while 37 degrees C. The aim of these researches is to provide an evidence for the prevention and cure of TWN3. According to the appearance of the diseased Silurus meridionalis Chen and results of artificial infection test on crucian carps, it is considered that Silurus meridionalis Chen is infected through digestive system and it is also recommended that Bdellovibrio should be used in biological control. Although the pathogenicity of P. vulgaris is extensive, there has been no report that P. vulgaris is considered as the pathogen of cultivated Silurus meridionalis Chen so far. In addition, the identification of strain TWN3 has positive effects on the future research on the taxonomy of Proteus.

Enteric bacteria mandibular osteomyelitis.

Osteomyelitis of the mandible is a relatively rare inflammatory disease that usually stems from the odontogenic polymicrobial flora of the oral cavity. We are reporting 2 unusual cases of mandibular osteomyelitis resulting from enteric bacteria infection. In one patient, abundant clinical evidence suggested a diagnosis of a chronic factitious disease, whereas in the second patient no obvious etiology was found.

Crystallization of urine mineral components may depend on the chemical nature of Proteus endotoxin polysaccharides.

Formation of infectious urinary calculi is the most common complication accompanying urinary tract infections by members of the genus Proteus. The major factor involved in stone formation is the urease produced by these bacteria, which causes local supersaturation and crystallization of magnesium and calcium phosphates as carbonate apatite [Ca(10)(PO(4))(6).CO(3)] and struvite (MgNH(4)PO(4).6H(2)O), respectively. This effect may also be enhanced by bacterial polysaccharides. Macromolecules of such kind contain negatively charged residues that are able to bind Ca(2+) and Mg(2+), leading to the accumulation of these ions around bacterial cells and acceleration of the crystallization process. The levels of Ca(2+) and Mg(2+) ions bound by whole Proteus cells were measured, as well as the chemical nature of isolated LPS polysaccharides, and the intensity of the in vitro crystallization process was compared in a synthetic urine. The results suggest that the sugar composition of Proteus LPS may either enhance or inhibit the crystallization of struvite and apatite, depending on its chemical structure and ability to bind cations. This points to the increased importance of endotoxin in urinary tract infections.

Venous sinus thrombosis after Proteus vulgaris meningitis and concomitant Clostridium abscess formation.

A 19-y-old woman presented with Proteus vulgaris meningitis as a complication of chronic otitis media. Despite treatment with ceftazidime and amikacin no clinical improvement was observed. Cranial MRI revealed right-sided mastoiditis/otitis media and venous sinus thrombosis. After mastoidectomy, repeat cranial MRI demonstrated abscess formation in the venous sinuses. The abscess was drained. Clostridium spp. was isolated from the abscess culture.

[Investigation of hydrophobicity of Proteus vulgaris strains and ability of Proteus vulgaris and Proteus penneri strains to penetrate bladder membrane HCV T-29 cells ]. / Badania hydrofobowosci szczepów Proteus vulgaris oraz zdolnosci szczepów Proteus vulgaris i Proteus penneri do penetracji komórek nablonka pecherza moczowego HCV T-29.

Proteus bacilli play a particularly important role in urinary tract infections (UTI). Fimbriae and adherence ability and hemolysins production (HpmA, HlyA) are one of the factors of pathogenicity of these bacteria. In this paper we describe the invasion of HCV T-29 transitional bladder urothelial cells carcinoma strains of P. penneri, as well as P. vulgaris strains belonging to different serogroups. The cytotoxic effect was observed at 8 hour of incubation of the tested cells with P. vulgaris O21 and the same effect (complete lysis) at 6 hours by P. vulgaris O4 (this strain manifests maximal activity in the production of HlyA hemolysin). P. penneri strains, produce different types of fimbriae, expressed similar bacterial invasiveness. The hydrophobic properties of 25 P. vulgaris strains were also tested and only 3 strains occur to have hydrophobic cell surface.

Biochemical identification and characterization of DNA groups within the Proteus vulgaris complex.

We placed 43 isolates belonging to the Proteus vulgaris complex into proposed DNA groups (genomovars) using five previously recommended tests (tests for esculin hydrolysis, production of acid from salicin, L-rhamnose fermentation, and elaboration of DNase and lipase). On the basis of the results of these five tests, 49% of the isolates fell into DNA groups 5 and 6, 37% fell into DNA group 2, and the remaining 14% fell into DNA groups 3 and 4. Sequencing of the 16S rRNA genes of 11 members of DNA groups 5 and 6 indicated that 10 of these isolates (91%) could be unambiguously assigned to one of these two genomospecies. Overall expression of selected enzymatic and virulence-associated characteristics did not differ significantly among DNA groups.

Biological activities of lipopolysaccharides of Proteus spp. and their interactions with polymyxin B and an 18-kDa cationic antimicrobial protein (CAP18)-derived peptide.

The saccharide constituents of lipopolysaccharides (LPS) of Proteus spp. vary with the strain and contain unique components about which little is known. The biological activities of LPS and lipid A from S- and R-forms of 10 Proteus strains were examined. LPS from all S-form Proteus strains was lethal to D-(+)-galactosamine (GalN)-loaded, LPS-responsive, C3H/HeN mice, but not to LPS-hypo-responsive C3H/HeJ mice. P. vulgaris 025 LPS evoked strong anaphylactoid reactions in N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP)-primed C3H/HeJ mice. LPS from S- and R-form Proteus strains induced production of nitric oxide (NO) and tumour necrosis factor (TNF) by macrophages isolated from C3H/HeN but not C3H/HeJ mice. Lipid A from Proteus strains also induced NO and TNF production, although lipid A was less potent than LPS. The effects of LPS were mainly dependent on CD14; LPS-induced NO and TNF production in CD14+ J774.1 cells was significantly greater than in CD14-J7.DEF.3 cells. All LPS from Proteus strains, and especially from P. vulgaris 025, exhibited higher anti-complementary activity than LPS from Escherichia coli or Pseudomonas aeruginosa. Polymyxin B inactivated proteus LPS in a dose-dependent manner, but these LPS preparations were more resistant to polymyxin B than E. coli LPS. CAP18(109-135), a granulocyte-derived peptide, inhibited proteus LPS endotoxicity only when the LPS:CAP18(109-135) ratio was appropriate, which suggests that CAP18(109-135) acts through a different mechanism than polymyxin B. The results indicate that LPS from Proteus spp. are potently endotoxic, but that the toxicity is different from that of LPS from E. coli or Salmonella spp. and even varies among different Proteus strains. The variation in biological activities among proteus LPS may be due to unique components within the respective LPS.

The structure of the capsular polysaccharide from a swarming strain of pathogenic Proteus vulgaris.

The structure was determined for the capsular polysaccharide (CPS) isolated from a swarming strain of Proteus vulgaris, CP2-96, which was obtained from the spleen of an infected mouse. The CPS was extracted from the cell pellet by hot water, precipitated with ethanol, and further purified by gel-permeation chromatography. The structure was established by glycosyl composition and linkage analyses, and by NMR spectroscopy. The sequence of the glycosyl residues was determined by a NOESY experiment. The CPS is composed of a tetrasaccharide repeating unit with the following structure: OAc [symbol: see text] 4 -->4)-beta-D-Glcp-(1-->3)-beta-D-GalpNAc-(1-->2)-alpha-D-Glcp-(1-->4)-al pha- D-GlcpA-(1-->.

Potential virulence factors of Proteus bacilli.

The object of this review is the genus Proteus, which contains bacteria considered now to belong to the opportunistic pathogens. Widely distributed in nature (in soil, water, and sewage), Proteus species play a significant ecological role. When present in the niches of higher macroorganisms, these species are able to evoke pathological events in different regions of the human body. The invaders (Proteus mirabilis, P. vulgaris, and P. penneri) have numerous factors including fimbriae, flagella, outer membrane proteins, lipopolysaccharide, capsule antigen, urease, immunoglobulin A proteases, hemolysins, amino acid deaminases, and, finally, the most characteristic attribute of Proteus, swarming growth, enabling them to colonize and survive in higher organisms. All these features and factors are described and commented on in detail. The questions important for future investigation of these facultatively pathogenic microorganisms are also discussed.

[The hydrophobicity of gram-negative bacteria isolated from patients with inflammatory and suppurative-destructive lung diseases]. / Gidrofobnost' gramotritsatel'nykh bakterii, vydelennykh u bol'nykh s vospalitel'nymi i gnoino-destruktivnymi zabolevaniiami legkikh.

Hydrophobic properties of 11 enterobacterial strains and 12 Pseudomonas aeruginosa strains isolated from patient with inflammatory and purulent destructive diseases of the lungs were studied. The hydrophobic properties of the isolated bacteria were determined by the method of salting out with ammonium sulfate. A high correlation (r = 0.56) between the above-mentioned properties of gram-negative bacteria and their isolation rate from patients with destructive diseases was established. The hydrophobic properties of bacteria were supposed to be of importance in the process of the colonization of pulmonary tissues with bacterial flora.

[The enterotoxigenic capacity of hemolysin-producing strains of Proteus isolated in acute intestinal infections in children]. / Enterotoksigennaia sposobnost' gemolizinprodutsiruiushchikh shtammov proteev, vydelennykh pri ostrykh kishechnykh infektsiiakh u detei.

The capacity of P. mirabilis and P. vulgaris strains isolated in acute enteric infections in children for producing enterohemolysin, a new type of hemolysin, has been shown. The relationship between the capacity of Proteus cultures for producing enterohemolysin and their capacity for inducing toxic secretory reaction on a ligated loop on the small intestine of rabbits in the absence of known thermostable and thermolabile antitoxin in bacteria.

Production and properties of haemolysins from clinical isolates of the Proteeae.

A collection of 198 clinical isolates of strains belonging to the tribe Proteeae was examined for haemolytic activity on blood agar and in Brain Heart Infusion Broth. The strains were of diverse bacteriocin and O-antigenic types and from a wide variety of sources. They included representatives of all species of Morganella, Proteus and Providencia. Approximately half of the M. morgani strains were haemolytic on blood agar. This activity was not associated with any particular bacteriocin type. The haemolysin was also produced during exponential growth in broth and was thermolabile and calcium dependent. All P. mirabilis strains and some P. vulgaris strains were non-haemolytic on blood agar. However, most strains of the Proteus spp., irrespective of their bacteriocin and antigenic type, produced, over a short period during exponential growth in broth, a heat-stable, cell-associated calcium-independent haemolysin. A smaller proportion of P. vulgaris and P. penneri strains produced, in addition, a thermolabile, calcium-dependent haemolysin which was associated with the formation of large haemolytic zones on blood agar. The relationship of these haemolysins to Escherichia coli haemolysin and their possible role in virulence is discussed. Haemolysin production was not found in any of the 74 strains of four species of Providencia.

[Effect of polyvinylpyrrolidone on microbial cells]. / Vliianie polivinilpirrolidona na mikrobnye kletki.

The oral administration of polyvinylpyrrolidone to rats produces no effect on the intestinal microflora. At the same time, polyvinylpyrrolidone solutions decrease the toxicity of Escherichia coli and Proteus vulgaris, produce a bactericidal and agglutinating effect.

Some biological features of proteus bacilli. 3. Comparison of haemolytic activity of Proteus and Serratia strains.

The haemolytic activity of Proteus mirabilis and P. vulgaris bacilli exhibited in young broth cultures was compared with the ability of Serratia marcescens strains to haemolyze human and sheep erythrocytes in the same conditions.

Cell invasiveness of Proteus mirabilis and Proteus vulgaris strains.

Cell penetration ability of haemolytic and non haemolytic Proteus rods was compared. Among four Proteus strains all were able to invade the tested cells (Vero 135, HeLa, L-929 and human blood lymphocytes) but the expression of this feature by haemolytic strains was markedly higher. The survival and multiplication of intracellular bacteria, especially in the case of fresh human blood lymphocytes may be of importance for the development of infection in higher organisms.