Blowing epithelial cell bubbles with GumB: ShlA-family pore-forming toxins induce blebbing and rapid cellular death in corneal epithelial cells.

Medical devices, such as contact lenses, bring bacteria in direct contact with human cells. Consequences of these host-pathogen interactions include the alteration of mammalian cell surface architecture and induction of cellular death that renders tissues more susceptible to infection. Gram-negative bacteria known to induce cellular blebbing by mammalian cells, Pseudomonas and Vibrio species, do so through a type III secretion system-dependent mechanism. This study demonstrates that a subset of bacteria from the Enterobacteriaceae bacterial family induce cellular death and membrane blebs in a variety of cell types via a type V secretion-system dependent mechanism. Here, we report that ShlA-family cytolysins from Proteus mirabilis and Serratia marcescens were required to induce membrane blebbling and cell death. Blebbing and cellular death were blocked by an antioxidant and RIP-1 and MLKL inhibitors, implicating necroptosis in the observed phenotypes. Additional genetic studies determined that an IgaA family stress-response protein, GumB, was necessary to induce blebs. Data supported a model where GumB and shlBA are in a regulatory circuit through the Rcs stress response phosphorelay system required for bleb formation and pathogenesis in an invertebrate model of infection and proliferation in a phagocytic cell line. This study introduces GumB as a regulator of S. marcescens host-pathogen interactions and demonstrates a common type V secretion system-dependent mechanism by which bacteria elicit surface morphological changes on mammalian cells. This type V secretion-system mechanism likely contributes bacterial damage to the corneal epithelial layer, and enables access to deeper parts of the tissue that are more susceptible to infection.

Oropharyngeal microbiome of an HIV-positive patient.

Studies on understanding the human microbiome continue to grow rapidly; nonetheless, reports on alterations in the microbiome post HIV infection are limited. Human microbiome is an aggregate of bacteria, fungi, viruses and archaea that have co-evolved with humans. These microbes have important roles in immune modulation, vitamin synthesis, metabolism etc. The human pharyngeal microbiome, which resides in the junction between digestive and respiratory tracts, might have a key role in the prevention of respiratory tract infections, akin to the actions of the intestinal microbiome against enteric infections. The respiratory tract is constantly exposed to various environmental and endogenous microbes; however, unlike other similar mucosal surfaces, there has been limited investigation of the microbiome of the respiratory tract. HIV infection is associated with alterations in the respiratory microbiome. The aim of this study was to use next-generation sequencing to determine the composition of the oropharyngeal microbiome in a HIV-positive individual. The bacterial composition was determined by illumina sequencing using MiSeq of partial 16S rRNA genes (V3-V4). A total of 3, 57,926 reads were analyzed. Overall, the genera Proteus, Enterococcus, Bacteroides, Prevotella and Clostridium were most prevalent bacterial populations in the oropharynx of an HIV positive patient.

Determination of epidemiology and antimicrobial susceptibility of extended spectrum beta lactamase producing uropathogens.

OBJECTIVE: To find the most suitable antibiotic against urinary tract infection caused by Extended Spectrum Beta Lactamase producing uropathogens, and the epidemiology of Extended Spectrum Beta Lactamase producers. METHODS: The cross-sectional study was conducted at Pir Mehr Ali Shah University of Arid Agriculture, Rawalpindi, Pakistan, from July 2014 to July 2015, and comprised urine samples of patients suffering from urinary tract infection which were cultured on Cysteine Lactose Electrolyte Deficient agar medium. Analysis was done on Muller- Hintonagar plates and optical density was set as 0.1 at 530nm. Antimicrobial sensitivity was tested using Kirby-Bauer disc diffusion method. Further confirmation was done through gram staining and biochemical tests. Extended Spectrum Beta Lactamase production was confirmed through phenotypic methods, including phenotypic confirmatory disc diffusion test, double disc synergy test and Epsilometer test. RESULTS: Of the 150 samples, 98(65%) showed growth of a total of 114 pathogenic isolates. Escherichia coli was the commonest organism in 94(82%) samples. Piperacillin Tazocin was the most suitable antimicrobial drug in 88(90%) cases. Overall, 23(20%) isolates were producers of Extended Spectrum Beta Lactamase. CONCLUSION: Piperacillin Tazocin was found to be the drug of choice for patient suffering from urinary tract infection.

Impact of pH regulation on multicopper oxidase production and swarming motility in the bacterium Proteus hauseri ZMd44.

Proteus hauseri ZMd44, a biodecolorizing bacterium, has been known to produce electricity and multicopper oxidase (Mco-laccase) under copper induction. However, optimization and regulation of production have not been explored. This study is the first attempt to evaluate several parameters on biomass and Mco-laccase production of P. hauseri ZMd44. Through orthogonal experiments with Taguchi's L9, it was found that P. hauseri ZMd44 was sensitive to pH value. The cells grew relatively quickly at pH 7, thus the biomass and Mco-laccase production reached 1.66 g/L and 1043.6 U/L, respectively. Higher pH values also influenced the swarming motility, which is an important characteristic of P. hauseri ZMd44 that affects urinary tract infection. The swarming circle and the diameter of the swarm, represented by the motility velocity, were found to be more controlled after 24 h of growth at pH 6. The swarming ability of P. hauseri was completely inhibited by the addition of 3 mM copper or zinc ions. Therefore, the Mco-laccase and swarming motility could be controlled by regulating pH and ion content.

Indigo- and indirubin-producing strains of Proteus and Psychrobacter are associated with purple rind defect in a surface-ripened cheese.

The rinds of surface-ripened cheeses have expected aesthetic properties, including distinct colors, that contribute to overall quality and consumer acceptance. Atypical rind pigments are frequently reported in small-scale cheese production, but the causes of these color defects are largely unknown. We provide a potential microbial explanation for a striking purple rind defect in a surface-ripened cheese. A cheese producer in the United States reported to us several batches of a raw-milk washed-rind cheese with a distinctly purple rind. We isolated a Proteus species from samples with purple rind defect, but not from samples with typical rind pigments, suggesting that this strain of Proteus could be causing the defect. When provided tryptophan, a precursor in the indigo and indirubin biosynthesis pathway, the isolated strain of Proteus secreted purple-red pigments. A Psychrobacter species isolated from both purple and normal rinds also secreted purple-red pigments. Using thin-layer chromatography and liquid chromatography-mass spectrometry, we confirmed that these bacteria produced indigo and indirubin from tryptophan just as closely related bacteria make these compounds in purple urine bag syndrome in medical settings. Experimental cheese communities with or without Proteus and Psychrobacter confirmed that these Proteobacteria cause purple pigmentation of cheese rinds. Reports of purple rinds in two other cheeses from Europe and the observation of pigment production by Proteus and Psychrobacter strains isolated from other cheese rinds suggest that purple rind defect has the potential to be widespread in surface-ripened cheeses.

Kinetic study of Reactive Black 5 degradation by Fe2+/S2O82- process via interactive model-based response surface methodology.

This study aimed to kinetically discover optimal conditions on characteristics of Reactive Black 5 decolorization/degradation via ferrous (Fe2+)-activated potassium persulfate (PS). Monod-like kinetics and interactive model-based response surface methodology (RSM) were applied to fitting and predict optimize treatment. Biodegradability of the intermediates was also tested by shaking culture with two species (Proteus hauseri ZMd44 and Shewanella sp. WLP72). Results showed that the optimal degradation efficiency was predicted (through RSM) as pH 3.72, (PS) = 0.39 mM, and (Fe2+) = 0.29 mM. The transformation products (dl-4-hydroxymandelic acid, benzoic acid, benzene, formic acid, oxalic acid and acetic acid) were less toxic than the original dye solution. According to those results, clean-up of dye pollutants by the Fe2+/S2O82- process is feasible as a pre-processing for the biodegradation, and the predicted optimal conditions are meaningful for further industry utilization.

Significance and Roles of Proteus spp. Bacteria in Natural Environments.

Proteus spp. bacteria were first described in 1885 by Gustav Hauser, who had revealed their feature of intensive swarming growth. Currently, the genus is divided into Proteus mirabilis, Proteus vulgaris, Proteus penneri, Proteus hauseri, and three unnamed genomospecies 4, 5, and 6 and consists of 80 O-antigenic serogroups. The bacteria are known to be human opportunistic pathogens, isolated from urine, wounds, and other clinical sources. It is postulated that intestines are a reservoir of these proteolytic organisms. Many wild and domestic animals may be hosts of Proteus spp. bacteria, which are commonly known to play a role of parasites or commensals. However, interesting examples of their symbiotic relationships with higher organisms have also been described. Proteus spp. bacteria present in soil or water habitats are often regarded as indicators of fecal pollution, posing a threat of poisoning when the contaminated water or seafood is consumed. The health risk may also be connected with drug-resistant strains sourcing from intestines. Positive aspects of the bacteria presence in water and soil are connected with exceptional features displayed by autochthonic Proteus spp. strains detected in these environments. These rods acquire various metabolic abilities allowing their adaptation to different environmental conditions, such as high concentrations of heavy metals or toxic substances, which may be exploited as sources of energy and nutrition by the bacteria. The Proteus spp. abilities to tolerate or utilize polluting compounds as well as promote plant growth provide a possibility of employing these microorganisms in bioremediation and environmental protection.

Novel approach for the ammonium removal by simultaneous heterotrophic nitrification and denitrification using a novel bacterial species co-culture.

Agricultural activities lead excessive emission of ammonia nitrogen in the environment and can profoundly interfere the equilibrium of the natural ecosystems leading to their contamination. Actually, the biological purification of wastewaters is the most adopted technique thanks to its several advantages such as high performance and low energy consumption. For this reason, two novel strains of Alcaligenes sp. S84S3 and Proteus sp. S19 genus were isolated from an activated sludge and applied in the treatment of ammonium and nitrite in aqueous solution. Under the optimum operating conditions of temperature (30 °C), pH (7), carbon substrate (2 g/L of glucose) and duration of incubation time (69 h), the strain Alcaligenes sp. S84S3 could oxidize 65% of the ammonium as high as 272.72 mg-NH4(+)/L. Moreover, during 48 h, the nitrate reduction rate performed by the strain Proteus S19 was about 99 % without production of nitrite intermediate (negligible concentration). Moreover, the coculture of the strains Alcaligenes sp. S84S3 and Proteus sp. S19 could eliminate 65.83% of the ammonium ions without production of toxic forms of nitrogen oxides during a short time of incubation (118 h) at the same operational conditions with providing the aeration in the first treatment phase. The coculture of our isolated strains is assumed to have a good potential for nitrification and denitrification reactions applied in the treatment of wastewater containing ammonium, nitrite and nitrate. As a result, we can consider that the mixed culture is a practical method in the treatment of high-strength ammonium wastewater with reducing of sludge production.

Taxonomic characterisation of Proteus terrae sp. nov., a N2O-producing, nitrate-ammonifying soil bacterium.

In the context of studying the influence of N-fertilization on N2 and N2O flux rates in relation to the soil bacterial community composition in fen peat grassland, a group of bacterial strains was isolated that performed dissimilatory nitrate reduction to ammonium and concomitantly produced N2O. The amount of nitrous oxide produced was influenced by the C/N ratio of the medium. The potential to generate nitrous oxide was increased by higher availability of nitrate-N. Phylogenetic analysis based on the 16S rRNA and the rpoB gene sequences demonstrated that the investigated isolates belong to the genus Proteus, showing high similarity with the respective type strains of Proteus vulgaris and Proteus penneri. DNA-DNA hybridization studies revealed differences at the species level. These differences were substantiated by MALDI-TOF MS analysis and several distinct physiological characteristics. On the basis of these results, it was concluded that the soil isolates represent a novel species for which the name Proteus terrae sp. nov. (type strain N5/687(T) =DSM 29910(T) =LMG 28659(T)) is proposed.

Simultaneous nitrification and denitrification by novel heterotrophs in remediation of fish processing effluent.

Three isolates viz. Lysinibacillus sp. HT13, Alcaligenes sp. HT15 and Proteus sp. HT37 isolated from fish processing effluent and having a C/N ratio of 2, removed 218, 169, and 400 µg cell(-1) day(-1) NH4(+)-N, respectively without subsequent build up of nitrite or nitrate. Ability of the selected isolates in removing NH4(+)-N, NO2(-)-N, and NO3(-)-N was checked in the presence of four commonly reported and tested effluent carbon sources viz. pyruvate, glycerol, methanol, and acetate. Further, when supplemented to fish processing wastewater containing 234 ppm total Kjeldahl's nitrogen, Lysinibacillus sp. HT13, Alcaligenes sp. HT15, and Proteus sp. HT37 could remediate 95.74, 86.17, and 76.6% nitrogen, respectively in 48 h. This is the first report of a Lysinibacillus sp. carrying out aerobically the process of simultaneous nitrification and denitrification. The results demonstrate the potential of the isolates for use in treatment of fish processing effluents and demonstrating the efficient removal of ammonia.

Role of plant growth promoting rhizobacteria in modulating the efficiency of poultry litter composting with rock phosphate and its effect on growth and yield of wheat.

The present study was aimed to evaluate the role of Plant Growth Promoting Rhizobacteria (PGPR) in P solubilisation from rock phosphate through composting with poultry litter, and further to study the effects of prepared enriched composts on growth, yield, and phosphorus uptake of wheat crop. Various phosphorus-enriched composts were prepared from rock phosphate and poultry litter (1:10) with and without inoculation of plant growth promoting rhizobacterias (Pseudomonas sp. and Proteus sp.). Results showed that the rock-phosphate-added poultry litter had higher total phosphorus, available (Mehlic-3 extracted) phosphorus, microbial biomass (carbon and phosphorus), and lower total organic carbon, total nitrogen, and carbon/nitrogen ratio over poultry litter alone. Inoculation of Pseudomonas sp. with rock phosphate-added poultry litter showed maximum increase in available phosphorus (41% of total phosphorus) followed by Proteus sp. inoculation (30% of total phosphorus) over uninoculated treatment (23% of total phosphorus) on the 120th day of composting. Microbial biomass (carbon and phosphorus) increased up to Day 45 and tended to decrease till the 120th day of composting, irrespective of the treatments. However, in pot experiments, wheat seeds receiving inoculation with plant growth promoting rhizobacterias, subsequently treated with rock phosphate-enriched compost proved highly stimulatory to plant height, phosphorus uptake, grain yield, and seed phosphorus content over uninoculated untreated control. The plant growth promoting rhizobacterias inoculation can be a sustainable source releasing phosphorus from low grade rock phosphate through composting and application of rock phosphate-enriched compost can be an alternative to chemical fertilisers for better crop production.

2-methylbutanal, a volatile biomarker, for non-invasive surveillance of Proteus.

Pathogen detection needs a paradigm shift from time-consuming conventional microbiological and biochemical tests to much simpler identification methods with higher sensitivity and specificity. In this regard, a simple detection method for frequently isolated nosocomial uropathogen, Proteus spp., was developed using the characteristic volatile 2-methylbutanal released in Luria Bertani broth. The instant reaction of the compound with 5-dimethylaminonaphthalene-1-sulfonylhydrazine (DNSH) has been adapted to develop a sensitive fluorescence assay named "ProteAl" (Prote, "Proteus" & Al, "Aldehyde"). The assay was performed by direct addition of the fluorescence reagent to the culture after 7 h of growth. The distinct green fluorescence by Proteus (other organisms show orange fluorescence) served as the simplest and quicker identification test available for Proteus. In the laboratory, it exhibited 100% specificity and 100% sensitivity during testing of 95 strains including standard and known clinical isolates representing frequently encountered uropathogens.

Decolorization of textile azo dye and Congo red by an isolated strain of the dissimilatory manganese-reducing bacterium Shewanella xiamenensis BC01.

Shewanella xiamenensis BC01 (SXM) was isolated from sediment collected off Xiamen, China and was identified based on the phylogenetic tree of 16S rRNA sequences and the gyrB gene. This strain showed high activity in the decolorization of textile azo dyes, especially methyl orange, reactive red 198, and recalcitrant dye Congo red, decolorizing at rates of 96.2, 93.0, and 87.5%, respectively. SXM had the best performance for the specific decolorization rate (SDR) of azo dyes compared to Proteus hauseri ZMd44 and Aeromonas hydrophila NIU01 strains and had an SDR similar to Shewanella oneidensis MR-1 in Congo red decolorization. Luria-Bertani medium was the optimal culture medium for SXM, as it reached a density of 4.69 g-DCW L(-1) at 16 h. A mediator (manganese) significantly enhanced the biodegradation and flocculation of Congo red. Further analysis with UV-VIS, Fourier Transform Infrared spectroscopy, and Gas chromatography-mass spectrometry demonstrated that Congo red was cleaved at the azo bond, producing 4,4'-diamino-1,1'-biphenyl and 1,2'-diamino naphthalene 4-sulfonic acid. Finally, SEM results revealed that nanowires exist between the bacteria, indicating that SXM degradation of the azo dyes was coupled with electron transfer through the nanowires. The purpose of this work is to explore the utilization of a novel, dissimilatory manganese-reducing bacterium in the treatment of wastewater containing azo dyes.

Comparative evaluations on bio-treatment of hexavalent chromate by resting cells of Pseudochrobactrum sp. and Proteus sp. in wastewater.

Two marine bacterial strains, B5 and H24, were isolated from long-term Cr(VI) contaminated seawater and identified as Pseudochrobactrum and Proteus, respectively, based on 16S rRNA gene sequence analyses. Both strains were examined for their tolerance to Cr(VI) and other metal salts and their abilities to reduce Cr(VI) to trivalent chromium [Cr(III)]. Growing cells of Pseudochrobactrum sp. B5 and Proteus sp. H24 could tolerate Cr(VI) at a concentration of 2000 and 1500 mg/l and completely reduce 1000 mg/l Cr(VI) in LB medium within 96 and 144 h, respectively. Resting cells of the two strains were able to reduce 200mg/l Cr(VI) in Tris-HCl buffer within 16 and 24h, respectively. Furthermore, resting cells of both strains were able to reduce Cr(VI) in industrial wastewaters three times consecutively. Overall, this study provides evidence of the potential for application of chromate-reducing bacteria to direct Cr(VI) decontamination of industrial effluents.

Detection and PFGE analysis of ESBL-producing isolates of Proteus species isolated from patients at Tehran hospitals.

BACKGROUND: The production of extended-spectrum ß-lactamases (ESBLs), in particular TEM and CTX-M type, by the strains of Proteus spp. is a contributing factor to the emergence of bacterial drug resistance. The aim of this study was to screen and determine of ESBLs genes among strains of Proteus spp. at Tehran hospitals. MATERIAL/METHODS: A total of 100 clinical isolates of Proteus species and 1 isolate of Morganella morganii were collected from hospitals in Tehran. Susceptibility of these isolates to various antibiotics was tested by disk diffusion method. Microbroth dilution assay was then used to determine the minimum inhibitory concentration (MIC) of ceftazidime. The phenotypic confirmatory test (PCT) was used for detection of ESBLs. Isolates showing MIC ≥4 µg/ml for ceftazidime were screened for detection of ESBLs genes by PCR. The genomic DNA from ESBL-producing isolates was extracted and analyzed by PFGE. RESULTS: During the study, 11.8% of the isolates were positive for ESBLs genes. The MICs of ceftazidime-resistant isolates were in the range of 4 to 8 µg/ml. The Frequencies of different genes encoding the ESBLs were as follows: blaCTX-M (n=13), blaTEM (n=10), blaVEB (n=7), blaSHV (n=1), and blaPER (n=0). Analysis of 12 ESBL-producing isolates by PFGE produced 5 distinct clusters designated as I-V. CONCLUSIONS: The Detection of ESBL-producing isolates of Proteus spp. with similar PFGE pattern is alarming and suggests the clonal outbreaks with these strains at Tehran hospitals is likely, necessitating control over prescription of new generation cephalosporins.

Anaerobic biodegradation of DDT to DDD in soil.

DDT labeled with carbon-14 was added to soil, and the mixture was incubated anaerobically for 2 weeks and 4 weeks. DDT and seven possible decomposition products were separated by thin-layer chromatography, and the radioactivity of material from individual spots was determined by liquid scintillation. The DDT was dechlorinated by soil microorganisms to DDD, and only traces of other degradation products were detected. No degradation of DDT was detected in sterile soil.

Growth and aroma contribution of Microbacterium foliorum, Proteus vulgaris and Psychrobacter sp. during ripening in a cheese model medium.

The growth and aroma contribution of Microbacterium foliorum, Proteus vulgaris and Psychrobacter sp., some common but rarely mentioned cheese bacteria, were investigated in a cheese model deacidified by Debaryomyces hansenii during the ripening process. Our results show that these bacteria had distinct growth and cheese flavour production patterns during the ripening process. P. vulgaris had the greatest capacity to produce not only the widest variety but also the highest quantities of volatile compounds with low olfactive thresholds, e.g. volatile sulphur compounds and branched-chain alcohols. Such compounds produced by P. vulgaris increased after 21 days of ripening and reached a maximum at 41 days. The three bacteria studied exhibited various degrees of caseinolytic, aminopeptidase and deaminase activities. Moreover, P. vulgaris had a greater capacity for hydrolysing casein and higher deaminase activity. Our results show that P. vulgaris, a Gram-negative bacterium naturally present on the surface of ripened cheeses, could produce high concentrations of flavour compounds from amino acid degradation during the ripening process. Its flavouring role in cheese cannot be neglected. Moreover, it could be a useful organism for producing natural flavours as dairy ingredients.

Analysis of CRISPR/Cas system of Proteus and the factors affected the functional mechanism.

BACKGROUND: The Proteus is one of the most common human and animal pathogens. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR/Cas) are inheritable genetic elements found in a variety of archaea and bacteria in the evolution, providing immune function against foreign invasion. OBJECTIVES: To analyze the characteristics and functions of the CRISPR/Cas system in Proteus genomes, as well as the internal and external factors affecting the system. METHODS: CRISPR loci were identified and divided into groups based on the repeat sequence in 96 Proteus strains by identification. Compared the RNA secondary structure and minimum free energy of CRISPR loci through bioinformatics, the evolution of cas genes, and the effects of related elements were also discussed. RESULTS: 85 CRISPR loci were identified and divided into six groups based on the sequence of repeats, and the more stable the secondary structure of RNA, the smaller the minimum free energy, the fewer base mutations in the repeat, the more stable the CRISPR and the more complete the evolution of the system. In addition, Cas1 gene can be a symbol to distinguish species to some extent. Of all the influencing factors, CRISPR/Cas had the greatest impact on plasmids. CONCLUSIONS: This study examined the diversity of CRISPR/Cas system in Proteus and found statistically significant positive/negative correlations between variety factors (the RNA stability, free energy, etc.) and the CRISPR locus, which played a vital role in regulating the CRISPR/Cas system.

Rheumatoid arthritis in smokers could be linked to Proteus urinary tract infections.

There is an interplay between genetic and environmental factors in the induction and development of rheumatoid arthritis (RA). The RA-associated HLA-DRB1 alleles which contain the shared epitope moiety as well as microbial triggers such as Proteus are involved in the aetiopathogenesis of this disease. Increased association between Proteus urinary tract infections (UTIs) and RA on one hand and the link between smoking and UTIs on the other hand could explain the increased frequency of RA among smokers. Novel therapeutic and prophylactic measures are proposed, which might help to treat and/or prevent the disease process in individuals who are susceptible to develop RA.