ABSTRACT:  Background: Aggregatibacter aphrophilus is a Gram-negative bacterium that is considered to be highly virulent in endocarditis and cerebral abscesses but has low virulence when it comes to periodontitis. One important mechanism for the virulence of A. aphrophilus is the serum resistance, which is partially obtained through the OmpA1 outer membrane protein. EmaA is an extracellular matrix protein adhesin that is found in the outer membrane of A. aphrophilus. This protein was earlier confirmed to contribute in autoaggregation and adhering of A. aphrophilus to human epithelial cells.   Aim:  The aim of our study has been to investigate if there is a possibility that EmaA contributes to serum resistance of A. aphrophilus, similar to OmpA1.  Methods: To examine if EmaA contributes to serum resistance of A. aphrophilus, a serum killing assay was executed on four different strains of A. aphrophilus. These strains included a wildtype strain HK83, strains with a knocked out emaA and ompA1 gene, respectively and double mutant (emaA ompA1). The construction of the double mutant was done through a DNA-transformation procedure. A two- tailed T-test was used to calculate the data’s statistical significance.  Results: As expected, the ompA1 mutant did not survive well in human serum. Interestingly, the ompA1 emaAdouble mutant had a clearly higher serum survival, suggesting that EmaA may negatively contribute to serum resistance.    Conclusion:  EmaA protein appears not to contribute to the serum resistance of A. aphrophilus but rather to serum sensitivity.

No description available.

Biogas is produced during anaerobic degradation of organic material and mainly consist of methane and carbon dioxide. During the anaerobic degradation, the products are fatty acids such as propionate, butyrate and acetate. In the biogas process a syntrophic cooperation between different microorganisms are important, this involves syntrophic acetate oxidizing bacteria (SAOB) which degrade acetate, syntrophic propionate oxidizing bacteria (SPOB) which degrade propionate and methanogens which produces methane. The propionate degradation is often the slowest step in the biogas process since the microorganism involved do not get much energy from it. It has been observed that FeSO4 and FeCl2 improved the propionate degradation rate in thermophilic SPOB enrichment cultures but no impact on mesophilic enrichment cultures was observed.  The aim of this study was to determine the impact by Fe/S-addition on the key species (SAOB, SPOB and methanogens) involved in the propionate degradation in mesophilic enrichment cultures and cultures with sludge with high ammonia concentration. Furthermore, cultivation studies are performed to reveal the optimal concentrations of iron (FeCl2), which boost the propionate degradation rate in mesophilic SPOB-cultures.  The cultivation study was performed in batches (<500 mL) with 50 mM propionate, 0.3 mM ammonium and different FeCl2 concentrations between 0 to 20 mM. To determine the optimal FeCl2 concentration, the cultures were analysed by measuring the propionate, methane and acetate concentration using High Performance Liquid Chromatography (HPLC). The mesophilic enrichment cultures were treated with FeSO4, FeCl2, NaSO4, FeS, FeCl3, Fe2(SO4)3 and Na2S. The mesophilic cultures with sludge were treated with FeSO4, FeCl2, FeCl3, and a combination of all three compounds. The impact of Fe/S compounds on SAOB, SPOB and methanogens were analysed on the mesophilic enrichment cultures and cultures with sludge by measuring the copies of 16S rRNA gene using quantitative PCR (qPCR). It could be concluded that in the mesophilic enrichment culture none of the Fe/S compounds affected the SPOB or methanogens, but FeS increased the concentration of SAOB. In the mesophilic culture with sludge, FeCl2 and FeCl3 increased the SPOB level, however, FeSO4 decreased the SPOB level. Moreover, the SAOB levels decreased for FeCl3 and when all three compounds were combined. The methanogen level decreased over time by the compounds FeCl2 and FeSO4. From the cultivation study, the optimal FeCl2 concentration was 20 mM.

No description available.

No description available.

<p>Conventional end-of-pipe solutions for wastewater treatment have been criticized from a sustainable view-point, in particular regarding recycling of nutrients. The integration of hydroponic cultivation into a wastewater treatment system has been proposed as an ecological alternative, where nutrients can be removed from the wastewater through plant uptake; however, cultivation of plants in a temperate climate, such as Sweden, implies that additional energy is needed during the colder and darker period. Thus, treatment capacity, additional energy usage and potential value of products are important aspects considering the applicability of hydroponic wastewater treatment in Sweden.</p><p>To enable the investigation of hydroponic wastewater treatment, a pilot plant was constructed in a greenhouse located at Överjärva gård, Solna, Sweden. The pilot plant consisted of several steps, including conventional biological processes, hydroponics, algal treatment and sand filters. The system treated around 0.56-0.85 m3 domestic wastewater from the Överjärva gård area per day. The experimental protocol, performed in an average of twice per week over a period of three years, included analysis and measurements of water quality and physical parameters. In addition, two studies were performed when daily samples were analysed during a period of two-three weeks. Furthermore, the removal of pathogens in the system, and the microbial composition in the first hydroponic tank were investigated.</p><p>Inflow concentrations were in an average of around 475 mg COD/L, 100 mg Tot-N/L and 12 mg Tot-P/L. The results show that 85-90% of COD was removed in the system. Complete nitrification was achieved in the hydroponic tanks. Denitrification, by means of pre-denitrification, occurred in the first anoxic tank. With a recycle ratio of 2.26, the achieved nitrogen removal in the system was around 72%. Approximately 4% of the removed amount of nitrogen was credited to plant uptake during the active growth period. Phosphorus was removed by adsorption in the anoxic tank and sand filters, natural chemical precipitation in the algal step induced by the high pH, and assimilation in plants, bacteria and algae. The main removal occurred in the algal step. In total, 47% of the amount of phosphorus was removed. Significant recycling of nitrogen and phosphorus through harvested biomass has not been shown. The indicators analysed for pathogen removal showed an achieved effluent quality comparable to, or better than, for conventional secondary treatment. The microbial composition was comparable to other nitrifying biological systems. The most abundant phyla were Betaproteobacteria and Planctomycetes.</p><p>In Sweden, a hydroponic system is restricted to greenhouse applications, and the necessary amount of additional energy is related to geographic location. In conclusion, hydroponic systems are not recommended too far north, unless products are identified that will justify the increased energy usage. The potential for hydroponic treatment systems in Sweden lies in small decentralized systems where the greenness of the system and the possible products are considered as advantages for the users.</p>

<p>Extensive use of the sulphonamide drugs against the bacterium <i>Neisseria meningitidis</i> has resulted in drug resistance development. Sulphonamide resistance in <i>N. meningitidis</i> is caused by alterations in the chromosomal <i>folP</i> gene, coding for DHPS (dihydropteroate synthase). One type of resistant DHPS has high sequence divergence compared to DHPS from susceptible strains. This divergent DHPS has a duplication of two amino acids, crucial for resistance, and an altered amino acid in position 68, important for both resistance and substrate binding. When introduced into a susceptible DHPS, these two alterations did not incur resistance and resulted in abnormal substrate binding properties. This indicated that the divergent DHPS was not directly developed by mutations, but rather had been acquired by horizontal transfer of <i>folP</i> from another species.</p><p>Commensal <i>Neisseria</i> species are implied as the origin of the horizontally transferred resistance. Sulphonamide-resistant commensal <i>Neisseria</i> isolates were detected in throat swabs from healthy individuals not exposed to these drugs; however, transformation of resistance from these commensals to <i>N. meningitidis</i> was restricted in the laboratory. A comparison of the genomic region surrounding <i>folP</i> revealed differences in gene organisation and in the DNA uptake sequence between <i>N. meningitidis</i> and distantly related commensals. These differences are likely to restrict transformation between distantly related <i>Neisseria</i> species.</p><p>DHPS participates in the folate biosynthesis pathway. The enzyme preceding DHPS in the pathway, HPPK (hydroxymethyl-dihydropterin pyrophosphokinase), from <i>N. meningitidis</i> was characterised and a method for studying substrate channelling from HPPK to DHPS was developed. The information gained could be exploited in the search for new antibiotics.</p><p>In conclusion, well-adapted sulphonamide-resistant strains of <i>N. meningitidis</i> and commensal <i>Neisseria</i> are established in the bacterial population and resistance can be horizontally spread by natural transformation. This may explain the abundance of sulphonamide-resistant <i>N. meningitidis</i>, although these drugs are no longer used against this bacterium.</p>

Extensive use of the sulphonamide drugs against the bacterium Neisseria meningitidis has resulted in drug resistance development. Sulphonamide resistance in N. meningitidis is caused by alterations in the chromosomal folP gene, coding for DHPS (dihydropteroate synthase). One type of resistant DHPS has high sequence divergence compared to DHPS from susceptible strains. This divergent DHPS has a duplication of two amino acids, crucial for resistance, and an altered amino acid in position 68, important for both resistance and substrate binding. When introduced into a susceptible DHPS, these two alterations did not incur resistance and resulted in abnormal substrate binding properties. This indicated that the divergent DHPS was not directly developed by mutations, but rather had been acquired by horizontal transfer of folP from another species. Commensal Neisseria species are implied as the origin of the horizontally transferred resistance. Sulphonamide-resistant commensal Neisseria isolates were detected in throat swabs from healthy individuals not exposed to these drugs; however, transformation of resistance from these commensals to N. meningitidis was restricted in the laboratory. A comparison of the genomic region surrounding folP revealed differences in gene organisation and in the DNA uptake sequence between N. meningitidis and distantly related commensals. These differences are likely to restrict transformation between distantly related Neisseria species. DHPS participates in the folate biosynthesis pathway. The enzyme preceding DHPS in the pathway, HPPK (hydroxymethyl-dihydropterin pyrophosphokinase), from N. meningitidis was characterised and a method for studying substrate channelling from HPPK to DHPS was developed. The information gained could be exploited in the search for new antibiotics. In conclusion, well-adapted sulphonamide-resistant strains of N. meningitidis and commensal Neisseria are established in the bacterial population and resistance can be horizontally spread by natural transformation. This may explain the abundance of sulphonamide-resistant N. meningitidis, although these drugs are no longer used against this bacterium.

Conventional end-of-pipe solutions for wastewater treatment have been criticized from a sustainable view-point, in particular regarding recycling of nutrients. The integration of hydroponic cultivation into a wastewater treatment system has been proposed as an ecological alternative, where nutrients can be removed from the wastewater through plant uptake; however, cultivation of plants in a temperate climate, such as Sweden, implies that additional energy is needed during the colder and darker period. Thus, treatment capacity, additional energy usage and potential value of products are important aspects considering the applicability of hydroponic wastewater treatment in Sweden. To enable the investigation of hydroponic wastewater treatment, a pilot plant was constructed in a greenhouse located at Överjärva gård, Solna, Sweden. The pilot plant consisted of several steps, including conventional biological processes, hydroponics, algal treatment and sand filters. The system treated around 0.56-0.85 m3 domestic wastewater from the Överjärva gård area per day. The experimental protocol, performed in an average of twice per week over a period of three years, included analysis and measurements of water quality and physical parameters. In addition, two studies were performed when daily samples were analysed during a period of two-three weeks. Furthermore, the removal of pathogens in the system, and the microbial composition in the first hydroponic tank were investigated. Inflow concentrations were in an average of around 475 mg COD/L, 100 mg Tot-N/L and 12 mg Tot-P/L. The results show that 85-90% of COD was removed in the system. Complete nitrification was achieved in the hydroponic tanks. Denitrification, by means of pre-denitrification, occurred in the first anoxic tank. With a recycle ratio of 2.26, the achieved nitrogen removal in the system was around 72%. Approximately 4% of the removed amount of nitrogen was credited to plant uptake during the active growth period. Phosphorus was removed by adsorption in the anoxic tank and sand filters, natural chemical precipitation in the algal step induced by the high pH, and assimilation in plants, bacteria and algae. The main removal occurred in the algal step. In total, 47% of the amount of phosphorus was removed. Significant recycling of nitrogen and phosphorus through harvested biomass has not been shown. The indicators analysed for pathogen removal showed an achieved effluent quality comparable to, or better than, for conventional secondary treatment. The microbial composition was comparable to other nitrifying biological systems. The most abundant phyla were Betaproteobacteria and Planctomycetes. In Sweden, a hydroponic system is restricted to greenhouse applications, and the necessary amount of additional energy is related to geographic location. In conclusion, hydroponic systems are not recommended too far north, unless products are identified that will justify the increased energy usage. The potential for hydroponic treatment systems in Sweden lies in small decentralized systems where the greenness of the system and the possible products are considered as advantages for the users. / QC 20101014

Actinomycosis is an infrequent bacterial infection involving Actinomyces spp and related organisms which may occur at many body sites. It can also be found in the microbiota. Actinomyces spp are described as gram-positive bacilli whereas some species grow strictly anaerobically and some facultative. Culturing is a standardized method when suspecting actinomycosis and can be a diagnostic challenge because of inhibition of microbiota and slow growth. Enriched agar plates are used when culturing fastidious bacteria and may be more selective when including antibiotics. The aim of this project was to evaluate if using antibiotic disc facilitates identification of Actinomyces spp when mixed with microbiota. A mix of microbiota was made by pooling together several species. The susceptibility of different isolates and microbiota was analysed using antibiotic discs to determine which disc to use in a trial. A trial was done by inoculating the isolates with the microbiota on agar plates, dispensing ciprofloxacin, and trimethoprim discs. A control group without antibiotic discs were also tested. Results showed variance for most isolates susceptibility. No disc performed superior in the trial, but ciprofloxacin on FAA plates incubated anaerobically gave slightly higher recovery. Both discs facilitated identification of some isolates by supressing much microbiota. Considering that the isolates had varying susceptibility it may be problematic to find one common disc. This study has given new insights on what may facilitate identification. Further studies are needed to determine if antibiotic discs could facilitate identification of Actinomyces and needs testing on clinical samples using larger sample size.