Evaluation of the new OxyPlate^(TM) Anaerobic System for the isolation of ocular anaerobic bacteria

AIM: Anaerobic bacteria can cause ocular infections. We tested the OxyPlateTM Anaerobic System (OXY) to isolate pertinent anaerobic bacteria that can cause ocular disease.METHODS: OXY, which does not require direct anaerobic conditions (i.e. bags, jars), was compared to conventional isolation of incubating culture media in anaerobic bags. Standard colonies counts were performed on anaerobic ocular bacterial isolates under aerobic and anaerobic conditions (anaerobic bags) using agar media: 1) OXY (aerobic only), 2) 5% sheep blood (SB), 3) Chocolate, and 4) Schaedler. The bacteria tested were de-identified ocular isolates cultured from endophthalmitis and dacryocystitis that include 10 Propionibacterium acnes and 3 Actinomyces species. The colony counts for each bacteria isolate, on each culturing condition, were ranked from largest to smallest, and non-parametrically compared to determine the best culturing condition.RESULTS: All anaerobic conditions were positive for all of the anaerobic isolates. SB and Schaedler’s agar under aerobic conditions did not support the growth of anaerobic bacteria. Sparse growth was noted on chocolate agar with Propionibacterium acnes . As an anaerobic system, SB in an anaerobic bag isolated higher colony counts than OXY (P=0.0028) and chocolate agar (P=0.0028).CONCLUSION: Although OXY did not test to be more efficient than other anaerobic systems, it appears to be a reasonable alternative for isolating anaerobic bacteria from ocular sites. The use of an agar medium in a specially designed plate, without the requirement of an anaerobic bag, rendered OXY as an advantage over other anaerobic systems.

Anaerobic bacteria and intrahepatic stones: detections of Clostridium sp. and Bacteroides fragilis

To detect anaerobic bacteria Clostridium sp . and Bacteroides fragilis in intrahepatic stones by molecular genetic method Methods DNA was extracted from 59 stone samples and subjected to polymerase chain reaction (PCR) amplification targeting the 16S rRNA gene of Clostridium sp . and the glutamine synthetase gene of Bacteroides fragilis Single-strand conformational polymorphism (SSCP) analysis was performed to identify the Clostridium sp Results 16S rRNA gene sequences for Clostridium sp. were identified in 49 stones (83%, 49/59) The two most common groups were detected in 19 (41%) and 17 (37%) of the 46 samples using SSPC analysis, and 25/59 (42%) stones were tested positive for Bacteroides fragilis Conclusions Anaerobes such as Clostridium sp and Bacteroides fragilis present in intrahepatic stones and may play a role in stone formation PCR is a useful technique to detect fastidious pathogens, which are difficult to culture SSCP of PCR products is a rapid method in differentiating bacterial species

Annual change of the number of anaerobic sulf ite reducing bacteria in sediment of the Daya Bay

The surface sediment samples were collected month by month at nine stations in the Daya Bay from January to December 1987, and the number of anaerobic sulfite reducing bacteria and their spores and the regularity of seasonal change were determined. The effect of environmental factors, water temperature and the resoluble oxygen concentration in the bottom of seawater on the number of them were discussed. The results show that the number of anaerobic su|fite reducing bacteria were low in sediment of the Daya Bay, indicating that the hay was less contaminated.

Phylogenetic analysis of aerobic anoxygenic phototrophic bacteria and their relatives based on farnesyl pyrophosphate synthase gene

The study aims to reveal phylogenetic and evolutionary relationship between aerobic anoxygenic phototrophic bacteria（AAnPB） and their relatives,anaerobic anoxygenic phototrophic bacteria（AnAnPB） and nonphototrophic bacteria（NPB,which had high homology of 16S rDNA gene with AAnPB and fell into the same genus）,and validate reliability and usefulness of farnesyl pyrophosphate synthase（FPPS） gene for the phylogenetic determination.FPPS genes with our modified primers and 16S rDNA genes with general primers,were amplified and sequenced or retrieved from GenBank database.In contrast to 16S rDNA gene phylogenetic tree,AAnPB were grouped into two clusters and one branch alone with no intermingling with NPB and AnAnPB in the tree constructed on FPPS.One branch of AAnPB,in both trees,was located closer to outgroup species than AnAnPB,which implicated that some AAnPB would be diverged earlier in FPPS evolutionary history than AnAnPB and NPB.Some AAnPB and NPB were closer located in both trees and this suggested that they were the closer relatives than AnAnPB.Combination codon usage in FPPS with phylogenetic analysis,the results indicates that FPPS gene and 16S rRNA gene have similar evolutionary pattern but the former seems to be more reliable and useful in determining the phylogenic and evolutionary relationship between AAnPB and their relatives.This is the first attempt to use a molecular marker beside 16S rRNA gene for studying the phylogeny of AAnPB,and the study may also be helpful in understanding the evolutionary relationship among phototrophic microbes and the trends of photosynthetic genes transfer.

Emphysematous pyelonephritis:Six case reports and review of literature

BACKGROUND Emphysematous pyelonephritis(EPN)is a severe acute necrotizing infection of the renal parenchyma and surrounding tissues that causes the presence of gas in the renal parenchyma,collecting system,or perinephric tissue and has a poor prognosis.EPN occurs primarily in people with diabetes mellitus(DM),but can occur in those without DM when the associated renoureteral unit is obstructed.CASE SUMMARY We describe our experience with six patients who developed EPN.Five patients had DM,including one with diabetic ketoacidosis,one with multisystem involvements,including eye,lung and brain.Bilateral urolithiasis was present in one case,along with emphysematous cystitis.Unilateral kidney stones were present in one patient.One patient was an older man in poor general health.Five individuals survived and underwent surgical procedures including ureteral stent installation(Double J stent placement),percutaneous nephrostomy and perinephric abscess puncture drainage,while one died because the patient’s family chose to terminate therapy.Klebsiella pneumoniae and Escherichia coli were the microorganisms implicated.CONCLUSION We conclude that EPN is a potentially fatal illness.A positive outcome necessitates early detection.Therapeutic measures should be implemented as soon as a diagnosis is made.

Disinfection Water Wells and Sterilization

Disinfection is an important step in ensuring that water is safe to drink. Well disinfection is used to inactivate or control bacteria populations in a well and the distribution system. Disinfection is the inactivation or destruction of pathogenic organisms through the disruption of the organisms’ normal life processes. The objective of this work is to clean and/or disinfect entire borehole to prepare wells for pump installation. The last water sample was collected after the well disinfec-tion on that time. The results of water chemical analysis are presented in Tables 2-4, and there was nothing found against the specification requirements. After a reaction time of approximately 12 h, the well was cleaned from the sterilization solution by airlifting until chlorine concentration in the water reached 0 mg/l. Then through analyzing the test for samples of water after disinfection and sterilization the wells showed the Iron Bacteria were not seen and the Bacteria Species/Anaerobic Bacteria were absent.

Roles of Thioredoxins in the Obligate Anaerobic Green Sulfur Photosynthetic Bacterium Chlorobaculum tepidum

Thioredoxin is a small ubiquitous protein that is involved in the dithiol-disulfide exchange reaction, byway of two cysteine residues located on the molecule surface. In order to elucidate the role of thioredoxin in Chlorobaculum tepidurn, an anaerobic green sulfur bacterium that uses various inorganic sulfur compounds and H2S as electron donors under strict anaerobic conditions for growth, we applied the thioredoxin affinity chromatography method （Motohashi et al., 2001）. In this study, 37 cytoplasmic proteins were captured as thioredoxin target candidates, including proteins involved in sulfur assimilation. Furthermore, six of the candidate proteins were members of the reductive tricarboxylic acid cycle （pyruvate orthophosphate dikinase, pyruvate flavodoxin/ferredoxin oxidoreductase, ^-oxoglutarate synthase, citrate lyase, citrate synthase, malate dehydrogenase）. The redox sensitivity of three enzymes was then examined： citrate lyase, citrate synthase, and malate dehydrogenase, using their recombinant proteins. Based on the information relating to the target proteins, the significance of thioredoxin as a reductant for the metabolic pathway in the anaerobic photosynthetic bacteria is discussed.