The aerobic flora of the perivaginal area: its relation to contraception and deodorant sprays.
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Adult↗
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Molecular survey of aeroplane bacterial contamination.
AIMS: To examine bacterial contamination of passenger aircraft and to identify aeroplane environments posing the greatest potential health risk. METHODS AND RESULTS: DNA was extracted from ten environmental samples collected on four different flights (three domestic, one international) from a variety of surfaces frequently touched by passengers. PCR clone libraries were made from the DNA samples using bacterial-specific 16S ribosomal DNA (rDNA) primers. A total of 271 bacterial rDNA sequences were obtained. We used BLAST analysis of the rDNA clone sequences to identify sequences in Genbank with the highest sequence similarity. The majority of the rDNA clones obtained from aeroplane environments were more than 97% identical to rDNA sequences from cultured bacterial species. Samples collected from the cabin surfaces (e.g., tray tables and arm rests) had undetectable levels of DNA and produced no PCR products. Bacterial diversity was highest on lavatory surfaces, including door handles, toilet handles, and sink faucets. Sequence data from these surfaces detected species from 58 different bacterial genera, and many of the best BLAST hits matched rDNA sequences of cultured species known to be opportunistic pathogens. The most frequently observed species came from five genera commonly associated with humans: Streptococcus, Staphylococcus, Cornybacterium, Proprionibacterium and Kocuria. CONCLUSIONS: The results show that there is a large diversity of bacterial contamination on aeroplanes, including organisms known to be opportunistic pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results indicate that aeroplanes have the potential to spread an enormous diversity of bacterial species among passengers and destinations. Aeroplane lavatories present an especially significant concern to public health.
Aircraft↗
Renibacterium salmoninarum bar forms: characterization, occurrence, and evidence of a host response to a R. salmoninarum infection.
Unique-staining Renibacterium salmoninarum (Rs) cells, termed bar forms, first observed in a coho salmon, Oncorhynchus kisutch (Walbaum), in 1983, could not be cultured, making their characterization difficult and significance obscure. They can be detected only by the fluorescent-antibody technique (FAT) and their numbers estimated only by a quantitative FAT (QFAT). Data collected over a 10-year period showed that bar forms were observed only in vivo and appeared associated with a host response. Bar forms were observed in 10 salmonid species from five countries and in fish from < 1 g to spawning adults. They were observed in 50.1% of kidney smears prepared from 10,061 Rs positive chinook, O. tshawytscha (Walbaum), coho, and Atlantic salmon, Salmo salar L. Bar forms were shown to be Rs cells based on absorption studies, their reaction with an Rs-specific FAT and enzyme-linked immunosorbent assay, and a transition from 'typical' Rs cells to bar forms in naturally and experimentally infected fish. Bar forms were determined to be non-virulent, damaged or dead Rs cells, based on fluorescence and electron microscopy observations, the inability to culture them, and mortality data. Bar forms appeared to represent visual markers of recovery from an Rs infection.
Actinomycetales Infections↗
Detection of Renibacterium salmoninarum in chinook salmon, Oncorhynchus tshawytscha (Walbaum), using quantitative PCR.
A quantitative polymerase chain reaction (QPCR) assay has been developed to detect varying levels of Renibacterium salmoninarum, the causative agent of bacterial kidney disease. This assay allows for the direct enumeration of bacterial DNA or RNA copy number within tissues and body fluids. The assay can be applied non-lethally and can be used to determine whether R. salmoninarum is transcriptionally active. The presence of R. salmoninarum in kidney tissues from 430 chinook salmon collected from five Idaho Fish and Game operated hatcheries was initially evaluated using the widely employed enzyme-linked immunosorbent assay (ELISA) with two sets of Kirkegaard and Perry Laboratories polyclonal antibodies, 'mother batches' 1 and 2. The same tissue samples were then analysed using the novel QPCR assay and the results compared. At moderate to high levels of infection [optical density (OD > 0.5)], ELISA values and estimated DNA copy number were highly correlated (r(2) > 0.80), although correlation to specific antibody batches varied. However, lower ELISA values (OD < 0.5) observed with either antibody batch did not correlate well with the QPCR assay (R(2) <or= 0.43). Negative controls, run concurrently with the PCR assay, did not indicate extraneous DNA contamination in the PCR samples.
Actinomycetales Infections↗
Atypical growth of Renibacterium salmoninarum in subclinical infections.
Two growth types of Renibacterium salmoninarum were isolated from subclinically infected rainbow trout, one producing the smooth colonies typical of R. salmoninarum and the other forming a thin film on the surface of the agar with no separate colonies. The atypical growth was present on kidney disease medium agar in primary cultures of the kidney but not on selective kidney disease medium (SKDM). Fluorescent antibody staining of the fresh isolate and polymerase chain reaction amplification were the most reliable techniques to identify the atypical growth of R. salmoninarum. The condition was reversible, with growth reverting from atypical to the smooth colony form in experimentally infected rainbow trout and under laboratory conditions. There was no mortality, or any clinical signs of bacterial kidney disease (BKD) in the fish challenged with the atypical growth, although small numbers of smooth colonies of R. salmoninarum were isolated from 8% of these fish. The atypical growth reported here may explain some of the failures of culture, when SKDM agar alone is used for the detection of BKD in subclinically infected fish.
Actinomycetales Infections↗
DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans.
Deinococcus radiodurans and other members of the same genus share extraordinary resistance to the lethal and mutagenic effects of ionizing and u.v. radiation and to many other agents that damage DNA. While it is known that this resistance is due to exceedingly efficient DNA repair, the molecular mechanisms responsible remain poorly understood. Following very high exposures to u.v. irradiation (e.g. 500 J m-2, which is non-lethal to D. radiodurans), this organism carries out extremely efficient excision repair accomplished by two separate nucleotide excision repair pathways acting simultaneously. One pathway requires the uvrA gene and appears similar to the UvrABC excinuclease pathway defined in Escherichia coli. The other excision repair pathway is specific for u.v. dimeric photoproducts, but is not mediated by a pyrimidine dimer DNA glycosylase. Instead, it is initiated by a second bona fide endonuclease that may recognize both pyrimidine dimers and pyrimidine-(6-4)pyrimidones. After high doses of ionizing-radiation (e.g. 1.5 Mrad), D. radiodurans can mend > 100 double-strand breaks (dsb) per chromosome without lethality or mutagenesis. Both dsb mending and survival are recA-dependent, indicating that efficient dsb mending proceeds via homologous recombination. D. radiodurans contains multiple chromosomes per cell, and it is proposed that dsb mending requires extensive recombination amongst these chromosomes, a novel phenomenon in bacteria. Thus, D. radiodurans may serve as an easily accessible model system for the double-strand-break-initiated interchromosomal recombination that occurs in eukaryotic cells during mitosis and meiosis.
Adenosine Triphosphatases↗
Biosynthesis of peptidoglycan in Gaffkya homari. The incorporation of peptidoglycan into the cell wall and the direction of transpeptidation.
Wall membrane enzyme preparations from Gaffkya homari catalyze the formation of peptidoglycan from the precursor pairs: UDP-N-acetylglucosamine + UDP-N-acetylmuramyl-pentapeptide (UDP-MurNAc-Ala-DGlu-Lys-DAla-DAla) and also from UDP-N-acetylglucosamine + UDP-N-acetylmuramyl-tetrapeptide (UDP-MurNAc-Ala-DGlu-Lys-DAla). Part of the reaction products is soluble in 2% sodium dodecylsfulfate whereas the other part is bound to pre-existing cell wall peptidoglycan. The incorporation into cell wall takes place by a transpeptidation reaction in which the D-alanyl-D-alanine sequences in the pre-existing cell wall function as donors and the epsilon-amino groups of the lysine residues in the newly synthesized peptidoglycan strands function as acceptors. Nepsilon-D-Alanyl-lysine linkages are formed. At saturating concentration of UDP-N-acetylglucosamine, the enzyme system exhibits similar apparent Km values (30--80 muM) for UDP-MurNAc-pentapeptide and UDP-MurNAc-tetrapeptide both for the formation of cell-wall bound peptidoglycan and total (i.e. soluble + cell-wall-bound) peptidoglycan. The V values are also in the same order of magnitude (270-650 pmol x min-1 x mg of protein -1). However, UDP-MurNAc-tetrapeptide was a slightly better substrate than UDP-MurNAc-pentapeptide for the formation of cell-wall-bound peptidoglucan. The synthesis of total and cell-wall-bound peptidoglycan from UDP-MurNAc-pentapeptide was competitively inhibited by UDP-MurNAc-tetrapeptide and vice versa. UDP-MurNAc-tripeptide and both UDP-Mur-NAc-pentapeptide and UDP-Mur-NAc-tetrapeptide in which the epsilon-amino group of the lysine residue was substituted by an acetyl group were utilized less efficiently than UDP-MurNAc-pentapeptide and UDP-MurNAc-tetrapeptide for the formation of soluble peptidoglycan; they were exceedingly poor substrates for the formation of cell-wall-bound peptidoglycan.
Acyltransferases↗
Biosynthesis of peptidoglycan in Gaffkya homari. The mode of action of penicillin G and mecillinam.
The effect of the beta-lactam antibiotics penicillin G and mecillinam on the incorporation of peptidoglycan into pre-formed cell wall peptidoglycan was studied with wall membrane enzyme preparations from Gaffkya homari. Using UDP-N-acetylglucosamine (UDP-GlcNAc) and UDP-N-acetylmuramyl-pentapeptide (UDP-MurNAc-pentapeptide) as precursors the incorporation of peptidoglycan into the pre-existing cell wall of G. homari was inhibited to an extent of 50% (ID50 value) at a concentration of 0.25 mug of penicillin G/ml. With UDP-GlcNAc and UDP-MurNAc-tetrapeptide as precursors the ID50 value was about 2500-fold greater (630 mug/ml). The inhibition by penicillin G of the incorporation of peptidoglycan from UDP-MurNAc-[14C]Lys-pentapeptide could be overcome by addition of non-radioactive UDP-MurNAc-tetrapeptide to the incubation mixture. In the presence of 5 mug of penicillin G/ml the incorporation of peptidoglycan formed from the mixture of UDP-MurNAc-Ala-DGlu-Lys-D-[14C]Ala-D[14C]Ala and non-radioactive UDP-MurNAc-tetrapeptide proceeded virtually without release of D-[14C]alanine by transpeptidase activity. The enzyme preparation also exhibited DD-carboxypeptidase activity which was only slightly more sensitive to penicillin G and mecillinam than was the incorporation of peptidoglycan into the cell wall. Since the ID50 values for the beta-lactam antibiotics are similar to the concentrations required to inhibit the growth of G. homari to an extent of 50%, the DD-carboxypeptidase must be the killing site of both penicillin G and mecillinam.
Azepines↗
The activities in vitro of DD-carboxypeptidase and LD-carboxypeptidase of Gaffkya homari during biosynthesis of peptidoglycan.
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Alanine↗
Rothia dentocariosa: taxonomy and differential diagnosis.
As recent external quality control results have shown, the diagnosis of Rothia dentocariosa infection still presents problems for clinical laboratories. This review describes the taxonomy, as well as the chemotaxonomic, morphological and biochemical characteristics, of this organism, and surveys bacteria that may be confused with Rothia dentocariosa.
Actinomycetales Infections↗
Cloning, functional expression and partial characterization of the glucose kinase from Renibacterium salmoninarum.
The complete glcK gene from the fish pathogen Renibacterium salmoninarum, encoding a glucose kinase, was analyzed and expressed. The partial characterization of the recombinant enzyme confirmed that it belongs to a group of glucose kinases involved in carbon catabolite repression. Multiple sequence alignments were used to deduce a new consensus sequence for this family of bacterial proteins, characterized by several conserved Cys residues. This sequence was more specific and allowed the detection of the first eukaryotic protein of this family. The recombinant enzyme was inhibited by N-ethylmaleimide and the substrates protected the enzyme from this inhibition, suggesting the presence of Cys residues in or close to the active site.
Amino Acid Sequence↗
Use of copper sulphate for the treatment of dermatophilosis ('brown spot' disease) in farmed crocodiles.
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Actinomycetales Infections↗
The conjunctival and contact lens bacterial flora during lens wear.
The conjunctival bacterial flora in wearers of hydrophilic, hard and silicone lenses and in a non-lens control group has been studied. Cultures were also taken from the posterior surface of the various contact lenses. Coagulase negative staphylococci were the most commonly occurring microorganisms both on the conjunctiva and on the lenses. Further identification of these bacteria showed Staph. epidermidis to be the predominant species. Compared with control group a significantly increased frequency of negative conjunctival cultures was found among the lens wearers (P = 0.020). The occurrence of coagulase negative staphylococci was reduced, but the difference was not statistically significant (P = 0.08). Gram-negative bacilli were more frequently found during lens wear (P = 0.012), but were mainly species which are traditionally regarded as apathogenic. The possible influence of lens disinfection, duration of lens wear and eye/lid hygiene on the conjunctival bacterial flora is discussed.
Actinomycetales↗
Antibacterial activities of ciprofloxacin, norfloxacin, oxolinic acid, cinoxacin, and nalidixic acid.
In vitro studies were performed comparing ciprofloxacin (Bay o 9867) and norfloxacin with three related organic acids. Ciprofloxacin was two to eight times more active than norfloxacin against 658 bacterial isolates representing 30 species. For all species tested, ciprofloxacin MICs for 90% inhibition were less than or equal to 2.0 micrograms ml. Additional tests with 5,994 isolates detected only 37 (0.6%) strains resistant to 2.0 micrograms of ciprofloxacin per ml and 106 (1.8%) resistant to 1.0 micrograms/ml. Only 6 (0.1%) of the 5,994 strains were resistant to 16 micrograms of norfloxacin per ml, and 129 (2.1%) were resistant to 4.0 micrograms/ml. The majority of resistant strains were streptococci or Pseudomonas spp. Resistance among the Enterobacteriaceae was extremely rare (i.e., greater than 99.8% susceptible to both drugs.
Anti-Bacterial Agents↗
Antimicrobial susceptibilities of Stomatococcus mucilaginosus and of Micrococcus spp.
The in vitro susceptibilities of 63 isolates of Stomatococcus mucilaginosus and of 188 isolates of Micrococcus spp. to 18 antimicrobial agents were determined by the agar dilution method. Many beta-lactams, imipenem, rifampin, and the glycopeptides were shown to be active in vitro against Stomatococcus and Micrococcus isolates, whereas the activities of antibiotics such as some aminoglycosides, erythromycin, and fosfomycin against an important number of these microorganisms are limited.
Anti-Bacterial Agents↗
Classification of bacteria from commercial egg washers and washed and unwashed eggs.
A total of 432 bacterial isolates from washed and unwashed eggs, egg-washer surfaces, and washwaters from five egg-grading plants in Maryland and southeastern Pennsylvania were classified. Counts on equipment surfaces showed considerable variation from plant to plant, reflecting care used in cleaning. Unwashed eggs had a higher percentage of gram-positive cocci (71%), and isolates included Streptococcus faecalis, Aerococcus, and Escherichia coli, which were not isolated from equipment surfaces and washwaters. Equipment surfaces had a higher proportion of actinomycetes than unwashed eggs, and predominant gram-negative rods were Alcaligenes and Moraxella, which were not found on unwashed eggs. Flavobacterium and Alcaligenes have been implicated in shell egg rots, Staphylococcus aureus has been implicated in food poisoning, and organisms resembling micrococci and actinomycetes have been found in broken-out egg products.
Animals↗
Effects of monovalent and divalent salts on the phospholipid and fatty acid compositions of a halotolerant Planococcus sp.
The phospholipid headgroup and fatty acid compositions of a halotolerant Planococcus sp. (strain A4a) were examined when cells were grown in the presence of high concentrations of a variety of salts. The fatty acid composition of Planococcus sp. strain A4a was altered primarily as a function of the osmolality of the growth medium. The phospholipid headgroup composition was influenced by both the osmolality of the growth medium and the nature of the cation species present. An increase in the cardiolipin/phosphatidylglycerol molar ratio was detected when cells were grown in the presence of high concentrations of monovalent cations.
Calcium Chloride↗
Molecular differentiation of Renibacterium salmoninarum isolates from worldwide locations.
Renibacterium salmoninarum is a genospecies that is an obligate pathogen of salmonid fish and is capable of intracellular survival. Conventional typing systems have failed to differentiate isolates of R. salmoninarum. We used two methods to assess the extent of molecular variation which was present in isolates from different geographic locations. In one analysis we investigated possible polymorphisms in a specific region of the genome, the intergenic spacer (ITS) region between the 16S and 23S rRNA genes. In the other analysis we analyzed differences throughout the genome by using randomly amplified polymorphic DNA (RAPD). We amplified the spacer region of 74 isolates by using PCR and performed a DNA sequence analysis with 14 geographically distinct samples. The results showed that the 16S-23S ribosomal DNA spacer region of R. salmoninarum is highly conserved and suggested that only a single copy of the rRNA operon is present in this slowly growing pathogen. DNA sequencing of the spacer region showed that it was the same length in all 14 isolates examined, and the same nucleotide sequence, sequevar 1, was obtained for 11 of these isolates. Two other sequevars were found. No tRNA genes were found. We found that RAPD analysis allows reproducible differentiation between isolates of R. salmoninarum obtained from different hosts and different geographic regions. By using RAPD analysis it was possible to differentiate between isolates with identical ITS sequences.
Animals↗