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Rapid identification of thermophilic Naegleria, including Naegleria fowleri using API ZYM system.

The suitability of the API ZYM system for identifying thermophilic Naegleria species, based on enzyme presence and activity, was investigated. Replicate testing on strains of N fowleri, N lovaniensis, and N australiensis cultured in a monoxenic and an axenic medium showed that the system could provide a rapid and reproducible means of identifying the species soon after primary isolation. No single enzyme was found specific for any one species, but considerable differences were found in the patterns of activity of acid phosphatase and leucine arylamidase. When these were compared the species could be differentiated. Use of the system in conjunction with a simple culture method is proposed as a readily available means of monitoring environmental and public bathing sites to prevent primary amoebic meningoencephalitis.

Acid Phosphatase

Comparison of identification of Enterobacteriaceae by API 20E and Sensititre Autoidentification System.

Of 251 isolates of the Enterobacteriaceae identified to species level by API 20E, 208 (83%) were similarily identified by the Sensititre Autoidentification System. Both systems shared a common problem in that discrimination between species of the genera Klebsiella, Enterobacter, and Serratia was poor. The eight digit biocode generated by the Sensititre system for individual isolates is not reproducible and therefore not of epidemiological value.

Bacteriological Techniques

Physical, chemical, and enzymatic studies on the major sucrase of honey bees (Apis mellifera).

A sucrase from honey bees (Apis mellifera) which precipitates between ammonium sulfate saturations of 50 and 70% (5 mg protein per millilitre) and which makes up the major portion of the sucrases of honey bees was purified to homogeneity as shown by several criteria. A large part of the sucrase was found in the head while most of the rest was in the abdomen (a small amount was in the thorax). The enzyme precipitated between the same values of ammonium sulfate saturation as did the sucrase in honey and honey sucrase exhibited kinetics very similar to those of this enzyme. The enzyme was found to be a relatively nonspecific alpha-glucosidase and was shown to have transglucosidase activity. The production of glucose from sucrose was rectilinear when plotted by the Hofstee method at low substrate concentrations but decreased at high sucrose concentrations. The production of fructose was rectilinear throughout the concentration range used. The production of both glucose and rho-nitrophenol when rho nitrophenyl alpha-D-glucoside was the substrate was linear by the Hofstee plot. These effects were found to be due to transglucolysis and a mechanism of action is proposed. Amino acid and amino sugar analyses indicated that the sucrase was a glycoprotein. The molecular weight was found to be between 51000 and 82000 by three different methods and an so20.w value of 4.0 S was obtained. There was no evidence for subunit structure. Tests of the enzyme under various denaturation conditions did not reveal any unusual stabilities. The sucrase bound very tightly to a hydrophobic column. Iodoacetic acid decreased the activity of the sucrase but a large concentration was needed to bring about a 50% activity loss. Reducing agents caused some activity declines. Diethyl pyrocarbonate activated the enzyme.

Amino Acids

Detoxified hymenoptera venoms: preliminary studies of in vitro cytotoxicity and antigenicity of Apis mellifera 'venomoid'.

The preparation of detoxified venom (venomoid) of Apis mellifera is described. The venomoid obtained by formaldehyde detoxification retained significant antigenicity. Radioallergosorbent test (RAST) inhibition studies indicated a loss of the affinity of the venomoid for the venom-specific human IgE antibody. In rabbits the venomoid was capable of producing precipitating antibodies which cross-reacted with the original venom. Cytotoxic activity of venomoid as measured by nonimmunological histamine release and hemolytic activity with human cells was not detectable in vitro. Further studies seem warranted for the establishment of a venomoid as a diagnostic and/or therapeutic agent in the management of the hypersensitivity reactions to venom.

Animals

Occurrence of intercellular bridges between follicle epithelial cells in the ovary of Apis mellifica queens.

Hitherto unknown intercellular bridges or fusomes between the follicle epithelial cells investing the oocytes of Apis mellifica queens have been observed both with light and electron microscopy. Usually each follicle cell has 2-3 intercellular bridges. In surfacial paraffin sections, the intercellular bridges can be seen to connect a series of follicle cells which may be branching. The intercellular bridges lie close to the egg cortex and this position is relatively constant. The width of the fusomal ring canal varies in different developmental stages. In stages 3 and 4 of oogenesis, which are the main vitellogenic stages, the intercellular bridges measure 0-5 micron, while in stages 1 and 2 they have a diameter ranging from 1-5 to 3-5 micron. In these stages the intercellular bridges are provided with numerous transverse microfilaments which disappear later. The fusomal lips are thickened and consist of electron-dense material and an additional layer of less electron-dense material both inside and outside. Ribosomes flow across the bridge. The intercellular bridges may serve to synchronize the differentiation and functional activity of the follicle epithelium during the course of oogenesis.

Animals

[Detection of extended-spectrum beta-lactamases by the rapid ATB E technique. Value of the API V2.1.1 expert system].

Twenty-two extended-spectrum betalactamase-producing strains of enterobacteriaceae recovered in the authors' hospital were tested using the Rapid ATB E coupled with the API V2.1.1. expert system. The expert system detected 90.9% of ESBL-producing strains. Two strains producing a SHV2 and a CTX1, respectively, escaped detection by the expert system despite concomitant resistance to aminoglycosides.

4-Quinolones

[Differentiation of glucidolytic mycoplasmas isolated from goats by the API 50 CH system and electrophoresis].

Carbohydrate metabolism of 62 glucidolytic strains of mycoplasma belonging to 4 species (M ovipneumoniae, M putrefaciens, M mycoides, M capricolum) has been studied using the API 50 Ch system for bacterial identification. This microtechnique and colony aspect were relevant in distinguishing M ovipneumoniae and M putrefaciens from the group M mycoides and M capricolum isolated from goats, but still presented a lack of specificity in distinguishing M mycoides from M capricolum. Similar results were obtained when the mycoplasma strains were tested by electrophoresis.

Animals

[Coliform bacteria in rinsed beer mugs--identification with the API 20 E system and resistance behavior].

653 11 beer mugs rinsed mechanically by conveyor belt dishwashing machines and 182 mugs cleaned in open vats were investigated for their content of faecal and total coliforms. Ten and more faecal coliform germs could be detected in 4.7% of the mechanically rinsed tankards and in 12.1% of those cleaned in open vats. Respectively, ten and more total coliforms were found in 30.3% and in 67.6%. Altogether 213 coliform strains as defined by the German drinking water regulation were characterised biochemically by the API 20 E-system and by their susceptibility to 15 antimicrobial agents. A total of 19 different species from eight genera could be differentiated, whereas 12 strains were unidentifiable by the applied system. Klebsiella oxytoca was the most frequently encountered germ, followed by Enterobacter cloacae, K. pneumoniae and Ent. sakazakii. 59 of the 213 strains were fully susceptible to all antibiotics used. 44 strains, however, revealed complete or moderate resistance against three and more agents. These results indicate that unhygienic dishwashing procedures may play a decisive role in the epidemiology of clinically relevant and multiresistant germs.

Beer

Comparison of the API Staph-Ident and DMS Staph Trac micromethods for the identification of coagulase-negative staphylococci.

Two rapid identification micromethods, the API Staph-Ident and the DMS Staph Trac, were compared for the identification of coagulase-negative staphylococci. Seventy-five isolates, mainly of bovine origin, were used as the test organisms. Species identification and profile numbers assigned to isolates by each system were compared. However, no clear correlation patterns emerged, indicating the two methods were not comparable.

Animals

Biotyping of clinical isolates of Escherichia coli of animal origin, using the Analytab API 20E system.

Using the Analytab (API 20E) Enterobacteriaceae system of biochemical identification, a total of 506 Escherichia coli isolates from different animal species were coded numerically or biotyped. Fifty-four different biotypes were identified, 11 accounting for 83.1% of the isolates examined. Three of these profiles accounted for 65.3% of the isolates and were found in almost all animal species. Some of the biotypes were found in only one animal species: six in cattle, five in horses, 15 in pigs, two in sheep, two in birds, one in dogs and one in a porpoise. Biotypes, as determined here, could not be related to a particular pathology and more work is needed to assess the extent and significance of this relative biotype specificity among animal species. The use of other, more sophisticated, typing systems, i.e. plasmid "fingerprinting", or restriction endonuclease analysis of chromosomal DNA, would have to be investigated.

Animals

Characterization of staphylococci using the API 20E system.

Fifty-four strains of coagulase-negative staphylococci, five isolates of coagulase-positive staphylococci, and seven species of Micrococcus were tested in the API 20E system. After 48 hours' incubation, results indicated that the various species and subgroups could be separated by their reactions. Staphylococcus saphrophyticus subgroup 3 was ONPG and mannitol positive. S. epidermidis subgroup 1 was positive for arginine dihydrolase (ADH), whereas S. epidermidis subgroup 4 was positive for ADH, mannitol, and arabinose. S. aureus coagulase negative was characterized by positive ONPG, ADH, and mannitol. In contrast, S. aureus coagulase positive was not positive for ONPG but was positive for ADH and mannitol. Most of these staphylococci were able to utilize nitrates, urea, glucose, and sucrose. The micrococci, on the other hand, gave varied reactions, with most species being only weakly reactive.

Bacteriological Techniques

Analysis of queen recognition by honey bee workers (Apis mellifera L.) in a metabolic bio-assay.

Queenless groups of honey bee workers (Apis mellifera L.) respond with typical increases in their metabolic activity when exposed to queens. Volatile queen odors release 46.8% of the reaction in worker groups compared to combinations of volatile odors and non-volatile factors. Worker groups which were allowed to learn the odor of a given queen were able to discriminate between volatile odors of related and unrelated queens. Although non-volatile queen compounds seem to be of minor importance for kin recognition they override effects releasable by volatile odors.

Animals

Lipophorin of the larval honeybee, Apis mellifera L.

Most insects have a major lipoprotein species in the blood (hemolymph) that serves to transport fat from the midgut to the storage depots in fat body cells and from the fat body to peripheral tissues. The generic name lipophorin is used for this lipoprotein. In larvae of the honeybee, Apis mellifera, a lipophorin has been found with properties that correlate well with those of the only other lipophorin reported for an immature insect, that of the tobacco hornworm, Manduca sexta. The honeybee lipophorin (Mr = 530,000) has a density of 1.13 g/ml, contains approximately 41% lipid and 59% protein, and contains two apoproteins, apoLp-I, Mr = 250,000 and apoLp-II, Mr = 80,000, both of which are glycosylated. The lipids consist predominantly of polar lipids, of which phospholipids and diacylglycerols represent 60% of the total. When the intact lipophorin is treated with trypsin, apoLp-I is rapidly proteolyzed, while apoLp-II is resistant, indicating a difference in exposure of the two apoproteins to the aqueous environment. Honeybee apoLp-II cross-reacts with antibodies to M. sexta apoLp-II, but not to anti-M. sexta apoLp-I. No cross-reactivity of honeybee apoLp-I to anti-M. sexta apoLp-I was observed.

Amino Acids

[Biochemical typing of salmonellae by a miniaturized system (API - 50 E) (author's transl)].

The biochemical properties of clinical isolates of Salmonella have been studied by means of a commercially available miniaturized system (API - 50 E), which permit an easier examination of 50 chemical reactions. This miniaturized system seems to be helpfull not only in epidemiological researches but also in characterization of "atypical strains".

Computers

Exposure scheme separates effects of electric shock and electric field for honey bees, Apis mellifera L.

Mechanisms to explain disturbance of honey bee colonies under a 765-kV, 60-Hz transmission line [electric (E) field = 7 kV/m] fall into two categories: direct bee perception of enhanced in-hive E fields, and perception of shock from induced currents. The same adverse biological effects previously observed in honey bee colonies exposed under a 765-kV transmission line can be reproduced by exposing worker bees to shock or E field within elongated hive entranceways (= tunnels). Exposure to intense E field caused disturbance only if bees were in contact with a conductive substrate. E-field and shock exposure can be separated and precisely defined within tunnels, eliminating dosimetric vagaries that occur when entire hives are exposed to E field.

Animals

Mechanism of biological effects observed in honey bees (Apis mellifera, L.) hived under extra-high-voltage transmission lines: implications derived from bee exposure to simulated intense electric fields and shocks.

This work explores mechanisms for disturbance of honey bee colonies under a 765 kV, 60-Hz transmission line [electric (E) field = 7 kV/m] observed in previous studies. Proposed mechanisms fell into two categories: direct bee perception of enhanced in-hive E fields and perception of shock from induced currents. The adverse biological effects could be reproduced in simulations where only the worker bees were exposed to shock or to E field in elongated hive entranceways (= tunnels). We now report the results of full-scale experiments using the tunnel exposure scheme, which assesses the contribution of shock and intense E field to colony disturbance. Exposure of worker bees (1,400 h) to 60-Hz E fields including 100 kV/m under moisture-free conditions within a nonconductive tunnel causes no deleterious affect on colony behavior. Exposure of bees in conductive (e.g., wet) tunnels produces bee disturbance, increased mortality, abnormal propolization, and possible impairment of colony growth. We propose that this substrate dependence of bee disturbance is the result of perception of shock from coupled body currents and enhanced current densities postulated to exist in the legs and thorax of bees on conductors. Similarly, disturbance occurs when bees are exposed to step-potential-induced currents. At 275-350 nA single bees are disturbed; at 600 nA bees begin abnormal propolization behavior; and stinging occurs at 900 nA. We conclude that biological effects seen in bee colonies under a transmission line are primarily the result of electric shock from induced hive currents. This evaluation is based on the limited effects of E-field exposure in tunnels, the observed disturbance thresholds caused by shocks in tunnels, and the ability of hives exposed under a transmission line to source currents 100-1,000 times the shock thresholds.

Animals