Actinobacillus actinomycetem-comitans: Fermentative Capabilities of 140 Strains

Fermentative capabilities of 140 strains of Actinobacillus actinomycetem-comitans were studied. Findings correspond closely with those reported previously by Heinrich and Pulverer (12 strains), and by King and Tatum (33 strains). All strains ferment glucose, levulose, and maltose and reduce nitrate to nitrite. Reactions with glycogen and starch are exceedingly diverse. Eight different biotypes have been identified on the basis of their reactions with galactose, mannitol, and xylose.     

The use of methanol extract of Leptospira interrogans in complement fixation tests for leptospirosis

Methanol extracts were obtained from L. interrogans serovars icterohaemorrhagiae and canicola and L. biflexa serovar patoc. Human sera from 167 normal individuals and 40 patients with different infectious diseases tested by complement fixation tests showed negative reactions. Sera from 100 patients with a suspicion of leptospirosis were tested by complement fixation tests and microscopic agglutination reactions. Agreement of 84% was found for those two reactions. Positive microscopic agglutination tests at a dilution 1:20-1:400 with negative complement fixation tests were observed in 5% of patients and negative microscopic agglutination with complement fixation tests in the range of 1:20-1:1280 were observed in 11% of the cases.     

Serotyping of mesophilic Aeromonas spp. on the basis of lipopolysaccharide antigens

A serotyping system has been developed for Aeromonas hydrophila, A. sobria and A. caviae based on lipopolysaccharide (LPS) antigens. Antigens are detected by slide agglutination of boiled cells and the serotype is confirmed by tube agglutination. The antigens involved in the serotyping reactions were shown to be capable of sensitizing chicken red blood cells and were extractable by ethyl-enediaminetetraacetate. Furthermore, the reactions could be prevented by absorb-ing antisera with purified LPS. Using 16 antisera, 63 of 137 (46%) strains isolated from human faeces could be serotyped.     

Monoclonal antibodies directed against the sexual binding site of Chlamydomonas eugametos gametes

Monoclonal antibodies were raised against the mt- sexual agglutinin of Chlamydomonas eugametos gametes. Those that blocked the agglutination site were selected. They were divided into two classes dependent upon whether they gave a weak (class A) or clear positive (class B) reaction with mt- flagellar membranes in an ELISA and an indirect immunofluorescence test using glutaraldehyde-fixed mt- gametes. Class A antibodies were shown to be specific for the agglutinin in an extract of mt- gametes, based on results from immunoblotting, immunoprecipitation, affinity chromatography, and the absence of a reaction with nonagglutinable cells. Surprisingly, class A mAbs also recognized two mt+ glycoproteins, one of which is the mt+ agglutinin. Class B antibodies were shown to bind to several glycoproteins in both mt- and mt+ gametes, including the mt- agglutinin. Fab fragments from class A mAbs blocked the sexual agglutination process, but those from class B did not, even though the parent antibody did. We conclude that the class A epitope lies in or close to the agglutination site of the mt- agglutinin, whereas the class B epitope lies elsewhere on the molecule. We also conclude that the mt- agglutinin is the only component on the mt- flagellar surface directly involved in agglutination. Class A mAbs were found to elicit several reactions displayed by the mt+ agglutinin. They bound to the mt- agglutinin on gamete flagella and induced most of the reactions typical of sexual agglutination, with the exception of flagellar tip activation. None of these reactions was induced by Fab fragments. High concentrations of class A mAbs completely repressed the sexual competence of live mt- gametes, but low concentrations stimulated cell fusion.     

Comparison of tests for the diagnosis of spontaneous encephalitozoonosis in rabbits

The effectiveness of immunofluorescence, complement fixation, microagglutination serologic tests, intradermal skin test, and detection of histologic lesions were compared for use in diagnosis of spontaneous encephalitozoonosis in rabbits. The India ink and microbead agglutination reactions were compared with immunofluorescence and complement fixation by testing 11 single or pooled sera. Serologic tests correlated best with each other and less well with intradermal tests or presence of lesions. Immunofluorescence, India ink reaction and microbead agglutination were equally useful in detecting antibodies to Encephalitozoon cuniculi. The intradermal test correlated best with the presence of detectable lesions.     

Commercially available histoplasmin sensitized latex particles in an agglutination test for histoplasmosis

Summary A commercial preparation of histoplasmin sensitized latex particles was tested in an agglutination test with sera from 50 culturally confirmed cases of histoplasmosis in varying stages of the infection. The reactions were superior to those obtained with collodion agglutination and complement fixation tests in which the antigen histoplasmin was also used. The latex agglutination test with the commercially available antigen is easy to do, and can be done in any laboratory equipped to carry out agglutination tests with the common bacterial antigens. It warrants more extensive trial in the general hospital laboratory as a screening test for histoplasmosis, especially the primary, pulmonary type.     

Reactions of antibodies against ferredoxin, ferredoxin-NADP+ reductase and plastocyanin with spinach chloroplasts

Purified antisera against ferredoxin, ferredoxin-NADP+ reductase and plastocyanin agglutinated osmotically shocked and washed spinach chloroplasts, prepared according to standard procedures. The monomeric antibody (immunoglobulin G fraction) of the reductase antiserum agglutinated chloroplasts specifically and directly, indicating that protruding structures (for example, the coupling factor) do not act as steric hindrances as has been suggested. With ferredoxin antiserum, the presence of a pentameric antibody (immunoglobulin M fraction) was obligatory to observe a positive agglutination reaction. Immunoglobulin G only inhibited ferredoxin-dependent reactions, like NADP+-photoreduction, but did not cause agglutination. Ferredoxin seems to be located in depressions of the membrane, possibly caused by a partial release of this protein in shocked chloroplasts. Similar results were obtained with purified immunoglobulins from a plastocyanin antiserum. Again the immunoglobulin G fraction inhibited electron transport reactions catalyzed by plastocyanin, whereas immunoglobulin M showed a positive agglutination, but had no influence on electron transport. It is concluded that ferredoxin, ferredoxin-NADP+ reductase and plastocyanin are peripheral electron transport components, located at the outer thylakoid membrane.     

Response and Specificity of Antibodies for Candida albicans

Rabbit antibodies for Candida albicans, reacting in agglutination and fluorescent-antibody reactions, were present in both IgM and IgG protein fractions. The two types of immune globulins were separated from ammonium sulfate-precipitated gamma-globulin either by filtration through a column of Sephadex G-200 or by diethylaminoethyl column chromatography performed by stepwise elution with various concentrations of sodium chloride. In the fluorescent-antibody test, initial separation of the IgG fraction, prior to its conjugation with dye, proved to be essential for the high specificity of this reaction. Investigation of the specificities of the two types of antibodies revealed that the IgG was highly specific, whereas the IgM was not very specific. Each antigen fraction, extracted by various methods, demonstrated its own characteristic antibody response. Only the IgG fraction yielded serotype-specific antibody useful for detection of a serotype of C. albicans in agglutination and fluorescent-antibody tests. The results indicate the importance of IgG for specific serological reactions with the Candida species.     

Modified Agglutination Test for Pasteurella tularensis

Several modifications in technique were incorporated into the standard agglutination test for Pasteurella tularensis. Reciprocal shaking of all tubes in a Kahn shaker was introduced to increase the rate of agglutination and quantity of agglutinated cell mass, making it possible to report preliminary results within 4 hr. Increased incubation time at a higher temperature was used to favor the rate of agglutination. A serum control for each serum tested was necessary to detect false positive tests. Finally, a verification procedure with 5% NaCl used as the diluent was instituted to prevent these false positive reactions.     

Trypanosoma rhodesiense: antigenic stability in embryonated chicken eggs

Trypanosoma rhodesiense was maintained for three serial passages in embryonated chicken eggs. Rapid passage in mice was used to prepare trypanosome suspensions, from each passage, and from the original population, for agglutination tests. Antisera to the various trypanosome populations studied were prepared in rats. The four antigens and all antisera were crosstested by the direct agglutination test. With a single exception all reactions were positive. It is concluded that no antigenic change in the trypanosomes had occurred during passage in eggs.     

Cross reactivity of Clostridium glycolicum with the latex particle slide agglutination reagent for Clostridium difficile identification

Three strains of unidentified clostridia and 15 known strains of Clostridium glycolicum were examined to investigate cross reactions with the latex particle agglutination reagent used to identify Clostridium difficile. The unknown strains were identified as Cl. glycolicum and cross reacting agglutination occurred in 15/15 (100%) stock Cl. glycolicum strains. Characteristics such as volatile fatty acid profile, propylene glycol fermentation, u.v. fluorescence and the production of p -cresol are required to distinguish between the two species.     

Antigenic characteristics of various strains of Campylobacter jejuni. Preliminary note: technic of extraction of soluble thermostable antigens

Immune serum from rabbit has been obtained against Campylobacter jejuni ATCC no 29428 from the same strain an antigen of the outer Envelope was prepared by EDTA extraction. The specificity of antibodies against the outer antigen has been observed by direct agglutination, C.F., passive haemoagglutination and houcherlony test in agar. In eight other strains of Campylobacter isolated from patients, cross reactions with the ATCC no 29428 strain have been stored, by direct agglutination: only one strain (H) cross-reacted with the immune serum in use.     

Trypanosoma cruzi: surface antigenic determinants

A fraction (FAd) capable of inhibiting specific agglutination reactions of anti-epimastigote sera (anti-LE) was obtained by extracting the sediment of lyophilized epimastigote lysates (LE) with 0.05 M phosphate buffered saline, at 37 degrees C for 1 h. These conditions favored the action of parasite proteinase whose presence was detected by tandem-crossed immunoelectrophoresis experiments. As expected from the proteinase properties, the addition of 2-mercaptoethanol or sodium iodoacetate to the extracting solution resulted, respectively, in either increased or decreased amounts of protein in the resulting FAd. FAd components could be precipitated by the addition of Concanavalin A, methylated albumins or 0.1 N HCl. This fraction presented a single component when subjected to electrophoresis in 1% agarose gel with an electrophoretic mobility 1.2 times higher than that of human albumin. FAd component(s) were unable to penetrate 15% polycrylamide gel matrix unless 1% SDS was used. Under this condition four glycopeptide components, with Rm of 0.5, 0.55, 0.6 and 0.86, were detected. The antigenic determinants present in FAd resisted heating at 100 degrees C for 30 min and the prolonged action of pronase. However, these determinants were completely destroyed by the action of 25 mM sodium periodate, thus suggesting polysaccharide characteristics. Immunization of rabbits with FAd induced the production of antibodies that were unable to precipitate with either FAd or with parasite proteinase. These antibodies exhibited positive agglutination reactions with epimastigote forms and positive immunofluorescence and immunoperoxidase reactions with trypomastigote and amastigote forms of the different strains tested. FAd was able to inhibit these reactions as well as those obtained with anti-LE and anti-FA immune sera, whereas purified proteinase was unable to inhibit any of these reactions.     

Detection of Bacillus cereus flagellar antigen by enzyme-linked immunosorbent assay (ELISA)

A serological typing scheme of Bacillus cereus has been developed by immunochemical analyses of flagellar antigen using an agglutination method. Enzyme-linked immunosorbent assay (ELISA) for the classification of flagellar serotype of Bacillus cereus had greater sensitivity. 10-500 times, than that of agglutination method. The specificity of flagellar antigen and antibody was determined by immunogold electron microscopy and ELISA inhibition assay. Application of ELISA is useful for the detection of the small amounts and many kinds of antigen-antibody reactions.     

Estudio comparativo de las reacciones serologicas cuantitativas con un antigeno metabolico de Aspergillus fumigatus

Summary The following quantitative serologic reactions: agar-gel immunodiffusion, complement-fixation, opposite electrophoresis and latex particle agglutination tests have been performed in 38 sera from mycologically proved pulmonary aspergillosis cases. A metabolic antigen from a strain ofAspergillus fumigatus according toAjello et al technic modified by us, has been employed. Sera from 120 subjects suffering from non-mycotic lung conditions, as well as 10 sera from histoplasmosis cases, 10 sera from S. A. blastomycosis and 2 sera from patients with lung aspergillosis produced byA. niger, gave negative results with the above mentioned seroligic reactions.One hundred per cent of positive results were obtained with the complement-fixation test (titre ranging from 1/20 to 1/1280), agar-gel immunodiffusion test (titre up to 1/64) and the opposite immunoelectrophoresis (titre ranging from 1/2 to 1/256). Twenty five per cent negative and 4 non-specific results were registered with the latex particle agglutination test.A correlation of the number of serum precipition bands obtained by the electrophoresis technic with the titre of the quantitative serologic reactions, as well as a correlation of the titre of the circulating antibodies with the severity of the clinical form of aspergillosis seems to be present.Electrophoretic motility of the specific antibody performed in 10 sera showed results like the IgM in 1 instance and an intermediate position between IgA and IgG in 9 samples.     

Technical note on the demonstration of rheumatoid factor by immunofluorescence

The authors present a new technic to point out the rheumatoid factor by immunofluorescence and compare this technic and the others to classical technics of agglutination. This technic allows to avoid the absorption of rheumatoid factor in the serological reactions using the immunofluorescence.     

Cultural, Biochemical, and Immunological Properties of Mima, Herellea, and Flavobacterium Species

From study of cultural and biochemical characteristics of 40 strains of Herellea, Mima, or Flavobacterium species, a proposed schema for identification was developed. The reactions observed by agglutination, gel diffusion, and immunofluorescence suggest antigenic heterogeneity of this group of organisms.     

Agglutination, Adherence, and Root Colonization by Fluorescent Pseudomonads

Two fractions of agglutination activity towards fluorescent pseudomonads were detected in root washes of potato, tomato, wheat, and bean. High-molecular-mass (>10⁶ Da) components in crude root washes agglutinated only particular saprophytic, fluorescent Pseudomonas isolates. Ion-exchange treatment of the crude root washes resulted in preparations of lower-molecular-mass (10⁵ to 10⁶ Da) fractions which agglutinated almost all Pseudomonas isolates examined. Also, components able to suppress agglutination reactions of pseudomonads with the lower-molecular-mass root components were detected in crude root washes of all crops studied. Pseudomonas isolates were differentially agglutinated by both types of root components. The involvement of these two types of root components in short-term adherence and in colonization was studied in potato, tomato, and grass, using Pseudomonas isolates from these crops. Short-term adherence of isolates to roots was independent of their agglutination with either type of root components. With agglutination-negative mutants, the high-molecular-mass components seemed to be involved in adherence of Pseudomonas putida Corvallis to roots of all crops studied. Short-term adherence to roots of four Pseudomonas isolates could be influenced by addition of both crude and ion-exchange-treated root washes, depending on their agglutination phenotype with these root wash preparations. Potato root colonization by 10 different isolates from this crop, over a period of 7 days, was not correlated with their agglutination phenotype. Agg^- mutants of P. putida Corvallis were not impaired in root colonization. It is concluded that the root agglutinins studied can be involved in short-term adherence of pseudomonads to roots but do not play a decisive role in their root colonization.     

Analysis of the agglutinating activity from unicellular algae

Agglutinating activity often varies both between and within the algal species assayed. However, it is difficulty to interpret such variation without further analysis. We report a statistical analysis of agglutinating activities against human, cow, sheep, and pig erythrocytes, using cell extracts from 43 taxa (strains) of freshwater microalgae. Most of the extracts agglutinated erythrocytes from at least one of the sources, but pig erythrocytes appeared to be most suitable for the detection of agglutination reactions. Chlorella cell extracts preferentially agglutinated human erythrocytes, whereas extracts of other taxa were less active against mammalian erythrocytes. Cluster analysis generated four distinct subclusters of taxa, characterized by different specificities for antigens or carbohydrate receptors on the erythrocytes. Principal component analysis further separated the agglutination characteristics of Chlamydomonas from Chlorella on the first two components. Specificity for pig erythrocytes accounted for most of the clustering or grouping of algal taxa in multivariate analysis. However, clustering or grouping patterns of Chlorella species on haemagglutinating activity resembled that based on DNA sequences, revealing a possible genetic connection of agglutinins and their biochemical characteristics in algal cells. Variability of agglutination reactions among the algae investigated is simplified and interpreted most easily using multivariate analysis.     

Isoimmunizations among marmosets to identify leucocyte antigens.

Saguinus fuscicollis ssp., S.o. oedipus, and S. nigricollis marmosets were immunized with white blood cells (WBC) from S.f. illigeri marmosets. Of the three genetic donor-host combinations attempted, only an interspecies immunization of S.o oedipus marmosets resulted in antibody formation which was detectable by agglutination or cytotoxicity tests. Analysis of one of the antisera with a panel of WBC from S.f. illigeri marmosets indicated varying specificities, reactions ranging from strong, intermediate, to completely negative. That the serum was detecting genotypic as well as phenotypic markers was suggested by the similarity of reactions of WBC from co-twin pairs and in preliminary testing of individual families.