Differentiation of three serovars of Malassezia furfur

Malassezia furfur strains were isolated from the clinically normal skin of 10 volunteers by swabbing four different sites (forehead, ear, back and chest). The strains could be divided into three basic groups on the basis of cultural characteristics. Both unabsorbed and absorbed specific rabbit antisera were prepared against nine of the strains, and both species and group specific antigens could be demonstrated. Serologically, three group specific surface antigens could be identified which corresponded to the three groups identifiable on cultural characteristics. The relevance of these findings to previous in vitro results is discussed.     

Glutamine synthetase content and immunochemical analysis of different strains of Bordetella pertussis

In this investigation 3 groups of strains isolated from pertussis patients have been studied: typical (group 1), atypical in their cultural properties (group 2), unidentified Gram-negative bacilli agglutinated by pertussis and parapertussis antitoxins (group 3). Besides, B. pertussis cultures, obtained by subculturing 2 museum strains and 2 newly isolated strains on synthetic casein-charcoal agar with subinhibiting doses of antibiotics or specific immune sera added, have been studied. As indicated by the results of this study, strains belonging to groups 1 and 2 contain glutamine synthetase, while in strains of group 3 this enzyme is absent. In immunoelectrophoresis strains of group 3 have been found to contain not a single antigen similar to the antigens of strains belonging to groups 1 and 2. Electrophoresis in polyacrylamide gel has revealed to differences in the protein spectrum of the strains of these 3 groups. The investigation has shown that the determination of glutamine synthetase and immunoelectrophoresis can be used for the differentiation of B. pertussis from similar Gram-negative bacilli. B. pertussis strains, changed as the result of experiments with antibiotics and specific immune sera, have also been shown to retain their antigenic composition and protein spectrum and to have no essential difference in the content of glutamine synthetase.     

Serology of Neisseria gonorrhoeae. Classification by co-agglutination

The co-agglutination (COA) method has been adapted for serological classification of Neisseria gonorrhoeae. COA reagents were prepared with selectively absorbed rabbit hyperimmune antibodies against gonoccal (GC) major outer membrane protein (MOMP) serotype strains. Using these reagents, the 16 MOMP reference strains could be referred to at least three antigen classes, tentatively named W, J and M. The GC antigens of class W were divided into three groups I, II and III, and they were in part sensitive to pronase. The antigens of class J reflected strain specific or serotype reactions, some sensitive and others resistant to proteolytic enzymes. The antigens of class M were sensitive to periodate and resistant to pronase. Strains used in serological studies by other authors were tested. The properties of class W correlated well with those of the so-called micro-immunofluorescence and immunotype systems, and class M with those of the so-called endotoxin and acid polysaccharide systems. Strains from three different laboratories could all be grouped by class W and M reagents. Identical strains obtained independently from different laboratories gave very similar reaction patterns with the reagents available. Repeated GC-isolates from patients infected with beta-lactamase producing strains showed stable reactions with class W and J reagents, while there was a time-related variation of the class M pattern. We have found that the COA method is rapid, easy and reproducible in the serological classification of Neisseria gonorrhoeae and all the 117 GC-strains tested could be classified.     

Differentiation of Two Groups of Corynebacterium acnes

One hundred and forty-three strains of Corynebacterium acnes, isolated from human skin and acne lesions, were compared with three strains of Propionibacterium acnes from the American Type Culture Collection. The 146 organisms could be separated into two groups. Members of the larger group (129 strains) hydrolyzed gelatin and usually produced indole, Gel-In(+), but were unable to ferment trehalose, maltose, or sucrose, TMS(-). The deoxyribonucleic acid from selected strains of this group had an average guanosine + cytosine (GC) content of 60.5%. The members of the smaller group (17) were Gel-In(-), TMS(+), and the deoxyribonucleic acid had an average GC content of 63.9%. Studies with absorbed and unabsorbed antisera to the smaller group showed that although there were antigens shared by the two groups, it was possible to distinguish them serologically. Members of each group produced propionic acid. The principal fatty acid component of members of each group was iso-C(15) fatty acid. Seventy per cent of the Gel-In(+) strains were lysed by phage 174, whereas only one of 15 Gel-In(-) strains was lysed. Pending further information on the genetics of the two groups, those Gel-In(-), TMS(+) strains are tentatively designated C. acnes, group II.     

Identification of Isolates of Clostridium perfringens Types C and D by Agglutination and Fluorescent-Antibody Methods

Agglutination and fluorescent-antibody methods were employed for screening Clostridium perfringens types C and D from 393 isolates of this organism. All of 50 strains which were isolated in Japan and were agglutinable with an antiserum prepared against a stock strain of type C no. 3182 toxigenically belonged to type C, but the antiserum showed no cross-agglutination with any of type C strains isolated in Denmark. All of the latter strains, however, were agglutinated by an antiserum prepared against a Danish strain, CWC11. Of 64 strains, showing heat-labile agglutinability by type D antiserum L9, 22 strains were toxigenically identified as type D strains which can be divided into three groups by the heat-stable antigens; no strains which were L-agglutination-positive but O-agglutination-negative were epislon-toxigenic. All of 13 strains, the heat-stable antigen of which was agglutinable by a type D antiserum VX81, were toxigenically type D strains. The results of fluorescent-antibody tests were almost in agreement with those of agglutination test with type C strains and completely with those of the O-agglutination test with type D strains. No beta-, epsilon- or delta-toxigenicity could be demonstrated in strains which were not agglutinated by our test sera for types C and D strains. Further examination of cultural properties of Japanese and Danish type C strains revealed that the two groups were considerably different in urease production, capsule formation, and delta- and alpha-toxigenicities.     

Characterization and Relatedness of Marine Vibrios Pathogenic to Fish: Physiology, Serology, and Epidemiology

Cultural characteristics and serological relationships of pathogenic marine vibrios isolated from fish in the Pacific Northwest were studied. These organisms were compared with cultures of Vibrio anguillarum, a known fish pathogen. On the basis of morphological and cultural characteristics, the Pacific Northwest strains of Vibrio were found to be closely related to V. anguillarum. Serological analyses of thermostable antigens served to distinguish three serotypes among the vibrios. Serotype 1 was composed of organisms isolated from Northwest salmonids; serotype 2 of strains of V. anguillarum from European waters; and serotype 3 of organisms isolated from Pacific herring. The epidemiology of vibrio disease among populations of fish in the Pacific Northwest is discussed.     

Aerobactin production and dulcitol fermentation in clonal groups of Escherichia coli O1: K1 strains from urinary tract infections

Abstract 70 urinary Escherichia coli O1:K1 strains were characterized for O1 antigen factors, mannose-resistant hemagglutination of human erythrocytes, flagellar and fimbrial antigens, dulcitol fermentation and aerobactin production. On the basis of their O1 and H antigens the strains could be assigned to 6 distinct groups. The most prevalent groups were: O1abcd: H⁻ :F9 (33 strains; pattern II), O1abc: H⁻ :F11 (9 strains; pattern IV), and O1abc: H7: F11 (19 strains; pattern V). Strains with patterns IV and V, both expressing fimbrial antigen F11, fermented dulcitol and produced aerobactin, whereas strains with pattern II were negative for both characteristics.     

Isolation of non-type-specific protein antigens of Streptococcus group B

Hydrochloride extracts obtained from group B streptococcal strains of different serotypes have proved to be the source of type-nonspecific protein antigens, precipitated with ethanol and studied by gel chromatography and spectrophotometric scanning in ultraviolet rays. Thus, 2 or 3 antigens, one of them found to be common for streptococci of groups A, B and G, as well as the admixture of group-specific polysaccharide, have been detected. In extracts obtained from group B streptococcal strains of different serotypes a common protein antigen, specific only for group B, has been detected. The suitability of gel chromatography with the use Toyopearl gel HW-55F for the preparative isolation of the specific fraction of protein type-nonspecific antigen with a view to the subsequent study of immune response to group B streptococci has been shown.     

Alloreactive bovine T lymphocyte clones: an analysis of function, phenotype, and specificity

A bovine alloreactive cell population was subjected to complement-dependent lysis with monoclonal antibody (mAb) IL-A11. The original population and the population depleted of cells bearing the determinant recognized by mAb IL-A11 were cloned. Parent cultures and 21 clones were examined for cytolytic function and for expression of determinants recognized by mAb IL-A11 and two additional mAb, IL-A12 and IL-A17. Clones could be classified according to maximal achievable levels of cytolysis by using Theileria parva-infected bovine lymphoblastoid target cells. In this way, three groups were identified--one capable of high level cytolysis, one of intermediate levels, and one group comprising apparently noncytolytic clones. The clones in the first group reacted with mAb IL-A17; those in the second and third groups, with mAb IL-A11 and IL-A12. It was shown that cytotoxicity effected by IL-A17+ clones could be inhibited by this mAb and also by a mAb directed to MHC class I determinants on target cells. Conversely, cytotoxicity effected by IL-A11+/IL-A12+ clones could be inhibited by mAb IL-A11 and by a mAb directed to MHC class II determinants on target cells. The levels of expression of class I and class II determinants on target cells correlated with the levels of killing by clones of the IL-A17+ phenotype and clones of the IL-A11+/IL-A12+ phenotype, respectively. The results indicate that cytotoxic bovine T lymphocyte clones specific for class I MHC antigens and both cytotoxic and noncytotoxic clones specific for class II MHC antigens can be obtained. Further, their specificity for class I or class II antigens can be determined by phenotyping with mAb.     

Analysis of Diversity among 3-Chlorobenzoate-Degrading Strains of <Emphasis Type="Italic">Rhodopseudomonas palustris</Emphasis>

The phenotypic and genetic characteristics of 14 strains of the purple nonsulfur bacterium Rhodopseudomonas palustris were studied to assess diversity within this species. While all strains had certain phenotypic characteristics in common, including the ability to metabolize benzoate and degrade 2- and 3-chlorobenzoate, there were also significant differences among the strains such as the rate of growth in media containing benzoate as a carbon source. Genetic characterization of the strains revealed there were three divergent lineages in the species. Based on 16S rRNA gene sequences, the 14 strains could be grouped into three distinct clusters (A, B, and C), and this clustering was congruent with that based on gene sequences of form II ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO). Although BOX-PCR genomic DNA fingerprints of all 14 strains exhibited differences, analysis of the fingerprint images and UPGMA/product-moment analysis of similarities showed there were three groupings that were entirely consistent with clusters based on other characteristics of the strains. Thus, regardless of the method of analysis used, strains in groups A and B consistently clustered together and were separate from those of group C. These results suggest that strains in groups A-B and C represent phylogenetically related clones that have diverged from one another. This indicates that at least three lineages of Rhodopseudomonas palustris exist among the strains included in this study, and that each may be particularly well adapted to a distinct ecological niche.     

Serological relationships among budding, prosthecate bacteria

The somatic antigens of 25 strains of budding bacteria were typed and 14 serologically distinct groups were identified, suggesting considerable antigenic diversity among hyphomicrobia. Ten of the groups were represented by a single isolate, 2 contained two isolates, 1 three isolates, and 1 eight isolates. The strains in the largest group of eight isolates each shared at least one common antigen. However, there was also considerable antigenic heterogeneity within this cluster. Serological activity resided in the lipopolysaccharide (LPS) portion of cell walls and also in a heat labile component of Hyphomicrobium neptunium. The amino acid utilizing isolates, H. neptunium and Hyphomonas polymorpha, were serologically unrelated and it is suggested that the two organisms could be grouped as members of the same genus but not the same species.     

Generation and characterization of a lipopolysaccharide-specific murine monoclonal antibody to <Emphasis Type="Italic">Proteus vulgaris</Emphasis> OX19

The three strains of non-pathogenic Proteus species namely, Proteus vulgaris OX2, P. vulgaris OX19 and Proteus mirabilis OXK used in the Weil–Felix test are the group-specific cross-reactive antigens for Rickettsia and Orientia species. Earlier studies have revealed that the group specific and cross-reactive antigens responsible for the Weil–Felix test lie mostly in the lipopolysaccharide (LPS) moiety of the bacterial cell wall [Amano et al. (1993a) Infect Immun 61:4350–4355, (1993b) Microbiol Immunol 37:927–933, (1998) Infect Immun 66:923–926]. The three Proteus strains (OX2, OX19 and OXK) were used to raise murine monoclonal antibodies (MAbs) by hybridoma technology. Several MAb-producing hybridomas could be stabilized following limiting dilution. Affinity and specificity of these MAbs were checked by indirect ELISA using a battery of homologous and heterologous antigens including LPS. Amongst these, one MAb was found to be specific for P. vulgaris OX19 LPS. Since the Weil–Felix reaction is based on the cross-reactivity between the LPS based epitopes, this MAb could be of potential use in mapping of epitopes on the cross-reactive LPS and may also be useful as a potential diagnostic reagent.     

Norovirus and histo-blood group antigens: demonstration of a wide spectrum of strain specificities and classification of two major binding groups among multiple binding patterns

Noroviruses, an important cause of acute gastroenteritis, have been found to recognize human histo-blood group antigens (HBGAs) as receptors. Four strain-specific binding patterns to HBGAs have been described in our previous report. In this study, we have extended the binding patterns to seven based on 14 noroviruses examined. The oligosaccharide-based assays revealed additional epitopes that were not detected by the saliva-based assays. The seven patterns have been classified into two groups according to their interactions with three major epitopes (A/B, H, and Lewis) of human HBGAs: the A/B-binding group and the Lewis-binding group. Strains in the A/B binding group recognize the A and/or B and H antigens, but not the Lewis antigens, while strains in the Lewis-binding group react only to the Lewis and/or H antigens. This classification also resulted in a model of the norovirus/HBGA interaction. Phylogenetic analyses showed that strains with identical or closely related binding patterns tend to be clustered, but strains in both binding group can be found in both genogroups I and II. Our results suggest that noroviruses have a wide spectrum of host range and that human HBGAs play an important role in norovirus evolution. The high polymorphism of the human HBGA system, the involvement of multiple epitopes, and the typical protein/carbohydrate interaction between norovirus VLPs and HBGAs provide an explanation for the virus-ligand binding diversities.     

Isolation and partial characterization of a genus common antigen and species specific antigen of Capnocytophaga

Sixteen strains of Capnocytophaga were isolated from the pocket of a localized juvenile periodontitis patient. These strains were divided into four groups on the basis of morphological and physiological traits. Strains from group I and group III were identified as C. ochracea and group II as C. sputigena. An antigen common to genus Capnocytophaga was purified utilizing immunoabsorbent chromatography from lysates obtained by sodium dodecyl sulfate treatment of C. ochracea strain S1. An antigen specific to C. ochracea was prepared by sequential gel filtration and preparative isoelectric focusing. The genus common and species specific antigens isolated were immunologically unique and pure when tested by immunoelectrophoresis and immunodiffusion against rabbit antisera prepared to Capnocytophaga and other gram-negative rods. The genus common antigen was susceptible to trypsin and pronase digestion, was soluble in chloroform-methanol, but was unaltered by ribonuclease and deoxyribonuclease treatments and periodate oxidation. Antigenicity of the species specific antigen was destroyed by periodate oxidation. The genus common antigen appeared to be lipid-associated protein, while the species specific antigen consisted mainly of carbohydrate. These specific immunological reagents would be valuable in diagnosing and monitoring diseases.     

Mutants of Group D1Salmonella Carrying the Somatic Antigen of Group A Organisms: Isolation and Serological Characterization

O antigen mutants were obtained from Salmonella durban, a group D(1) organism, by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. Serological studies demonstrated that the mutants lost the O-9 antigen factor of the parent organism but acquired the O-2 factor specific to group A Salmonella. Lipopolysaccharides of the mutant strains contained paratose which determines the specificity of O-2 factor. Tyvelose, present in the wild-type lipopolysaccharide, was not found in the mutants. H antigens and other biological characteristics of the mutant strains were the same as those of the wild-type organism. The present finding implies that group A Salmonella species might be derived from group D(1) organisms.     

四川海螺沟冰川土样芽胞杆菌资源分析

【背景】芽胞杆菌(Bacillus-like)是一类能形成具有强抗性芽胞且可在多种极端环境下存活的细菌,其产生的多种功能代谢产物在多种领域具有重要研究价值。由于冰川低温、寡营养的独特生态环境,其存在的芽胞杆菌可能具有特殊性,因此研究冰川芽胞杆菌有利于发掘新基因、丰富芽胞杆菌多样性。【目的】了解四川海螺沟冰川土壤芽胞杆菌资源,为挖掘芽胞杆菌新资源提供基础。【方法】采用纯培养法分离获得冰川土壤芽胞杆菌资源,利用16S r RNA基因进行系统发育分析,测定代表性菌株的生理生化特性并采用类平均法和欧氏距离模型进行聚类分析。【结果】共筛选到可培养细菌44株,经16S r RNA基因鉴定确定其中36株为芽胞杆菌,隶属于4个属的19个种,分别为芽胞杆菌属(Bacillus)11个种24株、类芽胞杆菌属(Paenibacillus)2个种3株、短芽胞杆菌属(Brevibacillus)4个种5株和赖氨酸芽胞杆菌属(Lysinibacillus)2个种4株,其中以芽胞杆菌属(Bacillus)为优势属。分离菌中仅3株芽胞杆菌可在4°C生存,7株能在50°C生长,大部分菌株在30°C下生长良好;有74%菌株能耐碱,有37%菌株能在无盐条件下生长。根据生理生化结果,采用类平均法和欧氏距离模型进行聚类分析,可分为3组,分别包含7种、4种和6种芽胞杆菌。第1组均可以水解七叶灵和利用葡萄糖,第2组均不能水解七叶灵和利用葡萄糖,第3组仅能共同利用葡萄糖。【结论】四川海螺沟冰川土壤蕴藏着较为丰富的芽胞杆菌资源,为芽胞杆菌新资源挖掘提供了资源保障。     

Characterization of envelope antigens of influenza A (H3N2) virus isolated during 1983-1985 epidemics and from sporadic cases of infection

Ten strains of influenza A (H3N2) virus isolated from an outbreak in 1983, and ten strains isolated in 1985 from sporadic cases of infection were included in the study. For characterization of envelope antigens were used the polyclonal and monoclonal antibodies tested in the reaction of haemagglutinin inhibition, neuraminidase inhibition, and by lectin test. The strains but slightly different in the tests with polyclonal antibodies could clearly be classified to 3-4 groups using 5 monoclonal antibodies to H antigen of A/Bangkok 1/79 and A/Philippines 2/82 strains. Strains from the 1983 epidemics represent a more homogeneous group of which only one of ten strains failed to react with monoclonals of the strains A/Bangkok and A/Philippines. Strains from sporadic cases of infection in 1985, except for two strains, did not react at all with the monoclonal discriminating A/Bangkok and A/Philippines. The other strains could be classified to three groups, i.e. whether they agreed with 4, 2 or none of the A/Philippines H antigen epitopes. Alterations of neuraminidase are less apparent, and cannot be defined by means of normal immune sera. With the use of monoclonal antibodies the strains under study do not react any more with the strains of 1968-1973 influenza virus; yet the monoclonals to A/Texas/77 strain still do recognize one or two epitopes of the 1983-1985 strains.     

Serological characteristics within the genus Desulfovibrio

Antisera were prepared against one strain each of Desulfovibrio desulfuricans, D. vulgaris and D. salexigens. The antisera were tested for cross reactivity against 36 heterologous Desulfovibrio strains by both agglutination titration and by double immunodiffusion precipitin plates.Generally no cross-reaction was demonstrated by agglutination even between heterologous strains of the same species, suggesting that the surface antigens of Desulfovibrio are highly specific. In immunodiffusion plates a single apparently genus-specific surface antigen could be shown to be present in all but two of the strains tested. Although other common precipitin bands showed the presence of some antigens common between heterologous strains these appeared to be randomly distributed among the strains tested, with the exception of one band shown to be generally specific to strains of D. salexigens. With this exception no other precipitin band could be shown to be consistently specific to any other species, nor consistently common to more than one species.     

Monosaccharide composition of the lipopolysaccharides of bacteria of the genus Citrobacter

The authors studied antigens obtained by Grasset's method from 13 strains of Citrobacter of the International collection. The strains possessed O- and H-antigens whose behaviur in the electric field differed. All the strains under study were divided into two groups (by the number of serologically-active components of their O-antigens); representatives of the second group had no cathode O-antigen component. Chemical composition of specific lipopolysaccharides (LPS) obtained by Westphal's method was determined. Fourteen different sugars were revealed. The strains under study were referred to the known chemotypes. Strain 16/52 (8a, 8c) was for the first time studied in respect to the monosaccharide composition of specific LPS, and was referred to chemotype designated as CC-L.     

Epitope analyses of pneumococcal surface protein A: a combination of two monoclonal antibodies detects 94% of clinical isolates

Immunisation of BALB/c mice with seven heat-treated Norwegian clinical isolates of Streptococcus pneumoniae of different serotypes elicited mainly monoclonal antibodies (mAbs) to pneumococcal surface protein A (PspA). It was remarkable that the fusions resulted only in a few mAbs directed against other protein antigens. Dot blot analysis with 16 mAbs using clinical isolates representing 23 different capsular types and the uncapsulated reference strain R36A showed that some of the mAbs bound to PspA epitopes expressed by a low number of strains whereas others bound to broadly distributed epitopes. On the basis of their reactivities, seven of these mAbs could be divided into two groups recognising different subsets of pneumococci. The three mAbs in the narrow reacting group bound to epitopes found in 21-25% of the strains whereas the four mAbs in the broad reacting group detected more than 57% of the analysed strains. The epitopes for these seven antibodies were surface exposed on live exponential phase grown pneumococci as shown by flow cytometry. The finding that a combination of mAb 180,C-1 (IgG2a) from the first group and mAb 170,E-11 (IgG2a) from the second group detected 94% of the examined strains is interesting because PspA has been reported by others to be a serological highly variable protein.