Globo-A--a new receptor specificity for attaching Escherichia coli

Uropathogenic Escherichia coli strains designated as ONAP, based on their O negative A positive agglutination of human P1 erythrocytes, were shown to prefer the globo-A glycolipid as a receptor structure. The dependence on both the A terminal and the globoseries chain was confirmed by agglutination of human AP1, but not Ap or OP1 erythrocytes and by binding to the globo-A glycolipid on TLC plates. Neither Gal alpha 1----4Gal beta nor the A trisaccharide GalNAc alpha 1----3(Fuc alpha 1----2)Gal beta alone functioned as receptors. The bacteria thus appeared to recognize an epitope resulting from the combination of the terminal and internal structures.     

Agglutination Typing of Vibrio anguillarum Isolates from Diseased Fish and from the Environment

Agglutinating activity was widely distributed among 101 Vibrio anguillarum strains of different origin and three Vibrio ordalii strains from salmonids. The spectrum of cells which were agglutinated comprised yeast cells and human (type O), poultry, guinea pig, and trout erythrocytes, whereas ovine, bovine, and tanned bovine erythrocytes were not affected. Mannose-sensitive hemagglutination, mannose-resistant hemagglutination, and non-agglutinating strains were recognized. The three V. ordalii strains showed mannose-resistant hemagglutination, whereas V. anguillarum exhibited either mannose-sensitive hemagglutination or was non-agglutinating. Among V. anguillarum, sensitivity to d-galactose and l-fucose occurred sporadically. An agglutination typing scheme was developed for strains of V. anguillarum based on the agglutination pattern of human, poultry, guinea pig, and trout erythrocytes and yeast cells. Eight different agglutination types (A through H) were defined. The distribution of these types among fish pathogenic and environmental V. anguillarum strains were studied. The application of the typing scheme in ecological and epidemiological studies and for preventive medical purposes is discussed.     

一株产凝集素的三叶半夏内生真菌的研究

研究三叶半夏内生真菌及其凝集素,旨在为半夏内生真菌及其凝集素的开发利用提供依据。对三叶半夏块茎内生真菌分离、纯化,液体发酵培养代谢产物,无水乙醇提取总蛋白,兔血红细胞检测其凝集活性,筛选出菌株gs1,其总蛋白对兔血红细胞凝集活性显著。使用甘露聚糖-Sepharose 4B亲和层析柱纯化菌株gs1总蛋白,得到凝集素。Brandford法定量检测分析表明,1000 ml gs1发酵培养液中含有9.58 mg 凝集素。SDS-PAGE 电泳分析显示该凝集素为单一条带,分子量约为12 kDa。凝集活性实验表明,该凝集素对兔、大鼠和小鼠的血红细胞具有凝集作用,对兔血红细胞效果最显著;而对人（A＼B＼O＼AB型）和鸡的血红细胞无凝集作用。糖结合活性实验表明,甘露糖对该凝集素的凝集活性具有抑制作用。通过初步分类鉴定,菌株gs1为半知菌亚门,丝孢纲,丛梗孢目,丛梗孢科,曲霉属。     

Study on the influence of cranberry extract ?uravit S·O·S(?) on the properties of uropathogenic Escherichia coli strains, their ability to form biofilm and its antioxidant properties

Consumption of cranberries is known to exert positive health effects, especially against urinary tract infections. For this reason, presumably, they are widely used in folk medicine. Different aspects of cranberry phenolics activity were studied in individual papers but complex study in this matter is missing. The aim of the present study is to provide complex data concerning various aspects of cranberry extract activity. We studied the effects of subinhibitory concentrations of commercially available extract (?uravit S·O·S(?)) against two Escherichia coli strains isolated from urine of patients with pyelonephritis. Additionally the main extract anthocyanins were characterized. The activity of extract against lipid peroxidation and its radical scavenging ability were also assessed. ?uravit S·O·S(?) decreased the hydrophobicity of one of the studied E. coli strains, reduced swimming motility and adhesion to epithelial cells of both studied strains, it also limited the ability of bacteria to form biofilm. Expression of curli was not affected by cranberry extract, the assessment of P fimbriae expression was not reliable due to extract-induced agglutination of erythrocytes. Cranberry extract caused filamentation in both studied E. coli strains. It also showed pronounced antioxidant and radical scavenging properties. The properties of the studied cranberry extract show that it could be effectively used in prevention and/or elimination of urinary tract infections, specially the recurrent ones.     

Hemagglutination (fimbriae) and hydrophobicity in adherence ofSerratia marcescens to urinary tract epithelium and contact lenses

The capacity of 59 isolates of Serratia marcescens, obtained from urinary tract infections, wounds, and contact lenses or their paraphernalia, to agglutinate erythrocytes from different animal species was tested. Three main patterns were found: mannose-sensitive agglutination of guinea-pig, fowl or horse erythrocyte; mannose-resistant agglutination of chicken or pigeon erythrocytes alone or in combination with mannose-sensitive agglutination; and no agglutination. Hemagglutination capacity was associated with isolates from urinary tract infection, but not with isolates associated with contact lenses. Adherence to human urinary tract epithelium did not correlate with the hemagglutination patterns nor with the origin of the isolates. Some strains of different hemagglutination pattern were selected for the study of hydrophobicity and adherence to contact lens polymers. Hydrophobicity, as determined by degree of partition in hexadecane and water (BATH-values), correlated neither with degree of adherence to contact lens polymers nor with the hemagglutination pattern. For a representative strain there was an excellent correlation (r2 = 0.98) between adherence and the water content (hydrophobicity) of the lens polymers. These results suggest that, as with tissues, other factors interact with hydrophobicity in causing adherence to plastics.     

A study of the candidate virulence factors of Bacteroides fragilis

Bacteroides fragilis strains were classified as virulent or avirulent on the basis of their clearance from the subcutaneous tissues of mice. To determine the factors which may contribute to the virulence of B. fragilis strains, we studied encapsulation, hydrophobicity, growth rate, serum sensitivity, agglutination with erythrocytes of different origin, and neuraminidase production. The strains of the virulent group displayed a higher growth rate in broth and a lower sensitivity to the bactericidal activity of serum than the strains of the avirulent group. They also agglutinated different types of erythrocytes more strongly than did the avirulent strains. No significant differences were found between the two groups of strains as regards encapsulation, hydrophobicity and neuraminidase activity.     

The use of xenoantigenic antisera for the identification of tilapiine species: comparative laboratory and field studies

Rapid and reliable identification of tilapiine taxa and strains is essential for selective breeding purposes and the conservation of natural genetic resources. There is evidence that antisera‐mediated erythrocyte agglutination assays can fit these requirements. We evaluated the applicability of agglutination tests by studying the capacity of species characteristic antisera to recognize erythrocytes from individuals of 10 natural Ghanaian populations of Oreochromis niloticus and Sarotherodon melanotheron. The vast majority of the 218 tested individuals could be identified based on antisera‐mediated erythrocyte recognition. Controls indicated the specificity of these reactions. Still, erythrocytes from 16% of all tested specimens did not respond to any antiserum (zero responders), indicating the possible existence of blood group properties in tilapias. We discuss the specificity of the antisera, the relevance of zero responses and the applicability of these tests in aquaculture and field studies.     

我国新城疫病毒分离株生物学特性的研究

本文系统地比较了我国不同地区分离的、有地区代表性的11株新城疫病毒分离物和北京株(国内攻毒株)的毒力范围,某些生物学特性,抗原性关系及对鸡的致病特征,结果所试验的分离株均属速发性嗜内脏型毒株。各株血凝素的热稳定性、凝集解脱速度、凝集哺乳动物红细胞的能力及对福尔马林的敏感性,均有明显差异。试验还表明,北京株和中监所分离株等对不同年龄鸡的致病表现也不同。 交叉血凝抑制试验和交叉鸡胚中和试验表明:各新城疫病毒株相互间的抗原性有较小程度的差异(r值0.66～1.11);其免疫原性也强弱不同,尤其是新疆株和内蒙株,其血清的血凝抑制抗体效价高于其它毒株,与部分毒株的差异极为显著(P<0.01),可能是制造新城疫灭能苗的优良毒株。     

Frequency among Enterobacteriaceae of the DNA sequences encoding type 1 pili.

Type 1 pili, characterized by mannose-inhibitable agglutination of fowl or guinea pig erythrocytes, have been found throughout the family Enterobacteriaceae. A radiolabeled probe was prepared from a restriction endonuclease-digested fragment of the Escherichia coli pil operon and used to detect homologous DNA sequences in 236 bacteria representing 11 genera of Enterobacteriaceae. Only isolates identified as E. coli or Shigella spp. exhibited homology. In contrast, mannose-sensitive hemagglutination was observed in nine genera. Probe DNA did not hybridize to plasmid DNA, indicating a chromosomal location for the pil operon. Analysis of restriction nuclease-digested whole-cell DNA from 60 E. coli and two Shigella sp. isolates indicated that internal sequences were conserved in most strains, but that changes in flanking sequences in the chromosome were common.     

Estimation of pyruvate decarboxylation in perfused rat skeletal muscle

A total of 46 E. coli strains showing mannose-resistant, P-blood-group independent hemagglutination of human erythrocytes were tested for binding to neuraminic acid. Nine of the strains completely lost their hemagglutination activity after the erythrocytes were treated with neuraminidase. To characterize the receptor structure, different neuraminic acid containing glycoproteins, their desialylated derivatives and neuraminyl oligosaccharides were tested for hemagglutination inhibition. These studies showed that the nine strains had binding specificity for alpha 2-3 linked neuraminic acid.     

Expression and detection of different biotype-associated cell-bound haemagglutinins of Vibrio cholerae O1

The expression of two cell-bound haemagglutinins, one sensitive to L-fucose (FSHA) and the other to D-mannose (MSHA), on Vibrio cholerae O1 strains of both the classical and the El Tor biotypes was studied by (i) agglutination of chicken and human group O erythrocytes in the presence of L-fucose or D-mannose, (ii) binding of the bacteria to L-fucose- and D-mannose-coated agarose beads, and (iii) agglutination of the bacteria by 'biotype-specific' antisera. All of the 12 classical strains studied that were isolated before 1979 gave FSHA of human O erythrocytes whereas only 6 of 17 classical strains isolated during recent epidemics expressed FSHA; a few of the classical strains expressed MSHA in addition to FSHA. All the El Tor strains gave MSHA of chicken erythrocytes and one strain also expressed FSHA. Both the cell-bound HAs were optimally expressed during the exponential phase of growth; FSHA markedly decreased during the late exponential phase while the MSHA usually persisted into the stationary phase. The expression of FSHA and MSHA correlated very well with the direct binding of vibrios to fucose- and mannose-coated agarose beads, respectively. Antiserum 'specific' for classical vibrios agglutinated classical strains expressing FSHA and also the El Tor strain exhibiting FSHA. Similarly, the anti-El Tor serum agglutinated all El Tor strains and also classical strains expressing MSHA, suggesting that the 'biotype-specific' sera were specific for the biotype-associated cell-bound HAs.     

Lectin activity from embryonic chick brain,heart, and liver: Changes with development

Extracts of embryonic chick brain, heart, and liver agglutinate glutaraldehyde-fixed trypsinized or pronase-treated rabbit erythrocytes. Agglutination activity of extracts from each organ was inhibited by a number of saccharides. Lactose was the most potent saccharide inhibitor of those tested. The specific agglutination activity of the extracts from each of the organs studied changed with development of the embryo. In general, specific agglutination activity declined later in embryogenesis, and after hatching. However, the pattern of developmental change differed for each of the organs tested. Liver was unusual in that, after hatching, agglutination activity rose again; and the agglutinin found at this time was apparently different from that found in the embryo.     

Occurrence of p-fimbriae in enteropathogenic E coli (EPEC) and in E. coli strains isolated from patients with urinary tract infections]

The aim of this study was to gain knowledge of prevalence of P+ clones among EPEC strains isolated from children with diarrhoea and E. coli strains isolated from urine. Three hundred eighty four E. coli strains isolated from children with diarrhoea were tested. They belonged to 11 serotypes (018, 025, 026, 044, 055, 0111, 0114, 0119, 0124, 0125, and 0128). Nine hundred thirty colonies of E. coli from Mac Conkey's agar plated quantitatively with urine samples of 178 individuals suffering from urinary tract infections were also tested. All strains were assayed by mannose-resistant active haemagglutination test (MRHA) and by slide agglutination using self prepared latex reagent for detection of P fimbriae. Out of 384 E. coli strains tested 122 (31.8%) showed presence of adhesins detected by mannose-resistant active haemagglutination test (MRHA) and in 90 (23.3%) out of all tested strains the presence of P fimbriae was found. The highest percentage of P fimbriae prevalence was found in E. coli belonging to the following serotypes: 018 (in 68.9% strains), 025 (in 29.2% strains), and 0125 (in 25.0% strains). This type of fimbriae was also detected in serotypes 026 (9.1%), 044 (8.7%), 055 (5.6%), and 0119 (in 2 strains out of 5 isolated). Out of 933 colonies of E. coli, isolated from 178 urine samples, 434 (46.5%) colonies gave positive results in MRHA test, including 133 positive in latex test for P fimbriae. These studies showed that for MRHA adhesins, including P fimbriae, a parallel examination of higher number of E. coli was necessary.(ABSTRACT TRUNCATED AT 250 WORDS)     

Reciprocal exchange of minor components of type 1 and F1C fimbriae results in hybrid organelles with changed receptor specificities.

Type 1 and F1C fimbriae are surface organelles of Escherichia coli which mediate receptor-specific binding to different host surfaces. Such fimbriae are found on strains associated with urinary tract infections. The specific receptor binding of the fimbriae is due to the presence of receptor recognition proteins present in the organelles as minor structural elements. The organization of the fim and foc gene clusters encoding these fimbriae, as well as the structures of the organelles, are very similar, although the actual sequence homology of the structural elements is not remarkable; notably, the sequence identity between the minor components of the type 1 and F1C fimbriae is only 34 to 41%. Type 1 fimbriae mediate agglutination of guinea pig erythrocytes, whereas F1C fimbriae do not confer agglutination of any types of erythrocytes tested. However, F1C fimbriae mediate specific adhesion to epithelial cells in the collecting ducts of the human kidney as well as to cells of various cell lines. This report addresses the question of fimbrial promiscuity. Our data indicate that minor fimbrial structural elements can be exchanged between the two fimbrial systems, resulting in hybrid organelles with changed receptor specificity. This is the first study on reciprocal exchange of structural components from two different fimbrial systems.     

Neuraminidase production by Bacteroides species

Abstract 128 strains of Bacteroides isolated from clinical specimens were surveyed for their ability to produce neuraminidase. All strains of Bacteroides fragilis and the B. fragilis group were neuraminidase-positive, as were strains of B. oralis and B. bivius . All strains of B. capillosus, B. ruminicola, B. disiens, B. multiacidus and B. uniformis did not produce a detectable neuraminidase. When human erythrocytes were exposed to cell extracts of neuraminidase-producing Bacteroides , and then tested with peanut ( Arachis hypogeae ) lectin, agglutination occurred. It was concluded that the production of neuraminidase by clinical isolates of Bacteroides may be associated with the pathophysiology of severe Bacteroides infections.     

The study of agglutination of trypsin-treated sheep red cells by Corynebacterium diphtheriae strains

Abstract 620 Corynebacterium diphtheriae strains from 472 sick and healthy persons were studied for their adhesive activity (AA) in direct agglutination of trypsin-treated sheep erythrocytes. Toxigenic strains had more active AA than non-toxigenic ones which was not dependent on the presence of toxin in the culture. Neither biotype nor serotype of the strains correlated with their AA. Several lysotypes among toxigenic and non-toxigenic strains were more active than others. Toxigenic strains from patients had higher AA than those from carriers. Both toxigenic and non-toxigenic strains isolated from the prolonged carriers possessed the highest AA. It was concluded that AA measured in this way was an important colonization factor for all diphtheria strains and a pathogenicity factor for toxigenic strains.     

Chemical characterization of the lipopolysaccharides from enteropathogenic Escherichia coli O142 and O158.

Lipopolysaccharides (LPS) from two enteropathogenic strains of E. coli O142 and O158 were isolated by hot phenol-water extraction procedure. Polyacrylamide gel electrophoretic pattern of the LPS showed the typical ladder like pattern of smooth type of LPS. The LPS of E. coli O158 was found to contain L-rhamnose, D-glucose and N-acetyl-D-galactosamine as major constituents together with D-galactose, N-acetyl-D-glucosamine, L-glycero-D-manno-heptose and 2-keto-3-deoxy-D-manno-octulosonic acid (KDO) whereas LPS from E. coli O142 contained L-rhamnose, N-acetyl-D-glucosamine and N-acetyl-D-galactosamine as major constituents together with D-glucose, D-galactose, N-acetyl-D-glucosamine, L-glycero-D-mannoheptose and 2-keto-3-deoxy-D-manno-octulosonic acid (KDO). LPS was degraded by mild acid hydrolysis to yield a degraded polysaccharide fraction and an insoluble lipid-A fraction. The main fatty acids of the lipid-A fraction of the LPS were C12:O, C14:O, and 3-OH C14:O for O158 strain whereas E. coli O142 lipid-A consisted of C12:O, C14:O, 3-OH C14:O, and C16:O. The degraded polysaccharide fraction on gel permeation chromatography gave a high moleculer weight O-chain fraction and a core oligosaccharide and a fraction containing degraded sugars. The chemical composition of LPS and its fragmented products are reported in this communication.     

Involvement of a sialic acid-binding lectin with hemagglutination and hydrophobicity of Flavobacterium psychrophilum

Strains of Flavobacterium psychrophilum were studied for their ability to adhere and cause agglutination of erythrocytes and yeast cells. Strains of the serotype Th showed low or no hemagglutinating (HA) properties toward human, avian, bovine, and rainbow trout erythrocytes, whereas strains of serotype Fd and Fp(T) exhibited distinct HA properties. None of the strains was able to cause agglutination of yeast cells. Greater adherence specificity toward rainbow trout blood cells was seen for the HA-positive strains. Growth at 5 degrees C, compared to that at 15 degrees C, induced an increase in the hemagglutination of some strains. HA activities of F. psychrophilum were inhibited only by sialic acid (N-acetyl-neuraminic acid), heat treatment at 65 degrees C, and proteinase K treatment and not by any of seven other carbohydrates, periodate oxidation, or treatment with trypsin. The supernatant from washed bacterial cells also showed some HA properties. All strains were shown to be highly hydrophobic by the hydrophobic interaction chromatography test, although some contradictions to the results of the salt aggregation test (showing some strains as less hydrophobic) were seen. These results indicate that the aggregation of F. psychrophilum and erythrocytes depend on a lectin present on the surface of HA-positive F. psychrophilum strains and absent on HA-negative strains. This lectin reacts specifically with sialic acid. The adhesion differences observed for F. psychrophilum strains do not appear to correlate with the virulence but still provide insights into the interaction of F. psychrophilum and rainbow trout.     

Incidence of enteritis of Enterobacteriaceae isolates possessing human colonization factor antigen. New human colonization factors

Of 462 Enterobacteriaceae strains including 435 Escherichia coli isolated from 250 patients, 298 haemagglutinating (HA) cultures were classified into 36 different HA groups. Sixteen of them belonged to Evan's I or II groups, although none possessed CF I or CF II antigen detectable by slide agglutination. Seventy-seven strains showed 4+ mannose resistant (MR) HA with human (53), bovine (2), chicken (6), guinea pig (7) or human and guinea pig (9) erythrocytes. These strains were significantly more frequent in patients under one year of age. Eighty-eight percent of the typable strains belonged to E. coli serogroups O1, O2, O4, O6, O18. HA positivity and fimbrial structures were correlated in 2 isolates (15/1, O18a, c:-K77: H-; 12/2/1 O1: K1: H .). Fimbriae of the two strains exhibited adhesive properties. Their fimbrial antigens differed serologically from each other and from those of the reference strains H 10407 and PB 176. Forty-nine of 4+ human MRHA strains showed variable reactions in the two sera for the new fimbrial antigens.     

Occurrence of P.fimbrii in strains from selected genera of Enterobacteriaceae]

This study was aimed at recognition of frequency of occurrence of P fimbriae in strains of Escherichia coli isolated from samples of feces of children with symptoms of diarrhoea and at search of these adhesions in strains representing other than Escherichia genera of Enterobacteriaceae strains. One hundred forty laboratory strains were investigated. They belonged to genus Citrobacter, Enterobacter, Hafnia, Klebsiella, Morganella, Proteus, Providencia, Salmonella, Shigella, and Yersinia. Also were tested 1277 colonies of enteric rods isolated from the MacConkey's medium inoculated with samples of feces from 163 children with symptoms of diarrhoea. Mannose-resistant active hemagglutination test was performed with human group O erythrocytes and guinea pig erythrocytes stabilized with glutaraldehyde. Presence of P fimbriae was detected by the slide latex test with latex covered by P1 glycoprotein. Among 140 laboratory strains of Enterobacteriaceae in 21 strains (3-E. cloaceae, 2-Hafnia, 13-K. pneumoniae, 2-P. rettgeri and in one strains of Providencia) presence of MRHA adhesins was demonstrated. Nine of these strains (2-Hafnia, 5-K. pneumoniae and 2-P. rettgeri) reacted specifically in the latex test. Among 1142 colonies of E. coli isolated from children with symptoms of diarrhoea, 326 colonies belonged to 13 EPEC serotypes. With 118 (36.2%) of EPEC colonies a positive result of MRHA reaction was found with human erythrocytes and 34 (10.4%) with guinea pig erythrocytes. Positive latex test was obtained with 77 (23.6%) colonies. All these colonies possessed MRHA adhesins. Remaining 816 colonies of E. coli strains did not represent microorganisms belonging to serotypes accepted as enteropathogenic. From 112 (13.7%) colonies out of 816 not belonging to EPEC, positive results was obtained in the MRHA test with human erythrocytes and this was the case also with 41 (5.0%) colonies in MRHA reaction with application of guinea pig erythrocytes. The latex test was positive in 65 (7.9%) colonies of E. coli. From remaining 135 colonies other than E. coli, positive result of latex test of presence of P fimbriae was obtained with 54 (40.0%) colonies, including 14 colonies of E. cloacae, 23-K. pneumoniae and 17-K. oxytoca. In all these strains presence of MRHA adhesins was demonstrated. These investigations demonstrated that among EPEC strains significantly more frequently, than not belonging to these serotypes of E. coli, MRHA adhesins, including P fimbriae was observed.(ABSTRACT TRUNCATED AT 400 WORDS)