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1.
Gesundheitswesen ; 78(11): 686-688, 2016 Nov.
Article in German | MEDLINE | ID: mdl-27756087

ABSTRACT

Public health is a population- and system-based approach that is needed to improve the health of societies and to decrease health inequalities. In the face of global challenges, the public health approach is essential. In Germany, the importance of public health is only partly reflected by its institutions and institutional arrangements. This applies equally to research, teaching and training, as well as to the public health service. Furthermore, the public health perspective is not sufficiently considered in cross-sectional topics that are relevant for health.There have been several initiatives to overcome structural deficits which can partly be traced back to historical circumstances. The White Paper presented here should encourage discussions about future policy options in public health. The authors represent public health in practice, research, and teaching in Germany.


Subject(s)
Delivery of Health Care/organization & administration , Health Policy , Health Services Accessibility/organization & administration , Models, Organizational , Organizational Objectives , Public Health Administration/methods , Germany , Quality Improvement
2.
Rev Sci Tech ; 33(3): 869-75, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25812210

ABSTRACT

The complement fixation test (CFT) is the only serological test prescribed by the World Organisation for Animal Health (OIE) for the diagnosis of glanders in international trading of equids. However, false-positive reactions have caused financial losses to the animal owners in the past, and false-negative tests have resulted in the introduction of glanders into healthy equine populations in previously glanders-free areas. Both warm (incubation at 37°C for 1 h) and cold (overnight incubation at 4°C) procedures are recommended by the OIE for serodiagnosis of glanders. In a comparison of the sensitivity and specificity of the two techniques, using the United States Department of Agriculture antigen, warm CFT was found to be significantly less sensitive (56.8%; p < 0.0005) than the cold CFT (83.6%). Cold CFT thus increases the detection rate of glanders but a lower diagnostic specificity has to be accepted. The immunoblot was used as the gold standard.


Subject(s)
Complement Fixation Tests/veterinary , Glanders/diagnosis , Temperature , Animals , Antigens, Bacterial , Complement Fixation Tests/methods , Glanders/microbiology , Sensitivity and Specificity
3.
J Clin Microbiol ; 51(4): 1099-109, 2013 Apr.
Article in English | MEDLINE | ID: mdl-23345291

ABSTRACT

Streptococcus canis is an animal pathogen that occasionally causes human infections. Isolates recovered from infections of animals (n = 78, recovered from 2000 to 2010 in three European countries, mainly from house pets) and humans (n = 7, recovered from 2006 to 2010 in Portugal) were identified by phenotypic and genotypic methods and characterized by antimicrobial susceptibility testing, multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and emm typing. S. canis isolates presented considerable variability in biochemical profiles and 16S rRNA. Resistance to antimicrobial agents was low, with the most significant being tet(M)- and tet(O)-mediated tetracycline resistance. MLST analysis revealed a polyclonal structure of the S. canis population causing infections, where the same genetic lineages were found infecting house pets and humans and were disseminated in distinct geographic locations. Phylogenetic analysis indicated that S. canis was a divergent taxon of the sister species Streptococcus pyogenes and Streptococcus dysgalactiae subsp. equisimilis and found evidence of acquisition of genetic material by S. canis from S. dysgalactiae subsp. equisimilis. PFGE confirmed the MLST findings, further strengthening the similarity between animal and human isolates. The presence of emm-like genes was restricted to a few isolates and correlated with some MLST-based genetic lineages, but none of the human isolates could be emm typed. Our data show that S. canis isolates recovered from house pets and humans constitute a single population and demonstrate that isolates belonging to the main genetic lineages identified have the ability to infect the human host, providing strong evidence for the zoonotic nature of S. canis infection.


Subject(s)
Genetic Variation , Multilocus Sequence Typing/methods , Phylogeny , Streptococcal Infections/diagnosis , Streptococcal Infections/veterinary , Streptococcus/classification , Zoonoses/microbiology , Animals , Anti-Bacterial Agents/pharmacology , Antigens, Bacterial/genetics , Bacterial Outer Membrane Proteins/genetics , Carrier Proteins/genetics , Humans , Microbial Sensitivity Tests , Molecular Epidemiology , Molecular Sequence Data , Pets , Recombination, Genetic , Sequence Analysis, DNA , Streptococcal Infections/microbiology , Streptococcus/genetics , Streptococcus/isolation & purification
4.
Appl Environ Microbiol ; 79(2): 655-62, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23160118

ABSTRACT

Staphylococcus aureus causes a wide range of infectious diseases in humans and various animal species. Although presumptive host-specific factors have been reported, certain genetic lineages seem to lack specific host tropism, infecting a broad range of hosts. Such Extended-Host-Spectrum Genotypes (EHSGs) have been described in canine infections, caused by common regional human methicillin-resistant S. aureus (MRSA) lineages. However, information is scarce about the occurrence of methicillin-susceptible S. aureus (MSSA) EHSGs. To gain deeper insight into EHSG MSSA and EHSG MRSA of human and canine origin, a comparative molecular study was carried out, including a convenience sample of 120 current S. aureus (70 MRSA and 50 MSSA) isolates obtained from infected dogs. spa typing revealed 48 different spa types belonging to 16 different multilocus sequence typing clonal complexes (MLST-CCs). Based on these results, we further compared a subset of canine (n = 48) and human (n = 14) strains, including isolates of clonal complexes CC5, CC22, CC8, CC398, CC15, CC45, and CC30 by macrorestriction (pulsed-field gel electrophoresis [PFGE]) and DNA-microarray analysis. None of the methods employed was able to differentiate between clusters of human and canine strains independently of their methicillin resistance. In contrast, DNA-microarray analysis revealed 79% of the 48 canine isolates as carriers of the bacteriophage-encoded human-specific immune evasion cluster (IEC). In conclusion, the high degree of similarity between human and canine S. aureus strains regardless of whether they are MRSA or MSSA envisions the existence of common genetic traits that enable these strains as EHSGs, challenging the concept of resistance-driven spillover of MRSA.


Subject(s)
Staphylococcal Infections/microbiology , Staphylococcal Infections/veterinary , Staphylococcus aureus/classification , Staphylococcus aureus/isolation & purification , Animals , Bacteriophages/genetics , Cluster Analysis , Dogs , Electrophoresis, Gel, Pulsed-Field , Genotype , Host Specificity , Humans , Methicillin Resistance , Microarray Analysis , Multilocus Sequence Typing , Staphylococcal Infections/epidemiology , Staphylococcus aureus/genetics , Staphylococcus aureus/physiology
5.
J Appl Microbiol ; 115(6): 1402-10, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24024971

ABSTRACT

AIMS: Lactobacilli strains with probiotic effects have been widely used in dairy products such as yoghurts as well as in food additives and pharmaceuticals. Despite their successful commercial application, the current species identification and quantification methods of the genus Lactobacillus are time-consuming and labour-intensive. METHODS AND RESULTS: To fulfil the requirements of a robust quality management, we have developed a quantitative real-time PCR assay based on the heat shock protein 60 gene (hsp60) for accurate identification and quantification of five commercially important Lactobacillus species. The developed assay allows an unambiguous species-specific detection of the species Lact. acidophilus, Lact. brevis, Lact. delbrueckii subsp. bulgaricus, Lact. helveticus and Lact. reuteri from bacterial cultures as well as directly from dairy products for instance yoghurt. CONCLUSIONS: With the assay, we were able to specifically detect lactobacilli strains down to 10(5)  CFU ml(-1) directly from yoghurt, which is a sufficient detection limit as commercial products usually contain 10(6) -10(12)  CFU ml(-1) of probiotic strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The real-time PCR assay developed here might become a convenient tool enabling an accurate, fast and sensitive detection of probiotic lactobacilli commercially used in food.

6.
Gut Pathog ; 12: 4, 2020.
Article in English | MEDLINE | ID: mdl-31988666

ABSTRACT

BACKGROUND: Following the ban on antimicrobial usage for growth promotion in animal husbandry in the EU, non-antimicrobial agents including heavy metal ions (e.g. zinc and copper), prebiotics or probiotics have been suggested as alternatives. Zinc has extensively been used in pig farming, particularly during weaning of piglets to improve animal health and growth rates. Recent studies, however, have suggested that high dietary zinc feeding during weaning of piglets increases the proportion of multi-drug resistant E. coli in the gut, contraindicating the appropriateness of zinc as an alternative. The underlying mechanisms of zinc effects on resistant bacteria remains unclear, but co-selection processes could be involved. In this study, we determined whether E. coli isolates from intestinal contents of piglets that had been supplemented with high concentrations of zinc acquired a higher tolerance towards zinc, and whether multi-drug resistant isolates tolerated higher zinc concentrations. In addition, we compared phenotypic zinc and copper resistance of E. coli isolates for possible correlation between phenotypic resistance/tolerance to different bivalent ionic metals. RESULTS: We screened phenotypic zinc/copper tolerance of 210 isolates (including antimicrobial resistant, multi-drug resistant, and non-resistant E. coli) selected from two, independent zinc-feeding animal trials by determining a zinc/copper minimal inhibitory concentration (Merlin, Bornheim-Hersel, Germany). In both trials, groups of piglets were supplemented either with high dietary zinc (> 2000 ppm) or control (50-70 ppm, background) concentrations. Our observations showed that high concentration zinc exposure did not have an effect on either zinc or copper phenotypic tolerance of E. coli isolates from the animals. No significant association was found between antimicrobial resistance and phenotypic zinc/copper tolerance of the same isolates. CONCLUSION: Our findings argue against a co-selection mechanism of antimicrobial drug-resistance and zinc tolerance after dietary zinc supplementation in weaning piglets. An explanation for an increase in multi-drug resistant isolates from piglets with high zinc dietary feeding could be that resistant bacteria to antimicrobial agents are more persistent to stresses such as zinc or copper exposure.

7.
Eur J Clin Microbiol Infect Dis ; 28(11): 1375-82, 2009 Nov.
Article in English | MEDLINE | ID: mdl-19701815

ABSTRACT

The aim of this study was to evaluate the prevalence and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) among pigs and estimate the impact of this animal reservoir on human healthcare. Nasal swabs were derived from 1,600 pigs at 40 German farms. The MRSA were characterized using S. aureus protein A (spa) typing, multilocus sequence typing (MLST) and detection of toxin genes. In a retrospective case control study, we compared risk factors for the carriage of MRSA between patients carrying spa types found among regional pigs and patients with other MRSA molecular types. Pigs carrying MRSA were identified on 70% of the farms (spa types t011, t034, t108, t1451 and t2510, all associated with MLST sequence type ST398). Contact to pigs and cattle were independent risk factors for the carriage of these spa types in patients at hospital admission. Our results indicate that livestock represents a relevant reservoir for the import of MRSA into regional German hospitals.


Subject(s)
Carrier State/veterinary , Cross Infection/microbiology , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/veterinary , Swine Diseases/microbiology , Zoonoses/microbiology , Zoonoses/transmission , Adolescent , Adult , Aged , Aged, 80 and over , Animals , Bacterial Typing Techniques , Carrier State/microbiology , Child , Child, Preschool , Cross Infection/epidemiology , DNA Fingerprinting , DNA, Bacterial/genetics , Female , Genotype , Germany/epidemiology , Hospitals , Humans , Infant , Male , Methicillin-Resistant Staphylococcus aureus/classification , Methicillin-Resistant Staphylococcus aureus/genetics , Middle Aged , Molecular Epidemiology , Nasal Cavity/microbiology , Prevalence , Risk Factors , Staphylococcal Infections/epidemiology , Staphylococcal Infections/microbiology , Swine , Swine Diseases/epidemiology , Young Adult , Zoonoses/epidemiology
8.
Vet Microbiol ; 124(1-2): 73-81, 2007 Sep 20.
Article in English | MEDLINE | ID: mdl-17498893

ABSTRACT

The aim of this study was to compare the in vitro antimicrobial activity of the veterinary fluoroquinolones against a panel of recently isolated porcine and bovine bacterial pathogens. The study used enrofloxacin as a benchmark against which other agents were compared, being the most common fluoroquinolone used in treatment of bovine and porcine infections. The activity of ciprofloxacin was also assessed as it is the main metabolite of enrofloxacin in cattle. Enrofloxacin and ciprofloxacin generally showed higher antibacterial activity, in terms of MIC(50) values, for most pathogen species when compared with marbofloxacin, difloxacin, danofloxacin and norfloxacin. Ciprofloxacin showed significantly greater in vitro antibacterial activity than enrofloxacin against M. haemolytica, P. multocida and E. coli, whereas enrofloxacin showed greater activity than ciprofloxacin against S. aureus. Marbofloxacin was significantly more active than enrofloxacin against M. haemolytica, E. coli and B. bronchiseptica but less active against P. multocida, S. aureus, coagulase negative Staphylococci, S. dysgalactiae, S. uberis, A. pleuropneumoniae and S. suis. Danofloxacin was significantly less active than enrofloxacin against P. multocida, E. coli, S. uberis, A. pleuropneumoniae and S. suis. Enrofloxacin and its metabolite ciprofloxacin showed the highest in vitro activities against most bovine pathogens tested and the porcine pathogens also showed a high degree of sensitivity to enrofloxacin. These data facilitate further pharmacokinetic/pharmacodynamic comparison of fluoroquinolones currently used in veterinary medicine.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteria/drug effects , Drug Resistance, Bacterial , Fluoroquinolones/pharmacology , Microbial Sensitivity Tests/veterinary , Animals , Bacterial Infections/drug therapy , Bacterial Infections/microbiology , Bacterial Infections/veterinary , Cattle , Cattle Diseases/drug therapy , Cattle Diseases/microbiology , Ciprofloxacin/pharmacology , Colony Count, Microbial/veterinary , Dose-Response Relationship, Drug , Enrofloxacin , Swine , Swine Diseases/drug therapy , Swine Diseases/microbiology
9.
J Comp Pathol ; 155(1 Suppl 1): S27-40, 2016 Jul.
Article in English | MEDLINE | ID: mdl-25958184

ABSTRACT

The close contact between household pets and people offers favourable conditions for bacterial transmission. In this article, the aetiology, prevalence, transmission, impact on human health and preventative measures are summarized for selected bacterial zoonoses transmissible by household pets. Six zoonoses representing distinct transmission routes were selected arbitrarily based on the available information on incidence and severity of pet-associated disease caused by zoonotic bacteria: bite infections and cat scratch disease (physical injuries), psittacosis (inhalation), leptospirosis (contact with urine), and campylobacteriosis and salmonellosis (faecal-oral ingestion). Antimicrobial resistance was also included due to the recent emergence of multidrug-resistant bacteria of zoonotic potential in dogs and cats. There is a general lack of data on pathogen prevalence in the relevant pet population and on the incidence of human infections attributable to pets. In order to address these gaps in knowledge, and to minimize the risk of human infection, actions at several levels are recommended, including: (1) coordinated surveillance of zoonotic pathogens and antimicrobial resistance in household pets, (2) studies to estimate the burden of human disease attributable to pets and to identify risk behaviours facilitating transmission, and (3) education of those in charge of pets, animal caretakers, veterinarians and human medical healthcare practitioners on the potential zoonotic risks associated with exposure to pets. Disease-specific recommendations include incentives to undertake research aimed at the development of new diagnostic tests, veterinary-specific antimicrobial products and vaccines, as well as initiatives to promote best practices in veterinary diagnostic laboratories and prudent antimicrobial usage.


Subject(s)
Bacterial Infections/epidemiology , Bacterial Infections/transmission , Pets/microbiology , Zoonoses/epidemiology , Zoonoses/transmission , Animals , Cats , Dogs , Humans , Public Health/legislation & jurisprudence
10.
Vet Immunol Immunopathol ; 105(1-2): 151-61, 2005 May 01.
Article in English | MEDLINE | ID: mdl-15797484

ABSTRACT

The influence of the probiotic bacterium Enterococcus faecium SF68 on the immune system and the intestinal colonization of pigs were determined in a feeding experiment with sows and piglets. Mucosal immunity of the developing piglets was monitored by isolation and detection of intestinal lymphocyte cell populations from the proximal jejunal epithelium and the continuous Peyers patches by the use of flow cytometry. The levels of intestinal IgA in both groups of piglets were compared, as well as total IgG in the serum of sows and piglets. Feces of the sows and intestinal contents of the piglets were taken for determination of total anaerobe and coliform bacterial counts in both probiotic and control groups. Villus length and depth of the crypts were measured in the jejunum of sacrificed piglets to monitor the development of the intestinal mucosal surface amplification. Total serum IgG of the sows appeared to be unaffected. Piglets of both groups showed similar IgG levels up to 5 weeks after birth with a slight tendency toward lower values in the probiotic group. At an age of 8 weeks the total IgG levels of the probiotic animals were significantly lower (p<0.01). No differences were observed in the populations of CD4+ and CD8+ T cells in the Peyers patches. However, the levels of cytotoxic T cells (CD8+) in the jejunal epithelium of piglets of the probiotic group were significantly reduced. The depth of the jejunal crypts and length of the villi were similar in both groups, suggesting the relative T-cell population differences were not due to alterations in the epithelial cell numbers. The total anaerobe and coliform bacterial populations were not significantly affected by the probiotic treatment, either in sows or in the piglets. However, a remarkable decline in the frequency of beta-haemolytic and O141 serovars of Escherichia coli was observed in the intestinal contents of probiotic piglets, suggesting an explanation for the reduction in cytotoxic T-cell populations.


Subject(s)
Enterococcus faecium , Immune System/drug effects , Probiotics/pharmacology , Swine/immunology , Animals , Animals, Suckling , Colony Count, Microbial/veterinary , Escherichia coli/growth & development , Feces/microbiology , Female , Immune System/growth & development , Immunity, Mucosal/immunology , Immunoglobulin A/analysis , Immunoglobulin G/blood , Immunophenotyping/veterinary , Intestinal Mucosa/immunology , Intestinal Mucosa/microbiology , Intestines/immunology , Intestines/microbiology , Lymphocytes/immunology , Lymphocytes/microbiology , Random Allocation , Serotyping/veterinary
11.
Arch Neurol ; 49(10): 1021-6, 1992 Oct.
Article in English | MEDLINE | ID: mdl-1417509

ABSTRACT

The neurocognitive effects of aluminum (Al) were studied in 35 hemodialysis patients. Higher Al levels were associated with a decline in visual memory. As Al levels increased, patients with lower vocabulary scores (a measure of premorbid intelligence) showed a decline in attention/concentration, frontal lobe functions, and on several neurocognitive measures, while those with higher vocabulary scores revealed no Al-related decline. These results suggest that individuals with lower verbal intelligence may possess less well-developed compensatory strategies to overcome the neurocognitive effects associated with Al. These data also indicate that Al is neurotoxic and, therefore, potential sources of environmental Al should be identified and eliminated.


Subject(s)
Aluminum/adverse effects , Cognition Disorders/chemically induced , Nervous System Diseases/chemically induced , Adult , Aged , Aluminum/metabolism , Female , Humans , Language Disorders/chemically induced , Male , Middle Aged , Neuropsychological Tests
12.
Neurology ; 41(11): 1806-9, 1991 Nov.
Article in English | MEDLINE | ID: mdl-1944913

ABSTRACT

Although increased levels of aluminum (Al) are present in patients with dialysis encephalopathy (DE), it is unclear if the association is causal. The enzyme dihydropteridine reductase (DHPR) plays a critical role in neurotransmitter formation and its activity. Elevated levels of Al are reported to decrease DHPR activity, which would alter neurotransmitter metabolism, thus producing DE. We examined the association between erythrocyte DHPR activity and Al levels, attention/psychomotor skills, and depression in a group of 21 patients with end-stage renal disease. DHPR activity was not related to Al level, mental status, psychomotor ability, or depression score. After administration of deferoxamine (an Al chelating agent), Al level increased significantly but DHPR activity remained the same. Our results suggest that the mechanism for the development for DE does not involve alterations of neurotransmitter metabolism caused by Al-mediated reductions in DHPR activity.


Subject(s)
Aluminum/blood , Cognition/physiology , Dihydropteridine Reductase/blood , Kidney Failure, Chronic/enzymology , Adult , Aged , Female , Humans , Kidney Failure, Chronic/blood , Kidney Failure, Chronic/parasitology , Male , Middle Aged , Regression Analysis
13.
FEMS Microbiol Lett ; 204(1): 75-9, 2001 Oct 16.
Article in English | MEDLINE | ID: mdl-11682182

ABSTRACT

We describe a locus of enterocyte effacement (LEE) which is part of a new pathogenicity island (PAI) detected in the bovine Shiga toxin-producing Escherichia coli strain RW1374 (O103:H2). This PAI is at least 80 kb in size and inserted in the vicinity of the pheV tRNA gene at 67 min of the E. coli chromosome. Furthermore, the PAI differs from the previously described LEEs by unique flanking regions at both sides, which harbor one copy each of an insertion element in an inverted orientation that is 96% identical to insertion site (IS)629. In addition, a 5-kb PAI-specific sequence downstream of the LEE core region and adjacent to the E. coli K12 region is duplicated upstream of the LEE core region as well. The duplicated sequences are more than 80% identical to each other and consist partially of prophage sequences.


Subject(s)
Cattle Diseases/microbiology , DNA Transposable Elements , Escherichia coli Infections/veterinary , Escherichia coli/pathogenicity , RNA, Transfer, Amino Acid-Specific/genetics , Animals , Cattle , Cattle Diseases/pathology , Enterocytes/microbiology , Enterocytes/pathology , Escherichia coli/genetics , Escherichia coli Infections/microbiology , Escherichia coli Infections/pathology , Nucleic Acid Hybridization , Phenylalanine , Restriction Mapping , Sequence Analysis, DNA , Shiga Toxins/biosynthesis , Shiga Toxins/genetics , Virulence/genetics
14.
FEMS Microbiol Lett ; 156(1): 49-53, 1997 Nov 01.
Article in English | MEDLINE | ID: mdl-9368360

ABSTRACT

The locus of enterocyte effacement pathogenicity island confers the attaching and effacing histopathology on epithelial cells infected with enteropathogenic and enterohemorrhagic Escherichia coli. We investigated the site of insertion of the locus of enterocyte effacement in E. coli strains in relation to their evolution based on conservation of housekeeping proteins in these strains. The results indicate that the insertion site of the locus of enterocyte effacement varies according to the evolutionary lineage, suggesting that it has inserted at multiple times and sites during the evolution of these pathogens.


Subject(s)
Escherichia coli/genetics , Escherichia coli/pathogenicity , Intestines/microbiology , Bacterial Adhesion , Base Sequence , Binding Sites , DNA Primers/genetics , DNA, Bacterial/genetics , Epithelial Cells/microbiology , Epithelial Cells/pathology , Escherichia coli/classification , Escherichia coli Infections/microbiology , Escherichia coli Infections/pathology , Evolution, Molecular , Gastrointestinal Hemorrhage/microbiology , Gastrointestinal Hemorrhage/pathology , Humans , Intestinal Diseases/microbiology , Intestinal Diseases/pathology , Intestines/pathology , Phylogeny , Polymerase Chain Reaction , Virulence/genetics
15.
Vet Microbiol ; 43(2-3): 131-41, 1995 Feb.
Article in English | MEDLINE | ID: mdl-7740752

ABSTRACT

Previous or present infection with Shiga-like toxin producing E. coli (SLTEC) was detected by an indirect neutralization assay of antibody titer. Bovine colostra and sera blocked the cytotoxic effects of Shiga-like toxin on Vero cell monolayers. SLT neutralizing antibodies were present in 84.0% (189/225) of the colostrum samples from randomly chosen cows in Bavaria, Germany. While all of the colostra with neutralizing activity reacted with SLT-I, only 14.7% neutralized both SLT-I and -II. Approximately 93.0% (37/40) of sera from heifers had SLT neutralizing activity. To quantify the neutralizing antibodies, colostra were tested in the Vero cell assay for their capability to reduce the 50% cytotoxic dose (CD50) of SLT standards, where the neutralizing units/ml (nu/ml) equal the log10 of CD50 reduction. Almost half of reactive colostra (48.7%) reduced the CD50 of the SLT-I standard by 10(4) to 10(5) (4-5 nu/ml). Higher reactivity (5-7 nu/ml) was found in 46.5% of positive colostra. The remaining colostra samples had over 7 nu/ml. To determine if the colostra were blocking receptors for SLT on Vero cells, cells were preincubated with colostra, and SLT was later added. No neutralizing activity was detected, indicating the reactivity of colostra was directed against SLT. When the colostra were subjected to ammonium sulphate precipitation and DEAE anion exchange chromatography, high levels of neutralizing activity were found in the IgG1 containing fractions. Colostrum fractions were tested for SLT-I binding antibodies in a capture ELISA, based on the binding of SLT-I to the toxin receptor analogue P1-glycoprotein. Only fractions from colostra with over 5 nu/ml were reactive in this assay, indicating the ELISA was less sensitive than the Vero cell assay. The results support the theory that SLTEC exposure of cows in Germany is more widespread than expected from epidemiological studies based on bacterial isolation. This possibly indicates a higher risk of human SLTEC infection via beef and milk products.


Subject(s)
Antibodies, Bacterial/analysis , Bacterial Toxins/immunology , Cattle Diseases/immunology , Colostrum/immunology , Escherichia coli Infections/veterinary , Animals , Antibodies, Bacterial/blood , Cattle , Cattle Diseases/blood , Cattle Diseases/microbiology , Chlorocebus aethiops , Escherichia coli Infections/blood , Escherichia coli Infections/immunology , Escherichia coli Infections/microbiology , Female , Neutralization Tests/veterinary , Shiga Toxin 1 , Shiga Toxin 2 , Vero Cells
16.
Vet Microbiol ; 43(1): 41-52, 1995 Jan.
Article in English | MEDLINE | ID: mdl-7716883

ABSTRACT

We constructed and purified recombinant B-subunits of the SLT-IIv as well as tested their usefulness in an immunoblot assay. The slt-IIvB gene amplified by PCR was ligated into the fusion vector pGEX-2T, and expressed in E. coli K 12 laboratory strains. Deletion of the signal sequence was necessary for optimal expression. High quantities of the fusion protein could be purified by affinity chromatography and subsequently used as antigen for immunoblot analysis with serum samples from diseased pigs and healthy controls. IgG antibodies against SLT-IIv were detected in the sera of 11 of 52 (21.15%) healthy pigs. By contrast, only in 1 of 28 (3.57%) serum samples of pigs with edema disease caused by SLT-IIv-producing E. coli we could demonstrate SLT-IIv-specific antibodies. During an outbreak of edema disease, sera from 10 pigs were taken at 4, 20, and 40 days after disease onset to investigate the immune response elicited by SLT-IIv. Immunoblot analysis with the recombinant SLT-IIv fusion protein revealed that the number of IgG-positive serum samples increased within this period of 40 days from one on day 4, to seven on day 20, to ten on day 40; the number of IgM-positive samples also increased from one after 4 days to eight after 20 days. Forty days after disease onset, IgM reactivity was no longer detectable. Since all animals seroconverted in the follow-up sera, the antigenicity of SLT-IIv during infection of pigs seems to differ from that of SLT-II in human hemolytic uremic syndrome where only a minority of patients are known to mount an immune response. The recombinant SLT-IIvB described here may be a possible candidate for vaccination trials.


Subject(s)
Antibodies, Bacterial/blood , Bacterial Toxins/biosynthesis , Bacterial Toxins/immunology , Edema/veterinary , Recombinant Proteins/biosynthesis , Swine Diseases/immunology , Animals , Base Sequence , Edema/immunology , Escherichia coli , Immunoglobulin G/blood , Immunoglobulin M/blood , Molecular Sequence Data , Polymerase Chain Reaction/veterinary , Shiga Toxin 2 , Swine
17.
Vet Microbiol ; 52(1-2): 153-64, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8914259

ABSTRACT

Naturally occurring enterohemolysin negative variants were observed during studies on bovine Shiga-like toxin-producing E. coli (SLTEC). Examination of three strains (413/89-1 and 332, 026:H-, and 570/89, O111:H-) and their isogenic variants (413/89-6, 332-I and 570/89-I, respectively) showed, that in each strain loss of the enterohemolytic phenotype correlated with the loss of a large plasmid ranging from 94 to 104 kb in size. The hemolysin determinant present on the 94 kb plasmid of strain 413/89-1 was cloned and discovered by DNA and N-terminal aminoacid sequence analysis to be highly homologous to the recently published EHEC-hemolysin (HlyEHEC; Schmidt et al., 1994; 1995). When a recombinant plasmid harboring this determinant was reintroduced into the enterohemolysin negative isogenic mutant 413/89-6, the enterohemolytic phenotype was restored. Southern blot hybridization analysis was used to demonstrate that the HlyEHEC is plasmid-borne in SLTEC-strains. Our cumulative data suggest that the enterohemolytic phenotype of SLTEC is encoded by the plasmid-borne HlyEHEC. These results further demonstrate the close similarity between SLTEC-isolates from bovine and human.


Subject(s)
Bacterial Toxins/biosynthesis , Cattle Diseases , Escherichia coli Infections/veterinary , Escherichia coli/genetics , Hemolysin Proteins/biosynthesis , Animals , Bacterial Toxins/genetics , Base Sequence , Cattle , DNA Primers , Enterotoxins/biosynthesis , Escherichia coli/growth & development , Escherichia coli/isolation & purification , Escherichia coli Infections/microbiology , Escherichia coli Proteins , Feces/microbiology , Genes, Bacterial , Genetic Variation , Hemolysin Proteins/genetics , Humans , Phenotype , Polymerase Chain Reaction , Shiga Toxin 1 , Shiga Toxin 2
18.
Vet Immunol Immunopathol ; 83(1-2): 19-36, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11604159

ABSTRACT

Neutral glycosphingolipids (GSLs) are considered activation markers on human lymphocytes, which are fundamental for studying the immune system. For cattle, only a limited number of activation markers has yet been identified. We recently showed that Shiga toxin 1, known to use globotriaosylceramide (Gb(3) syn. CD77) as a cellular receptor, depresses proliferation of activated bovine lymphocytes [Infect. Immunol. 67 (1999b) 2209]. In order to confirm the expression of Gb(3)/CD77 on bovine lymphocytes, we flowcytometrically examined a bovine B-lymphoma cell line (BL-3) and bovine peripheral blood mononuclear cells (PBMC) before and after mitogenic stimulation and biochemically characterized neutral GSLs extracted from PBMC. CD77 was detected on the surface of BL-3 cells and cultured PBMC essentially after mitogenic stimulation. Although expressed by all PBMC subpopulations identified, the portion of CD7+ cells was highest for BoCD8+ cells, followed by B-cells and BoCD4+ cells at day 4 of cultivation. Ceramide trihexoside of stimulated PBMC was structurally determined as Gal(alpha1-4)Gal(1-4)Glc(1-1)ceramide (Gb(3)). Biochemically, Gb(3) was also detected within unstimulated PBMC which contained ceramide monohexoside (CMH) and Gb(3) in a ratio of about 4:1. However, stimulation induced an increase of CMH and Gb(3) by a factor of 2.5 and 10, respectively, implicating that bovine lymphocytes regulate surface expression of Gb(3)/CD77 predominantly by quantitative changes in the Gb(3) metabolism. This report presents Gb(3)/CD77 as the first GSL identified on bovine immune cells and highly recommends this activation dependent antigen as a useful tool to investigate lymphocyte activation within the bovine immune system.


Subject(s)
B-Lymphocytes/metabolism , Trihexosylceramides/biosynthesis , Animals , B-Lymphocytes/immunology , Cattle , Enzyme-Linked Immunosorbent Assay/veterinary , Flow Cytometry/veterinary , Gas Chromatography-Mass Spectrometry/veterinary , Gene Expression Regulation , Immunohistochemistry/veterinary , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Lymphocyte Activation/physiology , Trihexosylceramides/chemistry , Tumor Cells, Cultured
19.
Vet Immunol Immunopathol ; 62(4): 309-21, 1998 Apr 30.
Article in English | MEDLINE | ID: mdl-9646436

ABSTRACT

To estimate the functional maturity of the phagocytic defence in neonatal calves, we analyzed the characteristics of blood phagocytes from calves (n = 10) 1 h post partum (p.p.) and 4 h p.p. At 1 h p.p., all calves were colostrum-deprived, while 5 calves had received colostrum before the 4 h p.p. sampling. The results were compared to those obtained from 3-9-week-old calves (n = 10). Phagocytic and oxidative burst activity of polymorphonuclear leukocytes (PMNL) and monocytes were determined in whole blood and separately analyzed by flow cytometry. In neonates prior to colostrum ingestion (1 h p.p.), phagocytic activity of PMNL against non-preopsonized E. coli was lower when compared to PMNL of 3-9-week-old calves. Opsonization of bacteria with pooled plasma from adult animals only partially restituted this lower PMNL phagocytic activity, indicating that humoral as well as cellular aspects of PMNL phagocytosis are altered in neonatal calves. In contrast to PMNL, monocytes of neonates exhibited an enhanced phagocytic activity. The oxidative burst activity of PMNL, as well as of monocytes was higher in newborn calves. During the first 4 h of life, the activities of blood phagocytes changed. Colostrum ingestion was accompanied by an increase in the percentage of phagocytizing PMNL and monocytes. This increase was absent in colostrum-deprived calves. In contrast, the oxidative burst activity of phagocytes decreased with age. In monocytes, the decrease of oxidative burst activity was only observed in colostrum-fed calves. In conclusion, some blood phagocyte functions in calves were found to be immature at birth, but these functions are presumably compensated by high absolute PMNL numbers and by other the more active mechanisms.


Subject(s)
Animals, Newborn/blood , Animals, Newborn/immunology , Cattle/blood , Cattle/immunology , Phagocytes/immunology , Aging/blood , Aging/immunology , Animals , Cell Differentiation , Colostrum/immunology , Escherichia coli/immunology , Female , In Vitro Techniques , Leukocyte Count , Male , Monocytes/cytology , Monocytes/immunology , Monocytes/physiology , Neutrophils/cytology , Neutrophils/immunology , Neutrophils/physiology , Opsonin Proteins/immunology , Phagocytes/cytology , Phagocytes/physiology , Phagocytosis , Respiratory Burst
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