Escherichia coli Nissle 1917 for probiotic use in piglets: evidence for intestinal colonization.

AIMS: This study was prompted to investigate the intestinal localization and colonization of orally administered Escherichia coli Nissle 1917 (EcN) in piglets. METHODS AND RESULTS: EcN was fed to ten EcN-negative piglets (3 months) over seven consecutive days. Faecal samples were collected repeatedly and tested for EcN-DNA by a combined culture/PCR assay and for viable EcN by culture methods, respectively. EcN-DNA was detectable in faeces of all piglets within the first 24 h after it was added to the feed. After the administration of EcN had been stopped, the presence of EcN-DNA in faecal samples indicated that all piglets shedded EcN with their faeces intermittently through up to 33 days. In addition, E. coli strains indistinguishable from EcN by all markers tested (rdar colony morphotype, multiplex PCR and GEI II-PCR analyses, XbaI-pattern, K5 phage susceptibility) were isolated from faecal samples and from mucosal swabs taken at euthanasia at the end of the experiment. CONCLUSIONS: EcN colonizes the intestine and persists in conventionally reared piglets for at least 4 weeks upon oral administration. SIGNIFICANCE AND IMPACT OF THE STUDY: Results of this study have implications for efficacy and safety assessments of EcN as a probiotic strain for use in pigs.

Incidence of Brucella species in marine mammals of the German North Sea.

In this study, organ samples from 426 common seals Phoca vitulina, 298 harbour porpoises Phocoena phocoena, 34 grey seals Halichoerus grypus and 10 other marine mammals were assessed for the presence of Brucella species. Forty-seven common seals, 2 harbour porpoises and 1 grey seal were found to be positive for these bacteria. A total of 91 Brucella strains were successfully isolated, due to the fact that Brucella spp. were found in more than one organ sample in 15 animals. The primary organ in which the bacteria were present was the lung. In addition, 2 strains were isolated from lungworms (Parafilaroides spp.). Forty-nine of the isolated strains were selected for further analysis using conventional phenotyping methods. Molecular characterisation was carried out by analysing the IS711 and omp2 loci. With respect to the distribution of the IS711 loci in the genome, the 49 field isolates differed strongly from the terrestrial Brucella species and marginally from the marine Brucella reference strain NCTC12890. Based on the results of the PCR restriction fragment length polymorphism (PCR-RFLP) investigation of the omp2 locus, the majority of the Brucella field isolates were classified as B. pinnipediae, recently proposed B. pinnipedialis, possessing 1 omp2a gene and 1 omp2b gene. Two field isolates revealed the presence of 2 omp2a genes, as has been described for Brucella ovis. To our knowledge, these results confirm for the first time the presence of Brucella species in the marine mammal population of the German North Sea. These findings highlight the need for additional research on the relevance of these Brucella species for marine hosts and their environment.

Neutralizing antibodies against Shiga-like toxins from Escherichia coli in colostra and sera of cattle.

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.

Construction of recombinant Shiga-like toxin-IIv (SLT-IIv) and its use in monitoring the SLT-IIv antibody status of pigs.

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.

Characteristics of alpha-hemolytic strains of Escherichia coli isolated from dogs with gastroenteritis.

Twenty-four hemolysin producing (Hly+) strains of Escherichia coli isolated from dogs with gastroenteritis were investigated for their virulence markers and their phenotypic properties. The strains were distributed over eleven known E. coli O-serogroups and most of them were heterogeneous for their phenotypes. All strains were found to produce alpha-hemolysin which was detected by Southern hybridization and colony immunoblotting using a specific gene probe and a monoclonal antibody. Eight strains were carrying plasmids encoding alpha-hemolysin sequences (hly-plasmids) and 16 strains carried chromosomal hly-determinants. Twelve of the strains showed enterotoxic activities which were tested for in different assays. Among these, three O42:H37 and two O70:H-strains carrying hly-plasmids were found to harbour other plasmids encoding the heat-stable enterotoxin STA1. The other seven strains showing enterotoxicity in the ileal loop or the suckling mouse assay were negative for STA1, STA2, or LT. None of the 24 strains were positive for invasiveness or for production of Vero (Shiga-like) toxins. The production of alpha-hemolysin was closely associated with the production of cytotoxic necrotizing factor (CNF), which was detected in 17 of 24 strains. Of these, 16 elaborated CNF1 and one strain produced an unknown CNF type. Surprisingly, all strains carrying ST-plasmids and six of eight strains carrying hly-plasmids were negative for CNF. Thus, in canine E. coli strains CNF production seems to be closely associated with production of chromosomally encoded alpha-hemolysin whereas hly-plasmids are more often associated with ST-producing, CNF negative isolates.

The enterohemolysin phenotype of bovine Shiga-like toxin-producing Escherichia coli (SLTEC) is encoded by the EHEC-hemolysin gene.

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.

Globotriaosylceramide (Gb(3)/CD77) is synthesized and surface expressed by bovine lymphocytes upon activation in vitro.

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.

Compensation of preliminary blood phagocyte immaturity in the newborn calf.

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.

[Animals as a source of infections for humans--diseases caused by EHEC]. / Tiere als Infektionsquelle für den Menschen--durch EHEC hervorgerufene Erkrankungen.

EHEC (enterohaemorrhagic E. coli) bacteria are new, only since 1982 recognized zoonotic pathogens. EHEC differ from E. coli intestinal commensales by the fact that they are lysogenic infected with bacteriophages, which carry the genetic information for the production of shigatoxins (Stx type 1 and/or 2). Due to the obligatory released Stx EHEC are classified also among the Shigatoxin producing E. coli (STEC). EHEC are capable of causing a Hemorrhagic Colitis and some sequelae of diseases such as the haemolytic uraemic syndrome. Due to their virulence factors they can be divided into typical and non-typical EHEC. Typical EHEC possess a pathogenicity island (Locus of Enterocyte Effacement) harboring genes, which apart from the characteristic necrotic activity of Stx enable the pathogens to closely attach to the epithelial cells of the intestinal mucosa and to destruct the microvilli. Additionally a so-called virulence plasmid codes for the production of a haemolysin, a peroxidase-katalase, an enterotoxin as well as a serine protease. EHEC are one of the world-wide most important causes of foodborne infections. Depending upon the country, most of the incidences in 1998 varied between 1 to 3 cases per 100,000 inhabitants. Since EHEC are only notifiable in a few countries, one must count however on substantially higher numbers. In Germany the estimated incidence is about 13 cases per 100,000 inhabitants. Since the first EHEC outbreaks were recognized in humans, studies investigating the prevalence of EHEC within animals were repeatedly performed. From the outset one assumed that cattle are a possible reservoir. Actually EHEC were isolated from fecal samples world-wide (typical and non-typical EHEC) from a large percentage of cattle (> 50%). Besides EHEC were isolated sporadically from fecal samples of other animals and healthy humans. The EHEC bacteria are shed by infected humans and animals, in particular by infected ruminants. They are spread over manure, slurry, sewage etc. Humans can get infected directly by contact with infected persons or animals or indirectly by contaminated food, water etc. The clinical outcome within humans appears as aqueous to bloody diarrhea. Beyond that approximately 5 to 10% of the patients develop the haemolytic uraemic syndrome. In contrast to humans, animals are mostly infected clinically inapparent. The therapy is based upon a symptomatic treatment. At present in man the control of EHEC infections concentrates on a particularly strict hand hygiene after the contact with infected humans and animals (above all ruminants). Since EHEC are heat sensitive, the prophylaxis by sufficient heating of risk food (raw milk, ground beef) is of special importance. In veterinary medicine above all EHEC infections must be controlled in ruminants, which are the primary reservoir. Due to the wide spread of EHEC in the ruminant population it is not realistic to demand an EHEC free cattle stock. Since EHEC are spread only via fecal excretion, at present it is most important to reduce the fecal shedding and to avoid fecal contamination of food of animal origin. In detail prophylactic hygienic measures concerning the farm management, the feeding hygiene, the food hygiene, the meat hygiene as well as the food hygiene are available.

[Prevalence of the Salmonella plasmid virulence gene "spvD" in Salmonella strains from animals]. / Häufigkeit des Salmonella-Plasmid-Virulenz-Gens "spvD" bei Salmonella-Stämmen von Tieren.

Strains of Salmonella isolated from animals in Germany (n = 878) were analysed for the presence of the spvD gene ("Salmonella plasmid virulence gene D") by DNA-DNA hybridization. The spvD gene was only detected in strains of serovars Typhimurium (93.3%), Enteritidis (97.1%), and Dublin (100%) as well as in two rough strains of Salmonella enterica. Salmonella isolates from mammals carried the gene more frequently (cattle 94.0%, horses 92.6%, pigs 73.7%) than those from birds (33.3%) or reptiles (4.5%). Due to its high prevalence in epidemiologically relevant salmonellae, the virulence factor spvD may represent a sensitive and specific target in various serovars for diagnostic or immunization strategies.

[Diagnosis of Lawsonia intracellularis using the polymerase chain reaction (PCR) in pigs with and without diarrhea and other animal species]. / Diagnostik von Lawsonia intracellularis mittels Polymerasekettenreaktion (PCR) bei Schweinen mit und ohne Diarrhoe und anderen Tierarten.

Lawsonia (L.) intracellularis, an obligately intracellular bacterium, causes proliferative enteropathy (PE) in swine and, occasionally, in other animals. To determine the spread of the agent among German pig herds pooled fecal samples of five animals each of clinically normal Hessian pig herds collected between november 1998 and february 1999 as well as feces (n = 1684) from individual animals representing 648 herds, sent to our laboratory by veterinarians from all parts of Germany, were tested for L. intracellularis using the polymerase chain reaction (PCR). In addition, fecal samples from diarrhoic foals (n = 46), dogs (n = 57), cats (n = 50), calves (n = 37), hedge hogs (n = 9), seals (n = 8) and one giraffe were also studied. DNA was extracted from feces using high concentrations of chaotropic salt and diatomaceous earth. For PCR, primers flanking a 279 bp fragment of L. intracellularis DNA were used (JONES, G. F., WARD, G. E., MURTAUGH, M. P., LINN, G. (1993), J. Clin. Microbiol. 31, 2611-2615). Amplificates were separated by agarose gel electrophoresis and visualized under UV-light. L. intracellularis was found in 26 (12.8%) samples from 21 (30.0%) of the Hessian pig herds without symptoms of diarrhoea. In feces of pigs with diarrhoea (n = 1684) the agent was present in 431 (25.6%) samples originating from 224 (34.6%) herds. Of the other animal species studied, L. intracellularis was detected in feces of 4 (7.0%) dogs, 2 (5.4%) calves, 3 (33.3%) hedge hogs and in the sample of the giraffe. The remaining species were all tested negative.

[The immune response in edema disease of weaned piglets measured with a recombinant B subunit of shiga-like toxin II]. / Untersuchungen zur Immunantwort bei der Odemkrankheit von Absetzferkeln mit einer rekombinanten B-Untereinheit des Shiga-like-Toxins-II.

An outbreak of edema disease (ED) was monitored in 80 piglets after weaning over a period of 4 weeks. The shedding of Shiga-like toxin-IIe) producing Escherichia coli strains, the serum bactericidal activity (SBA) against SLTEC-IIe, and the antibody response against SLT-IIe were investigated. The antibody response was monitored by utilizing a glutathione-S-transferase (GST) + SLT-IIe B/SUB fusion protein (FRANKE et al., in press) for immunoblot assays. E. coli-strain GO15III (0141:K85ac) was diagnosed as SLT-IIe-producing E. coli by polymerase chain reaction, DNA hybridization and cytotoxicity assays. Maximum excretion of GO15III appeared between days 8 and 15 after weaning. On day 1 after weaning no piglet shed GO15III, while the number increased on day 8 to 53 (66.2%) and on day 15 to 59 (73.8%) of the piglets. 4 week after weaning, GO15III was only isolated from 23 (28.8%) of the piglets. In parallel, serum bactericidal activity against GO15III increased significantly in the sera of 73 (91.2%) piglets, reaching a stable maximum from day 15 on. During the first two weeks after weaning, no piglet yielded detectable SLT-IIe-IgG. However, the number of SLT-IIe-IgG positive piglets increased steadily from day 15. On day 15, 5 (6.2%) piglets were positive in SLT-IIe immunoblot analysis and 29 days after weaning the number increased to 31 (38.8%). These data represent the first serological monitoring of a natural outbreak of edema disease in piglets after weaning by using a recombinant fusion protein (GST+SLT-IIe B/SUB). The recombinant protein proved to be a useful diagnostical tool for monitoring the specific antibody status of piglets.

[Pathogenesis, clinical aspects and diagnosis of the most important enteric bacterial diseases in animals]. / Pathogenese, Klinik und Diagnose der wichtigsten bakteriell bedingten Enteritiden beim Tier.

There are adhesive and invasive species among enteropathogenic bacteria for animals. Most frequently isolated adhesive bacteria are enterotoxigenic E. coli (ETEC). Salmonella, T. hyodysenteriae, M. paratuberculosis, Y. enterocolitica, Cl. perfringens and C. jejuni bacteria represent the most important invasive germs. The main clinical finding connected with bacterial enteritis is diarrhoea resp. dysentery. Evaluation of feces consistency, time of appearance and accompanying symptoms (e.g. fever, colic) allow a clinical suspective diagnosis, however a definitive diagnosis must be based on the demonstration of the causative agent. In addition to known cultural, microscopic, biochemical and serological diagnostic methods, the demonstration of virulence factors (enterotoxin formation, adhesive pili) plays an important role in diagnosis of enteropathogenic bacteria.

Oral immunization against tetanus.

Model experiments in mice were used for the investigation of oral immunization against tetanus. The efficacy of oral tetanus toxoid application was tested by subcutaneous challenge (10 LD50 tetanus toxin) and antitoxin determination by L+-method. Reasonable protection could only be achieved when toxoid was in intensive contact with the mucous membranes of the oral cavity, whereas immunization by way of the gastrointestinal tract failed completely. After a single oral vaccination with 200 Lf fluid tetanus toxoid onto mucous membranes of the oral cavity the efficiency index was 75. A 100% protection was obtained in challenge after 3 applications of 200 Lf toxoid. In comparison, a single subcutaneous dose of 2 Lf tetanus toxoid resulted in an efficiency index of 100 and a single intranasal immunization with 100 Lf in an efficiency index of 93. Development of immunity after oral immunization was much accelerated if compared with parenteral immunization. 20% of the orally treated animals were immune 5 days after vaccination. After the same period subcutaneously treated animals had failed to develop any protection at all. Antitoxin titers after one oral vaccination (200 Lf) were similar to the ones after one subcutaneous dose (2 Lf). A distinct increase in antitoxin titers could only be observed when revaccination was done subcutaneously. The results obtained with mice apply analogically to guinea pigs.

[Rabbit hemorrhagic disease (RHD)--comparative diagnostic studies using the hemagglutination test and electron microscopy]. / Rabbit Haemorrhagic Disease (RHD)--Vergleichende diagnostische Untersuchungen mittels Hämagglutinationstest und Elektronenmikroskopie.

A total of 56 liver specimens from rabbits with symptoms of rabbit haemorrhagic disease were tested for virus by electron microscopy (EM) and haemagglutination (HA). Both methods simultaneously gave positive or negative results in 28 or 22 cases, respectively. Divergent results were obtained in only 6 samples. Five of them were positive by EM but negative by HA and in one specimen with a HA-titer of 1:32 virus could not be detected.

Physical and genetic map of the obligate intracellular bacterium Coxiella burnetii.

Pulsed-field gel electrophoresis and PCR techniques have been used to construct a NotI macrorestriction map of the obligate intracellular bacterium Coxiella burnetii Nine Mile. The size of the chromosome has been determined to be 2,103 kb comprising 29 NotI restriction fragments. The average resolution is 72.5 kb, or about 3. 5% of the genome. Experimental data support the presence of a linear chromosome. Published genes were localized on the physical map by Southern hybridization. One gene, recognized as transposable element, was found to be present in at least nine sites evenly distributed over the whole chromosome. There is only one copy of a 16S rRNA gene. The putative oriC has been located on a 27.5-kb NotI fragment. Gene organization upstream the oriC is almost identical to that of Pseudomonas putida and Bacillus subtilis, whereas gene organization downstream the oriC seems to be unique among bacteria. The physical map will be helpful in investigations of the great heterogeneity in restriction fragment length polymorphism patterns of different isolates and the great variation in genome size. The genetic map will help to determine whether gene order in different isolates is conserved.

[Indirect hemagglutination for Eperythrozoon suis detection in experimentally and spontaneously infected swine]. / Untersuchungen mit der indirekten Hämagglutination zum Eperythrozoon suis-Nachweis in experimentell und spontan infizierten Schweinen.

Ten splenectomized and non-splenectomized pigs were experimentally infected with E. suis bearing red blood cells in order to determine the antibody response. All animals were monitored for antibody titer by indirect hemagglutination over a period of 80-290 days postinfection. Latent E. suis infection only yielded a detectable antibody titer in one pig. Acutely infected pigs had a titer ranging up to 1:640. Maximum antibody response lasted only 2 months and dropped below the level of detection of our assay within 2 to 3 months. At this time, the clinical symptoms could reappear and antibodies were again detectable. However, no booster effect was observed with this second outbreak. We also determined the antibody frequency in 138 pigs from 16 herds in Southern Germany. Pigs from only 4 out of 6 clinically positive herds had antibody titer against E. suis. 20 out of 78 pigs of the clinical positive herds demonstrated a detectable E. suis antibody titer. In 10 herds that were asymptomatic and presumed uninfected all 80 pigs were serologically negative for E. suis.

[Clinical experience with kennel-specific E. coli oral vaccines in dogs]. / Klinische Erfahrungen mit der zwingerspezifischen E.-coli Schluckimpfunbei Hunden.

Kennel-specific oral E. coli vaccines were tested for efficacy and side effects at breeding and boarding kennels with severe diarrhoea problems. Oral vaccines contained heat inactivated E. coli bacteria from specific kennels and were given once daily for 14 days. Oral vaccines were administered directly orally to puppies while for adult dogs vaccines were mixed with daily food rations. In breeding kennels with 4 to 10 week old puppies suffering from non-fatal diarrhoea, the oral immunization led to a decrease in morbidity from 86.5% to 0%. In kennels with some cases of fatal diarrhoea the rate of morbidity decreased from 45% to 21% and the mortality rate from 25% to 10.3% after using the vaccination. By vaccinating adult dogs in boarding kennels the morbidity rate dropped from 83.5% to 6.5% and the mortality rate from 4.1% to 0.5%. The kennel specific oral E. coli vaccine was found to be free of side effects. No adverse effects were observed in either puppies or adult dogs.