[Levobupivacaine vs. ropivacaine for continuous femoral analgesia after anterior cruciate ligament reconstruction]. / Levobupivacain und Ropivacain zur kontinuierlichen Nervenblockade des N. femoralis nach Rekonstruktion des vorderen Kreuzbandes.

BACKGROUND: Levobupivacaine and ropivacaine are both used for continuous femoral analgesia after anterior cruciate ligament reconstruction; however it is unknown whether both drugs are equally effective regarding pain control, preservation of mobility and patient satisfaction. METHODS AND MATERIALS: In this randomized, placebo-controlled trial 84 patients undergoing anterior cruciate ligament (ACL) reconstruction with quadruple hamstring tendons were studied. For postoperative pain therapy levobupivacaine 0.125%, ropivacaine 0.2% or placebo control with NaCl 0.9% at a rate of 6 ml/h were used for 48 h using a femoral nerve catheter. All patients also received an i.v. patient-controlled analgesia (IVPCA) pump with piritramide. RESULTS: Patient satisfaction was significantly higher and night rest was better in both treatment groups compared to the placebo group but there appeared to be no major differences between the two local anesthetics. Opioid consumption was significantly higher in the placebo group compared to the levobupivacaine group but not the ropivacaine group. The pain scores showed a trend towards higher scores in the placebo group throughout but the difference only reached statistical significance on postoperative day 1. No statistical significant differences in motor block were found between the three groups. CONCLUSION: Postoperative analgesia for ACL reconstruction during the first 48 h using femoral block with a continuous infusion of levobupivacaine 0.125% or ropivacaine 0.2% in combination with an IVPCA is similarly effective and better than a placebo. Both studied drugs seem to be equally appropriate for this purpose.

Pericarditis as primary manifestation of Mycobacterium bovis SSP. caprae infection.

A 76-year-old white male presented with progressive malaise, weight loss and dyspnea at rest. Echocardiography revealed a circular pericardial effusion and global hypokinesia. Pericardiocentesis showed a purulent exudate and microbiologic examination revealed Mycobacterium bovis fully sensitive to isoniazid, streptomycin, ethambutol, rifampin, and pyrazinamide. By spoligotyping the isolate could be further differentiated to M. bovis ssp. caprae. Antimycobacterial therapy was initiated but 3 weeks later the patient's circulation and renal function deteriorated and he died with clinical signs of sepsis despite intensive care treatment. Pericarditis is a rare manifestation of tuberculosis and can be fatal even when diagnosed and treated appropriately. In low incidence countries diagnosis is often delayed and even overlooked.

[The epizootiology of tuberculosis of cattle in the Federal Republic of Germany]. / Zur Epizootiologie der Rindertuberkulose in der Bundesrepublik Deutschland.

Mycobacterial strains from different outbreaks of tuberculosis of cattle in Germany from 1996 to 2001 were differentiated by two molecular biological methods (Spoligotyping, RFLP IS6110). The causative agent was in one case Mycobacterium (M.) africanum, in 10 cases M. bovis and in 17 cases M. bovis ssp. caprae, respectively. The results of the molecular biological methods are discussed from the perspective of epizootiology and the particular importance of infections by M. bovis ssp. caprae emphasized. Direct contact of the animals, purchase from infected stocks, infected zoo animals and wildlife, as well as livestock handlers are discussed as possible sources of infection.

Effects of a long-term exposure with OTA or OTC on functions of a human monocytic cell line.

An in-vitro model was developed to study the effects of a long-term exposure with a low concentration of ochratoxin A (OTA) or ochratoxin C (OTC) on a human monocytic cell line (THP-1).Cells were propagated in 24-well cell culture plates for 15 days. OTA and OTC preparations, respectively, at a concentration of 1 ng/ml were included in the cell culture medium during the whole cultivation period. At the end of the exposure time, parameters of cell viability and cell function were examined.After 15 days of exposure to ochratoxins, viability and function of the THP-1 cell line were modulated. Mitochondrial activity and the production of IL-6 were increased by all mycotoxin preparations. Cell membrane integrity was disturbed, proliferation and the production of TNF-α and IL-8 were inhibited. These parameters were most severely affected by mycotoxin preparations containing OTC.Our results show that long term exposure to OTA and especially OTC in low concentrations can cause subtle alterations of cell viability and function which may have remarkable consequences for human and animal health. In this context it seems to be necessary to study the contamination of food and feed stuffs with OTC more intensively.

[Colonisation studies using Campylobacter jejuni in chicks]. / Kolonisationsversuche mit Campylobacter jejuni bei Küken.

White Leghorn chicks used in this study were hatched from specific pathogen-free eggs. The colonizing capability of Campylobacter (C.) jejuni strains was investigated in 6 experiments. The formation of specific antibodies associated to colonization was also detected. In each experiment, day of hatch chicks were randomly separated into three groups of 24 birds each: two groups colonized experimentally and one control group. Chicks were reared on the floor in three separated, adjacent rooms with sterilized wood shavings as litter. At 2 or 8 days of age, respectively, the chicks in the experimentally colonized groups received between 3.3 x 10(7) and 2.0 x 10(8) colony-forming units (CFU) of C. jejuni via oesophageal gavage. Furthermore, 7, 14, 21, 28, 42 and 56 days after inoculation, 4 chicks of each group were sacrificed by cervical dislocation, at which time blood, liver and faeces were collected for processing. Serum was centrifuged and Campylobacter-specific IgG, IgA and IgM antibodies were measured by an indirect enzyme-linked immunosorbent assay (ELISA). Altogether, the colonizing capability of 11 C. jejuni strains was examined. Surprisingly, there were large differences between the C. jejuni isolates. After these experiments, we could divide the isolates into three groups. 4 out of 11 isolates could not be reisolated, 2 isolates caused weak or delayed colonization and 5 C. jejuni produced strong, long-lasting colonization. In the first days of life (9 days), the C. jejuni-free SPF chicks (control animals) had high IgG titres in sera, which decreased markedly up to the age of 15 days. During the experiments the IgM and IgA titres remained nearly at the same level, i.e., the amounts of maternal antibodies were low and there was no evidence for antibody formation in the chicks themselves. Two- and 8-day-old chicks were inoculated with C. jejuni strain Penner 1. Two-day-old chicks were colonized 3 weeks after inoculation. In comparison with these animals, 8-day-old chicks were colonized already 2 weeks after inoculation. There is the assumption, that the higher maternal antibodies in 2-day-old chicks could be responsible for this delay. In chicks the C. jejuni colonization resulted in a marked IgG (but not IgM and IgA) increase. Apparently, there is a positive relationship between the counts of this pathogen in caeca and the IgG increase.

[The use of the toxin binding inhibition test for potency testing of Clostridium novyi type B alpha toxoid vaccines]. / Anwendung des Toxin-Bindungs-Inhibitionstests zur Wirksamkeitsprüfung von Clostridium-novyi-Typ-B-Alphatoxoid-Impfstoffen.

The toxin binding inhibition test (ToBI) were developed for potency testing of C. novyi type B alpha toxoid containing veterinary vaccines to replace the currently used toxin neutralisation test in mice (TNT). The antitoxin titres of rabbit sera (AN-, HV- and SP sera) were determined with ToBI using the international reference serum with known antitoxin titre. In order to show the validity of the methods, the results were compared with those of the manufacturers/regulatory authorities and correlation coefficients were calculated. The correlation coefficients were r = 0.93 (AN sera), r = 0.73 (HV sera) and r = 0.85 (SP sera). All correlations were statistically significant. The specificity of the methods could be proved using heterologous antisera. The results of the ToBI were reproducible. Thus, the ToBI offers a suitable in vitro method for the determination of the antitoxin titre of rabbit antisera as an alternative to the toxin neutralisation in mice for potency testing of vaccines containing C. novyi type B alpha toxoid.

Effect of ochratoxin A and ochratoxin C on the monocyte and lymphocyte function.

The effect of practically relevant mycotoxin concentrations on functions of immune cells was studied in in vitro experiments. Porcine mononuclear cells were exposed to a crudeAspergillus-ochraceus toxin containing OTA, a HPLC fraction identical with OTC derived from the crude toxin (RE2), as well as pure OTA and OTC in a concentration range from 0.46 to 3000 ng/ml. The influence of mycotoxin exposure on metabolic activity, mitogen induced proliferation, expression of the activation marker CD25 and the cell cycle of lymphocytes and on the formation of free oxygen radicals as well as the production of the cytokines IL-6 and TNF-α by monocytes was determined. Exposure to high concentrations of all mycotoxin preparations lead to non-specific suppression of the immune cell functions, which was related to cytotoxic effects. Low concentrations caused ambivalent reactions, especially on monocyte function. In general, the HPLC fraction RE2 had an up to 100-fold stronger effect than pure OTA. Ochratoxin-induced suppression of lymphocyte proliferation was not abrogated by phenylalanine or aspartame. The results indicate that immunomodulation can be caused by very low mycotoxin concentrations which are not related to clinical symptoms or loss of performance.

Antibody reactions after aerogenous or subcutaneous immunization of pigs with Pasteurella multocida antigens.

Depending upon the antigens used and the initial titres, subcutaneous immunization of weaner pigs by means of a temperature-sensitive mutant of live Pasteurella multocida (serovar A), resulted in a significant rise of the level of specific IgG antibody already present in blood serum, but not in lung lavage fluid, and a specific stimulation of lung clearance as compared to non-immunized controls. Aerogenous immunization of a total of 108 animals in 18 experiments did not influence serum antibody titres but produced a significant rise in lung antibodies and P. multocida clearance as compared to an identical number of controls. In all 12 immunization experiments using the live mutant, increased specific IgA antibodies and in seven out of 10 experiments, elevated IgG antibodies were measured. Only aerogenous immunization was found to be capable of reducing the severity of pneumonia induced by intrabronchial infection.

A taxonomic study of bacteria isolated from grasses: a proposed new species Pseudomonas graminis sp. nov.

The taxonomic position of a yellow-pigmented group of bacteria, isolated from the phyllosphere of grasses was investigated. Results obtained from restriction analysis of amplified 16S rDNA with seven endonucleases (CfoI, HaeIII, AluI, HinfI, MspI, Sau3A and ScrFI) showed identical restriction patterns for each enzyme of all isolates studied, which suggests that all strains belong to the same species. The grass isolates displayed the characteristics of the genus Pseudomonas. They were Gram-negative, aerobic and rod-shaped with polar flagella. Isolates were catalase-positive and oxidase-negative, and unable to oxidize or ferment glucose with the production of acid. The isolates did not reduce nitrate to nitrite but were able to utilize a wide range of compounds individually as a sole carbon source, with preference being given to the utilization of monosaccharides. The disaccharides tested were not utilized as substrates. The DNA base compositions of the tested strains ranged from 60 to 61 mol% G+C. The major isoprenoid quinone of each was ubiquinone Q-9 and hydroxy fatty acids were represented by 3-hydroxydodecanoic acid and 2-hydroxydodecanoic acid. Comparison of 16S rDNA sequences showed that the bacteria were members of the genus Pseudomonas, with similarity values between 91.5 and 97.7%. DNA-DNA hybridization studies with closely related neighbours revealed a low level of homology (< 27%), indicating that the isolates represent an individual species. On the basis of phenotypic and phylogenetic analyses a new species, Pseudomonas graminis sp. nov. (type strain DSM 11363T), is proposed.

[Protein patterns of Pasteurella multocida strains in relation to the animal host and the geographic site of isolation]. / Proteinmustertypen von Pasteurella multocida in Abhängigkeit von der Herkunft der Stämme bezüglich Tierart und geographischer Lage der Bestände.

Protein patterns from 681 Pasteurella multocida strains of different animal species and distinct geographical regions were analyzed by PAGE. We found 9 protein types taking the most intense band as reference. This band represents a protein of the outer membrane (OMP). In assigning the strains to protein types a relation to animal hosts of the strains was established. The majority of isolates from rabbits (88%) belongs to type P6, which is not found in strains isolated from cattle or pigs. Strains from pigs did not include the protein types P2, P4 and P8, in cattle strains type P9 was absent. The distribution of protein types in bovine isolates was shown to be related to the geographical location. The results are discussed with particular emphasis on immuno-prophylaxis. It appears promising to develop P. m. vaccines specific for certain animal species.

[Detection of dermonecrotic toxin genes in Pasteurella multocida strains using the polymerase chain reaction (PCR)]. / Nachweis des Dermonekrotoxin-Gens in Pasteurella-multocida-Stämmen mittels Polymerase-Kettenreaktion (PCR).

A PCR method was developed which allows to distinguish between Pasteurella multocida strains carrying or lacking the dermonecrotic toxin gene. Specific primers were used to amplify a 1501-bp DNA fragment from the genomic dermonecrotic toxin gene region. Isolated DNA, broth cultures and swabs were used as samples. Detection of the toxin gene directly from swab samples accelerates considerably the diagnosis since cultivation steps can be omitted. The results of PCR corresponded to findings obtained by ELISA.

[Long-chain fatty acids of Pasteurella multocida nad Pasteurella haemolytica]. / Die langkettigen Fettsäuren von Pasteurella multocida und Pasteurella haemolytica.

The relative contents of long-chain fatty acids in P. multocida and P. haemolytica were investigated. A dependence on the composition of the broth was established. Accordingly, comparative quantitative studies on fatty acid contents have to be conducted using bacteria grown with the same lot of broth medium. As for P. multocida, there were significant differences between the serovars (C14 in TDHM and C16, delta 2C18 in BPL). These differences are, however, not significant to replace serotyping. Highly significant differences were also detected between P. multocida isolates from nasal swabs and pneumonic lungs (interims of C14, delta C16 on BPL and BRU). The largest differences were measured for strains grown on BRU, which is interpreted as an expression of virulence. Significant differences were found between biotypes A and T of P. haemolytica, namely for C14, C16 in TDHM, and C14, delta C16, C16, C18 in BPL medium.

[Investigation of outer membrane proteins of Pasteurella. 2: Iron-regulated outer membrane proteins of Pasteurella multocida and Pasteurella haemolytica]. / Untersuchungen zu den Proteinen der äusseren Membran von Pasteurellen 2. Mitteilung: Eisenregulierte Proteine der äusseren Membran bei Pasteurella multocida und Pasteurella haemolytica.

Pasteurella multocida and Pasteurella haemolytica produce specific proteins in the outer membrane under iron-depleted conditions. Pasteurella multocida serovar A expresses these proteins of molecular masses of 76 and 96 kDa as determined by electrophoresis. The analogous serovar D produces a further iron-regulated protein of 85 kDa. The Pasteurella haemolytica strains of serovar A1, A6 and T contain iron-regulated outer membrane proteins of molecular masses of 71, 77 and 100 kDa. These proteins possess binding positions for iron ions. Both Pasteurella multocida and Pasteurella haemolytica strains utilize iron from porcine and bovine transferrin, but not from haemin and haemoglobin.

[Immunization with potential Salmonella enteritidis mutants--1. Production and in vitro characterization]. / Untersuchungen zur Immunisierung mit geeigneten Salmonella Enteritidis-Mutanten--1. Herstellung und in-vitro-Charakterisierung.

Production and in vitro characterization of potential vaccine strains are the first steps leading to an efficient Salmonella Enteritidis oral live vaccine for homologous immunization of poultry. The paper presents the results of the production of adenine-amino acid auxotrophic mutants using N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis. The mutant strains were characterized using the following properties: auxotrophy, stability of mutation, reversion rate, generation time, metabolic properties, serotype, motility, plasmid content, phage type, SDS-PAGE patterns, as well as cell culture adhesion and invasion. Ten S. Enteritidis double auxotrophic mutants were obtained which are stable auxotrophically and where the risk of reversion was minimal. All strains were found to be plasmid-free. 5 mutants were selected for further investigations concerning their attenuation and immunological value.

[Proteins from the outer membrane of Pasteurella--1. Polyacrylamide gel electrophoresis of outer membrane proteins--a possibility for differentiation of Pasteurella haemolytica strains]. / Untersuchungen zu den Proteinen der äusseren Membran von Pasteurellen--1. Mitteilung: Polyacrylamidgelelektrophorese von Proteinen der äusseren Membran--eine Möglichkeit der Differenzierung von Pasteurella-haemolytica-Stämmen.

Using the sodium laurylsarcosinate method, outer membrane proteins (OMPs) of 63 Pasteurella haemolytica strains are isolated and their protein patterns obtained by SDS polyacrylamide gel electrophoresis (SDS-PAGE) are compared. A high degree of similarity became evident both within A and T biotypes in the molecular weight range of 25 to 50 KDa. Strain-to-strain variations are mainly limited to quantitative differences in individual protein bands. It is concluded that within both the A and T biotype groups equal sets of major proteins are present with differing amounts of individual constituents. Biotypes A and T can be clearly distinguished on the basis of marked variations in OMP profiles. OMPs of those AT strains included in comparative studies are exhibiting no general homogeneity, but a relative heterogeneity due to different protein compositions, which are, however, clearly distinct from the patterns of biotypes A and T. It is established unambiguously that isolates belonging to biotypes A or T, as well as AT strains, can be clearly distinguished from each other on the basis of SDS-PAGE analysis of OMP extracts. Thus, polyacrylamide gel electrophoresis of outer membrane proteins can be used for differentiation.

[The influence of cations on the letality and on the formation of the toxin of Pasteurella multocida]. / The influence of cations on the lethality and on the formation of the toxin of Pasteurella multocida.

The stability of the Pasteurella multocida toxin has proven as very pH dependent. Therefore, a pH lower than 6 should be avoided. A concentration of 1 mM Cu2+ decreased the lethal effect of the toxin but the equimolecular addition of chelators compensated this effect. Possible structure-effect mechanisms were discussed. Iron supplement (100 microM) to two culture media examined did not influence the production of the toxin but during the cultivation in the low-iron medium TDHM the toxin level increased. This increased toxin production was not due to a more intensive multiplication of bacteria.

Iron supply of Pasteurella multocida and Pasteurella haemolytica.

Pasteurella multocida and Pasteurella haemolytica do not produce hydroxamate- or phenolate type siderophores. However, transport- and utilization systems could be detected for the well known siderophores ferrioxamine B, E, G, rhizoferrin and the intermediate 2,3-dihydroxybenzoic acid by means of cross-feeding tests in both Pasteurella species. Enterobactin and ferrichrome did not feed any of the Pasteurella strains tested. Additionally, alpha-ketoacids and alpha-hydroxyacids such as pyruvic acid, alpha-hydroxyisovaleric acid and others acting as primary metabolites enabled growth of P. multocida and P. haemolytica under iron limitation.

[Detection of toxin-producing types of Pasteurella multocida--a comparison of methods]. / Nachweis toxinbildender Stämme von Pasteurella multocida--ein Vergleich von Methoden.

28 Pasteurella multocida strains were examined for production of toxin by 6 different methods. Identical results were obtained using a mice lethality test, a tissue cell culture assay and an ELISA. Different results were received with a dot-blot-immunoassay (1 strain), using a gene probe (6 strains) and a guinea pig skin test (8 strains). Corresponding differences were detected with 2 strains only. Tissue culture, ELISA and dot-blot-immunoassay are effective methods for the diagnosis of toxin-producing Pasteurella multocida strains. Animal experiments should be an exception.

[The effect of toxins from Pasteurella multocida type D in calves in vivo]. / Zur Wirkung des Toxins aus Pasteurella multocida Typ D beim Kalb in vivo.

By intratracheal injection of purified dermonecrotic toxin from a Pasteurella multocida type D strain, pneumonitis in calves could be produced. This demonstrates the important role of this toxin in the pathogenesis of pneumonitis in calves.

[The localization of toxin in the cells of Pasteurella multocida]. / Die Lokalisation des Toxins in Zellen von Pasteurella multocida.

The application of the degradation procedure for Gram-negative bacteria according to Bewick and Lo to Pasteurella multocida indicates that the obvious localization of the toxin is in the periplasm. The stability of the outer membrane and of the substances adhering to it is essential for the release of the toxin. The production of the toxin clearly depend on the media used.