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Summary Most of the porcine enterotoxigenic and drug resistant E. coli isolated in Hungary carry a conjugative R plasmid. In R and Ent plasmid transfer experiments 4 out of 11 donors (36%) transferred both R and Ent plasmid simultaneously when the J-53 strain was used as recipient. Treatment of E. coli diseases caused by these strains is quite difficult. The epidemiological importance of these observations is discussed. Zusammenfassung Vorkommen und Ubertragung von R- und Ent-Plasmiden bei enterotoxinbildenden E. coli-Stammen vom Schwein Die meisten der in Ungarn vom Schwein isolierten E. coli-Stamme weisen ein konjugiertes R-Plasmid auf. Durchgefuhrte R- und Ent-Plasmid Transfer-Experimente zeigten, das bei 4 von 11 Stammen (36%) gleichzeitig R- und Ent-Plasmid ubertragen wurden, wenn der Stamm J-53 als Empfanger diente. Die Behandlung von Erkrankungen, die durch solche Stamme hervorgerufen werden, ist ziemlich schwierig. Diskutiert wird die epidemiologische Bedeutung dieser Beobachtungen. Resume Presence et transmission de plasmides R et Ent chez des souches d'E. coli productrices d'enterotoxines d'origine porcine La plupart des souches d'E. coli isolees chez le porc en Hongrie presentent un plasmide R. conjugue. Des essais de transfert experimental de plasmides R et Ent ont montre que 4 des 11 souches (36%) etaient porteuses simultanement des plasmides R et Ent lorsque la souche J-53 servait de recepteur. Le traitement des affections dues a de telles souches est assez difficile. La signification epidemiologique de ces observations est discutee. Resumen Prevalencia y transmision de plasmides R y Ent en las estirpes E. coli enterotoxinogenas del cerdo La mayor parte de las estirpes E. coli aisladas en Hungria de los cerdos ostentan un plasmide R conjugado. Las experiencias de traspaso efectuadas con plasmides R y Ent muestran que en 4 de 11 estirpes (36%) se transmitieron al mismo tiempo plasmides R y Ent cuando la cepa J-53 servia como receptora. El tratamiento de las enfermedades ocasionadas por tales cepas es bastante dificil. Se discute la importancia epidemiologica de estas observaciones.

Balogh E., M. Toth, G. Bolcshazi, Zs. Abonyi-Toth, E. Kocsis, G. Semjen: Oral Hypoglycaemic Drugs in Alloxan-Induced Diabetes Mellitus in Dogs. Acta Vet. Brno 2008, 77: 363-371. The effect of the alpha-glycosidase inhibitor acarbose, the insulin sensitiser metformin and the insulin secretiser gliclazide on blood glucose level of dogs were examined in experimental diabetes. Dogs were randomly divided into three groups. During the first week of the experimental period control blood glucose data were determined. During the subsequent five weeks, the first group (n = 6) was administered acarbose (200 mg/dog/day), the second (n = 5) was treated with metformin (1700 mg/dog/day), and the third was given gliclazide (160 mg/dog/day). The drug was administered twice daily when feeding at 7:00 and 15:00 h. The average difference between postprandial and fasting blood glucose was determined for the treated and the control period, and the statistical significance of their difference (mean decrease) was evaluated by two-sampled t-test. The mean decrease in blood glucose was 0.49 mmol/l (P = 0.09) for acarbose, 1.15 mmol/l (P = 0.01) for metformin, and 0.08 mmol/l (P = 0.88) for gliclazide. According to the results of statistical evaluation, metformin was the only drug that caused a significant decrease in postprandial blood glucose. Acarbose, metformin, gliclazide, blood glucose level, diabetes mellitus, therapy

Norfloxacin was administered orally to chickens and turkeys at 15 mg/kg body weight by pulse dosing at 24 h intervals and by continuous dosing at 100 mg/L in drinking water for five days. Blood samples were taken serially. Plasma norfloxacin concentrations were determined by high-performance liquid chromatography. The plasma norfloxacin concentrations increased slowly during continuous dosing and reached the MIC 90 (250 ng/mL) for Gram-negative pathogens by 12 h in chickens and 18 h in turkeys. The steady-state plasma concentration was attained in 36 h and remained at approximately 776.67 ± 33.23 ng/mL in chickens and 682.50 ± 28.55 ng/mL in turkeys. After pulse dosing, the plasma norfloxacin concentrations increased rapidly and exceeded the MIC 90 at 2 h in both species and remained above MIC 90 for 8 h in chickens and 6 h in turkeys. Pulse dosing provided half the steady-state concentration that was achieved by continuous dosing, 365.32 ± 39.31 ng/mL in chickens and 306.03 ± 32.26 ng/mL in turkeys, during the dosing interval of 24 h. Data for daily pulse dosing suggested that every administration corresponded to a single, daily repeated bolus administration although pulse dosing produced higher plasma concentrations more readily. Continuous and pulse dosing are both rational for the administration of norfloxacin to flocks of chickens and turkeys. We recommend that treatment be commenced with a pulse oral dose administered over a 4 h period and maintained by continuous oral medication for three to five consecutive days.

Animal experiments were carried out with osteoconductive bone substitute beta-tricalcium phosphate (beta-TCP), with the aim of assessing the effects of the growth factors synthesised by thrombocytes on the speed of beta-TCP incorporation and on the quality of newly formed bone. The question to be answered was the extent to which platelet-rich plasma (PRP) accelerated the resorption of beta-TCP and the formation of new bone. Two teeth were removed symmetrically from each side of the mandible of 12 Beagle dogs; the resulting cavities were filled on one side with beta-TCP alone, and on the other side with a mixture of beta-TCP + PRP (obtained from autologous blood). The quality of the newly formed bone and the effects of this PRP were studied by histological and histomorphometric methods. In week 6, bone formation was already more effective when PRP was applied in comparison with beta-TCP alone, and in week 12 the growth was significantly greater. The results demonstrate that the use of PRP accelerates the remodelling of new bone created by beta-TCP.

Experimental colibacillosis was produced in 40 healthy, 7-day-old broiler chickens and turkeys by intratracheal injection of 1 × 108 CFU/chick and 1.23 × 109 CFU/poult bacteria of an O1:F11 strain of Escherichia coli, respectively. Two days before E. coli challenge all chicks were vaccinated with a live attenuated strain of infectious bronchitis virus (H-52). This model of infection - at least in chicken - proved to be useful for evaluating the efficacy of antimicrobial medication, by recording mortality, body weight gain, pathological alterations and frequency of reisolation of E. coli. Using this model, the efficacy of two different dosing methods of norfloxacin (continuous and pulse dosing) was evaluated. The once-per-day pulse dosing of norfloxacin administered via the drinking water at 15 mg/kg body weight proved to be more efficacious than the continuous dosing method of 100 mg/L for 5 days in chickens, while there were no convincing differences between the two treatment regimens in turkeys. The results confirmed earlier observations on the pharmacokinetic properties of norfloxacin in chicks and turkeys (Laczay et al., 1998).

Summary Light microscopy showed that bacteria attach in greater number to the normal rumen mucosa than to the parakeratotic ruminal epithelium. Various species of facultative bacteria were studied for adherence to isolated rumen epithelial cells and it was found that more bacteria attached to fully keratinized cells than to partially cornified ones. It is suggested that: (i) keratinization is a factor in bacterial adherence to the rumen mucosa, (ii) disturbances in the process of cornification (parakeratosis) result in decreased bacterial colonization on the rumen mucosa. Zusammenfassung Haftung von Bakterien am Pansenepithel bei Parakeratose Lichtmikroskopische Untersuchungen von Pansenepithel ergaben, das an gesunder Pansenmukosa mehr Bakterien anhaften als an einer durch Parakeratose geschadigten. Unterschiedliche in Betracht kommende Bakterienarten wurden bezuglich ihres Anhaftungsvermogens an isolierten Epithelzellen getestet. Dabei konnte festgestellt werden, das sich mehr Bakterien an vollstandig keratinisierten Zellen anlagern als an unvollstandig keratinisierten. Diese Beobachtungen lassen folgende Schlusse zu: 1. Die Keratinisierung von Zellen ist ein wichtiger Faktor fur die Anhaftung von Bakterien an der Pansenmukosa. 2. Storung im Keratinisierungsprozes (Parakeratose) fuhrt zu einer Abnahme von Bakterien an der Pansenmukosa.

The attachment to fully characterized primary rumen epithelial cell cultures of Escherichia coli strains isolated from different animal species and expressing F1–F4 or F17 fimbriae was examined. As the cell cultures contained stratified (keratinized) and non-stratified (non-keratinized) cells which grew either confluently or non-confluently, the strength of attachment of the different bacterial strains was assessed in relation to the differentiation state of the cells. Thus, strains having F1 fimbriae attached to all types of cultured cells, while strains with F2 and F3 fimbriae did not bind at all. E. coli strains having F4 or F17 fimbrae attached only to non-keratinized cells, particularly to confluent areas. As membrane glycosylation is known to change with differentiation (keratinization), our results suggest that the attachment of fimbriated E. coli strains which were capable of binding to rumen cells was more likely to be dependent on differentiation than the host specificity of the bacteria.

The therapeutic efficacy of sulphachlorpyrazine and toltrazuril against experimentally induced Eimeria tenella infection was compared in battery and floor pen raised broiler chickens. In the battery studies, both drugs prevented coccidiosis-related mortality and decrease of weight gain to a similar degree, but toltrazuril was more effective in reducing intestinal lesions and faecal scores, when treatments were initiated 24 h postinfection. When medication was delayed until 72 h after inoculation, the sulphonamide proved to be more effective in preventing reduction of weight gain and intestinal lesions caused by the parasites. Under simulated use conditions both drugs showed an appropriate anticoccidial efficacy without major differences between them.

Two strains of Streptococcus bovis (A1 and A5) and one strain of Escherichia coli (0141: H28) isolated from the surface of bovine ruminal mucous epithelium were examined for adherence to isolated and cultured ruminal epithelial cells. The E. coli adhered to the target cell by means of fimbriae, which had several common properties with type 1 common fimbriae and caused mannose-sensitive haemagglutination. The A1 strain of S. bovis was devoid of fimbriae and its adherence to the epithelial surface was not inhibited by treatment with sugars or phenol-treated bacterial membrane from the same organism. It was therefore postulated that the bacterial glycocalyx of the S. bovis organisms acted as ligand. The extent of bacterial adherence depended on the state of differentiation of the target cell in both the isolated and the cultured ruminal cell systems. The receptors for both adherent bacterial species were in all probability associated with the glycocalyx of the target cells.

Because of the rapid development and spread of antimicrobial resistance it is important that a system be established to monitor antimicrobial resistance in pathogenic zoonotic and commensal bacteria of animal origin. Susceptibility testing of bacteria from carcasses and different samples of animal origin has been carried out in veterinary institutes for a long time but by an inconsistent methodology. The disc diffusion method proposed by the National Committee for Clinical Laboratory Standards (NCCLS) was introduced in all institutes in 1997. In order to obtain a coherent view of the antimicrobial resistance of bacteria a computer system was consulted, consisting of a central computer to store all data and some local computers attached to it through the network. At these local measuring stations computers are connected to a video camera, which displays the picture of Petri dishes on the monitor, and inhibition zone diameters of bacteria can be drawn with the mouse by the inspector. The software measures the diameters, evaluates whether or not the bacteria are sensitive, and stores the data. The evaluation is based upon the data of the NCCLS. The central computer can be connected to as many local computers with measuring stations as we wish, so it is suitable for an integrated system for monitoring trends in antimicrobial resistance of bacteria from animals, food and humans, facilitating comparison of the occurrence of resistance for each circumstance in the chain. It depends on the examiners which antibiotics they want to examine. Thirty-two different antibiotic panels were compiled, taking into consideration the active ingredients of medicinal products permitted for veterinary use in Hungary, natural resistance and cross-resistance, the mechanism of resistance and the animal species, i.e. which drugs were recommended for treatment in the given animal species, and the recommendations of the OIE Expert Group on Antimicrobial Resistance. The members of the panels can be changed any time, even during the measuring process. In addition to the inhibition zone diameters of bacteria the database also includes information about bacterial and animal species, the age of animals and the sample or organ where the bacteria are from. Since January 2001 the antibiotic susceptibility of E. coli, Salmonella, Campylobacter and Enterococcus strains isolated from the colons of slaughter cows, pigs and broiler chickens has also been examined. Each of the 19 counties of Hungary submits to the laboratory three tied colon samples from a herd of the above-mentioned animals every month.

Experimental fowl cholera was induced in 60 healthy 10-week-old broiler chickens and 8-week-old turkeys by intramuscular inoculation with approximately 80 colony-forming units (cfu) of Pasteurella multocida X-73 strain and with approximately 70 cfu of P. multocida P-1059 strain, respectively. This method of infection proved to be useful for evaluating the efficacy of anti-microbial medication, by measuring mortality, weight gain, pathological responses and frequency of re-isolation of P. multocida. The efficacies of two different dosing methods, continuous and pulse dosing, were compared. Using the continuous-dosing method, norfloxacin was administered to drinking water at 100 mg/l for 5 days in chickens. Efficacies were slightly improved compared with pulse dosing at 15 mg/kg bodyweight for the same length of time. The opposite was observed in turkeys, to the degree of control of mortality and maintenance of weight gain.

The pharmacokinetics and the influence of food on the kinetic profile and bioavailability of doxycycline was studied after a single intravenous (i.v.) and oral dose of 10.0 mg/kg body weight in 7-week-old broiler chickens. Following i.v. administration the drug was rapidly distributed in the body with a distribution half-life of 0.21 ± 0.01 h. The elimination half-life of 6.78 ± 0.06 h was relatively long and resulted from both a low total body clearance of 0.139 ± 0.007 L/h·kg and a large volume of distribution of 1.36 ± 0.06 L/kg. After oral administration to fasted chickens, the absorption of doxycycline was quite fast and substantial as shown by the absorption half-life of 0.39 ± 0.03 h, the maximal plasma concentration of 4.47 ± 0.16 —g/mL and the time to reach the Cmax of 1.73 ± 0.06 h. The distribution and the final elimination of the drug were slower than after i.v. administration. The absolute bioavailability was 73.4 ± 2.5%. The presence of food in the intestinal tract reduced and extended the absorption (t1/2a = 1.23 ± 0.21 h; Cmax = 3.07 ± 0.23 µg/mL; tmax = 3.34 ± 0.21 h). The absolute bioavailability was reduced to 61.1% ± 4.4%.

The efficacy of doxycycline was investigated in two sets of experiments. In the first experiment 40, in the second experiment 60, hence altogether 100 five-week-old Ross broilers of both sexes were used. The birds were randomly allocated into groups (A and B in experiment 1; A, B and C in experiment 2) of 20 birds in each. All birds were infected intramuscularly with approx. 2 x 10(3) colony forming units of Pasteurella multocida strain X-73 (serotype A:1). Birds in groups A were non-medicated controls. Chickens in groups B were given doxycycline via the drinking water at a dose of 10 mg/kg body weight for 5 days, while group C was treated with chlortetracycline at a dose of 20 mg/kg body weight for 5 days. The trial lasted for 9 days, then the surviving chickens were sacrificed. Clinical symptoms, number of deaths, post mortem lesions and bacteriological findings were recorded using a special score system. Acute fowl cholera developed in broilers within a few hours after infection, as evidenced by the clinical symptoms, the high mortality rate (90% of the birds died within 4 days after infection), the pathological lesions and the recovery of P. multocida from the challenged birds. Doxycycline reduced the number of deaths (30% and 5% of birds died in experiments 1 and 2, respectively) and the severity of the clinical symptoms, and P. multocida could be re-isolated only from one of the survivors. In contrast, chlortetracycline slightly influenced the mortality; however, it delayed death and reduced the severity of clinical symptoms. These data indicate that doxycycline is highly effective for the treatment of experimental pasteurellosis in chickens.

The tolerance of chickens to monensin (12.5 mg/kg of feed) and maduramicin (3.0 mg/kg of feed) fed at a reduced dose in the presence of the antioxidant duokvin was studied in two experiments including 2 x 200 Tetra-82 broiler chickens. Tolerance was assessed by the appearance of clinical signs indicative of a toxic effect, the number of deaths, the groups' body weight gain, feed and drinking water intake, the aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities, calcium ion, inorganic phosphate and total protein content of the blood plasma, the haematocrit value, and haemoglobin concentration. When applied at a dose that had proved to be optimum in the efficacy studies, neither the monensin-duokvin combination (12.5 mg monensin per kg of feed + 120 mg duokvin per kg of feed) nor the maduramicin-duokvin combination (3.0 mg maduramicin per kg of feed + 120 mg duokvin per kg of feed) exerted a statistically significant influence on the parameters tested.

Two trials were carried out on a total of 2 x 360 Tetra-82 broiler chickens to study how the presence of the antioxidant duokvin as potentiating agent influenced the compatibility of reduced doses of monensin (12.5 mg/kg of feed) or maduramicin (3.0 mg/kg of feed) with other chemotherapeutic agents (tiamulin, erythromycin, sulfaquinoxaline, sulfachlorpyrazine, flumequine, tylosin, kitasamycin) widely used in broiler rearing. Compatibility was assessed on the basis of the appearance of clinical signs suggestive of toxic interaction, the mortality rate, body mass gain, feed consumption and drinking water intake, and changes in AST and LDH activities of the blood plasma. The monensin-duokvin combination (12.5 mg monensin/kg of feed + 120 mg duokvin/kg of feed) was found to be compatible with erythromycin, sulfaquinoxaline, sulfachlorpyrazine, flumequine, tylosin and kitasamycin. For tiamulin, a slight incompatibility was observed; however, this was much less severe than that found for monensin administered at a dose of 100 mg/kg of feed. The maduramicin-duokvin combination (3.0 mg maduramicin/kg of feed + 120 mg duokvin/kg of feed) was compatible with all the compounds tested; thus, it can be safely applied also in combination with tiamulin.

In two experiments, the compatibility of the anticoccidial combinations of monensin and duokvin, as well as that of maduramicin and duokvin, with some antimicrobials widely used in the broiler production was studied in cockerels. The monensin-duokvin combination was found to be fully compatible with erythromycin, sulphachlorpyrazine, and sulphaquinoxaline. With tiamulin, a slight interaction was observed, but it was far less severe than the toxic interaction between monensin and the diterpene antibiotic. The maduramicin-duokvin combination proved to be compatible with all of the chemotherapeutics tested, including tiamulin. The results of the studies indicate that the adverse interactions of monensin and maduramicin with certain antimicrobials can be considerably diminished or even abolished by using them in reduced doses in combination with the dihydroquinoline compound duokvin.

Summary Oxoid VET-RPLA, ST-EIA and Pharmacia Phadebact ETEC-LT enterotoxin tests were compared to find a simple but reliable method for detecting enterotoxigenic Escherichia coli (ETEC) in Hungary. In the Oxoid tests, all six reference LT-or ST-producing strains, except one ST-producer, gave positive results. Of 11 reference porcine enterotoxigenic strains, all four LT-producers gave positive reactions for LT but three of 10 ST-producers gave negative reactions for ST. Thirteen of 50 strains from culture collections of H. Steinruck (Germany) were LT+ and nine of 33 were ST+. When 31 isolates were tested simultaneously with the Oxoid and the Pharmacia LT tests, 12 strains were LT+ by the Oxoid LT test but by the Phadebact LT test only seven of these strains were LT+ and, of the remainder, three gave uncertain results and two gave negative results. Of 69 porcine strains, seven were LT+ and three ST+. Of 901 human strains isolated in Hungary, 10 were LT+ and one of 24 tested was ST+. In two cases, ETEC strains were isolated from contacts of travellers returning from Mongolia and Bangladesh. Results of comparative studies with reference strains corresponded well to those of the classical toxin detection tests. The Oxoid test was rapid, sensitive, specific and easy to perform and is recommended for use in screening ETEC isolates.

Protective activities of heat-inactivated (60°C for 30 min) merthiolate preserved Bordetella bronchiseptica and B. pertussis bacterins were compared in intraperitoneally immunized mice challenged intracerebrally (i.p./i.c.) or intraperitoneally (i.p./i.p.). In the i.p./i.c. assay (Kendrick test), a B. pertussis bacterin protected mice against challenge with B. pertussis 18-323, as well as against phase I cytotoxic and non-cytotoxic strains of B. bronchiseptica. A B. bronchiseptica bacterin, prepared from a phase I cytotoxic strain, gave protection against two phase I B. bronchiseptica strains, irrespective of their cytotoxin-production. A non-cytotoxic phase I strain of B. bronchiseptica elicited protection against the homologous strain only. Neither cytotoxic nor non-cytotoxic B. bronchiseptica strains protected mice challenged with B. pertussis 18-323. Vaccines prepared from phase III strains of B. bronchiseptica were not protective at all against any of the challenge strains. No such differences in the protective activities of the bacterins could be detected by the i.p./i.p. method. They seem to cross-protect equally well. The results indicate that the Kendrick test may be useful in testing potency of different B. bronchiseptica bacterins.

The effect of sub-minimal inhibitory concentrations of gentamicin on the haemagglutinating activity of Bordetella bronchiseptica was studied. Gentamicin exerted dissimilar effect on the production of haemagglutinin to calf and dog erythrocytes. The drug significantly reduced the haemagglutinating titres of calf-negative strains with dog red blood cells while it caused only a slight decrease in the haemagglutinating titres of calf-positive strains with bovine erythrocytes. The results support the view that bovine haemagglutinin is an adhesin of B. bronchiseptica.

Escherichia coli O78: K80 strains isolated from an outbreak among premature and newborn infants with meningitis, sepsis and enteritis, from sporadic cases of enteritis and from healthy carriers were compared with one another and with different E. coli serogroups. The O78: K80 cultures uniformly failed to give the rabbit intestinal loop test and the guinea pig eye reaction and none of them contained L1 antigen. After intraperitoneal injection into mice, the organisms multiplied in the peritoneal cavity and caused bacteriaemia lasting at least 2 weeks. E. coli strains originating from septicaemia (O78: K80, O18a,c: K?, O83: K?) showed significantly lower LD50 values for mice (9 x 10(3)--7 x 10(5)) than did E. coli serogroups associated with infantile enteritis only (3 x 10(8)--7 x 10(8)). It is assumed that the isolates differ in pathogenicity not only from E. coli strains associated with "cholera-like" disease and with "dysenteriform" infection, but also from L1 antigen-containing cultures described in neonatal meningitis, and constitute a separate group characterized by an ability to cause meningitis, sepsis and enteritis within the same outbreak.

The various sulphonamides show marked differences in disposition characteristics after administration to ruminants. For use in combination with a diaminopyrimidine derivative such as trimethoprim or baquiloprim, it is essential that a sulphonamide has similar pharmacokinetic properties in order to obtain optimal synergy. In the present study the pharmacokinetics of sulphamethoxazole, sulphatroxazole, and sulphamerazine were investigated in dwarf goats (n = 6) after IV and intraruminal administration at a dose of 30 mg/kg bodyweight. In addition, the in vitro binding of sulphamerazine to ruminal contents was studied as a possible explanation for a reduced absorption rate. Sulphamethoxazole showed the most rapid absorption after intraruminal administration (mean tmax +/- SD : 0.8 +/- 0.2h). However, the drug was rapidly eliminated from the plasma (t1/2 beta : 2.4 +/- 1.5 h) and the bioavailability was only 12.4 +/- 4.7%, most likely due to an extensive 'first-pass' effect. The bioavailability of orally administered sulphamerazine and sulphatroxazole was much higher (67.6 +/- 13.5% and 70.2 +/- 32.3%, respectively). After intraruminal administration, sulphatroxazole showed the highest plasma peak concentration (26.1 +/- 6.3 mg/l) and the longest plasma half-life (4.7 +/- 1.8h) and mean residence time (13.9 +/- 4.5 h). Sulphamerazine showed considerable binding to rumen contents in vitro. Based on its pharmacokinetic properties sulphatroxazole appears to be a suitable candidate to be used in combination with the more recently developed diaminopyrimidines such as baquiloprim.