The beneficial effects of rosmarinic acid on a non-tumorigenic epithelial cell line

The aim of this study was to investigate the effects of rosmarinic acid (RA) on a porcine, non-tumorigenic IPEC-J2 enterocyte cell line. The cell viability was tested with MTS assay and the transepithelial electrical resistance (TER) values were measured to determine the changes of the membrane integrity. The extracellular H2O2 production was analyzed with Amplex Red assay. The results show that the RA was not cytotoxic in low concentrations up to 100 uM. At 50 uM concentration RA promoted cell proliferation and did not cause significant increase in H2O2 production

Investigation of sphingosin-1-phosphate-triggered matriptase activation using a rat primary hepatocyte model

Sphingosine-1-phosphate (S1P) has been reported as a matriptase activator. The aim of this study was to reveal if S1P can influence hepcidin production. Furthermore, we investigated how S1P can affect the viability and the redox status of primary hepatocytes. Rat primary hepatocytes were cultivated for 72 h and were treated with 50, 200, 1000 ng/ml S1P. Cell-free supernatants were collected every 24 h. Cell viability was tested by a colorimetric method using tetrazolium compound (MTS). The hepcidin levels in the cell-free supernatants were examined with hepcidin sandwich ELISA to determine the effect of S1P on the hepcidin-modulating ability of matriptase. In order to estimate the extent of S1P-generated oxidative stress, extracellular H2O2 measurements were performed by the use of fluorescent dye. Based on the findings, S1P treatment did not cause cell death for 72 h at concentrations up to 1000 ng/ml. S1P did not influence the extracellular H2O2 production for 72 h. The hepcidin levels were significantly suppressed in hepatocytes exposed to S1P treatment. Further studies would be needed to explore the exact mechanism of action of S1P

Seroprevalence of bovine viral diarrhoea virus in Hungary — situation before launching an eradication campaign

Bovine viral diarrhoea (BVD) is a viral disease appearing in various forms and causing high economic losses in the cattle stocks of Hungary. The aim of the present study was to determine the prevalence of bovine viral diarrhoea virus (BVDV) in Hungary through a monitoring survey carried out on samples collected in cattle-keeping units throughout the country. Since no such survey had been carried out in Hungary during the last thirty years, our study may serve as a basis for later monitoring investigations aimed at following the progress of an expected eradication campaign of BVD. The tests were carried out using an ELISA method, on a total of 1200 blood samples submitted from 54 cattle herds. The herds had not been vaccinated against BVDV before the sampling. Out of the 1200 samples, 521 proved to be positive (43.4%), 40 gave doubtful result (3.3%) and 639 were negative (53.3%). In some stocks the samples were collected from cows having completed several lactation periods, and therefore the seronegativity indicates the BVDV-free status of the given stock. Moreover, among the positive herds we found a few where the seropositivity rate was rather low (< 5%). According to the results of the survey, a rather high portion (about one third) of the cattle-keeping units of Hungary can be regarded as BVDV free, which ratio is much higher than had been expected on the basis of surveys carried out on a lower number of samples and in smaller regions of the country. Hence, the chances of an eradication campaign launched in the near future, or carried out parallel to the IBR eradication programme, are better than previously expected

Evaluating synergy between marbofloxacin and gentamicin in Pseudomonas aeruginosa strains isolated from dogs with otitis externa

The aim of this study was to determine antimicrobial susceptibility of Pseudomonas aeruginosa strains to marbofloxacin and gentamicin, and investigate the possible synergistic, additive, indifferent or antagonistic effects between the two agents. P. aeruginosa strains can develop resistance quickly against certain antibiotics if used alone, thus the need emerges to find synergistic combinations. A total of 68 P. aeruginosa strains isolated from dogs were examined. In order to describe interactions between marbofloxacin and gentamicin the checkerboard microdilution method was utilized. The MICs (minimum inhibitory concentrations) for marbofloxacin and gentamicin were in the range 0.25–64 mg/L and 0.25–32 mg/L, respectively. The combination of marbofloxacin and gentamicin was more effective with a MIC range of 0.031–8 mg/L and a MIC90 of 1 mg/L, compared to 16 mg/L for marbofloxacin alone and 8 mg/L for gentamicin alone. The FIC (fractional inhibitory concentration) indices ranged from 0.0945 (pronounced synergy) to 1.0625 (indifference). Synergy between marbofloxacin and gentamicin was found in 33 isolates. The mean FIC index is 0.546, which represents a partial synergistic/additive effect close to the full synergy threshold. In vitro results indicate that marbofloxacin and gentamicin as partially synergistic agents may prove clinically useful in combination therapy against P. aeruginosa infections. Although marbofloxacin is not used in the human practice, the interactions between fluoroquinolones and aminoglycosides may have importance outside the veterinary field

Oxidatív stressz okozta sejtkárosodás megelőzésének lehetőségei in vitro sejtmodellen

The oxidative stress via causing cell and tissue damage can lead to formation of acute and chronic inflammation. Intestinal epithelium acts as a strong physical and chemical barrier against invading bacteria, toxins, oxidative stress and various chemical agents. Malfunction of the epithelial defense mechanisms as a result of damaged gut mucosa and altered intestinal microbial homeostasis can easily lead to leaky gut syndrome which can influence the general health condition of animals even on optimal nutritional regimens. The first aim of this experimental work was the development of an in vitro system mimicking intestinal epithelium, where oxidative stimuli can be introduced by peroxide treatment and the regulatory effect of acute oxidative stress can be monitored continuously. The prerequisite for finding the optimal dose and treatment time of peroxide administration was the maintenance of cell viability whereas the changes in relative gene expression level of proinflammatory cytokines could indicate the acute phase of inflammatory processes. IPECJ2 cells isolated from the jejunum of a neonatal piglet were selected for in vitro study as their glycosylation pattern, proliferation rate and colonisation ability can characterize better the in vivo conditions in the gut ecosystem. The high transepithelial electrical resistance value of IPEC-J2 monolayers grown on collagen-coated polyester membrane inserts demonstrated the functional integrity of the continuous cell association, acting as a single-layer tight physical barrier separating apical and basolateral compartments in the 3D model system

Treatment options and new perspectives in iron homeostasis disturbances Literature review

SUMMARY The authors summarize in their study the causes and the symptoms of iron toxicosis and iron deficiency anaemia and the diagnostic and treatment options of disturbances in iron homeostasis. Plasma iron level can be a reliable indica tor of iron deficiency anaemia, however, the low iron level can also be caused by chronic inflammation, infections and cancers. The iron supplementation of neonatal suckling piglets is of high veterinary importance, which is the most appropriate treatment for prevention of iron deficiency anaemia. The authors point out that iron toxicosis developed after administration of excessive iron can be compensated through parenteral deferoxamine, however other chelators applied mainly in human medicine such as deferiprone or deferasirox might be also effective in veterinary field. In this study the role of hepcidin in iron homeo stasis of the body is also discussed. The maintenance of physiological iron lev els in spite of rapid turnover of the iron suggests the regulation of hepcidin by plasma iron. In case of iron deficiency, the hepcidin amount is lowered, which then facilitates the rate of iron transport into the blood. In contrast, if the iron storage is saturated, the level of hepcidin produced by liver becomes elevated. Hepcidin oversecretion can also be induced by infection and inflammation with excessive interleukin-6 production. The authors also describe the compounds acting via hepcidin regulation which can be therapeutically beneficial in the treatment of iron homeostatic disturbances. The pharmacological intervention of the interplay between hepcidin and matriptase-2 has been one of the most recently discovered research fields which might involve the introduction of mat riptase modulators into the drug therapy of iron disorders

The impact of quercetin on a porcine intestinal epithelial cell line exposed to deoxynivalenol

Quercetin (Que) is present in many vegetables and fruits as a secondary antioxidant metabolite. Deoxynivalenol (DON) produced by various Fusarium mould species can induce cytotoxicity and oxidative stress in the gastrointestinal tracts of humans and farm animals. The aim of this study was to investigate the effects of Que on DON-induced oxidative stress in a non-tumourigenic porcine IPEC-J2 cell line. Two experimental designs were used in our experiments as follows: (a) pretreatment with 20 mu mol/L Que for 24 h followed by 1-h 1 mu mol/L DON treatment and (b) simultaneous application of 20 mu mol/L Que and 1 mu mol/L DON for 1 h. Cell cytotoxicity, transepithelial electrical resistance (TER) of cell monolayers and extracellular/intracellular redox status were studied. It was found that DON significantly decreased TER and triggered oxidative stress, while Que pretreatments were beneficial in maintaining the integrity of the monolayers and alleviated oxidative stress. However, co-treatment with Que was unable to preserve the integrity and redox balance of the cells exposed to DON. These results indicate that only the 24-h preincubation of cells with 20 mu mol/L Que was beneficial in compensating for the disruption caused by DON in extracellular oxidative status