Preliminary Characterization of the Transcriptional Response of the Porcine Intestinal Cell Line IPEC-J2 to Enterotoxigenic Escherichia coli, Escherichia coli, and E. coli Lipopolysaccharide

IPEC-J2, a promising in vitro model system, is not well characterized especially on the transcriptional level, in contrast to human counterparts. The aim of this study was to characterize the gene expression in IPEC-J2 cells when coincubated with enterotoxigenic Escherichia coli (ETEC), nonpathogenic E. coli, and E. coli endotoxin. Apical infection of polarized IPEC-J2 monolayers caused a time-dependent decrease in transepithelial electrical resistance (TEER). Microarray analysis showed up-regulation of interleukins when IPEC-J2 were cocultured with E. coli strains this has so far never been measured in this cell line. Highest IL8 expression was found with the ETEC strain possessing the F4 fimbrium, suggesting IPEC-J2 cells to be F4 receptor positive, confirmed in a brush border membrane adhesion assay. It is concluded that the innate immune responses to pathogens and LPS makes the IPEC-J2 cell line a suitable model for research on intestinal host pathogen interaction

Non-invasive intestinal biomarkers: a new ELISA test for Pancreatitis Associated Protein detection in pig

Feed additives are commonly used to improve pig performance and health, but they need to be tested so new biomarkers for intestinal health, non- or minimally invasive, are under investigations.The quantification of Myeloperoxidase (MPO) and Pancreatitis Associated Protein (PAP) in feces could prove useful to non-invasively monitor intestinal health (Niewold, 2015). MPO is an enzyme that permits to quantify the number of inflammatory cells present in tissues and feces (Prokopowicz et al., 2012) , while PAP is a protein mainly produced in the small intestine with anti-inflammatory and bactericidal activity (Cash et al., 2006; Mukherjee et al., 2014). Because of the lack of a commercial ELISA kit for porcine PAP detection, the main aim of this study was to develop and validate a new sandwich ELISA test for the quantification of PAP in pig fecal samples. Our study consisted of two phases: test development and test validation. During the development phase we used polyclonal antibodies previously immunized from rabbit serum with a pure peptide containing the N-terminus of pig PAP (Soler et al., 2015). The validation of the test was then performed with fecal extraction samples derived from animals with known high or low growth performance.Moreover, the temperature stability of PAP in feces and the optimal extraction method was tested. Even if only preliminary, our results seem to show a fair relationship between fecal consistency, used as health indicator, and PAP fecal concentrations. Furthermore, no relevant differences in PAP concentration after 24h of incubation at 37 °C, 4°C or room temperature were detected.To date, the present results suggest that PAP seems to be exceptionally stable in feces and is a very promising candidate as a non-invasive (fecal) biomarker for intestinal health and growth

The acute phase protein, haptoglobin : a potential parameter in welfare assessment?

Physiological parameters are important measures in animal welfare assessment. To assess the amount of stress an animal experiences, stress hormones like cortisol are frequently used. However, measuring cortisol has major disadvantages due to its rapid reactivity and decline and many influencing factors. Other potential alternative markers are acute phase proteins, since stress is known to affect the immune system. A pilot study was conducted to investigate the response of the acute phase protein, plasma haptoglobine (HP), in pigs subjected to a stressor (food deprivation) and to examine the correlation between HP levels and average daily growth (ADG). Forty grower pigs (25.1 ± 4.4 kg, mean ± SD) (sex and former pen mates balanced), were allocated to 4 conventional pens, 2 treatment (T) and 2 control (C) groups (10 pigs per pen). After 10 days of adaptation the experiment started and ran for 3 weeks. In the 2nd week, T groups were repeatedly subjected to an 8-hour food deprivation (day 1, 3, 5 and 7 of week 2), C groups had normal, unrestricted, access to food. Pigs were weighed twice a week and blood was collected once a week (every 5th day). Mean levels of plasma HP of C and T groups showed large variation between individuals (C groups, week 2: 1.84 ± 3.11 mg/ml; T groups, week 2: 1.40 ± 1.16 mg/ml). No significant differences (Kruskal-Wallis test) in HP levels or growth were found between the C and T groups or between the different weeks within the T groups. Significant negative weak to moderate correlations were found between ADG and HP levels (HP week 1 and ADG week 1: rs = -0.47, p=0.005; HP week 2 and ADG total; rs= -0.60, p=0.015; HP week 3 and ADG total: rs = -0.43, p=0.025; average HP total and ADG total: rs= -0.41, p=0.017). Large variations in HP levels between individuals were shown and no effect of treatment on HP levels or growth was found. Possibly, food deprivation had no apparent stress eliciting effect. Despite these results, interesting correlations between the level of HP and ADG were found, corroborating the inverse relationship between the acute phase response and growth. To further investigate the relation of the acute phase response and stress a successive experiment will be conducted in which we apply a stronger stressor (mixing pigs) and combine the physiological data with behavior

Занятие-общение - коммуникативно-ориентированная форма обучения научному стилю речи

The aim of the study was to determine the intraarticular set-Urn amyloid A (SAA) response pattern in horses with inflammatory arthritis. Inflammatory arthritis was induced by injection of lipopolysaccharide (LPS) into the radiocarpal joint of four horses. Serum and synovial fluid (SF) samples were collected before and at 4, 8, 12, 24, 48, 72, 96, and 144 h after injection. Concentrations of SAA were measured by immunoturbidometry, and expression of SAA isoforms was visualized by denaturing isoelectric focusing and Western blotting. The LPS injection caused systemic and local clinical signs of inflammation. Serum amyloid A appeared in serum and SF within 8 h after LPS injection. Isoelectric focusing showed three major SAA hands with apparent isoelectric points (pI) of 7.9, 8.6, and > 9.3 in serum and SF. Synovial fluid contained two additional isoforms with highly alkaline apparent pI values (apparent pl value extrapolated from standard curve = 10.0 and 10.2), which were not present in any of the serum samples. In conclusion, intraarticular injection of LPS induced systemic and local inflammatory responses in the horses. By demonstrating SF-specific SAA isoforms the results of the present Study suggest that SAA is synthesized locally in the equine inflamed joint, similar to what has been demonstrated in humans previously. The marked local SAA synthesis suggests an important pathophysiological role in inflammatory arthritis

Growth promotion in broilers by both oxytetracycline and Macleaya cordata extract is based on their anti-inflammatory properties

The non-antibiotic anti-inflammatory theory of antimicrobial growth promoters ( AGP) predicts that alternatives can be selected by simple in vitro tests. In vitro, the known AGP oxytetracycline (OTC) and a Macleaya cordata extract (MCE) had an anti- inflammatory effect with a half-maximal inhibitory concentration of 88 and 132 mg/l, respectively. In vivo, chickens received three different concentrations of MCE in drinking-water, OTC in feed and a control. Body weight (BW), feed intake (FI) and gain: feed (G:F) ratio were determined on days 14, 21 and 35. On day 35, body composition was determined. Plasma alpha(1)-acid glycoprotein (alpha(1)-AG) concentration was measured on days 21 and 35, and the expression of several jejunal inflammatory genes was determined on day 35. OTC-fed chickens showed a significantly higher BW, FI and G:F ratio compared with the control group at all time points. MCE had a significant linear effect on BW on days 21 and 35, and the G:F ratio was improved only over the whole period, whereas FI was not different. Only MCE but not OTC decreased the percentage of abdominal fat. Plasma alpha(1)-AG concentration increased from day 21 to 35, with the values being lower in the treatment groups. Both OTC and MCE significantly reduced the jejunal mucosal expression of inducible NO synthase. For most parameters measured, there was a clear linear dose-response to treatment with MCE. In conclusion, the results are consistent with the anti- inflammatory theory of growth promotion in production animals

Proteomic approaches to study the pig intestinal system

One of the major challenges in pig production is managing digestive health to maximize feed conversion and growth rates, but also to minimize treatment costs and to warrant public health. There is a great interest in the development of useful tools for intestinal health monitoring and the investigation of possible prophylactic/ therapeutic intervention pathways. A great variety of in vivo and in vitro intestinal models of study have been developed in the recent years. The understanding of such a complex system as the intestinal system (IS), and the study of its physiology and pathology is not an easy task. Analysis of such a complex system requires the use of systems biology techniques, like proteomics. However, for a correct interpretation of results and to maximize analysis performance, a careful selection of the IS model of study and proteomic platform is required. The study of the IS system is especially important in the pig, a species whose farming requires a very careful management of husbandry procedures regarding feeding and nutrition. The incorrect management of the pig digestive system leads directly to economic losses related suboptimal growth and feed utilization and/or the appearance of intestinal infections, in particular diarrhea. Furthermore, this species is the most suitable experimental model for human IS studies. Proteomics has risen as one of the most promising approaches to study the pig IS. In this review, we describe the most useful models of IS research in porcine and the different proteomic platforms available. An overview of the recent findings in pig IS proteomics is also provided. © 2014 Bentham Science Publishers.Peer Reviewe

Synergistic toxicity of dietary aflatoxin B1 (AFB1) and zearalenone (ZEN) in rainbow trout (Oncorhynchus mykiss) is attenuated by anabolic effects

Although several studies have reported co-occurrence of mycotoxins in fish feed limited information is available on the physiological effects. In the current study, the single and combined effect of added dietary AFB1 and ZEN in various concentrations for 60 days was tested in rainbow trout on growth parameters, mortality, intestinal digestive enzymes activity and on serum and intestinal oxidative stress and immunological parameters and on intestinal histopathology, versus untreated controls. Both mycotoxins groups irrespective of concentration, showed a significant deterioration of SGR and FCR as compared to control, whereas no consistent differences in mortality were present. The small intestinal enzyme activity was significantly reduced by ZEN in particular at the higher mycotoxin concentrations and when combined with AFB1. Serum and intestinal immunological parameters were significantly affected largely consistent with additive and synergistic negative effects on the (innate) immune system, in particular enhanced inflammation. The latter was also reflected in the serum and intestinal oxidative stress parameters. Histopathological examination showed decreasing villus length and Goblet cell density with increasing mycotoxin load. It is concluded that the combination of AFB1 and ZEN clearly increased the toxic effect on the fish as far as the serum and intestinal parameters are concerned. This is in some cases merely additive, but clear synergistic toxicity was seen too, in particular at the higher mycotoxin concentrations. Surprisingly, the increasing intestinal impairment did not translate into increased growth retardation and mortality since those were similar in all non-control groups. This may be consistent with anabolic effects of mycotoxin (metabolites) as has been described before