Immunohistochemical Expression (IE) of Oestrogen Receptors in the Intestines of Prepubertal Gilts Exposed to Zearalenone.

This study was conducted to determine if a low monotonic dose of zearalenone (ZEN) affects the immunohistochemical expression (IE) of oestrogen receptor alpha (ERα) and oestrogen receptor beta (ERß) in the intestines of sexually immature gilts. Group C (control group; n = 18) gilts were given a placebo. Group E (experimental group; n = 18) gilts were dosed orally with 40 µg ZEN /kg body weight (BW), each day before morning feeding. Samples of intestinal tissue were collected post-mortem six times. The samples were stained to analyse the IE of ERα and Erß in the scanned slides. The strongest response was observed in ERα in the duodenum (90.387-average % of cells with ERα expression) and in ERß in the descending colon (84.329-average % of cells with ERß expression); the opposite response was recorded in the caecum (2.484-average % of cells with ERα expression) and the ascending colon (2.448-average % of cells with ERα expression); on the first two dates of exposure, the digestive tract had to adapt to ZEN in feed. The results of this study, supported by a mechanistic interpretation of previous research findings, suggest that ZEN performs numerous functions in the digestive tract.

Exposure to Low Zearalenone Doses and Changes in the Homeostasis and Concentrations of Endogenous Hormones in Selected Steroid-Sensitive Tissues in Pre-Pubertal Gilts.

This study was undertaken to analyze whether prolonged exposure to low-dose zearalenone (ZEN) mycotoxicosis affects the concentrations of ZEN, α-zearalenol (α-ZEL), and ß-zearalenol (ß-ZEL) in selected reproductive system tissues (ovaries, uterine horn-ovarian and uterine sections, and the middle part of the cervix), the hypothalamus, and pituitary gland, or the concentrations of selected steroid hormones in pre-pubertal gilts. For 42 days, gilts were administered per os different ZEN doses (MABEL dose [5 µg/kg BW], the highest NOAEL dose [10 µg/kg BW], and the lowest LOAEL dose [15 µg/kg BW]). Tissue samples were collected on days seven, twenty-one, and forty-two of exposure to ZEN (exposure days D1, D2, and D3, respectively). Blood for the analyses of estradiol and progesterone concentrations was collected in vivo on six dates at seven-day intervals (on analytical dates D1-D6). The analyses revealed that both ZEN and its metabolites were accumulated in the examined tissues. On successive analytical dates, the rate of mycotoxin accumulation in the studied tissues decreased gradually by 50% and proportionally to the administered ZEN dose. A hierarchical visualization revealed that values of the carry-over factor (CF) were highest on exposure day D2. In most groups and on most exposure days, the highest CF values were found in the middle part of the cervix, followed by the ovaries, both sections of the uterine horn, and the hypothalamus. These results suggest that ZEN, α-ZEL, and ß-ZEL were deposited in all analyzed tissues despite exposure to very low ZEN doses. The presence of these undesirable compounds in the examined tissues can inhibit the somatic development of the reproductive system and compromise neuroendocrine coordination of reproductive competence in pre-pubertal gilts.

The Effect of Low Doses of Zearalenone (ZEN) on the Bone Marrow Microenvironment and Haematological Parameters of Blood Plasma in Pre-Pubertal Gilts.

The aim of this study was to determine whether low doses of zearalenone (ZEN) influence the carry-over of ZEN and its metabolites to the bone marrow microenvironment and, consequently, haematological parameters. Pre-pubertal gilts (with a body weight of up to 14.5 kg) were exposed to daily ZEN doses of 5 µg/kg BW (group ZEN5, n = 15), 10 µg/kg BW (group ZEN10, n = 15), 15 µg/kg BW (group ZEN15, n = 15), or were administered a placebo (group C, n = 15) throughout the entire experiment. Bone marrow was sampled on three dates (exposure dates 7, 21, and 42-after slaughter) and blood for haematological analyses was sampled on 10 dates. Significant differences in the analysed haematological parameters (WBC White Blood Cells, MONO-Monocytes, NEUT-Neutrophils, LYMPH-Lymphocytes, LUC-Large Unstained Cells, RBC-Red Blood Cells, HGB-Haemoglobin, HCT-Haematocrit, MCH-Mean Corpuscular Volume, MCHC-Mean Corpuscular Haemoglobin Concentrations, PLT-Platelet Count and MPV-Mean Platelet Volume) were observed between groups. The results of the experiment suggest that exposure to low ZEN doses triggered compensatory and adaptive mechanisms, stimulated the local immune system, promoted eryptosis, intensified mycotoxin biotransformation processes in the liver, and produced negative correlations between mycotoxin concentrations and selected haematological parameters.

The Effect of Deoxynivalenol on Selected Populations of Immunocompetent Cells in Porcine Blood-A Preliminary Study.

Deoxynivalenol (DON) is one of the most prevalent mycotoxins in Europe. Pigs are an animal species that is most susceptible to this mycotoxin. Deoxynivalenol causes significant losses in pig production by lowering feed intake, decreasing daily weight gains, disrupting immune responses, and increasing susceptibility to diseases. The aim of this experiment was to determine the influence of feed contaminated with DON at concentrations insignificantly higher than recommended by the European Commission (900 µg/kg). The experimental feed contained 1008 µg DON/kg. The experiment was performed on eight weaners from the same litter. The animals were randomly divided into two groups: an experimental group (M, n = 4) fed contaminated feed and a control group (C, n = 4) administered feed free of mycotoxins. The experiment lasted for six weeks, and peripheral blood samples were collected from the animals for analyses of selected morphological parameters and changes in the percentages of CD4⁺8-, CD4-8⁺, and CD4⁺8⁺ lymphocytes and antigen-presenting cells (APC) with CD14⁺172⁺ (monocytes), CD172ahigh4-14- (conventional dendritic cells, cDC), and CD172adim4⁺14- (plasmacytoid dendritic cells, pDC) phenotypes. The morphological parameters of porcine blood samples were determined by flow cytometry with non-fluorescent particle-size calibration standards, and no differences were observed between groups M and C. An immunophenotyping analysis of lymphocytes and dendritic cells (DC) revealed an increase in the percentage of CD4⁺8-, CD172ahigh4-14-, and CD172adim4⁺14- cells, and a decrease in the number of CD4-8⁺ cells in group M. The results of this experiment suggest that prolonged exposure to low doses of DON can change the proportions of immunocompetent cells (a shift towards humoral immunity), without affecting their overall counts.

Changes in the metabolic profile and body weight of pre-pubertal gilts during prolonged monotonic exposure to low doses of zearalenone and deoxynivalenol.

The aim of this study was to determine whether exposure to low doses of ZEN + DON induces changes in serum biochemical and hematological parameters in pre-pubertal gilts. In the evaluated groups, minor but statistically significant changes were noted in selected serum biochemical parameters, including glucose, total cholesterol, ALT, AST, AP, total bilirubin, Pin, Fe, K and Cl, and in hematological parameters, including WBC, eosinophils, basophils, monocytes, Ht, Hb, MCHC, HDW and PLT. A statistical analysis of the results revealed significant differences between groups in the values of WBC, eosinophils, basophils, Hb, Ht, PLT, glucose, ALT, AP, total bilirubin, Fe and K. Change trends were noted mainly in weeks II and V-VI. An analysis of the metabolic profile of pre-pubertal gilts exposed to ZEN + DON indicates that homeostasis and biotransformation of ZEN + DON can be toned down at the expense of the animals' energy reserves. Body weight gains were lower in group E, and BW gains were not observed in weeks II and VI. The activity levels of gilts decreased in the first weeks of exposure (I and II), but the drop was minimized by a compensatory effect, or in the last two weeks of exposure due to nutrient deficiency or insufficient supply of protein and energy with feed and feed additives, which decreased BW gains. Low doses of mycotoxins induce completely different changes in the metabolic test than higher doses. The above can probably be attributed to: (i) a negative compensatory effect, (ii) initiation of adaptive mechanisms and stimulation of the immune system, probably due to the allergizing properties of mycotoxins, (iii) excessive loss of energy and protein due to more effective feed utilization, or (iv) involvement in detoxification processes which leads to fatigue. Depending on the body's energy stores, the above processes tend to tone down the biotransformation of low doses of the examined mycotoxins but in the present study, the BW of gilts did not increase under exposure to a combination of ZEN + DON.

Changes in the Subpopulations of Porcine Peripheral Blood Lymphocytes Induced by Exposure to Low Doses of Zearalenone (ZEN) and Deoxynivalenol (DON).

Zearalenone and deoxynivalenol are secondary metabolites of fungi of the genus Fusarium. The presence of mycotoxins in cereals and the resulting contamination of feeds and foods pose health risks for animals and humans. The dangers associated with high doses of mycotoxins have been extensively researched but very little is known about NOAEL (No Observed Adverse Effect Level) doses or exposure to a combination of mycotoxins (mixed mycotoxicoses). The aim of this study was to determine the effects of six-week exposure to NOAEL doses of individual and combined mycotoxins on the subpopulations of CD4⁺8(-), CD4(-)8⁺ and CD4⁺8⁺ lymphocytes in the peripheral blood of pigs. The experiment was performed on 72 gilts with average body weight of 25 kg, divided into three experimental groups (E1, E2 and E3, administered zearalenone (ZEN), deoxynivalenol (DON) and ZEN + DON, respectively, on a daily basis) and a control group (C) receiving placebo. Changes in lymphocyte subpopulations were evaluated by flow cytometry at weekly intervals (experimental days 7, 14, 21, 28, 35 and 42). A linear increase in the percentage of CD4⁺8⁺ lymphocytes was highly correlated with time (r = 0.682) in group C. The correlations and linear increase in the above subpopulation were disrupted in the remaining groups. In group E3, a statistically significant (p < 0.05) decrease in CD4⁺8⁺ counts was observed in week 5, which could point to a transient depletion of regulatory mechanisms of immune responses. The noted results also suggest that in mixed mycotoxicosis, ZEN and DON exerted stronger immunomodulatory effects.