Anna Breitholtz-Emanuelsson

Effects of ochratoxin A on the mouse immune system after subchronic exposure.

The effects on the immune system of oral, subchronic exposure to ochratoxin A (OA) at 6, 250 or 2600 micrograms/kg diet were studied in female Balb/c mice. After 28 days of exposure, antibody production plague-forming cells/spleen, was suppressed in a dose-dependent manner which was significant in the two highest exposure groups. In addition, a decrease in thymocyte cell counts was seen in the 250-micrograms/kg group. After 90 days exposure, flow cytometry analysis of thymic lymphocyte subpopulations revealed a decreased proportion of mature (CD4+ or CD8+) cells. Furthermore, the mitogenic responsiveness of thymocytes and splenocytes to concanavalin A (Con A) was significantly decreased. This effect was observed in all three treatment groups. Interleukin-2 production of Con A-stimulated lymphocytes, natural killer cell activity, and humoral antibody titres to a viral antigen were not affected by OA treatment. The present results indicate that subchronic, oral exposure to OA affects certain immune functions in mice at exposure levels that may be found in contaminated food products.

Placental and lactational transfer of ochratoxin A in rats

The placental and lactational transfer of ochratoxin A (OA) was investigated in a cross-fostering study in rats. Dams were given 50 microg OA(-1) kg body weight by gastric intubations 5 times a week for 2 weeks before mating, during gestation and then 7 days a week during lactation. Neonates from OA-treated dams were cross-fostered at birth to control dams treated with only vehicle. In the same way, neonates from control dams were cross-fostered to OA-treated dams. Treatment with OA did not result in any effects on birth weight or growth development of the pups during the first 21 days of life. There were no effects on milk quality as measured by milk lipids, protein or lactose concentrations, or on milk production, assessed by the mammary gland content of RNA and DNA. A mean milk:blood ratio of approximately 0.6 was found. The dose of OA from milk to the suckling pup at 14 days of age can be calculated to about 50 microg kg(-1) body weight(-1) day, which is similar to the dose given to the dams. Pups exposed to OA only via milk had blood and kidney levels of OA approximately 3 times higher than their dams, indicating a high absorption and/or a low excretion of OA in the sucklings. At 14 days of age the highest blood and kidney levels of OA were found in offspring exposed both via placenta and milk, with the highest contribution from milk. Offspring exposed only via milk had about 4-5 times higher levels of OA in blood and kidney compared to offspring exposed only via placenta. As milk could be a significant source of OA exposure in newborns, adverse health effects resulting from postnatal exposure should be studied and evaluated in the risk assessment of OA.

Effects of Ochratoxin A on the rat immune system after perinatal exposure

Effects on the immune system after perinatal exposure to ochratoxin A (OA) were studied in Sprague-Dawley rats after single or repeated exposure of the dams. In a short-term study, dams with litters were given a single dose of OA (0, 10, 50 or 250 micrograms/kg body weight) on day 11 of lactation. The effects on cell numbers in spleen and thymus añd on the mitogen responses of lymphocytes were evaluated in the suckling pups on day 14 of lactation. The proliferative response of splenocytes to the T-cell mitogen Concanavalin A (Con A) was significantly stimulated in pups from dams given 10 or 50 micrograms OA/kg body weight as compared to control pups. In addition, proliferation of thymocytes in response to Con A was stimulated in pups from dams exposed to 50 micrograms OA/kg body weight. In a long-term, cross-fostering study comparing pre- and postnatal exposure, half of the dams received 50 micrograms OA/kg body weight 5 days a week by gastric intubation 2 weeks before mating, during gestation and then 7 days a week until weaning. Effects on immune parameters were studied in the pups on day 14 of lactation and at 13 weeks of age. Suppressed mitogenic responses were seen to both lipopolysaccharide (LPS) and Con A in prenatally exposed pups sampled on day 14 of lactation. At 13 weeks the response of splenocytes to LPS was still impaired. The primary antibody response to a viral antigen was also lower in the prenatally exposed pups than in the control pups. These effects on the immune response were not seen in the groups of pups exposed postnatally or both pre- and postnatally, although blood concentrations of OA were higher in these groups at the time of the first sampling. This indicates that the decrease in proliferation and antibody production resulted from prenatal modulation of the immune system. The results show that prenatal exposure to relatively low doses of OA may induce immunosuppression. In contrast, short-term exposure of suckling pups to OA via the milk stimulates the proliferative responses of lymphocytes to polyclonal activation.

Prenatal exposure of balb/c mice to ochratoxin A: Effects on the immune system in the offspring

Effects of prenatal exposure to the mycotoxin ochratoxin A (OA) on the immune system were evaluated in Balb/c mice. Dams were exposed to OA in their diet at doses of 0.18 (control), 30 or 200 micrograms/kg before and during gestation. At birth, pups were cross-fostered to non-exposed dams. OA exposure of the dams did not influence reproductive outcome, that is, the numbers of litters, litter sizes and body weight of the pups. Flow cytomety analysis of T-lymphocyte subpopulations on days 14 and 28 postpartum revealed a decrease in the percentages of splenic CD4+ and CD8+ cells in offspring from the high-dose group (200 micrograms/kg diet), but no significant alterations in absolute numbers of these cell populations nor in the total numbers of splenocytes were observed. In the thymus, a relative as well as an absolute increase in the CD4+ subpopulation was seen in exposed pups on day 14. On day 28, the absolute numbers of CD4+, CD8+ and CD4+CD8+ (double positive) cells were increased, reflecting an elevated number of thymocytes in the high-dose group. No significant differences were found in the proliferative responses of splenic or thymic lymphocytes to mitogens, or in the production of interleukin-2 in concanavalin A-stimulated cell cultures. Further, the plaque-forming cell response to sheep red blood cells and the humoral antibody response to the viral antigen PR8 were not affected by prenatal exposure to OA. No significant differences in natural killer cell activity were observed. The results indicate that exposure of dams to relatively low levels of dietary OA alters absolute and relative numbers of lymphocyte subpopulations in lymphoid organs, but does not suppress immune functions in the offspring.

Penicillium verrucosum in feed of ochratoxin A positive swine herds.

Ochratoxin A contamination of cereal feed grain was monitored during October 1989–September 1990 by analysis of blood samples from slaughter swine in Sweden. The detection of ochratoxin A in swine blood was used as a method to identify swine herds fed ochratoxin A contaminated feed. The contamination level of ochratoxin A in the blood of the positive herds was in the range 2–45 ng/ml with the mean concentration 5.2 ng/ml. Feed samples for mycological analysis were collected from both ochratoxin A positive herds (⩾2 ng/ml blood) and ochratoxin A negative herds (<2 ng/ml blood). From the ochratoxin A positive herds and the ochratoxin A negative herds 22 and 21 feed samples were collected, respectively. No quantitative differences in mould content, as determined by colony forming units, were observed between the two groups. However, there were differences in the mycoflora. The incidence of storage fungi (Penicillium and Aspergillus spp.) was significantly higher (p < 0.05) in feed from ochratoxin A positive herds. Particularly, Penicillium verrucosum was found to be significantly more common (p < 0.001). Altogether 274 isolates were screened for their ability to produce ochratoxin A. Ochratoxin A producers were found only within P. verrucosum; 38% of the 63 isolates produced detectable amounts of ochratoxin A. Ochratoxin A producing isolates of P. verrucosum were found in 60% of the feed samples collected from ochratoxin A positive swine herds and in one sample (5% ) of the feed samples collected from the ochratoxin A negative herds.

Influence of perinatal ochratoxin A exposure on the immune system in mice

The mycotoxin ochratoxin A (OA) is a well-documented immunotoxic agent which affects both cellular and humoral immunity. In the present study, the effects of maternal exposure to single doses of OA during gestation or lactation were studied in Balb/c offspring. A single dose exposure of the dams to OA (500 micrograms/kg body weight) on day 16 of gestation resulted in decreased proliferation of splenic and thymic lymphocytes in response to mitogens in the pups at 15 days of age. Flow cytometry analysis of thymocyte subpopulations revealed lower percentages of mature CD4+ cells and higher percentages of immature, double-positive (CD4+CD8+) cells in the exposed pups. In contrast, a single exposure of the dams of OA on day 10 postpartum significantly increased the proliferative responsiveness of lymphocytes in the offspring when stimulated with B or T cell mitogens 3 days after the exposure. This effect was most prominent in the highest dose group (500 micrograms/kg body weight). The present results are in accordance with previous observations in rats, and show that the time of exposure significantly influences the immunotoxic effects of OA on the developing immune system in rodents.