Effect of aqueous extract from Scutellaria baicalensis Georgi roots on CD4+ and CD8+ T cell responses during experimental infection with Trichinella spiralis in mice.

The aim of this study was to investigate the effect of aqueous extract from Scutellaria baicalensis Georgi roots (SB) on blood parameters and immune response during an experimental trichinellosis. A total of 60 mice infected with 200 Trichinella spiralis larvae were assigned into two groups. One of them served as a control and the second received SB extract orally from day 5 before infection to day 28 after infection (dpi). Blood was sampled at 7, 14, 21 dpi. Lymphocytes obtained from the spleen and mesenteric lymph nodes (MLN) at 7, 14, 21, and 28 dpi were counted, CD4+ and CD8+ subpopulations were analyzed by flow cytometry, and lymphocyte proliferation was estimated with colorimetric (MTT) assay. The intensity of intestinal and muscle invasion was also studied. SB caused a remarkable elevation of banded neutrophils in the blood at 7 dpi. SB increased ConA-stimulated splenocyte proliferation and CD4+ and CD8+ splenocyte subsets at 14 and 21 dpi, whereas MLN lymphocyte subset stimulation involved only CD4+ at 14 dpi. After administration of SB a downward trend in the number of T. spiralis larvae in the muscle was observed. These results suggest that Scutellaria baicalensis root extract stimulates murine cellular immune response during intestinal phase of T. spiralis infection.

In vitro study of the effect of corn dried distillers grains with solubles on rumen fermentation in sheep.

The aim of the in vitro study was to determine the effect of corn dried distillers grains with solubles (corn DDGS), used as a replacement for the concentrate ingredients of sheep diet, on rumen fermentation. The material for the study was the ruminal fluid of Polish Merino sheep which was incubated during 4-, 8- or 24-hour periods. Five groups of samples were prepared for in vitro fermentation: C - control, incubated with the substrate consisting of the concentrate ingredients; D1, D2 and D3, where DDGS was used as a substrate added in proportions of 10, 20 and 30% of dry matter of the concentrate; and D4, where 100% DDGS was used as a substrate. After fermentation, the gas and short chain fatty acids (SCFAs) analyses were performed using gas chromatography. The ammonia concentration and pH were also determined, and the SCFA utilization index (NGR), the fermentation efficiency (FE) and the index of cell yield of ruminal microorganisms (CY) were calculated. This research showed no effect of DDGS on the methane emission. The positive correlations between the amount of methane and ammonia concentrations in the 8- and 24-hour fermentation periods were found. DDGS addition increased propionate proportion, but decreased production of acetate (p<0.01). Additionally, D1, D2, D3 and D4 substrates lowered isobutyrate (p<0.05) and isovalerate (p<0.01) production. Based on the results obtained, it can be stated that partial substitution of the concentrate ingredients with DDGS did not have deleterious effect on sheep rumen fermentation processes.

The effect of humic-fatty acid preparation on selected haematological and biochemical serum parameters of growing rabbits.

In this study, we assessed the effect of a humic-fatty acid preparation (HFA) used in rabbit feed on certain haematological and biochemical serum parameters (the lipid profile and the Ca, P and Fe contents). A higher RBC, HGB and HCT values were observed in the groups that were given HFA. An increase in Fe concentrations were also noted. The total and LDL cholesterol were lower in the groups which received HFA than in the control group.

Trichinella spiralis: the influence of short chain fatty acids on the proliferation of lymphocytes, the goblet cell count and apoptosis in the mouse intestine.

This study was carried out to determine the influence of short chain fatty acids (SCFA) on spleen and mesenteric lymph node lymphocyte proliferation, goblet cells and apoptosis in the mouse small intestine during invasion by Trichinella spiralis. BALB/c mice were infected with 250 larvae of T. spiralis. An SCFA water solution containing acetic, propionic and butyric acids (30:15:20 mM) was administered orally starting 5 days before infection and ending 20 days post infection (dpi). Fragments of the jejunum were collected by dissection 7 and 10 dpi, and were examined for apoptotic cells in the lamina propria of the intestinal mucosa, and for goblet cells. The proliferation index of the cultured spleen and mesenteric lymph node lymphocytes with MTT test was also determined. The orally administered SCFA solution decreased the proliferation of mesenteric lymph node lymphocytes in the mice infected with T. spiralis at both examination times, but did not influence the proliferative activity of the spleen cells. Seven dpi, both in the spleen and mesenteric lymph nodes, the highest proliferation index of concanavalin A (Con A)-stimulated lymphocytes was found in the group of uninfected animals receiving SCFA animals. This tendency could still be seen 10 dpi in the mesenteric lymph nodes but not in the spleen, where the proliferation index in this group had significantly decreased. In vitro studies revealed, that butyric and propionic acids added to the cell cultures suppressed the proliferation of Con A-stimulated mesenteric lymph nodes and spleen lymphocytes taken from uninfected and T. spiralis-infected mice. Acetic acid stimulated proliferation of splenocytes taken from uninfected mice but did not affect lymphocyte proliferation in mesenteric lymph nodes from uninfected or infected mice. Orally administered SCFA increased the number of goblet cells found in the epithelium of the jejunum 7 dpi, but this number had decreased 10 dpi. The number of apoptotic cells in the lamina propria of the intestinal mucosa of animals infected with the T. spiralis and receiving SCFA was also lower, particularly 10 dpi. The above results show that SCFA can participate in the immune response during the course of trichinellosis in mice.

Effects of a skullcap root supplement on haematology, serum parameters and antioxidant enzymes in rabbits on a high-cholesterol diet.

We studied the effects of Scutellaria baicalensis root on blood parameters and antioxidant enzyme activities in rabbits fed a high-cholesterol diet. Thirty-two New Zealand White rabbits were divided into four groups of eight animals each. They were fed a standard diet (C group), a diet with a 1% pure cholesterol supplement (CH group), a diet with a 1% pure cholesterol supplement and a 9% skullcap root supplement (CH+SR group), or a diet with a 9% skullcap root supplement (SR group). After 6 weeks, the rabbits fed the high-cholesterol diet had significantly decreased RBC and Hb levels and significantly increased MCV, MCH and Fe levels (p≤ 0.05). The skullcap root supplement had no adverse effects on the haematological parameters. The values for RBC, Hb, HCT, MCV, MCH and MCHC were similar in the skullcap root-treated and control rabbits. We also observed a remarkable elevation in the serum TC, LDL and TG levels at the end of the 6-week period. The rabbits fed the cholesterol diet showed decreased activity of the erythrocyte GSH-Px compared with the rabbits fed the basal diet. The GSH-Px activity was significantly higher in the rabbits fed the CH+SR diet than in those on the CH diet. The erythrocyte SOD activity was also significantly decreased in the rabbits on the CH diet. However, the CH+SR group rabbits showed significantly enhanced erythrocyte SOD activity. The SOD level in the CH+SR rabbits was 34.91 U/ml, which was a 23% increase (p≤ 0.05) in relation to the results for the CH group and only 15% diminished in relation to the control group. These results suggest that the dietary supplementation of skullcap root may improve rabbit antioxidant systems and protect against the risks from a high-cholesterol diet.

The effect of Linola and W92/72 transgenic flax seeds on the rabbit caecal fermentation--in vitro study.

The effect of W92/72 transgenic flax seeds taken from a variety of Linola on the production of SCFA, ammonia and methane by bacteria inhabiting rabbit caecum was studied. The in vitro method was used where caecal contents from rabbits was incubated with W92/72 transgenic or Linola flax seeds, or without any additives (control samples). The total concentration of SCFA was higher in samples with the addition of flax seeds than in the control samples. The increase in concentrations of acetic, propionic and butyric acids was the highest in samples with Linola seeds added. A higher percentage of propionic and butyric acids was observed in the contents incubated with addition of flax seeds as compared to the control samples. This increase was the result of a percentage decrease in acetic acid. No differences were observed in the concentration of ammonia between fermented samples. Moreover, the addition of flax seeds resulted in slight decrease of pH in incubated samples. In gas samples, the methane level was higher in samples with flax seeds added, although the highest level was found in samples with transgenic seeds. In addition, gas pressure was significantly higher in samples with flax seeds added as compared to control samples, and this may indicate a higher intensity of microbiological fermentation processes. These studies suggest that neither Linola nor W92/72 flax seeds have any unfavorable effect on the caecal microflora activity of rabbits. A beneficial influence of flax seeds on the microbiological fermentation process in rabbit caecum was observed, based on an increase in percentage ratio of propionic acid in samples with flax seeds added.

CD4+ and CD8+ cells in mice infected with Trichinella spiralis and treated with cyclosporine A.

The mice infected with 200 Trichinella spiralis larvae were injected intraperitoneally with Cyclosporine A (CyA) between 14-18 days post infection (dpi). The drug was administered in a dosis of 50 mg/kg/day. The animals were killed at 21, 28, 35, 42 and 60 dpi and the fragments of spleen, mesenteric lymph node, jejunum and musculus masseter were sectioned in a cryostat and fixed in acetone. The slides were examined with monoclonal sera by the immunofluorescent or immunoenzymatic method. It was found that the number of CD4+ cells in the control and in the CyA-treated mice was similar but in the animals receiving the drug the reaction was less intensive. The stimulation of CD8+ cells of CyA treated mice--especially in the jejunum--was stronger than in the control animals. This fact is important because the CD8+ cells are the APC cells in this organ.

The influence of adriamycin on the CD4+ and CD8+ cells in the course of trichinellosis in mice.

The effect of adriamycin on the behaviour of CD4+ and CD8+ cells in the course of trichinellosis in mice has been studied. The animals infected with 200 larvae per mouse were administered intraperitoneally adriamycin (Adriblastin from Farmitalia) at 1st and 28th day post infection (dpi), in a dosis of 0.2 mg. The mice were killed weakly for 6 weeks and then at the 60th dpi. The examinations were made on histologic sections from the spleen, mesenteric lymph node, jejunum and masseter muscle using immunofluorescent and immunoenzymatic methods with monoclonal antibody. The mice receiving adriamycin exhibited more CD8+ cells in the intestinal mucosa and by the end of the experiment also in the muscles in comparison with the control animals, which, however, did not affect the course of the infection.