Research Note: Immunocompetent cells in blood and intestine after administration of Lacto-Immuno-Vital in drinking water of broiler chickens.

The understanding of the synbiotics´ impact on the host is incomplete. To improve the knowledge, we study the effect of Lacto-Immuno-Vital synbiotic preparation in chickens on local and systemic immune response by evaluation of immunocompetent cells in the peripheral blood and jejunal mucosa. Hematological method was used for determination of white blood cell count, and flow cytometry for measurement the functions of phagocytes and subpopulation of lymphocytes (CD3, CD4, CD8, IgM, and IgA). Cell Qest programme (Germany) was used for analysing of data obtained from flow cytometer and GraphPad Prism version 4.0 for comparison by paired t test between control and experimental groups. The experiment was conducted in a commercial broiler chicken fattening farm, the birds were handled and sacrificed in a humane manner. A flock of 64,400 one-day-old Hybrid ROSS 308 chickens were included in the 42-d experiment. The chickens were randomly divided into 2 equal groups, experimental and control, and each group of chickens was housed in a different hall while maintaining the same conditions. The chickens in the experimental group (Lactovital) received 500 g of Lacto-Immuno-Vital (Hajduvet Kft., Hungary) in 1,000 L of drinking water. Lacto-Immuno-Vital was administered daily from the first day (D1) to D7 of the experiment. From D 7 to D 22 it was given in a pulsed manner (every third day) at a dose of 300 g in 1,000 L of drinking water. Control group received only the standard diet. For immune analyses 6 randomly chosen chickens from experimental and control group were taken from the halls. The sampling days were set at D 8 and D 22 of the experiment. Samples of peripheral blood were collected from vena subclavia. The chickens were euthanized and whole jejunum was taken during necropsy into Hanks ice solution (pH 7.2-7.3). Administration of Lacto-Immuno-Vital in drinking water of nonstressed broilers during fattening period in commercial production increased phagocytic activity and phagocytic index. The number of IgA+ and CD8+ cells in lamina propria of intestine was decreased in chickens fed diet supplemented with Lacto-Immuno-Vital in drinking water. We suggest that increased phagocytic activity and decreased number of immunocompetent cells in mucosa of intestine was caused by improved systemic and local immune system function.

Influence of Lacto-Immuno-Vital on growth performance and gene expression of IgA, MUC-2, and growth factor IGF-2 in the jejunum of broiler chickens.

The effects of Lacto-Immuno-Vital synbiotic preparation on gene expression of IgA, MUC-2, and growth factor IGF-2 in the jejunum and on BW gain in broiler chickens were studied. A flock of 64,400 1-day-old Hybrid ROSS 308 chickens was inducted in the 42-day experiment. The chickens were divided into 2 equally size groups in separate halls. The chickens in the experimental (E) group received 500 g of Lacto-Immuno-Vital in 1,000 L of drinking water. The preparation was administered daily from the first day (day 1) to day 7 of the experiment. From day 7 to day 22, it was given in pulsed manner (every third day) at a dose of 300 g in 1,000 L of drinking water. The broiler chickens in the E group gained more weight (P < 0.001) compared with control from day 10 to day 42. Death of animals during feeding period was 1,078 chickens in the E group compared with 1,115 dead chickens in the control group. Feed conversion ratio was 1.61 kg of supplemented diet/kg of BW in the E group compare with 1.67 kg of nonsupplemented diet/kg of BW in control. The relative expression of IgA gene in the jejunum was upregulated on day 22 in the E group compared with control (P < 0.05), whereas relative expression of MUC-2 gene was upregulated in the E group compared with control on day 8 and day 22 (P < 0.05; P < 0.001). Similarly, relative expression of IGF-2 gene was upregulated in the E group compared with control on both samplings (P < 0.01). The composition of Lacto-Immuno-Vital synbiotic preparation showed beneficial effects on growth performance, feed conversion ratio, morbidity, mortality, and selected parameters of mucosal immunity in the chicken jejunum.

The Influence of Chestnut Wood and Flubendazole on Morphology of Small Intestine and Lymphocytes of Peripheral Blood, Spleen and Jejunum in Broiler Chickens.

The study examined subpopulations of lymphocytes in peripheral blood, spleen, and jejunum including morphology of that segment in broiler chicken farm after treatment with flubendazole (Flimabend) and natural extract from chestnut wood (Farmatan). A total of 24 forty-day-old Kalimero-Super Master hybrid chickens were divided into 4 groups (n=6): the Fli group received Flimabend per os, 100 mg/g suspension in 1.43 mg of active substance/kg body weight during 7 day of experiment, Far group received Farmatan per os at 0.2 % concentration for 6 hours per day during 5 day (experimental days - from 3 to 7); the Far+Fli group received a combination of doses administered in the same way as for the first two groups; and control -C group with no active substance administration. The results demonstrated mild increase of leukocytes, lymphocytes, monocytes, leucocyte common antigen CD45, IgM+ and IgA+ cells in peripheral blood after administration of Flimabend. Similarly, subpopulations of followed lymphocytes (CD3+, CD4+, CD8+, IgM+) were increased in the jejunum after application of that drug. On the other hand, administration of Farmatan revealed opposite effect on determined immunocompetent cells what proves anti-inflammatory effect. Morphology of villi was also negatively influenced by administration of Flimabend. Administration of Farmatan suggests also its preventive administration in chickens. This tanin-containing drug as plant natural product may be used due to its antibacterial activity and as promising alternative to conventional drug with possible antihelminthic effect.

Determination of carcinoembryonic antigen and cancer antigen (CA 15-3) in bitches with tumours on mammary gland: preliminary report.

The aim of this work was to determine levels of carcinoembryonic antigen (CEA) and cancer antigen (CA 15-3) in the blood serum of 45 bitches. A modified procedure was used to determine the CEA and CA 15-3 markers with the human kits using the radioimmunoassay method. Samples collected from extirpated tumour of mammary glands were histologically processed and classified as per WHO guidelines. The average age of animals with tumour was 10.00 ± 2.2 years; for healthy bitches average age was 4.2 ± 3.2 years. Values of CEA and CA 15-3 were considered positive, if they exceeded 0.23 ng mL(-1) and 7 IU mL(-1) , respectively. Average levels of CEA in the tumour group were 0.25 ± 0.06 versus 0.20 ± 0.03 in healthy bitches (P = 0.0001). The average CA 15-3 value in bitches with tumour was 8.58 ± 1.27 versus 5.14 ± 1.34 in healthy animals (P < 0.0001).

[Experimental use of the recipient's epithelial cells in tracheal allotransplantation--initial outcomes]. / Vyuzitie epiteliálnych buniek príjemcu pri allotransplantácii trachey v experimente--prvé výsledky.

The management of long tracheal lesions requires development of tracheal implants, which would enable resection combined with anastomosis. The authors' scientific study is based on tracheal allotransplantation on an animal model (sheep), using tracheal epithelial cells of the recipient. The project covers preparation of the graft, so that all components of the major histocompatibility complex (MHC), which participate in graft rejection, are removed. Histological examination of the allograft with cultivated epithelial cells showed its good healing with revasculatization and with no signs of graft rejection.

Leukocytic responses of broilers following dietary contamination with deoxynivalenol and/or treatment by dietary selenium supplementation.

1. This experiment was to investigate the effects of natural dietary contamination with a mycotoxin product (deoxynivalenol: DON) and/or with dietary selenised yeast (Se-yeast), on respiratory burst and phagocytic activity of granulocytes and the frequency of B- and T-lymphocytes in peripheral blood of broilers. 2. Sixty one-day-old chicks of both sexes were divided into 4 groups, each of 15 birds, fed on a control diet that contained 0.2 mg DON/kg and 0.4 mg Se/kg (CON group), a diet supplemented with 1 mg Se-yeast/kg (Se-yeast group), a diet contaminated with 3 mg DON/kg (DON group) or a diet contaminated with DON and supplemented with Se-yeast (DON plus Se-yeast group). 3. Blood samples collected from the birds at the age of 4 weeks showed that neither B- and T-cell numbers nor granulocytic respiratory burst were influenced by 3 mg DON/kg. Blood granulocyte phagocytic activity was not reduced by DON but numbers of heterophils were increased. In the DON plus Se yeast group phagocytic activity was the same as in the CON group. The Se-yeast and DON plus Se-yeast groups had increased numbers of CD3(+), CD4(+), and CD8(+) T-cells as well as IgM(+) B-cells in their blood compared to both CON and DON-groups. 4. The results show there is no significant effect of dietary DON up to 3 mg/kg on leukocytes apart from the compromised blood granulocytes phagocytic activity and increased numbers of heterophils. The increased numbers of B- and T-lymphocytes in blood of birds fed on diets with supplementation of organic Se indicates some positive effects of this essential microelement on poultry lymphoid cells.

Spreading of Salmonella enteritidis in the cecum of chickens.

Adhesion and colonization of high (2 x 10(8) CFU) and low doses (2 x 10(2) CFU) of Salmonella enteritidis (phage type 4) was determined in the ceca collected 6 h-4 weeks after inoculation (pi), of 1-d-old White Plymouth Rock orally-inoculated chickens. S. enteritidis was associated with the epithelial surface of the villi in the low-dose group 18 h-7 d pi, the penetration in the cecal lamina propria was observed on day 1 and 10 pi. In the high-dose group, adhesion and colonization was observed in all birds killed 6 h-14 d pi; penetration of the bacteria into the cecal lamina propria was seen 1-21 d pi. Large numbers of macrophage-like cells containing S. enteritidis were observed in the cecal lamina propria on days 3-21 pi. Colonization and migration by S. enteritidis in the intestinal tract of chickens was shown to be dose dependent.

Effect of aflatoxin B1 on CD3 T cells and alkaline phosphatase in the intestine of mice.

The aim of the study was to determine the effect of repeated applications of aflatoxin B1 (AFB1) on immunocompetent cells (CD3 T cells) and alkaline phosphatase in the intestinal mucosa. Mice were orally treated with AFB1 for 24 days. The mucosa of the intestine showed a significant decrease in the number of CD3 T cells and a significantly lower level activity of alkaline phosphatase on day 24 in AFB1 treated mice. Similarly, with changes in the small intestine, qualitative haematological parameters were modified in systemic immunity as lymphopenia, and neutropenia, monocytopenia. AFB1 treated animals showed reduction in body weight gain and increased liver weight. We supposed that changes found in the small intestine are secondary to primary systemic haematological lesions. The decrease in CD3 T cells suggests a connection with the decrease in the host's resistance to infectious diseases.

[The histopathologic picture after chemical eye injury and the role of limbal stem cells]. / Histopatologický obraz po chemickom poranení oka, úloha limbálnych kmenových buniek.

The aim of this experimental study was to evaluate the role of limbal stem cells on corneal wound healing. The experimental objects were New Zealand white rabbits. Damage to the cornea was caused by 2.5 N NaOH solution for 50 sec. and was subsequently washed out by saline solution. The rabbits were euthanased 24 hours after alkaly burn and the tissue was fixated in 10% formalin. The samples were embedded in paraffin and stained with haematoxilin and eosin. The specimen showed focal inflammatory reaction with the presence of macrophages an round--nucleated cells in the limbal area.

Effect of low and high doses of Salmonella enteritidis PT4 on experimentally infected chicks.

Chicks (1-d-old, three groups, each containing 50 chicks) were inoculated with 2 x 10(2) and 2 x 10(8) CFU of Salmonella enteritidis; the third group were kept as uninoculated control. Five birds from each group were euthanized at intervals from 6 h to 4 weeks post-inoculation (pi). In the low-dose group S. enteritidis was isolated from 60% cecal samples at 18 h pi, and from 20% of livers at 3 d pi. Individual variation in the frequency of S. enteritidis recovery was observed in this group. The clearance of salmonella from the organs was faster in the low-dose group, and salmonella was not isolated from the liver and cecum at 21 and at 27 d pi, respectively. However, in the high-dose group, S. enteritidis was isolated from all ceca and 80% of liver 6 h pi, and salmonella was detected in the cecum and liver throughout the experiment. Serous typhlitis and unabsorbed yolk sac were the most prevalent lesions in both groups. Granulomatous nodules in the cecum were found occasionally in some cases in both inoculated groups, which can play a role as reservoirs in carrier chicks.

CD3 cells in the intestine of broilers inoculated with intestinal homogenate.

1. Reactivity of polyclonal rabbit CD3 antiserum to human T cells and density of CD3 cells in chicken intestine were studied after the inoculation of 1-d-old broilers with intestinal homogenate to develop the Runting and Stunting Syndrome. 2. Reactivity of CD3 antiserum in chicken intestine was very good. 3. Increased numbers of epithelial lymphocytes as well as infiltration of CD3 cells into the lamina propria were demonstrated. 4. Intestinal homogenate caused impaired growth and bad feathering in broiler chickens until 21 d of age.