Lactoferrin (LF) is an iron-binding protein from the transferrin family, present in mucosal secretions, granules of neutrophils and the serum of mammals. Many biological functions have been attributed to lactoferrin, including immunomodulatory and anti-inflammatory properties. Its presence has not been detected in poultry and its influence on their mechanisms of resistance has not been investigated. The aim of present in vitro study was to investigate the influence of bovine lactoferrin on the activity of chicken blood leukocytes. After isolation cells were cultured in complete RPMI - 1640 medium containing 0 (control), 2, 1, 0.5, 0.25 and 0.1 mg/ml of bovine LF, lymphocytes for 48 h and granulocytes for 18 h, respectively. Then the effect of LF on the activity of phagocytes (RBA and PKA tests) and lymphocytes (MTT test) was evaluated. The obtained results suggest that bovine lactoferrin has significant influence on the activity of chicken leukocytes in vitro. The use of LF increased the ability of chicken granulocytes for respiratory burst, however no stimulation of the killing activity was observed. Bovine lactoferrin also increased the proliferative response of lymphocytes T stimulated by ConA, whereas a similar effect on lymphocytes B stimulated by LPS was not noticed. The observed stimulation of the investigated parameters is encouraging for further research concerning the possibility of the use of LF as immunostimulant in poultry husbandry.
Hepatic oval cells (OC) are considered to be the stem cells of the liver and have been linked to the development of hepatocellular carcinoma HCC, one of the most lethal cancers. The objective of this study was to analyse the proliferative response of OC obtained from rats receiving diethylnisamine DEN. In addition, we investigated the effects of resveratrol (Res) on the proliferation and oxidative markers of OC in vitro. OC isolated by in situ collagenase liver perfusion methods were treated with different concentrations of Res for 24, 48 and 72 hours. At the end of these periods the proliferative activity of OC, the release of superoxide anion by OC and the medium concentration of malonylodialdehyd were analyzed . The proliferation of OC cultured without Res increased from IP = 0.945 ± 0.02 to IP = 1.525 ± 0.031 after 24 and 72 hours respectively. A marked (p ≤ 0.05) inhibition of OC proliferation was observed in the presence of 1.0 µM, 25 µM and 100 µM of Res. A significant decrease in the release of O₂⁻. by OC, observed throughout the experimental period, was attributed to the presence of 25 µM of Res. Exposition of OC to 100 µM of Res inhibited the release of O₂⁻. only after 48 and 72 hours of incubation. The presence of 25 µM of Res resulted in the depletion of MDA level, which did not exceed 0.19±0.009 nM. Proliferative activity of OC isolated from carcinogenic rats intensified throughout the culture period. When subjected to the carcinogenic effect of DEN, Res exerts anti-proliferative influence on OC. Moreover, our results provide evidence that Res attenuates oxidative stress during hepatocarcinogenesis.
Silver nanoparticles (SNP) have been recently one of the most widely utilized nanomaterials, mostly because of their antimicrobial activity. For some time there has been a great interest in SNP of unconventional medicine, which recommends their use not only as an antimicrobial, but also as an immunostimulant. However, little is known about SNP’s impact on immunocompetent cells in vitro. The aim of a present study was to investigate the influence of the colloidal nano-silver solution on the viability and proliferative response of mice peripheral blood mononuclear leukocytes and splenocytes in vitro. After isolation cells were cultured in complete RPMI-1640 medium containing 0 (control), 20, 10, 5, 2, 1, 0.5, 0.2, 0.1 and 0.05 ppm of SNP, for 24 and 48 h to investigate SNP’s impact on cell viability, and for 72 h to evaluate their effect on the proliferative response of cells. Both parameters were assessed using MTT assay. Obtained results suggest that SNP have significant influence on the both investigated parameters. High doses of SNP significantly decreased (p < 0.01) the viability (concentrations of 5-20 ppm for splenocytes and 2-20 ppm for leukocytes, respectively) and proliferative response of cells (splenocytes: 5-20 ppm, leukocytes: 10-20 ppm) whereas low SNP’s doses slightly increased (p > 0.05) the viability of cells (splenocytes: 1 ppm, leukocytes: 0.2 ppm) and significantly increased stimulation index of both cell types induced by mitogens (splenocytes: 0.2-0.5 ppm, p < 0.05; leukocytes: 0.1-0.5 ppm, p < 0.01). These experimental data constitute an encouragement for further investigations concerning the possibility of therapeutic use of SNP’s low doses.
The purpose of this experiment was to estimate the most effective doses of TFX, used in injection and in bath, for the stimulation of the activity of lymphocytes in common carp The study was performed on 9-month-old carps obtained after spring catches (in March). The fish were divided into 10 groups. Fish in groups I-IV were given TFX by intramuscular injection at doses of 20, 10, 0.5, and 0.2 mg/fish. Fish in groups VI-IX were bathed for 24, 48, and 96 hours in TFX at concentrations of 200, 100, 50, and 20 mg/l of water. Groups V and X were control. Blood samples were taken immediately after the administration of TFX in bath and after 2, 7, 14, and 28 days in groups I-IV and VIII-IX. The following parameters were determined: the number of leukocytes and lymphocytes in 1 μl of blood, the percentage of ANAE+ and E-rosette-forming cells, and the proliferative activity of lymphocyte cell cultures stimulated with Con A. Results: TFX at doses of 100, 200 mg/l of water and 10, 20 mg/fish administered by injection showed a suppressive effect on the parameters investigated. TFX at doses of 50, 20 mg/l of water and 0.5, 0.2 mg/fish caused an increase in the number of leukocytes and lymphocytes, a higher percentage of ANAE+ cells and rosette-forming cells, as well as a multiple increase in the proliferative activity of lymphocytes stimulated with Con A. The highest values were obtained after the administration of TFX in a 96-hour bath at a concentration of 50 mg/l of water.
The experiment studied 32 crossbred fatteners. The animals were fed in a two-stage system with the addition of feed antibiotic (control group - C) or prebiotic BIO-MOS (Saccharomyces cerevisiae, strain 1026; 0.1% during the first period of fattening) (experimental group - E). From 16 animals chosen at random (8 fatteners each from groups E and C) the following sections of small intestine were sampled post mortem and preserved for histological, histochemical and immuno-histochemical evaluation: duodenum, jejunum and ileum. Staining by the review (H-E) and paS-Alcjan method was carried out. Antibodies bax, bcl-x, bcl-2 and PCNA and Ki-67 were used to evaluate apoptosis and proliferation. The preliminary studies in situ revealed the suitability of the employed antibodies: bax, bcl-x and bcl-2 in evaluating apoptosis. Histological, histochemical and immuno-histochemical evaluation of the mucosa in the small intestines of the animals receiving feed antibiotic or its replacer - prebiotic, showed that the effect of the employed additives differentiated in terms of morphological traits, apoptosis and proliferation abilities of crypts’ epithelium. It was concluded that there were no negative effects of BIO-MOSS on the mucosa of the small intestine in fatteners.
The aim of the study was to determine the effect of adding an antibiotic or zinc oxide together with the feed on the histological picture as well as on the apoptotic and proliferation activity of enterocytes in the mucus of the small intestine of fatteners. The investigations were carried out on 32 crossbred fatteners. The animals were fed in a two-stage system, with the addition of feed antibiotic (control group - C) or with the 0.1-percent addition of zinc oxide (the first stage of fattening; experimental group - E). Samples of small intestine segments (duodenum, jejunum and ileum) were collected post mortem and secured from 16 randomly chosen animals (8 animals from group E and 8 from C) fr histological, histochemical and immuno-histochemical analysis. Staining was conducted using the review method (H-E) and paS-Alcjan. For evaluation of apoptosis and proliferation, the antibodies bax, bcl-x bcl-2 and Ki-67 and PCNA were used. The histological, histochemical and immuno-histochemical evaluation of the mucus of the small intestine of the animals, receiving feed antibiotic or ZnO, showed a comparable influence of the used additives on the morphological traits as well as differentiated the apoptosis and proliferation abilities of the crypt epithelium (cells Ki-67 positive - duodenum and jejunum: P ≤ 0.05 K vs D; PCNA positive duodenum: P ≤ 0.01 K vs D). No negative influence of the zinc oxide additive on the mucus of the small intestine in fatteners was found, which indicates the potential possibility of employing it in the first stage of the fattening period.
The aim of this study was to evaluate the effect of flumequine on the percentage of peripheral blood leukocytes of carp, alpha naphthyl acetate esterase (ANAE) activity and proliferative activity of lymphocytes stimulated with concanavalin A (Con A) and lipopolysaccharide (LPS). Flumequine was administered to 40 carp, weighing 150 ± 10 g, at a dose of 12 mg/kg, once (group I) and four times, every 2 days (group II). Among white blood cells in flumequine, treated fish (group II) observed a decrease in percentage of lymphocytes and an increase of neutrophiles. Identification of the ANAE esterase activity in fish lymphocytes of the control group showed the advantage of positive cells over the negative ones and amounted to 62.65 ± 3.22%. After administration of flumequine in group II fish, this value decreased to 44.75 ± 3.70%. The present study clearly demonstrated that both Con A and LPS induced lymphoid cell proliferation in vitro in group I. There was an increase in activity after stimulation of LPS and its reduction after Con A in group II. This indicates a stimulating effect of flumequine on B lymphocytes. The results of this study are not conclusive as to the positive effect of the drug on the immune system of fish and indicate the need for caution in its use.