scholarly journals The influence of dietary supplementation with the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on chemotaxis, phagocytosis and respiratory burst of peripheral blood granulocytes and monocytes in calves

2020 ◽  
Author(s):  
Roman Wójcik ◽  
Joanna Małaczewska ◽  
Grzegorz Zwierzchowski ◽  
Jan Miciński ◽  
Edyta Kaczorek-Łukowska

Abstract Background A healthy immune system plays a particularly important role in newborns, including in calves that are far more susceptible to infections (viral, bacterial and other) than adult individuals. Therefore the objective of this study was to evaluate the effect of HMB on the chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and respiratory burst (BURSTTEST® kit) of peripheral blood granulocytes and monocytes in calves by flow cytometry. Results An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of both granulocytes and monocytes after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-β-myristate-13-acetate), expressed in terms of the percentage of oxidative metabolism and MFI. Conclusion HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.

2020 ◽  
Author(s):  
Roman Wójcik ◽  
Joanna Małaczewska ◽  
Grzegorz Zwierzchowski ◽  
Jan Miciński ◽  
Edyta Kaczorek-Łukowska

Abstract Background A healthy immune system plays a particularly important role in newborns, including in calves that are far more susceptible to infections (viral, bacterial and other) than adult individuals. Therefore, the present study aimed to evaluate the influence of HMB on the chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and oxidative burst (BURSTTEST® kit) of monocytes and granulocytes in the peripheral blood of calves by flow cytometry. Results An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of monocytes and granulocytes stimulated with PMA (4-phorbol-12-β-myristate-13-acetate) and Escherichia coli bacteria, expressed as MFI values and the percentage of oxidative metabolism. Conclusion HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.


2020 ◽  
Author(s):  
Roman Wójcik ◽  
Joanna Małaczewska ◽  
Grzegorz Zwierzchowski ◽  
Jan Miciński ◽  
Edyta Kaczorek-Łukowska

Abstract Background A healthy immune system plays a particularly important role in newborns, including in calves that are far more susceptible to infections (viral, bacterial and other) than adult individuals. Therefore, the present study aimed to evaluate the influence of HMB on the chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and oxidative burst (BURSTTEST® kit) of monocytes and granulocytes in the peripheral blood of calves by flow cytometry. Results An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of monocytes and granulocytes stimulated with PMA (4-phorbol-12-β-myristate-13-acetate) and Escherichia coli bacteria, expressed as MFI values and the percentage of oxidative metabolism. Conclusion HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.


2020 ◽  
Author(s):  
Roman Wójcik ◽  
Joanna Małaczewska ◽  
Grzegorz Zwierzchowski ◽  
Jan Miciński ◽  
Edyta Kaczorek-Łukowska

Abstract Background The objective of this study was to evaluate the effect of HMB on the chemotactic activity, phagocytic activity and respiratory burst of peripheral blood granulocytes and monocytes in calves. Method The experiment was performed on 14 calves aged 30±2 days, divided into two equal groups of control (group I) and experimental (group II) animals. The feed administered to experimental group calves was supplemented with HMB (Metabolic Technologies Inc. Ames, IA, USA) at 40 mg/kg BW, whereas control calves were administered standard farm-made feed without supplementation. Blood was sampled from the jugular vein immediately before the experiment (day 0) and on experimental days 15, 30 and 60 to determine: chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and respiratory burst (BURSTTEST® kit) of peripheral blood granulocytes and monocytes by flow cytometry. Results An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of both granulocytes and monocytes after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-β-myristate-13-acetate), expressed in terms of the percentage of oxidative metabolism and MFI. Conclusion HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.


2020 ◽  
Author(s):  
Roman Wójcik ◽  
Joanna Małaczewska ◽  
Grzegorz Zwierzchowski ◽  
Jan Miciński ◽  
Edyta Kaczorek-Łukowska

Abstract Background A healthy immune system plays a particularly important role in newborns, including in calves that are far more susceptible to infections (viral, bacterial and other) than adult individuals. Therefore, the present study aimed to evaluate the influence of HMB on the chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and oxidative burst (BURSTTEST® kit) of monocytes and granulocytes in the peripheral blood of calves by flow cytometry.Results An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of monocytes and granulocytes stimulated with PMA (4-phorbol-12-β-myristate-13-acetate) and Escherichia coli bacteria, expressed as MFI values and the percentage of oxidative metabolism.Conclusion HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.


2020 ◽  
Author(s):  
Roman Wójcik ◽  
Joanna Małaczewska ◽  
Grzegorz Zwierzchowski ◽  
Jan Miciński ◽  
Edyta Kaczorek-Łukowska

Abstract Background The objective of this study was to evaluate the effect of HMB on the chemotactic activity, phagocytic activity and respiratory metabolism of peripheral blood granulocytes and monocytes in calves.Method The experiment was performed on 14 calves aged 30±2 days, divided into two equal groups of control (group I) and experimental (group II) animals. The feed administered to experimental group calves was supplemented with HMB (Metabolic Technologies Inc. Ames, IA, USA) at 40 mg/kg BW, whereas control calves were administered standard farm-made feed without supplementation. Blood was sampled from the jugular vein immediately before the experiment (day 0) and on experimental days 15, 30 and 60 to determine: chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and respiratory metabolism (BURSTTEST® kit) of peripheral blood granulocytes and monocytes by flow cytometry.Results An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of both granulocytes and monocytes after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-β-myristate-13-acetate), expressed in terms of the percentage of oxidative metabolism and MFI.Conclusion HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.


Animals ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 1031 ◽  
Author(s):  
Roman Wójcik ◽  
Katarzyna Ząbek ◽  
Joanna Małaczewska ◽  
Stanisław Milewski ◽  
Edyta Kaczorek-Łukowska

The objective of this study was to determine the effect of β-hydroxy-β-methylbutyrate (HMB) on the chemotactic activity, phagocytic activity, and oxidative metabolism of peripheral blood granulocytes and monocytes in goats. Goat kids aged 30 ± 3 days were divided into two groups of 12 animals each: I—control, and II—experimental. Experimental group animals were fed a diet supplemented with HMB in the amount of 50 mg/Kg BW; whereas the diets of control goats were not supplemented. At the beginning of the experiment (day 0) and on experimental days 15, 30, and 60, blood was sampled from the jugular vein to determine and compare chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit), and oxidative metabolism (BURSTTEST® kit) of peripheral blood granulocytes and monocytes by flow cytometry. The analyses of the chemotactic and phagocytic activity of granulocytes and monocytes revealed statistically higher levels of phagocytic activity in the experimental group than in the control group, as expressed by the percentage of phagocytic cells and mean fluorescence intensity. HMB also enhanced the oxidative metabolism of both granulocytes and monocytes, expressed by the rate of oxidative metabolism and mean fluorescence intensity after stimulation with Escherichia coli bacteria and PMA (4-phorbol-12-β-myristate-13-acetate).


2014 ◽  
Vol 83 (4) ◽  
pp. 347-354 ◽  
Author(s):  
Roman Wójcik

The effect of the Leiber Beta-S (1,3-1,6-β-D-glucan) dietary supplement on the phagocytic activity (Phagotest) and oxidative metabolism (Phagoburst) of peripheral blood granulocytes and monocytes in calves was determined by flow cytometry. Fourteen animals were divided into two groups: a control group without dietary supplementation and an experimental group administered 50 mg/kg body weight/day of 1,3-1,6-β-D-glucan for 60 days. At the beginning of the experiment (day 0) and on days 15, 30 and 60, blood was sampled from the jugular vein to determine and compare immunological indicators. Leiber Beta-S significantly influenced (P ≤ 0.05, P ≤ 0.01, P ≤ 0.001) the percentages of phagocytic granulocytes (days 15 and 30) and monocytes (days 15, 30 and 60), and the percentage of bacteria engulfed by granulocytes (days 15 and 30) and monocytes (day 30). Leiber Beta-S increased the percentage of cells generating a respiratory burst in the population of granulocytes stimulated with fMLP (N-formyl-met-leu-phe) (day 15), PMA (4-phorbol-12-β-myristate-13-acetate) and Escherichia coli bacteria (days 15 and 30), and in the population of monocytes stimulated with fMLP (day 30), PMA (days 30 and 60) and E. coli (days 15, 30 and 60). The analyzed supplement increased mean fluorescence intensity in granulocytes stimulated with E. coli and fMLP (days 15 and 30) and PMA (days 30 and 60) as well as in monocytes stimulated with PMA, E. coli and fMLP (day 60). The findings of this study contribute to a better understanding of the effects of Leiber Beta-S on phagocytosis in calves, which have not been investigated to date.


2014 ◽  
Vol 17 (2) ◽  
pp. 281-291 ◽  
Author(s):  
R. Wójcik

AbstractThe objective of this study was to determine the effect of Biolex-MB40 on the phagocytic activity and oxidative metabolism of peripheral blood granulocytes and monocytes in lambs. The experimental material comprised 32 lambs aged 30 ± 3 days, divided into two equal groups: control and experimental. Experimental group animals were fed a diet supplemented with the Biolex-MB40 (Saccharomyces cerevisiae) in the amount of 3 g/kg of the concentrate. At the beginning of the experiment (day 0) and on experimental days 15, 30 and 60, blood was sampled from the jugular vein to determine and compare the phagocytic activity (PHAGOTEST) and oxidative metabolism (BURSTTEST) of peripheral blood granulocytes and monocytes by flow cytometry. Based on the results of an analysis of granulocyte and monocyte phagocytic activity, statistically higher levels of phagocytic activity were observed in the group of lambs administered Biolex-MB40 than in the control animals, expressed in terms of the percentage of phagocytic cells as well as mean fluorescence intensity. Biolex-MB40 also had a positive effect on the oxidative metabolism of both granulocytes and monocytes after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-β-myristate-13-acetate), expressed in terms of the percentage of oxidative metabolism as well as mean fluorescence intensity


2020 ◽  
Vol 10 (3) ◽  
pp. 551-557
Author(s):  
O. A. Kolenchukova ◽  
N. I. Sarmatova ◽  
A. V. Moshev

Current study performed to estimate the phagocytic activity of blood monocytes of varying phenotypes exposed to MRSA and MSSA strains.  Objects: Blood monocytes were collected from 25 healthy adults (age: 25–45 years). Live suspensions of MRSA/MSSA strains were used at concentration of 106 colony-forming units (CFU)/mL.  Metods. Phagocytic functions were estimated by using fluorescein isothiocyanate (FITC)-labelled MRSA and MSSA strains followed by running flow cytometry on FC 500 series flow cytometer (Beckman Coulter, USA). Whole peripheral blood cells were directly labelled with immunofluorescently tagged monoclonal CD14-PE/CD45-ECD/HLA-DR-PC5/CD16-PC7 antibodies (Beckman Coulter, USA). Respiratory burst intensity was evaluated in monocytes by measuring activity of lucigeninand luminol-dependent spontaneous and induced chemiluminescence. Monocytes were induced by using live suspension of MRSA/MSSA strains at a concentration of 106 CFU/mL. Results and discussion. While studying luminol-dependent monocyte activities after exposure to MRSA vs. MSSA, it was observed a 3.5-fold decreased curve square, whereas lucigenin-dependent chemiluminescence was increased by 6-fold. Compared to MSSA exposure, index of activation (IA) was decreased by 1.1-fold in response to MRSA exposure that was confirmed by lowered release of reactive oxygen species (ROS) from monocytes in response to MRSA exposure. Moreover, IRSS increased by 1.3-fold upon MRSA exposure. Examining monocyte oxygen-independent phagocytosis against MRSA vs. MSSA revealed significantly increased phagocytic number and concomitantly decreased phagocytic index. An evaluation of the activities of various monocyte subsets in response to MRSA vs. MSSA revealed increased phagocytic index by 1.5-fold for CD14lowCD16+ and CD14+CD16+ monocyte subsets as well as 3-fold for CD14+CD16– monocytes. Counts for all phagocytic subsets were decreased (1.4-, 1.5- and 4-fold for CD14lowCD16+, CD14+CD16+ and CD14+CD16– monocytes, respectively). To summarize, intensity of the respiratory burst was lowered upon MRSA exposure and percentage of monocyte subsets. Overall deficiency of superoxide anion production was observed in response to MRSA. In contrast, oxygen-independent event revealed phenotypic changes in frequency of peripheral blood monocytes upon MRSA exposure. We observed that CD14+CD16– classical monocytes were more rapidly activated. Conclusion. Thus, we concluded that CD14+CD16– monocytes became more rapidly activated but exhibited less effective phagocytosis, whereas CD14+CD16+ and CD14lowCD16+ monocytes were more slowly activated and demonstrated stronger phagocytic activity.


2021 ◽  
Author(s):  
Nerea Montes ◽  
Èlia Domènech ◽  
Silvia Guerrero ◽  
B&aacuterbara Oliv&aacuten-Bl&aacutezquez ◽  
Rosa Magallón-Botalla

Introduction: The objective of this study is to analyse the specific immune response against SARS-CoV-2 in those affected by Long Covid (LC), attributable to T cells (cell-mediated immunity) and to carry out a parallel analysis of the humoral response and lymphocyte typing. Methodology: Descriptive cross-sectional study of 74 patients with LC for at least 4 months since diagnosis. The collected data were: information on the COVID-19 episode and the persistent symptoms, medical history and a specific cell-mediated immunity to SARS-CoV-2 through flow cytometry, assessing the release of interferon-gamma (IFN-Ɣ) by T4 lymphocytes, T8 lymphocytes and NK cells. Descriptive and comparative analyses were carried out. Results: Patients with LC had negative serology for Covid-19 in 89% of cases but 96% showed specific cellular immunity to SARS-CoV-2 an average of 9.5 months after infection: 89% of this response corresponded to T8 lymphocytes, 58% to NK cells, and 51% to T4 lymphocyte (20% negligibly positive). Most of them had altered immune cell typing and we found that T4 lymphocyte counts were low in 34% of cases and NK cell high in 64%. Macrophage populations were detected in the peripheral blood of 7% of them. Patients displayed a higher percentage of illnesses related to ″abnormal″ immune responses, either preceding SARS-CoV-2 infection (43%) or following it in 23% of cases. Conclusion: The immune system appears to have an important involvement in the development of LC and viral persistence could be the cause or consequence of it. Further analysis with a control group should be performed.


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