scholarly journals Isolation and identification of Bengle (Zingiber cassumunar roxb) as a stimulant in phagocytic activity of macrophages

10.5219/1238 ◽  
2020 ◽  
Vol 14 ◽  
pp. 328-335
Author(s):  
Muhamad Fauzi Ramadhan ◽  
Nurkhasanah Mahfudh ◽  
Nanik Sulistyani

Immunomodulators are pharmacological agents that modify or regulate the immune system through stimulating the functioning of the immune system and, at the same time, inhibiting excessive immune responses. This study was conducted to determine the active compound in Z. cassumunar that is responsible for increasing the immune system based on the parameters of phagocytic activity. The isolation method began with fractionation, which involved extraction with ethanol and successive fractionation with hexane and chloroform. Z. cassumunar extract, hexane fraction, and chloroform fraction were tested on mice macrophage cells for their phagocytic functions. The phagocytic activity of macrophages was measured by active phagocytic cells (averagely 39.194 ±1.597, 27.923 ±2.941, and 62.090 ±6.947) and phagocytic index (in a row, averagely 47.513 ±2.844, 41.129 ±7.195, and 101.527 ±10.555). The results showed that the Z. cassumunar extract,  hexane fraction, and chloroform fraction exhibited more significant phagocytic activities of macrophages (p <0.05) compared with the normal group. Since the chloroform fraction showed the best result, this fraction was further separated by column chromatography. This procedure yielded five sub-fractions, namely F1, F2, F2C, F3, and F4. Based on the phagocytic activity testing, the results were as follows: (1) the active phagocytic cells of F1, F2, F2C, F3 and F4 were 18.860 ±3.191, 27.077 ±4.482, 15.749 ±3.026, 64.333 ±1.780, and 44.943 ±2.944, respectively, and (2) the phagocytic indices were 30.0249 ±3.4231, 44.5969 ±8.3646, 24.5597 ±5.4487, 102.7447 ±1.0806, and 76.5007 ±4.7293. Because F3 produced the best result, this subfraction was then identified using 1H-NMR and 13C-NMR. The identification results showed that F3 was (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-ol as an active compound.

Author(s):  
L. Bondarenko

The immune system is central to ensuring the consistency of the body's homeostasis. The state of the body's natural resistance is determined by a set of non-specific protective mechanisms. Lymphocytes and phagocytes are actively involved in maintaining immunity. Lymphocytes recognize the antigens of pathogenic microorganisms, and phagocytes absorb and destroy the pathogens themselves. During the weaning of piglets from sows there is a decrease in the protective forces of their body. During this period, the natural resistance of the piglets is reduced due to the stressful situation caused by changing conditions of confinement, the transition to full feed and lack of sows. The immune system of weaning pigs is relatively weak, so when exposed to environmental and technological stressors, they become susceptible to various diseases. The use of probiotic drugs stimulates the activity of the immune system, prevents stress and immunodeficiency. One of these probiotics is the probiotic of domestic production Protecto-active. It w observed the the influence of the probiotic Protecto-active on the indices of nonspecific resistance of the young pigs organism to the growth. An increase in bactericidal activity of blood serum by 12.10% (P <0.05) and lysozyme activity of blood in the piglets of the experimental group was increased by 3.71% compared to control, which indicates the activation of the body's defenses and the increase in adaptive capacity. An important step in the study of the influence of the probiotic Protekto-active on the state of the immune system is to determine the phagocytic activity of neutrophils, phagocytic index and phagocytic number. In the experimental group of piglets that were fed the probiotic Protecto-active, we found an increase in leukocyte phagocytic activity by 9.0% (P <0.001), a phagocytic index by 51.7% (P<0.001) and a phagocytic number by 24.8% ( P <0.01) compared with the control group. Thus, using a probiotic Protecto-active, all indicators of phagocytosis increase: the number of phagocytes increases, their ability to capture microorganisms and increases their digestive capacity, it increases the bacterial and lysozyme activity of blood serum, which is positively reflected in the immunobiosity. Key words: probiotic, phagocytosis, phagocytic index, phagocytic number, phagocytic activity of leukocytes, cellular immunity, piglets


2020 ◽  
Vol 3 (3) ◽  
pp. 20-24
Author(s):  
W. S. Said ◽  
V. V. Stybel ◽  
B. V. Gutyj ◽  
O. B. Pryima ◽  
A. G. Sobolta ◽  
...  

The aim of the study was to determine the state of the dogs' immune system in experimental toxocariasis. The work was performed during 2017-2020 at the Department of Parasitology and Ichthyopathology in Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv. The 12 dogs aged 2 to 4 months were used for research studies. Two groups were formed of six animals in each: control and experimental. Puppies of the experimental group (E) were infected with the causative agent of toxocariasis at a dose of 5000 invasive eggs of Toxocara canis per kg of body weight. The control puppies were clinically healthy. At the clinical display of toxocariasis invasion at dogs the cellular, humoral and nonspecific links of the immune system are suppressed and the secondary immunodeficiency comes. Suppression of cellular immunity was accompanied by a decrease in the T- and B-lymphocytes count in the blood of dogs of the (E) group, which indicates the suppression of the lymphoid immune system and a reduction in the resistance of animals. Along with the fall in cellular immunity in infected puppies, suppression of the nonspecific immune system was established, which is manifested by a decline in the phagocytic activity of neutrophils and a decrease in the phagocytic index. Thus, on the 15th and 20th days of the test, the phagocytic activity of neutrophils reduced to 1.6 and 3.9 %, while the phagocytic index – to 5.4 and 6.9 %, respectively. In the study of antimicrobial activity of serum of infected dogs, T. canis found inhibition of bactericidal and lysozyme activity, which reflects the suppression of the physiological state of the humoral immune system of animals with the development of toxocariasis. On the 25th day of the research, a deccline in BABS to 24.0 ± 2.7 %, while in the (C) group of dogs this figure was 30.6 ± 1.9 %. The lowest LABS was on the 25th day of the (E) in animals that were experimentally infected with the pathogen toxocariasis, where compared with the (C) group, this figure decreased by 6.0 %, respectively. The high level of CIC in the serum of dogs infected with T. canis indicates the suppression of the immune system of their body due to the accession of specific antibodies to the products of metabolism of toxocara, which act as antigens.


1996 ◽  
Vol 151 (1) ◽  
pp. 113-118 ◽  
Author(s):  
M Sakai ◽  
M Kobayashi ◽  
H Kawauchi

Abstract The activation of rainbow trout (Oncorhynchus mykiss) phagocytic cells by chum salmon GH, prolactin (PRL) and somatolactin (SL) was investigated in vitro. Rainbow trout kidney leucocytes were cultured in RPMI 1640 medium containing 1, 10, 50 or 100 ng of each hormone/ml and the production of superoxide anion was measured using reduction of nitroblue tetrazolium (NBT) and ferricytochrome C. Macrophages incubated with 10–100 ng GH or PRL/ml showed significantly enhanced production of superoxide anion in both the NBT and ferricytochrome C tests compared with control macrophages (without hormone). SL did not induce enhanced production of superoxide anion in macrophages. The phagocytic cells treated with GH or PRL also showed increased phagocytic activity and phagocytic index. However, the cells treated with SL showed no enhancement of phagocytic activity or index. These results indicate that GH and PRL stimulate macrophages in vitro. Journal of Endocrinology (1996) 151, 113–118


Author(s):  
Ida N Jamal ◽  
Reiny A Tumbol ◽  
Remy E.P Mangindaan

Motile Aeromonas Septicaemia disease (MAS) attacking tilapia has increased in recent years as a consequence of intensive aquaculture activities, which led to losses in aquaculture industry. The agent causing MAS disease is Aeromonas hydrophila. The disease can be controlled with the β-glucan. As immunostimulants, β-glucans can also increase resistance in farmed tilapia. Studies on the use of β-glucan extracted from baker's yeast Saccharomyces cerevisiae was intended to evaluate the non-specific immune system of tilapia that were challenged with Aeromonas hydrophila. The method used was an experimental method with a completely randomized design consisting of four treatments with three replicats. The dose of β-glucan used as treatments were 0 mg.kg-1 fish (Control), 5 mg.kg-1 fish (B), 10 mg.kg-1 fish (C) and 20 mg.kg-1 fish (D), each treatment as injected three times at intervals of 3 days, the injection volume of 0.5 ml/fish for nine days and resistance surveillance for seven days. The results showed that the difference in the amount of β-glucan and the frequency of the injected real influence on total leukocytes, phagocytic activity and resistance. Total leukocytes, phagocytic activity and resistance to treatment was best achieved by the administration of C a dose of  10 mg.kg-1 of the fish© Penyakit Motil Aeromonas Septicaemia (MAS) yang menyerang ikan nila mengalami peningkatan selama beberapa tahun terakhir sebagai konsekuensi dari kegiatan akuakultur intensif, yang menyebabkan kerugian dalam industri budidaya. Agen utama penyebab penyakit MAS adalah Aeromonas hydrophila. Untuk mengendalikan penyakit tersebut dapat dilakukan dengan pemberian β-glukan. Sebagai imunostimulan, β-glukan juga dapat  meningkatkan resistensi pada ikan nila yang dibudidayakan. Pengkajian mengenai pemanfaatan β-glukan yang diekstrak dari ragi roti Saccharomyces cerevisiae dimaksudkan untuk menguji sistem imun non spesifik ikan nila yang diuji tantang dengan bakteri Aeromonas hydrophila. Metode yang digunakan yaitu metode eksperimen dengan rancangan acak lengkap yang terdiri dari empat perlakuan dan tiga ulangan. Dosis β-glukan  yang digunakan sebagai perlakuan sebesar 0 mg.kg-1 ikan (Kontrol), 5 mg.kg-1 ikan (B), 10 mg.kg-1 ikan (C) dan 20 mg.kg-1 ikan (D), masing-masing perlakuan diinjeksi sebanyak 3 kali dengan interval waktu 3 hari selama 9 hari, volume injeksi 0,5 mL/ekor ikan dan pengamatan resistensi selama tujuh hari. Hasil penelitian menunjukkan perbedaan jumlah β-glukan dan frekuensi pemberian yang diinjeksikan memberikan pengaruh nyata terhadap total leukosit, aktivitas fagositosis dan resistensi. Total leukosit, aktivitas fagositosis dan resistensi terbaik dicapai pada perlakuan C dengan dosis 10 mg.kg-1 ikan©


Author(s):  
Ch. N. Sambyla ◽  
N. M. Bessonova ◽  
R. B. Chysyma

The Republic of Tyva is a region in the geographical center of Asia, which located at the junction of the Siberian taiga and Central Asian desert-steppe landscapes, in a wide band of mountains and intermountain plains. The mountain-forest area of Tyva has long been considered a natural habitat for antler deer, one of which is the maral (Cervus elaphus sibiricus). In order to restore maral breeding and increase the number of marals in the former limits, marals of the Altai-Sayan breed have been imported to the Republic of Tyva from the Republic of Altai. The preservation of productive traits, reproductive function and the realization of the genetic potential of animals introduced to new climatic conditions largely depends on the ability of these animals to adapt to existing environments. We have assessed in this paper the natural resistance of the marals of Altai-Sayan breed during introduction in the Tyva Republic in comparison with the indicators of the marals have been bred in the Altai Republic. The researches have been carried out in 2019. The research material was blood samples of marals of Altai-Sayan breed imported to the Republic of Tuva (n=27) and marals of the same breed bred in the Republic of Altai (n=17). Studies have shown some deviations in the blood leukogram of imported marals, such as a decrease in the number of eosinophils and rod nuclear neutrophils (P < 0,001), the increase in the number of segmented nuclear neutrophils and lymphocytes (P < 0,05). The number of monocytes have been increased in 18,6 times compared to the Altai marals. The change in the number of monocytes exceeded the physiological norm by 24,7 %. The indicator of adaptation evaluation in imported marals had higher values (6,8), which were in 1,7 times higher than in marals bred in the Altai Republic (4,1), which indicates the intensity of adaptive mechanisms in imported animals during adaptation. Analysis of phagocytic activity and phagocytic index revealed intensive phagocytosis in imported marals: phagocytic activity – by 12,4 % (P < 0,05), phagocytic index – by 5,1 %, and the increase in the content of lysosomal and cationic proteins by 12,8 % (P < 0,05). Analysis of the bactericidal activity of blood serum has shown its lower level in imported animals (45,97±1,36 %), compared with marals of the same breed bred in the Altai Republic (52,19±2,15 at P < 0,05). Thus, according to most indicators of natural resistance marals of Altai-Sayan breed imported to Tyva have a fairly high level of natural protection, which indicates that they can be adapted to the natural climatic and feeding conditions of the Republic of Tyva.


2018 ◽  
Vol 18 (2) ◽  
pp. 401-416 ◽  
Author(s):  
Ewelina Kulak ◽  
Iwona Sembratowicz ◽  
Anna Stępniowska ◽  
Katarzyna Ognik

AbstractThe purpose of the study was to verify the hypothesis that there are doses of silver nanoparticles (Ag-NPs) that improve immune defence in chickens without compromising their health. To verify this hypothesis, an attempt was made to determine which doses of Ag-NPs (5 nm) consumed by chickens as a result of administration of hydrocolloids during varying time periods at a concentration of 5 or 10 mg Ag/l stimulate immune defence. The experiment was performed on 296 chickens assigned to 3 treatments. Chickens from the control treatment received drinking water without experimental additives. Chickens from the T-5 and T-10 treatments received a hydrocolloid of Ag-NPs at a concentration of 5 mg/l (treatment T-5) or 10 mg/l (treatment T-10) from their second week of life. Blood for analysis was collected at the age of 42 days from 8 birds per treatment. Ceruloplasmin (Cp), leukocyte count (WBC), erythrocyte sedimentation rate (ESR), interleukin IL-6, immunoglobulins IgA, IgY, phagocytic cells (% PC), phagocytic index (PI), nitroblue tetrazolium reduction (NBT), and lysozyme content in the blood was determined. Application of Ag-NPs at a concentration of 5 mg/l or 10 mg/l in the dose range of 2.87-12.25 mg/bird (administration of Ag-NPs in concentration 5 mg/l in weeks: 2; 2 and 3; 2 and 4; 2 and 5; 2 and 6 or concentration 10 mg/l in week 2) resulted in an immunostimulatory effect expressed as an increase in heterophil respiratory burst and an increased concentration of lysozyme. Higher doses of Ag-NPs exerted a pro-inflammatory effect, as indicated by elevated levels of IL-6 and ceruloplasmin, as well as a high ESR. They also stimulated B lymphocytes to produce IgA and IgA immunoglobulins.


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.


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