• Journal of fish pathology
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• v.30 no.2
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• pp.97-105
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• 2017

Parasitic leeches could directly (through causing poor growth, anemia and wound in the fish) and indirectly (by predisposition of the fish to secondary bacterial and fungal infections) affects their hosts. In the present study, fishes that were attacked by leeches in natural and experimental environment were studied. Pathologic samples were obtained from damages at the site of leech bite, as well as kidney and liver of the fish. Histopathological examination revealed numerous lesions at the site of leech bite including tissue demolition, detachment at the site of leech bite in the epidermis of epithelial tissue in the skin, destructed nucleus in epithelial cells of the skin plus necrosis in the damaged skin and weak inflammatory penetration to acute necrotic damages along with piercing dermis layer. Pathologic lesions in the kidney included some changes such as proliferation by increasing glomerular cells and membrane cells in capillary vein of the kidney, blood cell necrosis in kidney with infiltration of white blood cells mainly mononuclear and less polymorphonuclear which are the symptoms of anemia due to blood feeding and sucking by leeches. There was also a chronic kidney infection probably originated from another part of body such as skin. Moreover, leeches caused hemorrhagic anemia due to blood consumption of the hosts, which led to observation of immature red blood cells. Also results showed that diseases induced by leeched in fish could be acute or chronic, which depends on size of fish, species of leech and severity of infection.

• Journal of Veterinary Clinics
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• v.37 no.2
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• pp.61-66
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• 2020

Nickel is a nutritionally essential trace element that plays an important role in the immune system of several animal species. The aim of this study was to examine the effect of nickel chloride on chemotactic activity of peripheral blood polymorphonuclear cells (PMNs) and whether this effect is associated with interleukin (IL)-8 and a nuclear factor-kappa B (NF-κB)-dependent pathway. Peripheral blood mononuclear cells (PBMCs) and PMNs were isolated by Percoll solution (Specific gravity; 1.080) and 1.5% dextran treatment, respectively. A modified Boyden chamber assay was used to measure the chemotactic activity of PMNs. The level of IL-8 in culture supernatant from PBMCs was measured by enzyme-linked immunosorbent assay (ELISA). Both of PBMCs and PMNs exhibited a low viability when cultured with concentration of greater than 1,000 μM of nickel chloride for 24 h. Thus, nickel chloride was used at concentration of 500 μM, which preserved cell viability. Treatment with nickel did not directly affect the chemotactic activity of PMNs. However, the chemotactic activity of PMNs was remarkably increased by culture supernatant from PBMCs treated with nickel chloride (500 μM) for 24 h. Recombinant porcine IL-8 polyclonal antibody (pAb) neutralized the enhancing effect on the chemotactic activity of PMNs by culture supernatant from PBMCs treated with nickel and this culture supernatant had higher IL-8 levels than the culture supernatant from untreated PBMCs. In addition, n-tosyll-phenylalanine chloromethyl ketone (TPCK), a NF-κB inhibitor, antagonized the enhancing effect on the chemotactic activity of PMNs by the culture supernatant from PBMCs treated with nickel. These results suggested that nickel stimulates porcine PBMCs to produce IL-8, which increases the chemotaxis of PMNs via NF-κB-dependent pathway.

• Journal of Veterinary Clinics
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• v.37 no.5
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• pp.249-254
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• 2020

Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.6
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• pp.1267-1272
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• 1998

In order to investigate the preventive and therapeutic of royal jelly on diabetes, the levels of blood glucose and serum lipids as well as the number of blood cells were determined in streptozotocin(STZ) diabetic rats. Rats were divided into seven groups. The RJ group was administered royal jelly and the STZ group was treated with streptozotocin to induce diabetes. To determine the preventive effect, diabetes was induced after administration of royal jelly for 2 weeks in group RS1/RS2. In group SR1/SR2 diabetic rats were administered royal jelly for 2 weeks to investigate the therapeutic effect. After 3 weeks, the body weight was reduced in STZ and SR1 groups and food intake was increased in the STZ, RS1 and SR1 groups. The blood glucose level was similar to the control group in the RJ, RS1 and RS2 groups and there was no effect in the other groups. The total lipid and triglyceride level were lower in the SR1 group as compared to STZ, and the total cholesterol level was higher in the STZ group. The index of atherogenesis was lower in the RJ and SR1 groups compared to the normal group. The number of red blood cells and hemoglobin was higher in the RJ and SR1 groups and the number of white blood cells was higher in the RJ and SR2 groups.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.4
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• pp.471-478
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• 2014

Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

• Environmental Mutagens and Carcinogens
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• v.21 no.1
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• pp.34-43
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• 2001

Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.14 no.1
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• pp.154-166
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• 2001

Naesosan(NSS) has been used in Oriental Medicine as a drug that treated carbuncle and cellulitis. So, the purpose of this Study was to investigate effects of NSS on the cytotoxicity of cancer cell lines and lymphocytes in vitro, proliferation of Ll210 cells and lymphocytes in L1210 cells transplanted mice, improvement of blood count in Ll210 cells transplanted mice, tumor weight and body weight in sarcoma-180 cells transplanted mice, survival prolongation in sarcoma-180 cells transplanted mice. We used NSS extract with freeze-dried, 8wks-old male mice(balb/c and ICR mouse $18{\pm}2g$). Ll210 cell lines, and sarcoma-180 cell lines for this Study, The proliferation of cells was tested using a colorimetric tetrazoliun assay(MTT assay). The results of this Study were obtained as follows ; 1. NSS showed significantly cytotoxicitic effects of cancer cell lines, did not show cytotoxicitic effects of lymphocytes. 2, Proliferation of lymphocytes in L1210 cells transplanted mice did not effects by NSS. 3. NSS inhibited significantly the proliferation of L1210 cells in L1210 cells transplanted mice. 4. NSS improved significantly the blood count in Ll210 cells transplanted mice. 5. NSS increased significantly th body weight in sarcoma-180 cells transplanted mice. 6. NSS dereased significantly the tumor weight in sarcoma-180 cells transplanted mice. 7. NSS prolonged significantly the survival time in sarcoma-180 cells transplanted mice.

• Development and Reproduction
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• v.23 no.2
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• pp.139-148
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• 2019

Rising of water temperature due to global warming is a great concern to aquaculturists and fishery biologists. Hence, the present study aimed to investigate the effects of high water temperature on juvenile red spotted grouper, Epinephelus akaara based on the evaluation of stress responses in blood. E. akaara juveniles were exposed to different thermal conditions ($25^{\circ}C$, $28^{\circ}C$, $31^{\circ}C$, and $34^{\circ}C$) for 6 weeks following 2 weeks of acclimation at $25^{\circ}C$. Blood cell morphology and number were examined at three sampling points (2, 7, and 42 days) from a total of 180 fish. Major erythrocytic cellular abnormalities (ECA) observed in blood smears of thermally stressed groups ($31^{\circ}C$ and $34^{\circ}C$) after 6 weeks were echinocytes, teardrop-like cells, swollen cells and vacuolated cells. Both red and white blood cell number (RBC and WBC) were significantly (p<0.05) elevated in $31^{\circ}C$ and $34^{\circ}C$ group after 6 weeks thermal exposure. Differential leucocytes number showed significant increases in neutrophil (N) and decreases in lymphocytes (L) in the highest temperature ($34^{\circ}C$). Different N:L ratio was observed at different thermal conditions which can be used as a reliable alternative to measure stress response. Taken together, these results suggest that higher temperature ($31^{\circ}C$ and $34^{\circ}C$) can interfere the immune system of red spotted grouper by altering the blood cell morphology and number.

• Asian-Australasian Journal of Animal Sciences
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• v.7 no.4
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• pp.459-466
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• 1994

Primordial germ cells (PGCs) in aves are the progenitor cells for the gametes. These cells first appear in the epiblast (Eyal-Giladi et al.. 1981). Then translocate and concentrate to endoderm of germinal crescent area in the junction of the area opaca and area pellucida lateral to the primitive streak in stage 4 through 7. They separate from the endoderm, temporarily circulate via the blood vascular system, leave the blood vessels, and finally settle down in the gonadal anlagen at stage 20-24 where they rapidly proliferate to form germ cells. Recently, several attempts have been made to introduce foreign gene into the avian genome to form a transgenic chicken. The stem cells most readily available as vehicles for genetic manipulation of germline in avian species are the PGCs. PGCs have recently been manipulated genetically and used successfully as a vector for gene transfer.

• Journal of Microbiology and Biotechnology
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• v.18 no.10
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• pp.1709-1716
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• 2008

The porcine reproductive and respiratory syndrome Virus (PRRSV) is an infectious disease that causes abortions and respiratory disorders in swine. In this study, the interaction between PRRSV and porcine dendritic cells generated from $CD14^{+}$ monocytes in the presence of GM-CSF and IL-4 was examined. As a result, it was shown that immature and mature dendritic cells can be productively infected with PRRSV. When the expression of surface MHC molecules on infected dendritic cells was determined, MHC classes I and II were found to be downregulated when compared with un infected dendritic cells. With the exception of the IL-4 and IFN-$\gamma$ cytokines, the induction of the IL-10, IL-12, and TNF-$\alpha$ cytokines all increased in dendritic cells infected with PRRSV. A mixed lymphocyte reaction showed that peripheral blood mononuclear cells cocultured with PRRSV-infected dendritic cells were less stimulated than peripheral blood mononuclear cells cocultured with dendritic cells treated with PBS, LPS, or UV-inactivated PRRSV. Therefore, these results suggest that PRRSV would appear to modulate the immune stimulatory function of porcine dendritic cells.