In vitro effects of plant and mushroom extracts on immunological function of chicken lymphocytes and macrophages

1.?The present study was conducted to examine the effects of organic extracts from milk thistle (Silybum marianum), turmeric (Curcuma longa), reishi mushroom (Ganoderma lucidum), and shiitake mushroom (Lentinus edodes) on innate immunity and tumor cell viability.

2.?Innate immunity was measured by lymphocyte proliferation and nitric oxide production by macrophages, and the inhibitory effect on tumor cell growth was assessed using a non-radioactive assay. For measuring the cytokine levels in the HD11 macrophages which were treated with extracts of turmeric or shiitake mushroom, the levels of mRNAs for interferon-α (IFN- α), interleukin-1β (IL-1β), IL-6, IL-12, IL-15, IL-18, and tumor necrosis factor superfamily 15 (TNFSF15) were quantified by real time RT-PCR.

3.?In vitro culture of chicken spleen lymphocytes with extracts of milk thistle, turmeric, and shiitake and reishi mushrooms induced significantly higher cell proliferation compared with the untreated control cells. Stimulation of macrophages with extracts of milk thistle and shiitake and reishi mushrooms, but not turmeric, resulted in robust nitric oxide production to levels that were similar with those induced by recombinant chicken interferon-γ. All extracts uniformly inhibited the growth of chicken tumor cells in vitro at the concentration of 6·3 through 100 µg/ml. Finally, the levels of mRNAs encoding IL-1β, IL-6, IL-12, IL-18, and TNFSF15 were enhanced in macrophages that were treated with extracts of turmeric or shiitake mushroom compared with the untreated control.

4.?These results document the immunologically-based enhancement of innate immunity in chickens by extracts of plants and mushrooms with known medicinal properties in vitro. In vivo studies are being planned to delineate the cellular and molecular mechanisms responsible for their mechanism of action.     

Infectious aetiologies of mastitis on Korean dairy farms during 2008

The objective of this study was to determine the frequency of different mastitis pathogens in 1255 milk samples collected from 368 lactating cows on 24 dairy farms in Korea during 2008. The proportion of cows and quarter milk samples having SCC ? 200,000 cells/ml, an indicator of udder infection, was 54.3% (200/368 cows) and 35.5% (446/1255 samples), respectively. Of the 446 milk samples subjected to bacteriological examination, 16.5% (74) showed no bacterial growth and 3.5% (16) were contaminated. In total, 356 of 1255 (28.3%) samples were bacteriologically positive, from which 415 bacteria were isolated. The most frequently isolated pathogen was coagulase-negative staphylococci (40.7%), followed by Gram-negative bacteria (19.5%) other than Escherichia coli, Staphylococcus aureus (12.2%), Streptococcus uberis (5.3%), Enterococcus spp. (4.8%), E. coli (4.5%), and environmental streptococci (3.1%) other than S. uberis. This study demonstrates that environmental pathogens were the vast majority of bacteria isolated from mastitic bovine milk samples in Korea.     

Ameliorative Effects of Melatonin against Hydrogen Peroxide‐Induced Oxidative Stress on Boar Sperm Characteristics and Subsequent In Vitro Embryo Development

Melatonin, the major secretory product of the pineal gland, scavenges a variety of reactive oxygen and nitrogen species in vivo and in vitro, indicating that melatonin is a potent function as an antioxidant. The objective of this study was to investigate the effect of melatonin in the presence or absence of hydrogen peroxide (H₂O₂) on sperm characteristics (motility, viability, survival rate, membrane integrity, lipid peroxidation (LPO) and mitochondria activity) and also to examine the developmental rates to the blastocysts stage of porcine oocytes fertilized in vitro with semen treated with or without melatonin (100 nm ) in the presence or absence of H₂O₂ (250 μm ). The sperm were treated with melatonin in the presence or absence of H₂O₂ for 3, 6, 9 and 12 h at 37°C and then analysed for the sperm characteristics. The porcine embryos were produced by in vitro maturation and in vitro fertilization (IVM/IVF) using semen treated with or without melatonin (100 nm ) in the presence or absence of H₂O₂ (250 μm ) for 6 h. The semen characteristics, including motility, viability, survival rate, membrane integrity and mitochondria activity, were higher in the groups that were treated with melatonin in comparison to other groups, irrespective of incubation periods. Malondialdehyde levels in control, melatonin and melatonin + H₂O₂ groups were lower than H₂O₂ only group. A positive correlation was shown among motility, viability, survival rate and membrane integrity, but a negative correlation was observed between LPO and the other evaluation methods. The developmental rates to blastocysts of IVM/IVF porcine oocytes fertilized by semen treated with melatonin were significantly increased compared with any other groups, with the cell number of blastocysts shown to have a similar trend to the developmental rates. These results demonstrate that melatonin can improve the semen characteristics during in vitro storage and support the developmental ability of IVM/IVF embryos in pigs.     

Antioxidative Effects of Astaxanthin against Nitric Oxide‐Induced Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct Epithelial Cell and the Developmental Competence of Bovine IVM/IVF Embryos

The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl‐2, Caspase‐3 and Bax) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μm ) in the presence or absence of sodium nitroprusside (SNP, 1000 μm ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μm ) lowered, while astaxanthin addition (50–500 μm ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μm ) gradually recovered according to the increase in astaxanthin additions (100–500 mm ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μm astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes.     

64-Channel multi-detector row CT angiographic evaluation of the micropigs for potential living donor lung transplantation

Micropigs are the most likely source animals for xenotransplantation. However, an appropriate method for evaluating the lung of micropigs had not been established. Therefore, this study was performed to evaluate the feasibility of 64-channel multi-detector row computed tomography (MDCT) to measure the diameter of the pulmonary arteries and the lung volume in micropigs. The mean diameters of the trachea, and left and right bronchi were 1.6 ± 0.17, 1.18 ± 0.14, and 1.1 ± 0.11 cm, respectively. The mean diameters of the main, right, and left pulmonary arteries were 1.38 ± 0.09, 1.07 ± 0.26, and 0.98 ± 0.13 cm and the diameters of right, left, and common inferior pulmonary veins were 0.97 ± 0.20, 0.76 ± 0.20, and 1.99 ± 0.26 cm, respectively. The mean lung volume was 820.3 ± 77.11 mL. The data presented in this study suggest that the MDCT may be a noninvasive, rapid, and accurate investigational method for pulmonary anatomy in living lung donors.     

Anti-inflammatory effects of a Houttuynia cordata supercritical extract

Anti-inflammatory effects of Houttuynia cordata supercritical extract (HSE) were investigated in a carrageenan-air pouch model. HSE (200 mg/kg, oral) suppressed exudation and albumin leakage, as well as inflammatory cell infiltration. Dexamethasone (2 mg/kg, i.p.) only decreased exudation and cell infiltration, while indomethacin (2 mg/kg, i.p.) reduced exudate volume and albumin content. HSE lowered tumor-necrosis factor (TNF)-α and nitric oxide (NO), as well as prostaglandin E₂ (PGE₂). Dexamethasone only reduced TNF-α and NO, while indomethacin decreased TNF-α and PGE₂. The suppressive activity of HSE on NO and PGE₂ production was confirmed in RAW 264.7. These results demonstrate that HSE exerts anti-inflammatory effects by inhibiting both TNF-α-NO and cyclooxygenase II-PGE₂ pathways.     

Arsenic accumulation by rice grown in soil treated with roxarsone

Poultry litter is widely used as a fertilizer for lowland rice in Taiwan and China. However, the organic‐arsenic compound roxarsone (additive of poultry feed) in poultry litter can be absorbed by the plants and the resulting arsenic (As) contamination may pose a serious threat to human health. This study used various amounts of poultry litter contaminated with roxarsone in pot experiments to evaluate the effect of roxarsone on rice agronomic parameters and the bioaccumulation of total and inorganic As in rice‐plant tissues. Rice‐grain yield decreased significantly with increasing As content of the soil, and the critical threshold that killed rice was 200 mg roxarsone (kg soil)^–1. The As concentrations in root, straw, leaf, husk, and grain increased with increasing soil As (p < 1%). At 100 mg roxarsone per kg of soil, the As concentration in the rice grain exceeded the statutory permissible limit of 1.0 mg As (kg dry weight)^–1 and at 25 mg roxarsone (kg soil)^–1, the inorganic As concentrations in grains exceeded the statutory limit of 0.15 mg of inorganic As kg^–1 in China. For all treatments, the As concentrations in various plant tissues at maturity follow the order: root > stem > leaf > husk > grain. Arsenite was the predominant species in root, straw, and grain, while arsenate was the predominant species in leaf and husk. No significant difference existed between the amounts of arsenite and arsenate when various amounts of poultry litter were applied. This result illustrates that large amounts of added roxarsone are not only toxic to rice but also accumulate in grains in the inorganic As forms, potentially posing a threat to human health via the food chain.     

Assessing the risk of recurrence of porcine epidemic diarrhea virus in affected farms on Jeju Island,South Korea

BackgroundPorcine epidemic diarrhea virus (PEDV) is a swine enteropathogenic coronavirus that has devastated the swine industry in South Korea over the last 30 years. The lack of an effective method to control the endemics has led to a surge in PEDV recurrences in affected farms throughout the country.ObjectivesIn the first step toward establishing systematic monitoring of and active control measures over the swine populations, we constructed an assessment model that evaluates the status of (1) biosecurity, (2) herd immunity, and (3) virus circulation in each of the PEDV-infected farms.MethodsA total of 13 farrow-to-finish pig farms with a history of acute PEDV infection on Jeju Island were chosen for this study. The potential risk of the recurrence in these farms was estimated through on-site data collection and laboratory examination.ResultsOverall, the data indicated that a considerable number of the PEDV-infected farms had lax biosecurity, achieved incomplete protective immunity in the sows despite multi-dose vaccination, and served as incubators of the circulating virus; thus, they face an increased risk of recurrent outbreaks. Intriguingly, our results suggest that after an outbreak, a farm requires proactive tasks, including reinforcing biosecurity, conducting serological and virus monitoring to check the sows’ immunity and to identify the animals exposed to PEDV, and improving the vaccination scheme and disinfection practices if needed.ConclusionsThe present study highlights the significance of coordinated PEDV management in infected farms to reduce the risk of recurrence and further contribute towards the national eradication of PEDV.     

Preparation of poly(vinyl alcohol)/ZrO2 composite nanofibers via co-axial electrospinning with higher ZrO2 particle content

Poly(vinyl alcohol) (PVA)/zirconium oxide (ZrO₂) composite nanofibers with a skin-core structure were prepared and the effect of ZrO₂ particle content on uniform web formation was investigated. The optimized polymer concentration, tip to collector distance, and applied voltage for electrospinning were 11 wt%, 12 cm, and 20 kV, respectively. Skin-core PVA/ZrO₂ composite nanofibers containing up to 12 wt% ZrO₂ were successfully prepared, but it was difficult to obtain PVA/ZrO₂ composite nanofiber webs via conventional electrospinning. Increasing the amount of ZrO₂ caused the morphology of the PVA/ZrO₂ composite nanofibers to become a non-uniform nanoweb with irregular nanofiber diameters. While it was difficult to obtain a uniform nanofiber web containing a content of ZrO₂ over 6 wt% for conventional electrospinning, a more uniform nanofiber web could be obtained at up to 9 wt% ZrO₂ using a skin-core dual nozzle. More uniform webs could also be obtained when ZrO₂ was in the skin rather than the core.