Isolation and Characterization of Efficient Active Compounds Using High-Performance Centrifugal Partition Chromatography (CPC) from Anti-Inflammatory Activity Fraction of Ecklonia maxima in South Africa

Ecklonia maxima is a brown seaweed, which is abundantly distributed in South Africa. This study investigated an efficient approach using high-performance centrifugal partition chromatography (HPCPC), which has been successfully developed for the isolation and purification of phlorotannins, eckmaxol, and dieckol from the ethyl acetate fraction of E. maxima (EEM). We evaluated EEM for its inhibitory effect against lipopolysaccharide (LPS)-induced inflammatory responses in zebrafish embryos. The separation of eckmaxol and dieckol from samples of EEM using HPCPC was found to be of high purity and yield under an optimal solvent system composed of n-hexane:ethyl acetate:methanol:water (2:7:3:7, v/v/v/v). To evaluate the anti-inflammatory efficacy of EEM containing active compounds, zebrafish embryos exposed to LPS were compared with and without EEM treatment for nitric oxide (NO) production, reactive oxygen species (ROS) generation, and cell death two days after fertilization. These evaluations indicate that EEM alleviated inflammation by inhibiting cell death, ROS, and NO generation induced by LPS treatment. According to these results, eckmaxol and dieckol isolated from brown seaweed E. maxima could be considered effective anti-inflammatory agents as pharmaceutical and functional food ingredients.     

Antihypertensive Effects of IGTGIPGIW Peptide Purified from Hippocampus abdominalis: p-eNOS and p-AKT Stimulation in EA.hy926 Cells and Lowering of Blood Pressure in SHR Model

The aim of this study was to assess the potential hypertensive effects of the IGTGIPGIW peptide purified from Hippocampus abdominalis alcalase hydrolysate (HA) for application in the functional food industry. We investigated the antihypertensive effects of IGTGIPGIW in vitro by assessing nitric oxide production in EA.hy926 endothelial cells, which is a major factor affecting vasorelaxation. The potential vasorelaxation effect was evaluated using 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate, a fluorescent stain. IGTGIPGIW significantly increased the expression of endothelial-derived relaxing factors, including endothelial nitric oxide synthase and protein kinase B, in EA.hy926 cells. Furthermore, oral administration of IGTGIPGIW significantly lowered the systolic blood pressure (183.60 ± 1.34 mmHg) and rapidly recovered the diastolic blood pressure (143.50 ± 5.55 mmHg) in the spontaneously hypertensive rat model in vivo. Our results demonstrate the antihypertensive activity of the IGTGIPGIW peptide purified from H. abdominalis and indicate its suitability for application in the functional food industry.     

STRAIN ELASTOGRAPHY USING DOBUTAMINE‐INDUCED CAROTID ARTERY PULSATION IN CANINE THYROID GLAND

Thyroid disease is common in dogs and conventional ultrasonography is a standard diagnostic test for diagnosis and treatment planning. Strain elastography can provide additional information about tissue stiffness noninvasively after applying external or internal compression. However, natural carotid artery pulsations in the canine thyroid gland are too weak to maintain sufficient internal compression force. The objective of the present study was to describe the feasibility of strain elastography for evaluating the canine thyroid gland and the repeatability of dobutamine‐induced carotid artery pulsation as an internal compression method. In seven healthy Beagle dogs, strain on each thyroid lobe was induced by external compression using the ultrasound probe and internal compression using carotid artery pulsation after dobutamine infusion. The thyroid appeared homogeneously green and the subcutaneous fat superficial to the thyroid lobe appeared blue. Strain values and strain ratios did not differ among dogs or between the left and right lobes. Interobserver repeatability was excellent for both compression methods. Intraobserver repeatability of the strain ratio measured using the carotid artery pulsation method (intraclass coefficient correlation = 0.933) was higher than that measured using the external compression method (0.760). Mean strain values of thyroid lobes for the external compression method (142.93 ± 6.67) differed from the internal method (147.31 ± 8.24; P < 0.05). Strain ratios between the two methods did not differ. Strain elastography was feasible for estimating thyroid stiffness in dogs. Carotid artery pulsation induced by dobutamine infusion can be used for canine thyroid strain elastography with excellent repeatability.     

Chitosan as an edible invisible film for quality preservation of herring and atlantic cod

The effect of chitosan with different molecular weights as coatings for shelf-life extension of fresh fillets of Atlantic cod (Gadus morhua) and herring (Clupea harengus) was evaluated over a 12-day storage at refrigerated temperature (4 +/- 1 degrees C). Three chitosan preparations from snow crab (Chinoecetes opilio) processing wastes, differing in viscosities and molecular weights, were prepared; their apparent viscosities (360, 57, and 14 cP) depended on the deacetylation time (4, 10, and 20 h, respectively) of the chitin precursor. Upon coating with chitosans, a significant (p < or = 0.05) reduction in relative moisture losses of 37, 29, 29, 40, and 32% was observed for cod samples coated with 360 cP chitosan after 4, 6, 8, 10, and 12 days of storage, respectively. Chitosan coating significantly (p < or = 0.05) reduced lipid oxidation as displayed in peroxide value, conjugated dienes, 2-thiobarbituric acid reactive substances and headspace volatiles, chemical spoilage as reflected in total volatile basic nitrogen, trimethylamine, and hypoxanthine, and growth of microorganisms as reflected in total plate count in both fish model systems compared to uncoated samples. The preservative efficacy and the viscosity of chitosan were inter-related; the efficacy of chitosans with viscosities of 57 and 360 cP was superior to that of chitosan with a 14 cP viscosity. Thus, chitosan as edible coating would enhance the quality of seafoods during storage.     

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.     

An unusual case of concomitant infection with chicken astrovirus and group A avian rotavirus in broilers with a history of severe clinical signs

A molecular study of intestinal samples from 21 broiler flocks with a history of enteritis revealed that 23.8% and 14.3% were positive for chicken astrovirus (CAstV) and avian rotavirus (ARV), respectively. CAstV and group A ARV were simultaneously detected in only one broiler flock. Birds in this group developed the significant intestinal lesions characterized by frothy contents, paleness, and thin intestinal walls. In this report we present an unusual case of runting stunting syndrome (RSS) with a history of high mortality and growth retardation in broiler chickens. We also make the first identification of CAstV and group A ARV in broiler chickens in Korea.     

Effects of chilled drinking water on the performance of lactating sows and their litters during high ambient temperatures under farm conditions

Heat stress has detrimental effects on lactating sows and their litters, including decreased feed intake and milk production in sows and decreased weight gain in their offspring. It also increases respiration rate, rectal temperature, and weaning to estrus interval in sows. This study was conducted to investigate the effects of chilled water (CW) on the performance of lactating sows and their litters exposed to high ambient temperatures for 21 days. Ninety multiparous sows (Yorkshire × Landrace; parity range: 2 to 5) and their litters (Yorkshire × Landrace × Duroc) were divided into 3 blocks (30 each). Sows and their litters within each block were randomly assigned to one of the three treatment groups (n = 10 each). Each group was provided with drinking water at 10, 15, or 22 °C (control) under farm conditions where ambient temperature above 25 °C was consistent during the experimental period. Sows on CW (both 10 and 15 °C) consumed more feed and water than did control (P < 0.01) and higher estimated milk production compared with control (P < 0.01), but had lower rectal temperature and respiration rate compared with control (P < 0.01). Mean weaning weight (P < 0.01) and average daily gain (P < 0.01) of litters in CW groups were higher than those in control group. There were no differences in the measured variables, with the exception of the respiration rates, between the two CW treatment groups. These results suggest that CW may improve the performance of sows and their litters exposed to high ambient temperatures and the 15 °C water, from an energy viewpoint, may be more effective.     

A study of experimental autoimmune encephalomyelitis in dogs as a disease model for canine necrotizing encephalitis

In the present study, the use of dogs with experimental autoimmune encephalomyelitis (EAE) as a disease model for necrotizing encephalitis (NE) was assessed. Twelve healthy dogs were included in this study. Canine forebrain tissues (8 g), including white and grey matter, were homogenized with 4 mL of phosphate-buffered saline for 5 min in an ice bath. The suspension was emulsified with the same volume of Freund''s complete adjuvant containing 1 mg/mL of killed Mycobacterium tuberculosis H37Ra. Under sedation, each dog was injected subcutaneously with canine brain homogenate at four sites: two in the inguinal and two in the axillary regions. A second injection (booster) was administered to all the dogs using the same procedure 7 days after the first injection. Clinical assessment, magnetic resonance imaging, cerebrospinal fluid analyses, necropsies, and histopathological and immunohistochemical examinations were performed for the dogs with EAE. Out of the 12 animals, seven (58%) developed clinically manifest EAE at various times after immunization. Characteristics of canine EAE models were very similar to canine NE, suggesting that canine EAE can be a disease model for NE in dogs.     

Evaluation of ultrasonic sensor for variable-rate spray applications

Automatic variable-rate sprayers require accurate measurement of canopy size. An estimate of canopy size is made by measuring the distance to the canopy at several elevations above the ground; an ultrasonic sensor was used to determine canopy distance in this study. It is sometimes necessary to conduct spray operations during harsh operating conditions. In this study ultrasonic sensors were subjected to simulated environmental and operating conditions to determine their durability and accuracy. Conditions tested included exposure to extended cold, outdoor temperatures, cross winds, temperature change, dust clouds, travel speeds and spray cloud effects. The root mean square (RMS) error in a series of measurements of the distance to a simulated plant canopy was used to test for significant difference among treatments. After exposure to outdoor cold conditions for 4 months, the RMS error in distance measured by the ultrasonic sensor increased from 3.31 to 3.55 cm, which was not statistically significant. Neither the presence of dust cloud nor the changes in cross-wind speeds over a range from 1.5-7.5 m/s had significant effects on the mean RMS errors. Varying sensor travel speed from 0.8 to 3.0 m/s had no significant influence on sensor detection distances. Increasing ambient temperature from 16.7 to 41.6 °C reduced the detection distance by 5.0 cm. The physical location of the spray nozzle with respect to the ultrasonic sensor had a significant effect on mean RMS errors. The mean RMS errors of sensor distance measurements ranged from 2.3 to 83.0 cm. The RMS errors could be reduced to acceptable values by proper controlling the sensor/spray nozzles spacing on a sprayer. In addition, multiple-synchronized sensors were tested for their measurement stability and accuracy (due to possible cross-talk errors) when mounted on a prototype sprayer. It was found that isolating the pathway of the ultrasonic wave of each sensor reduced detecting interference between sensors during multiple sensor operation. Test methods presented herein may be useful in the design of standardized testing protocols for field use distance sensors.     

Therapeutic trial of granulocyte-colony stimulating factor for dilated cardiomyopathy in three dogs

Three dogs were presented to us for evaluation of cardiac problems. Electrocardiographic recordings revealed severe tachyarrhythmia and atrial fibrillation with ventricular tachycardia in 2 of the 3 dogs. The echocardiographic findings of the 3 dogs revealed markedly decreased fractional shortening and a marked increase in E-point septal separation. Based on the results of electrocardiographic and echocardiographic evaluation, the 3 dogs were diagnosed as dilated cardiomyopathy (DCM). The dogs were treated with conventional cardiac medication, but cardiac function did not improve and the clinical signs remained. We subsequently attempted treatment with granulocyte-colony stimulating factor (G-CSF; 10 microg/kg, subcutaneously). The specific purpose of G-CSF therapy for DCM was to improve cardiac function and a significant improvement in cardiac function was confirmed. The three dogs had no treatment side effects. This case report suggests that G-CSF might have therapeutic effects for medically refractory DCM in dogs.