Animal welfare has been gradually gathering more attention from consumers over time, making it increasingly important to assess the level of stress experienced by livestock. Traditionally, stress has been measured by collecting blood to assess cortisol levels, an action that can be considered distressing for the animal. Therefore, we aimed to explore the feasibility of using hair as an alternative medium to blood for stress assessment. We utilized B/F (blood collected at the farm), B/A (blood collected after transport to the auction), and H/A (hair collected at the auction after blood sampling) from calves at the age of 7-9 months transported from the farm to the auction. Hair underwent a washing and extraction process to utilize hair extracts, while blood was centrifuged to analyze using ELISA. The cortisol concentration in the blood was significantly higher in B/A compared to B/F (p<0.05), confirming that the calves experienced stress during transportation. Additionally, H/A was significantly lower than both B/A and B/F (p<0.0001). These results emphasized that cortisol in hair is not suitable for investigating short-term stress in livestock, as it is with blood. While measuring stress indices using hair may not be appropriate for replacing blood, it is considered a highly suitable practice for animal welfare, and further research in this area should be continued.
Flavobacterium sp. strain DS5(NRRL B-14859) was used to convert oleic acid to 10-ketostearic acid(10-KSA) via 10-hydroxystearic acid(10-HSA). Increase in cell concentration by centrifuging, collecting cells grown in two flasks, and resuspending in one flask, improved 10-KSA production to 6.5 g/L from 3.5 g/L in a usual flask culture. Tween-80 addition to the culture did not greatly affect the production of 10-KSA and 10-HSA. When culture broth was centrifuged after fermentation, it was observed that pellets were separated into two parts(yellow and white). Gas chromatography analysis showed that 10-KSA and 10-HSA were detected only in a white pellet, suggesting that the bioconversion products of strain DS5 are extracellularly produced and can be easily recovered from cells by a simple centrifugation step.
Pig oocytes contain high levels of lipids in the ooplasm, which reduces the visibility of the germinal vesicle (GV) under microscopic examination. Therefore, the purposes of this study were to investigate the effects of relative centrifugation force (RCF) on the visibility and maturation rates of the GV stage oocytes after centrifugation. In Experiment 1, cumulus-oocyte-complexes (COCs) were collected from slaughterhouse ovaries and randomly allocated to different RCFs (3,000 rpm: 970 g; 6,000 rpm: 3,900 g; or 10,000 rpm: 10,840 g) for 10 or 20 min. Percentages of visible GV were 76-79% in the oocytes centrifuged with 10,000 rpm, which were significantly higher (p<0.01) than those with 3,000 and 6,000 rpm. No significant differences in GV visibility were observed among oocytes with different lengths of centrifugation (p<0.05) regardless of the RCFs. In esperiment 2, the maturation rate of the oocyte was found significantly lower in the 20 min than in the 10 min group received 10,840 g of RCF (30 vs. 75%, p<0.05). In conclusion, the GV of porcine oocytes can be clearly visible by centrifugation at 10,840 g for 10 min without compromising their subsequent maturation rates and a longer centrifugation time (20 min) had no beneficial influence on the visibility of GV stage pig oocytes.
In previous studies, we reported that sow which was transferred OPS-freezing embryos not able to deliver a piglet (Kim et al, 2004). This study was conducted to investigate a possibility of gilt as recipients which produce piglets after transfer of OPS-freezing embryos. All transferred embryos were prepared by in vitro production (IVP) system. In vitro culture (IVC) medium used glucose-free NCSU23 supplemented with 5mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$. From day 3 of IVC, 10% fetal bovine serum albumin was added to the culture medium. In preparing of freezing embryos, embryos were treated with 7.5 $\mu g/ml$ cytochalasin-B for 30 min and centrifuged at $13,000{\times}g$ for 13 min. And then, embryos were exposed sequentially to an ethylene glycol (EG) solution, aspirated into open pulled straw (OPS), and plunged or thawed into the liquid nitrogen. In embryo transfer (ET), we used two kinds of type (surgical method vs. non-surgical method). In surgical method of embryo transfer, $55\sim65$ embryo were transferred in both uterine horn of two recipient gilts by plastic straw. Non-surgical method which is like artificial insemination was performed on three gilts. Each 140 frozen embryos were transferred to two gilts and 40 fresh embryos to one gilt. Pregnancy establishment was shown one recipient at 45 days after ET. However, the one recipient was also aborted at 58 days after ET. These results suggest that gilts can be considered as a candidate of recipients for OPS-freezing embryo transfer.
(3-mercaptopropyl)trimethoxysilane (MPTMS) was used as a silylation agent, and modified silica nanoparticles were prepared by solution polymerization. 2.0 g of silica nanoparticles, 150 ml of toluene, and 20 ml of MPTMS were put into a 300 ml flask, and these mixtures were dispersed with ultrasonic vibration for 60 min. 0.2 g of hydroquinone as an inhibitor and 1 to 2 drops of 2,6-dimethylpyridine as a catalyst were added into the mixture. The mixture was then stirred with a magnetic stirrer for 8 hrs. at room temperature. After the reaction, the mixture was centrifuged for 1 hr. at 6000rpm. After precipitation, 150 ml of ethanol was added, and ultrasonic vibration was applied for 30 min. After the ultrasonic vibration, centrifugation was carried out again for 1 hr. at 6000rpm. Organo-modification of silica nanoparticles with a ${\gamma}$-methacryloxypropyl functional group was successfully achieved by solution polymerization in the ethanol solution. The characteristics of the ${\gamma}$-mercaptopropyl modified silica nanoparticles (MPSN) were examined using X-ray photoelectron spectroscopy (XPS, THERMO VG SCIENTIFIC, MultiLab 2000), a laser scattering system (LSS, TOPCON Co., GLS-1000), Fourier transform infrared spectroscopy (FTIR, JASCO INTERNATIONL CO., FT/IR-4200), scanning electron microscopy (SEM, HITACHI, S-2400), an elemental analysis (EA, Elementar, Vario macro/micro) and a thermogravimetric analysis (TGA, Perkin Elmer, TGA 7, Pyris 1). From the analysis results, the content of the methacryloxypropyl group was 0.98 mmol/g and the conversion rate of acrylamide monomer was 93%. SEM analysis results showed that the organo-modification of ultra-fine particles effectively prevented their agglomeration and improved their dispensability.
Experimental tracheal ligation (TL) has been shown to reverse the pulmonary hypoplasia associated with congenital diaphragmatic hernia (CDH) and to normalize gas exchange. The purpose of this study was to determine whether the TL would correct the surfactant deficiency present in the fetal rabbit model of CDH by using lamellar body count. Lamellar bodies are synthesized and secreted by the type II pneumocytes of fetal lung. The phospholipids present in these bodies constitute the major component of pulmonary surfactant. Twenty-one pregnant New Zealand rabbits underwent hysterotomy and fetal surgery on gestational day 24. Two fetuses of each pregnant rabbit were operated. In the fetus of one end of bicornuate uterus, left DH was created by excision of fetal diaphragm through open thoracotomy (DH Group). In the fetus of the other end of bicornuate uterus, left DH and TL were created (TL Group). The fetuses were delivered by Cesarean section on gestational day 31. Fourteen in control group, 12 in the DH group and 13 in TL group were born alive. En bloc excision of lungs, bronchi and trachea was done in all newborn rabbits. A five Fr catheter was inserted through trachea and repeated irrigations with 10 cc normal saline were done. The irrigated fluid was centrifuged at $280{\times}g$ for 5 minutes and the lamellar bodies were counted with the upper level fluid in platelet channel of electronic cell counter. The average lamellar body counts were $37.1{\pm}14.2{\times}10^3/{\mu}L$ in control group, $11.5{\pm}4.4 {\times}10^3/{\mu}L$ in DH group, and $6.5{\pm}0.9{\times}10^3/{\mu}L$ in TL group. Lamellar body count in DH group was lower than in control group and did not increase after TL. This study shows TL has no therapeutic effect on decreased surfactant level of CDH and the pregnant rabbit is appropriate for the animal model of CDH.
Objective : To evaluate the relationship between the dose of Clodronate and serum level of alkaline phosphatase (ALP), calcium (Ca), and phosphate (PO4) during orthodontic tooth movement MaterialS and MethodS: A total of 18 sex-matched Wistar rats (weight=180~230g, mean age=8 weeks) were allocated into the 2.5mM Clodronate (2.5C) group, 10mM Clodronate (10C) group, or control group (n=6 for each group). After the application of a nickel-titanium closed coil spring (force of 60g) between the upper central incisors and first molars (UFM), 2.5C, 10C, or saline was injected every third day into the subperiosteum of the alveolar bone adjacent to UFM for the experimental and control groups. The animals were sacrificed 17 days later. Trunk blood was quickly collected into a heparinized tube and centrifuged at 2,000 rpm for 20 min. The plasma was used for the biochemical assays of the serum level of ALP, Ca, and PO4. Kruskall-Wallis test and Mann-Whitney test with Bonferroni correction were performed for the statistical analyses. Results : Dose-dependent increase in the level of ALP (P<0.01) and decrease in the level of Ca (P<0.001) were observed among the control, 2.5C, and 10C groups. Although there was no significant difference in PO4 between the 2.5C and 10C groups, the 10C group showed a significantly higher level of PO4 than the control group (P<0.01). Conclusion : Since Clodronate induced significant dose-dependent change in the serum level of ALP, Ca, and PO4 during orthodontic tooth movement, orthodontists should consider these biochemical markers not only as a diagnostic tool for bone turnover rate but also as a monitoring tool for orthodontic tooth movement.
Objectives: This study evaluated the antibacterial effect and mechanical properties of composite resins ($L_{CR}$, $M_{CR}$, $H_{CR}$) incorporating chitosan with three different molecular weights (L, Low; M, Medium; H, High). Materials and Methods: Streptococcus (S). mutans 100 mL and each chitosan powder were inoculated in sterilized 10 mL Brain-Heart Infusion (BHI) solution, and was centrifuged for 12 hr. Absorbance of the supernatent was measured at $OD_{660}$ to estimate the antibacterial activities of chitosan. After S. mutans was inoculated in the disc shaped chitosan-containing composite resins, the disc was cleansed with BHI and diluted with serial dilution method. S. mutans was spread on Mitis-salivarius bacitracin agar. After then, colony forming unit (CFU) was measured to verify the inhibitory effect on S. mutans biofilm. To ascertain the effect on the mechanical properties of composite resin, 3-point bending and Vickers hardness tests were done after 1 and 3 wk water storage, respectively. Using 2-way analysis of variance (ANOVA) and Scheffe test, statistical analysis was done with 95% significance level. Results: All chitosan powder showed inhibition effect against S. mutans. CFU number in chitosan-containing composite resins was smaller than that of control resin without chitosan. The chitosan containing composite resins did not show any significant difference in flexural strength and Vickers hardness in comparison with the control resin. However, the composite resin, $M_{CR}$ showed a slightly decreased flexural strength and the maximum load than those of control and the other composite resins $H_{CR}$ and $L_{CR}$. Conclusions: $L_{CR}$ and $H_{CR}$ would be recommended as a feasible antibacterial restorative due to its antibacterial nature and mechanical properties.
An experiment was conducted to quantify the flow of soluble non-ammonia nitrogen (SNAN) in the liquid phase of ruminal (RD) and omasal digesta (OD), and to investigate diurnal pattern in SNAN flow in OD. Five ruminally cannulated Finnish-Ayrshire dairy cows in a $5{\times}5$ Latin square design consumed a basal diet of grass silage and barley grain, and that supplemented with four protein feeds (kg/d DM basis) as follows: skimmed milk powder (2.1), wet distiller' solubles (3.0), untreated rapeseed meal (2.1) and treated rapeseed meal (2.1). Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 1.0 h interval during a 12 h feeding cycle. Both RD and OD were acidified, centrifuged to remove microbes and precipitated with trichloroacetic acid followed by centrifugation. The SNAN fractions (free amino acid (AA), peptide and soluble protein) in RD and OD were assessed using ninhydrin assay. Free AA, peptide and soluble protein averaged 60.0, 89.4 and 2.1 g/d, respectively, for RD, and 81.8, 121.5 and 2.5 g/d, respectively, for OD. Although free AA flow was relatively high, mean peptide flow was quantitatively the most important fraction of SNAN, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis. Diurnal pattern in flow of peptide including free AA in OD during a 12 h feeding cycle peaked 1 h post-feeding, decreased by 3 h post-feeding and was relatively constant thereafter. Protein supplementation showed higher flow of peptide including free AA immediately after feeding compared with no supplemented diet. There were no differences among protein supplements in diurnal pattern in flow of peptide including free AA in OD.
The purpose of this study was to investigate dietary taurine intakes, plasma taurine levels and urinary taurine excretions of women in Seoul(Kangbuk-ku) and Kyunggi(Yeoju) area, Korea. Seventy married women aged 39.7$\pm$8.9 have volunteered for this study: 36 from Seoul area and 34 from Kyunggi area. Diet samples were collected from the participants and the samples included three meals (breakfast, lunch and supper), snacks, drinks and whatever the participants had eaten for 24 hours. The plasma was obtained by allowing a 5 $m\ell$ fasting blood sample to be in a heparinized tube for 30 min and centrifuging it at 11,000${\times}$g for 20 min. The collected diets were blended, centrifuged and deproteinized. Taurine levels in the diet and plasma were determined as the dabsyl derivative using HPLC with Rf-detector. The intake of taurine ranged from 6.8 to 837.8 mg/day and its mean value was l45.5$\pm$164.0 mg/day (mean$\pm$SD). The 90th, 50th and 10th percentile values of the taurine intake were 280.0, 94.3 and 26.8 mg/day, respectively. There was a significant difference between two groups: 202.0$\pm$204.9 for Seoul area and 85.5$\pm$67.2 mg/day for Kyunggi area(p<0.00l). The taurine level in plasma ranged from 42.1 to 201.9 $\mu$mol/L and its mean value was 74.9$\pm$22.8 $\mu$mol/L. The 90th, 50th and 10th percentile values of the plasma taurine were 101.1, 70.7 and 54.6 $\mu$mol/L, respectively. There was no significant difference between Seoul area and Kyunggi area in plasma taurine level.
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