• Animal Bioscience
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• v.36 no.10
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• pp.1530-1535
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• 2023

Objective: Semen cryopreservation result in decreased sperm parameters and fertilization ability. Sericin exhibits antioxidant activity by reducing lipid peroxidation resulting from free radicals, which can potentially improve cryopreservation outcomes. The present study aimed to examine the efficacy of various sericin concentrations supplemented with a rooster semen-freezing extender on post-thaw semen quality and fertilizing ability of sperm after cryopreservation. Methods: Semen samples were collected from 40 roosters (5 reps), then were pooled, and divided into four groups by the levels of sericin supplementation (0%, 0.25%, 0.50%, and 0.75%) in a freezing extender. Semen suspensions were loaded in medium straw (0.5 mL) and cryopreserved with the traditional liquid nitrogen vapor method. Post-thawed semen was evaluated for sperm motility, sperm viability, and lipid peroxidation. Also, the fertility test was determined. Results: The results showed that supplementation of the freezing extender with 0.50% to 0.75% sericin resulted in greater total motility and progressive motility and lower malondialdehyde levels than the other groups after cryopreservation (p<0.05). However, the viability of 0.75% decreased compared with the value of 0.50% sericin supplementation (p<0.05). Moreover, the fertility and hatchability of total eggs were significantly higher in the 0.50% sericin group than in the other groups (p<0.05). Conclusion: In conclusion, 0.50% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved rooster semen.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.275-279
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• 2009

The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.21 no.1
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• pp.67-83
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• 1995

Skin irritations accompany the series of complex, physical and chemical changes. Alternative methods which reflect the biological phenomenon more sensitively are necessary for the purpose of reducing the animal tests and improving the accuracy and reliability. Chorioallantoic membrane (CAM) vascular assay is a in vitro method which utilizes the chorioallantoic membrane of hen's fertilized egg of about 10 days old. Test substances are placed directly onto the surface of the CAM and 1 hour later the CAM vasculature is subjectively evaluated to determine with a chemically related-injurious reponse including the appearance of haemorrage, congestion, coagulation, and so on. In this research, using the various surfactants, the correlations of CAMVA with in vivo models (intradermal safety test and human primary irritation test) were investigated. And CAMVA closely correlated to intradermal safety test (r=0.89) and human primary irritation test (r=0.90). From the result, it seems that CAMVA can also be used as a method for predicting the skin irritaions.

• Journal of Veterinary Clinics
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• v.27 no.1
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• pp.66-70
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• 2010

Reference interval is critical for interpreting laboratory results, monitoring response to therapy and predicting the prognosis of the patients in clinical settings. The aim of the present study was to update established reference intervals for routine hematologic and serum chemistry values for a population of clinically healthy dogs (range, 1-8 years) seen in an animal hospital. Blood was obtained by venipuncture while animals were physically restrained, and samples were analyzed for 9 chemistries on MS9-5H (Melot Schloesing Lab, France) and 6 hematology on Vet Test 8008 (IDEXX, USA). Data from 105 dogs (52 males and 53 females) for hematology and 113 dogs (37 males and 76 females) for chemistry were used to determine reference intervals using the parametric, nonparametric and bootstrap methods. Prior to analysis, all parameters were tested for normal distribution using Anderson-Darling criterion. Of the 9 biochemical analytes, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, creatinine, total protein, and glucose concentrations did not fit normal distribution for both original and transformed data. All but eosinophil count satisfied normal distribution for either original or transformed data. Parametric method can be used for original cholesterol concentrations, RBC, WBC, and neutrophil counts. This technique can also be used for power-transformed values of blood urea nitrogen concentrations and for logarithm of lymphocyte and monocyte counts. Non-parametric or bootstrap method was the preferred choice for the remaining 7 biochemical parameters and eosinophil count as they did not follow normal distributions. All three statistical techniques performed in similar reference intervals. When establishing reference intervals for clinical laboratory data, it is essential to assess the distribution of the original data to increase the accuracy of the interval, and non-parametric or bootstrap methods are of alternative for the data that do not fit normal distribution.

• The Korean Journal of Pesticide Science
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• v.16 no.3
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• pp.261-266
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• 2012

It is common to use many experiment animals to evaluate the toxicity of chemicals including pesticides. For protecting animal, the concepts of 3R (Reduction, Replacement, Refinement) were introduced and in vitro alternatives methods actively have been developed all over the world. Many experimental animals for toxicological tests have been used, so that it is important to establish the alternative methods. In this study, the alternative method using reconstituted human skin model (Keraskin$^{TM}$) was conducted for classification of skin irritation on pesticides. Sixteen formulations selected on the basis of the degree of irritation were treated by Keraskin$^{TM}$ test. The percent of cell viability was measured into the culture medium collected after treatment of the pesticides for 24-72 hrs. The skin irritations of formulations were evaluated by the cell viability. In this study, The 4 formulations with mild irritation in rabbits were evaluated as nonirritant, the 6 formulations with moderate and severe irritation were evaluated as irritant in human skin model test. We suggest that the alternative test using Keraskin$^{TM}$ model could be used as toxicity evaluation for primary irritation index (P.I.I.) score of greater than or equal to 2.1 of pesticides. The further studies should be required to apply for hazardous assessment of pesticides on alternative skin irritation methods because of the interindividual variability of the sensitivity of skin irritation on pesticides.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.707-718
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• 2004

This study was conducted to evaluate the adequacy of an alternative a.m. - p.m. testing scheme for milk yield in comparison with the official test method based on weighing two milkings within 24 h. A total of 8,309 p.m. milking weights and 6,767 am. milking weights from 72 Holstein cows raised at N.L.R.I. were collected between October 2000 and November 2001. Seven statistical models were fitted to the data to derive formulas for estimating daily milk yields from morning or evening yields. In general, use of evening milkings less accurately estimated than did use morning rnilkings. Although the models do not differ much in the correlations between estimated and true daily milk yields, systematic under- and overestimation of daily milk yields were observed in all models with the exception of model 7, which accounted for heterogeneous variances by parity class, milking interval class, and lactation stage by fitting separate regression formulas within each combination of three factors.

• Toxicological Research
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• v.33 no.2
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• pp.107-118
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• 2017

Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.77-81
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• 2007

Diagnostic performance of polymerase chain reaction (PCR) for detecting Dirofilaria immitis in dogs was evaluated when no gold standard test was employed. An enzyme-linked immunosorbent assay test kit (SnapTM, IDEXX, USA) with unknown parameters was also employed. The sensitivity and specificity of the PCR from two-population model were estimated by using both maximum likelihood using expectation-maximization (EM) algorithm and Bayesian method, assuming conditional independence between the two tests. A total of 266 samples, 133 samples in each trial, were randomly retrieved from the heartworm database records during the year 2002-2004 in a university animal hospital. These data originated from the test results of military dogs which were brought for routine medical check-up or testing for heartworm infection. When combined 2 trials, sensitivity and specificity of the PCR was 96.4-96.7% and 97.6-98.8% in EM and 94.4-94.8% and 97.1-98% in Bayesian. There were no statistical differences between estimates. This finding indicates that the PCR assay could be useful screening tool for detecting heartworm antigen in dogs. This study was provided further evidences that Bayesian approach is an alternative approach to draw better inference about the performance of a new diagnostic test in case when either gold test is not available.

• Journal of the Korean Applied Science and Technology
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• v.30 no.1
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• pp.35-48
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• 2013

Biodiesel is an alternative diesel fuel which can be obtained from the transesterification of vegetable oils, animal fats and waste cooking oil. The objective of this study is to evaluate the properties of biodiesel obtained from different feedstocks (soybean, waste cooking, rapeseed, cottonseed and palm oils). The biodiesel derived from different feedstocks was analyzed for FAME (fatty acid methyl esther) content, kinematic viscosity, flash point, CFPP (cold filter plugging point) and glycerin content. The quality of biodiesel was tested according to the Korean and European standard (EN14214, requirements and test method for biodiesel fuel). The biodiesels derived from soybean, waste cooking, rapeseed and cottonseed oils contain high amount of unsaturated fatty acid, while palm biodiesel is dominated by saturated fatty acid. The fuel properties of biodiesel, such as low temperature performance, kinematic viscosity and oxidation stability are correlated with the FAME composition components in biodiesel.

• Journal of Marine Life Science
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• v.9 no.1
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• pp.9-21
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• 2024

Paralytic shellfish poisoning (PSP) including Saxitoxin (STX) is caused by harmful algae, and poisoning occurs when the contaminated seafood is consumed. The mouse bioassay (MBA), a standard test method for detecting PSP, is being sanctioned in many countries due to its low detection limit and the animal concerns. An alternative to the MBA is the Neuro-2a cell-based assay. This study aimed to establish various test conditions for Neuro-2a assay, including cell density, culture conditions, and STX treatment conditions, to suit the domestic laboratory environment. As a result, the initial cell density was set to 40,000 cells/well and the incubation time to 24 hours. Additionally, the concentration of Ouabain and Veratridine (O/V) was set to 500/50 μM, at which most cells died. In this study, we identified eight concentrations of STX, ranging from 368 to 47,056 fg/μl, which produced an S-shaped dose-response curve when treated with O/V. Through inter-laboratory variability comparison of the Neuro-2a assay, we established five Quality Control Criteria to verify the appropriateness of the experiments and six Data Criteria (Top and Bottom OD, EC₅₀, EC₂₀, Hill slop, and R² of graph) to determine the reliability of the experimental data. The Neuro-2a assay conducted under the established conditions showed an EC₅₀ value of approximately 1,800~3,500 fg/μl. The intra- & inter-lab variability comparison results showed that the coefficients of variation (CVs) for the Quality Control and Data values ranged from 1.98% to 29.15%, confirming the reproducibility of the experiments. This study presented Quality Control Criteria and Data Criteria to assess the appropriateness of the experiments and confirmed the excellent repeatability and reproducibility of the Neuro-2a assay. To apply the Neuro-2a assay as an alternative method for detecting PSP in domestic seafood, it is essential to establish a toxin extraction method from seafood and toxin quantification methods, and perform correlation analysis with MBA and instrumental analysis methods.