• 한국동물생명공학회지
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• 제36권2호
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• pp.91-98
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• 2021

The objective of this study was to establish a selection process for high quality sperm in bovine semen using sperm separation by magnetic activation (MACS). For this, semen from 21 Nellore bulls was collected using an artificial vagina. To guarantee the presence of pathologies in the ejaculate, animals previously declassified in four consecutive spermiogram were used. Semen was analyzed in five statuses: (1) fresh semen (fresh); (2) density gradient centrifugation (DGC), percoll column; (3) non-apoptotic fraction after separation by MACS (MAC); (4) apoptotic fraction from the separation (MACPOOR); and (5) MAC followed by DGC (MACDGC). Using a computerized analysis system (CASA), motility was measured. The sperm morphology was evaluated by phase contrast, and the supravital test was completed with eosin/nigrosin staining. For DGC, 20 × 10⁶ cells were used in a gradient of 90% and 45% percoll. MACS used 10 × 10⁶ cells with 20 μL of nanoparticles attached to annexin V, and filtered through the MiniMACS magnetic separation column. Membrane integrity was assessed with SYBR-14/IP and mitochondrial potential with JC-1 by flow cytometry. Processing sperm by MACDGC, was more effective in obtaining samples with high quality sperm, verified by the total of abnormalities in the samples: 35.04 ± 2.29%, 21.50 ± 1.47%, 17.30 ± 1.10%, 30.68 ± 1.94% and 10.50 ± 1.46%, respectively for fresh, DGC, MAC, MACPOOR, and MACDGC. The subpopulation of non-apoptotic sperm had a high number of live cells (82.65%), membrane integrity (56.60%) and mitochondrial potential (83.98%) (p < 0.05). These findings suggest that this nanotechnological method, that uses nanoparticles, is efficient in the production of high-quality semen samples for assisted reproduction procedures in cattle.

• 한국환경보건학회지
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• 제48권4호
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• pp.244-253
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• 2022

Background: The indiscriminate use of pesticides in Nigeria may have harmful effects on reproductive health of farmers. Objectives: This study assessed the awareness of reproductive health, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone, estradiol, progesterone and sperm characteristics of male farmers occupationally exposed to pesticides. Methods: Eighty four male farmers were recruited for the study. Structured questionnaire was used to obtain the socio-demographic data. Blood and semen samples were collected from the subjects in the morning for hormonal assays and semen analysis using enzyme linked immunosorbent assay (ELISA) method and SQAV sperm quality analyzer. Data were analyzed using chi square, Student's-t-test, and Regression analysis. Results: Serum FSH (p<0.01), LH (p<0.005) and Estradiol (p<0.001) were significantly higher while prolactin (p<0.02) and testosterone (p<0.001) were significantly lower among pesticides exposed farmers than nonexposed subjects. Some 34/84 (40.5%) of the pesticides exposed farmers had serum testosterone levels below the lower limit of the reference range. Those with low testosterone levels (p<0.001), also had FSH (p<0.05), LH (p<0.001) and Estradiol (p<0.002) significantly lower than those with normal testosterone levels. The sperm count among pesticides exposed farmers; total motility and percentage morphology were significantly lower than non-pesticides exposed subjects. Some 14/84 (16.7%) of the pesticides exposed farmers had sperm count below 15 million/mL (oligozoospermia). More than 70% of the farmers were not aware of the reproductive health risks associated with pesticides and only 23.8% of the farmers were using protective devices. Conclusions: Deliberate efforts to improve awareness, knowledge, personal hygiene, and interventions necessary to lessen both pesticides exposure and health risks by adopting safe practices are suggested.

• Journal of Microbiology and Biotechnology
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• 제33권5호
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• pp.668-679
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• 2023

Edwardsiella tarda is one of the most significant fish pathogens, causes edwardsiellosis in a variety of freshwater fish species, and its antibiotic resistance against multiple drugs has made it a health risk worldwide. In this study, we aimed to investigate the antibiotic resistance (ABR) genes of E. tarda and establish its antibiotic susceptibility. Thus, 540 fish (299 Oreochromis niloticus, 138 O. mossambicus, and 103 O. aureus) were collected randomly from twelve fish farms in three districts of Punjab in Pakistan. E. tarda was recovered from 147 fish showing symptoms of exophthalmia, hemorrhages, skin depigmentation, ascites, and bacteria-filled nodules in enlarged liver and kidney. Antimicrobial susceptibility testing proved chloramphenicol, ciprofloxacin, and streptomycin effective, but amoxicillin, erythromycin, and flumequine ineffective in controlling edwardsiellosis. Maximum occurrence of qnrA, blaTEM, and sul3 genes of E. tarda was detected in 45% in the liver, 58%, and 42% respectively in the intestine; 46.5%, 67.2%, and 55.9% respectively in O. niloticus; 24%, 36%, and 23% respectively in summer with respect to fish organs, species, and season, respectively. Motility, H₂S, indole, methyl red, and glucose tests gave positive results. Overall, E. tarda infected 27.2% of fish, which ultimately caused 7.69% mortality. The Chi-squared test of independence showed a significant difference in the occurrence of ABR genes of E. tarda with respect to sampling sites. In conclusion, the misuse of antibacterial agents has led to the emergence of ABR genes in E. tarda, which in association with high temperatures cause multiple abnormalities in infected fish and ultimately resulting in massive mortality.

• Animal Bioscience
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• 제37권1호
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• pp.50-60
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• 2024

Objective: Testicular fat deposition has been reported to affect animal reproduction. However, the underlying mechanism remains poorly understood. The present study explored whether sperm meiosis and testosterone synthesis contribute to mouse testicular fat deposition-induced reproductive performance. Methods: High fat diet (HFD)-induced obesity CD1 mice (DIO) were used as a testicular fat deposition model. The serum hormone test was performed by agent kit. The quality of sperm was assessed using a Sperm Class Analyzer. Testicular tissue morphology was analyzed by histochemical methods. The expression of spermatocyte marker molecules was monitored by an immuno-fluorescence microscope during meiosis. Analysis of the synthesis of testosterone was performed by real-time polymerase chain reaction and reagent kit. Results: It was found that there was a significant increase in body weight among DIO mice, however, the food intake showed no difference compared to control mice fed a normal diet (CTR). The number of offspring in DIO mice decreased, but there was no significant difference from the CTR group. The levels of follicle-stimulating hormone were lower in DIO mice and their luteinizing hormone levels were similar. The results showed a remarkable decrease in sperm density and motility among DIO mice. We also found that fat accumulation affected the meiosis process, mainly reflected in the cross-exchange of homologous chromosomes. In addition, overweight increased fat deposition in the testis and reduced the expression of testosterone synthesis-related enzymes, thereby affecting the synthesis and secretion of testosterone by testicular Leydig cells. Conclusion: Fat accumulation in the testes causes testicular cell dysfunction, which affects testosterone hormone synthesis and ultimately affects sperm formation.

• 한국임상수의학회지
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• 제32권1호
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• pp.36-40
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• 2015

산통은 이환률과 치명률이 높아 말 산업에 가장 우려되는 질병이다. 많은 나라에는 산통마의 내과적, 외과적 치료에 따른 치명률과 예후인자 평가를 위한 연구 결과가 있지만 한국에서는 산통의 예후 판단을 위한 객관적 연구가 이루어지지 않았다. 본 논문에서 한국 산통마의 치명률을 확인하고, 잠재적 예후 인자들을 평가를 하고자 하였다. 2011년 1월부터 2013년 12월까지 한국마사회에 내원한 119마리의 산통마를 대상으로 조사하였다. 유의성을 평가하기 위하여 카이제곱검정, T-test와 Mann-Whitney test를 사용하였다. 그 결과, 전체 치명률은 15%였으며 내원한 산통마 중 78%가 내과적 치료를 받았다. 개체적 인자인 성별, 나이, 내원 시기는 예후와 유의성은 없었으나 3세 이하 (59%)의 수컷 (50%)에서 발생률이 높았다. 탈수의 대표적 지표인 구강점막의 색깔과 PCV는 생존집단과 폐사집단사이에 유의적인 차이를 보였다. 그 외 신체적 인자인 심박수, 장운동성 그리고 직장온도도 예후와 유의적으로 관련이 있었다. 임상병리인자 중에는 적혈구수, 혈색소농도, 충진세포용적, 혈소판수, 크레아틴키나아제(CK), 젖산탈수소효소(LDH), 혈당치가 산통의 예후인자로서 통계적으로 유의성이 있음을 확인하였다.

• 한국동물위생학회지
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• 제18권1호
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• pp.41-56
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• 1995

The present study was conducted to investigate results of B. anthracis isolated from Anthrax in the Kyong-Ju of Feb. 12. 1994. 1. In biochemical feature, B. anthracis was a gram-positive rod, non-motility, sporulation, capsulation. It was positive in gelatinase, starch hydrolysis, glucose. But negative in urease, arabinose, mannitol, xylose. 2. B. anthracis grew well on B4 Br A TSA after incubation for 24 hours. The organisim grew well on BA, Br. A, NA, TSA after incubation for 72 hours. The media grew well on Br A instead of BA. 3. On 5% blood agar by laboratory animal, ${\beta}$ -hemolysis was produced from 36 hours to 48 hours incubation. There was perfect ${\beta}$-hemolysis after incubation for 48 hours. On the other side ${\beta}$-hemolysis was begun on 5% goat blood agar after incubation for 60 hours. 4. In the test of antimicrobial susceptibility, B. anthracis was very sensitive to AM, CF, TE, ENR, GM, AN, DFX, S, P, TYLO, N, KM, C, E, Lins＋Sp, NN, CC, CFP, CB were sensitive one by one. B. anthracis was no-sensitive to L, XNL, TIA, CL, SXT 5. B. anthracis had never sensitivity to direct inoculation of rat and chicken, after subcutanous inj. It was very sensitive to mouse and goat, hamster, guinea pig, rabbit had a sensibility one by one. 6. The dead laboratory animal which had been inoculated with B. anthracis preserved at $37^{\circ}C$ incubation, B. anthracis didn't cultivate on non-dissected animal after 80 hours but cultivate on dissected animal after 360 hours. 7. The rapidly death could cause high concentration, died from 420 after S. C. 8. The blood smeared samples of hamster from inoculation with B. anthracis, spore germinated In 37$^{\circ}C$ after 5 hours, in $32^{\circ}C$ after 6 hours, in room temperature after 9 hours, in $-4^{\circ}C$ to $-20^{\circ}C$ after 10 hours. 9. B, anthracis inoculated to laboratory animal after SC or PO. Mice and rats feces didn't cultivated with B. anthracis after SC, but did cultivated with B. anthracis after PO. 10. In the test of disinfectant, B. anthracis was high effective to $HgC1_2$, formalin, effect phenol, cresol, but non-effect NaOH, ethanol.

• 대한물리치료과학회지
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• 제12권3호
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• pp.47-58
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• 2005

The purpose of this study is to estimate how much the motility of affected and unaffected lower limb respectively would have effects on their ambulation through comparing the difference between their affected and unaffected lower limb and studying the correlation between such difference and their ambulation. In addition, the study also intends to find out remedial measures suitable for improving their ambulation with relevant physical treatment. To do this, a quantitative electromyogram(QEMG) test was done to both quadriceps and hamstring of affected and unaffected lower limb so as to yield IP values. Based on such IP values, RMS(root mean square) values as the total sum of IP values were found with QEMG analysis system (made by Medelec Co.) and then the ambulation depending on the difference in muscular strength were analyzed by ink-foot print method as well as the corresponding statistics were processed by T-test through SPSS. The effect of differences in muscular strength of quadriceps in affected and unaffected limb of hemiplegic patients on their ambulation was also different in each element of ambulation. However, there was not any statistically significant difference (p>.05). The differences in muscular strength of hamstring in unaffected limbs of hemiplegic patients only affect stride length. It was statistically significant. The effect of difference in muscular strength of the hamstring in affected limb of hemiplegic patients on their ambulation was very different between strong and weak group in terms of walking velocity, step per minute, stride and step length, showing statistically significant difference(p<.05). Although there was a difference in the step length of the affected limb, it was not statistically significant(p>.05). For the unaffected hamstring, there were significant differences of stride length between in unaffected limb and in affected limb. For the affected hamstring, there were also significant differences in walking velocity, step per minute, stride and step length. Besides, it was found that the difference of the muscular strength between strong and weak group was at the highest. Thus, the effect of difference in muscular strength on ambulatory factors such as walking velocity, step per minute, stride and step length did not show any statistical significance but differences in affected and unaffected quadriceps. However, it was found that both the difference of stride in unaffected hamstring and the difference of all ambulatory factors except from step length with affected limb had effects on ambulation depending upon muscular strength.

• 생명과학회지
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• 제15권3호
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• pp.387-396
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• 2005

The aims of the study were to establish a short-term screening test for detecting testicular toxicity of chemicals in rats and to determine whether a 2-week administration period is sufficient to detect testicular toxicity of 2-bromopropane (2-BP) as an example. Male Sprague-Dawley rats were subcutaneously administered with 1000 mg/kg/day of 2-BP or its vehicle for 2 weeks. Ten male rats each were sacrificed on days 3, 7 and 14 after the initiation of treatment. Parameters of testicular toxicity included genital organ weights, testicular sperm head counts, epididymal sperm counts, motility and morphology, and qualitative and quantitative histopathologic examinations. The early histopathological changes observed on day 3 of treatment included degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, and decreased number of spermatogonia in stages II and V. On day 7 of treatment, atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in stages VII and XII. On day 14 after treatment, a significant decrease in the weights of testes and seminal vesicles was found. Atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in all spermatogenic stages were also observed. In addition, a slight non-significant decrease in testicular sperm head counts, daily sperm production rate and epididymal sperm counts was found. The results showed that 2 weeks of treatment is sufficient to detect the adverse effects of 2-BP on male reproductive organs. It is considered that the short-term testicular toxicity study established in this study can be a useful tool for screening the testicular toxic potential of new drug candidates in rats.

• Toxicological Research
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• 제33권2호
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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.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제20권3호
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• pp.153-159
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• 2017

Purpose: Acid neutralization during chemical clearance is significantly prolonged in children with cystic fibrosis, compared to symptomatic children without cystic fibrosis. The absence of available reference values impeded identification of abnormal findings within individual patients with and without cystic fibrosis. The present study aimed to test the hypothesis that significantly more children with cystic fibrosis have acid neutralization durations during chemical clearance that fall outside the physiological range. Methods: Published reference value for acid neutralization duration during chemical clearance (determined using combined impedance/pH monitoring) was used to assess esophageal acid neutralization efficiency during chemical clearance in 16 children with cystic fibrosis (3 to < 18 years) and 16 age-matched children without cystic fibrosis. Results: Duration of acid neutralization during chemical clearance exceeded the upper end of the physiological range in 9 of 16 (56.3%) children with and in 3 of 16 (18.8%) children without cystic fibrosis (p=0.0412). The likelihood ratio for duration indicated that children with cystic fibrosis are 2.1-times more likely to have abnormal acid neutralization during chemical clearance, and children with abnormal acid neutralization during chemical clearance are 1.5-times more likely to have cystic fibrosis. Conclusion: Significantly more (but not all) children with cystic fibrosis have abnormally prolonged esophageal clearance of acid. Children with cystic fibrosis are more likely to have abnormal acid neutralization during chemical clearance. Additional studies involving larger sample sizes are needed to address the importance of genotype, esophageal motility, composition and volume of saliva, and gastric acidity on acid neutralization efficiency in cystic fibrosis children.