• Title/Summary/Keyword: Semen analysis

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Effects of Different Concentrations of Escherichia coli and Days of Preservation on Boar Sperm Quality

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.213-217
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    • 2013
  • The objective of this study was to investigate the effect of bacterial contamination on elapsed time after preservation on boar semen. Known numbers of Escherichia coli (E. coli) were inoculated to freshly ejaculated semen and sperm parameters such as viability, motility, agglutination, acrosome integrity and hypo-osmotic swelling test were performed during 7 days of liquid preservation. Semen samples were prepared using antibiotic free BTS extender and 4 different levels of E. coli were treated to semen with following concentrations; 3,000, 5,000, 7,000, 10,000 CFU/ml of sperms. Semen samples were preserved at $17^{\circ}C$ for 7 days in semen storage until analyzed. Aliquots were subjected to measure the sperm viability, motility and agglutination using computer assisted sperm analysis (CASA) system, acrosome integrity was performed using chlortetracycline (CTC) staining method and hypo-osmotic swelling test was performed using hypotonic solution from day 1 (day of semen collection) to 7. Detrimental effects on sperm motility and viability were observed 3 days after preservation at the level of 5,000 CFU/ml (p<0.05). Percentage of sperm abnormality was higher (p<0.05) in over 5,000 CFU/ml groups. Sperm agglutination rate was also significantly higher (p<0.05) in groups of 5,000 and 7,000 CFU/ml. The rate of acrosome reacted sperm was higher as preservation time goes in all the samples but the pattern was clearly higher among E. coli contaminated groups (p<0.05). The sperm membrane integrity in terms of hypo-osmotic test, E. coli affects little compared to other sperm parameters. The deleterious effects observed due to the bacterial contamination in semen suggest that importance of hygiene protocol to minimize the bacterial contamination during semen collection and processing.

Study on Quality Changes Caused by Rancidity and Methods to Reduce Rancidity for Domestically Distributed Herbal Medicines (유통 한약재의 산패에 따른 품질변화 및 산패 저감화 연구)

  • Young-Ae Park;Suk-Kyung Ko;Hyun-Kyung Lee;Eun-Jung Choi;Sung-Cho Hong;Yun-Seon Park;Ji-Hun Jung;Ju-Sung Park;Yong-Seung Shin
    • Korean Journal of Pharmacognosy
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    • v.54 no.2
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    • pp.80-87
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    • 2023
  • Rancidity changes were examined for 6 herbal medicines, namely Persicae Semen, Armeniacae Semen, Lini Semen, Trichosanthis Semen, Arecae Semen, Myristicae Semen known to have relatively high fat content. In order to reduce rancidity of herbal medicines, samples were stored at 3 different conditions of room, refrigerating and freezing temperatures, and the rancidity was measured for 10 months with every 2 month interval. Fat content was extracted by using ethyl ether, and acid values and peroxide values, which are generally accepted indicators of fat rancidity, were measured. When storing Persicae Semen, Lini Semen and Arecae Semen at room temperature, the acid values increased as the storage period increased, and it was higher than when stored in refrigeration or freezing. The measurement of peroxide value showed more significantly higher initial degree of rancidity when Persicae Semen, Trichosanthis Semen, Arecae Semen and Myristicae Semen were stored at room temperature. It was observed that storing herbal medicines in refrigeration or freezing inhibited their rancidity compared to storing them at room temperature. To investigate the quality changes according to rancidity, the analysis of aflatoxins and indicator components showed that aflatoxins B1 and B2 were detected in Armeniacae Semen, Arecae Semen and Myristicae Semen, and the amount of amygdalin was well maintained within the specification standard.

Effect Of Nelumbinis Semen On The Recovery Of The Cardiac Muscle Activity by Proteome Analysis (연자육(蓮子肉)의 심근 경색 모델에 대한 Proteom 분석)

  • Ahn, Chang-Joon;Lee, Gi-Hyun;Kim, Yang-Seok;Hong, Moo-Chang;Bae, Hyun-Su;Kim, Jong-Hoon;Shin, Min-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.6
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    • pp.962-969
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    • 2010
  • The purpose of this investigation was to confirm the effect of Nelumbinis Semen on the recovery of the cardiac muscle activity. We studied the effect of Nelumbinis Semen on the recovery of ischemic SD rat hearts perfused with Nelumbinis Semen, using a model of ex-vivo perfusion (Non-working Langendorff perfusion system) and working heart perfusion system at the same time. To explore the effect of Nelumbinis Semen at the level of proteome, two-dimensional electrophoresis and MALDI-TOF analysis were performed. We found out that the proteins increased after perfusion of Nelumbinis Semen are Mitochondrial aconitase, ATP synthase alpha chain, Lactate dehydrogenase B, Creatine kinase, Glyceraldehyde 3-phosphate dehydrogenase, Alpha B-crystallin, Myosin and Heart fatty acid binding protein. Almost, all of them are concerned with ATP production in the cardiac muscle with glucose metabolism.

Profiling of differentially expressed proteins between fresh and frozen-thawed Duroc boar semen using ProteinChip CM10

  • Yong-Min Kim;Sung-Woo Park;Mi-Jin Lee;Da-Yeon Jeon;Su-Jin Sa;Yong-Dae Jeong;Ha-Seung Seong;Jung-Woo Choi;Shinichi, Hochi;Eun-Seok Cho;Hak-Jae Chung
    • Journal of Animal Science and Technology
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    • v.65 no.2
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    • pp.401-411
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    • 2023
  • Many studies have been conducted to improve technology for semen cryopreservation in pigs. However, computer-assisted analysis of sperm motility and morphology is insufficient to predict the molecular function of frozen-thawed semen. More accurate expression patterns of boar sperm proteins may be derived using the isobaric tags for relative and absolute quantification (iTRAQ) technique. In this study, the iTRAQ-labeling system was coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis to identify differentially expressed CM10-fractionated proteins between fresh and frozen-thawed boar semen. A total of 76 protein types were identified to be differentially expressed, among which 9 and 67 proteins showed higher and lower expression in frozen-thawed than in fresh sperm samples, respectively. The classified functions of these proteins included oxidative phosphorylation, mitochondrial inner membrane and matrix, and pyruvate metabolic processes, which are involved in adenosine triphosphate (ATP) synthesis; and sperm flagellum and motile cilium, which are involved in sperm tail structure. These results suggest a possible network of biomarkers associated with survival after the cryopreservation of Duroc boar semen.

An Experimental Study on the Effect of the Epididymal Biopsy on the Obstruction of the Epididymal Ducts (부고환 생검이 부고환의 폐색에 미치는 실험적 연구)

  • Kim, Du-Cheun;Kim, Sae-Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.11 no.2
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    • pp.77-81
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    • 1984
  • Epididymal biopsy has been performed without consideration of the possibility of epididymal ductal severance and obstruction which result in obstructive azoospermia. An attempt was made to study the effect of the epididymal biopsy on the obstruction of the epididymal ducts. Bilateral epididymal biopsies were done in 8 healthy rabbits (New Zealand White strain) weighing over 3 kg, and then ejaculated semens have been analyzed 5 times every other week from 1 month after biopsies. Microscopic examination of the biopsied epididymides was also done after the 5th semen analysis. The results were as follows. 1. Semen analysis: 6 out of 8 rabbits showed azoospermia from the 4th semen analysis and 2 cases showed normal number of the sperms in the 5th semen analysis. 2, Microscopic examination: 6 cases of azoospermia showed complete obstruction of the biopsied sites of the epididymides, and abscence of sperms in epididymal ducts distal to the biopsied sites of the epididymides. However, recanalization of the epididymal ducts was noted in 2 cases showing normal sperm count. Therefore, it is concluded that the epididymal biopsy should be avoided in patients who want to be fertile, because it may cause the epididymal severance and obstruction of the epididymal ducts.

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Effect of Storage in Different Commercial Semen Extenders on the Motility, Viability and Fertility In Vitro of Boar Spermatozoa (수퇘지 정자의 운동성, 생존성 및 체외수정 능력에 대한 시판 액상 정액 보존액과 보존 기간의 영향)

  • Sa, Soo-Jin;Kim, Myung-Jick;Cho, Kyu-Ho;Kim, Du-Wan;So, Kyoung-Min;Chung, Ki-Hwa;Son, Jung-Ho;Kim, In-Cheul
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.203-207
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    • 2011
  • The objective of this study was to determine the effect of semen extenders on the motility, viability and fertility in vitro of spermatozoa during storage of fresh boar semen diluted in different commercial extenders used for pig artificial insemination (AI). In this experiment, semen were diluted in Androhep plus, Beltsville Thawing Solution (BTS), Modena, Seminark and Vitasem LD. Five ejaculates were collected from three Duroc boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different extenders. Semen was stored at $170^{\circ}C$ for 10 days. Sperm motility and viability was assessed using Computer-Assisted Semen Analysis (CASA) and flow-cytometry on 1, 3, 5 and 10 day post collection The motility of spermatozoa stored in different extenders was gradually decreased by increasing the duration of storage of semen. However, there was not significant1y different in the sperm motility and viability among other extenders. On the other hand, the in vitro-matured oocytes were fertilized and cultured in vitro to assess the fertility of boar spermatozoa stored for 3 and 10 days in different extenders. The percentage of morula and blastocyst were taken as indicators of fertility in vitro of spermatozoa. Therefore, there were no differences in the rate of embryos developed to the molular and blastocyst stage. There were no differences in the motility and fertility in vitro among 5 kinds of commercial boar semen extenders.

Molecular authentication of Lepidii seu Descurainiae Semen by the development of matK amplification primers and analysis of sequences (matK 증폭용 primer 개발 및 염기서열 분석을 통한 정력자(葶藶子) 유전자 감별)

  • Moon, Byeong Cheol;Kim, Wook Jin;Yang, Sungyu;Park, Inkyu;Yeo, Sang Min;Noh, Pureum
    • The Korea Journal of Herbology
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    • v.33 no.3
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    • pp.25-35
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    • 2018
  • Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

Protective Effect of Carthamus tinctorius L. Semen on Hepatotoxicity by Carbon Tetrachloride in Rats (사염화탄소에 의한 간독성에 미치는 홍화자의 보호작용)

  • 정기화;정춘식
    • Biomolecules & Therapeutics
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    • v.4 no.4
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    • pp.428-436
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    • 1996
  • The protective effect of Carthamus tinctorius L. Semen on the carbon tetrachloride induced liver damaged rats were studied. First, methanol extract was prepared and the extract was fractionated with hexane, $CHCl_3$, BuOH and $H_2O$ respectively. Animals were divided into 6 groups and each group was treated with each fraction respectively. To investigate the hepato-protective effect of Carthamus tinctorius L. Semen AST, ALT, albumin, TP, cholesterol, TG, creatinine and total bilirubin values were measured in each treated group and compared with those of control group. GST activity was increased in BuOH group compared with the control group. In malondialdehyde levels, all fractions was decreased compared with the control group. In histopathologic examination, hexane and $H_2O$ fractions of Carthamus tinctorius L. Semen observed mild degree of ballooning degeneration. The results show the protective effect of Hexane,$CHCl_3$, BuOH and $H_2O$ fractions on hepatotoxicity of $CHl_4$ by decreasing ALT, AST, bilirubin, cholesterol, TG and BUN. It seems that the decrease of MDA are related to the recovery effect. The protective effects of Carthamus tinctorius L. Semen fractions in hepatotoxic pathogenesis by $CHl_4$ was suggested in blood chemistry analysis and histopathologic examination.

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The Effects of Supplementation of BSA or Fatty Acid Free BAS on the Motility of Fresh or Cryopreserved Rooster Spermatozoa (BSA 또는 Fatty Acid Free BSA 첨가가 닭 희석 정자와 동결 정자의 생존성에 미치는 영향)

  • Kim, Sung Woo;Kim, Min Soo;Yu, Yeonhui;Kim, Chan-Lan;Jeon, Ik Soo;Kim, Chongdae
    • Korean Journal of Poultry Science
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    • v.44 no.1
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    • pp.59-65
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    • 2017
  • This study examined factors affecting the analysis of motility of chicken semen. The viability of spermatozoa was estimated using varying dilution ratios and supplementation with BSA or fatty acid free (FAF)-BSA as protein sources in semen diluent. Fresh semen was examined after preparing dilutions in beltsvile poultry semen extender (BPSE) of 1/8, 1/16 and 1/32 at $25^{\circ}C$. The motility of incubated semen at each dilution was observed at 3 min (89.9%, 69.9% and 53.2%), 30 min (86.7%, 71.4% and 51.7%), 1 h (89.5%, 74.0% and 53.5%) and 3 h (78.5%, 66.5% and 45.7%), respectively. The addition of BSA or FAF-BSA to BPSE diluent significantly increased the viability of semen in 1/32 dilution with results of 53.2% (control), 84.8% (BSA) and 92.9% (FAF-BSA) (p<0.05). This phenomenon was also observed in the dilution of frozen semen, where FAF-BSA treatment increased the viability of thawed semen from 17.6% to 34.0% in a 1/8 dilution (p<0.05). When the protein sources were used in the dilution, the survival rates of diluted chicken semen were also increased with time lapse. These results show that FAF-BSA may act to protect chicken semen and is suitable as a basic component of chicken semen diluent for the method of analyzing rooster semen after freezing.

Sperm chromatin structure assay results in Nigerian men with unexplained infertility

  • Faduola, Paul;Kolade, Charles Oluwabukunmi
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.3
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    • pp.101-105
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    • 2015
  • Objective: Several publications have established a relationship between sperm DNA damage and male factor infertility, based on data from America, Europe, and Asia. This study aimed to compare the extent of sperm DNA damage in sperm samples from Nigerian men with unexplained infertility and in sperm samples from a fertile group composed of sperm donors who had successfully impregnated a female partner naturally or through assisted conception. Methods: A total of 404 men underwent male fertility evaluation at Androcare Laboratories and Cryobank participated in this study. Semen analysis and a sperm chromatin structure assay (SCSA) were performed on all subjects. Results: The men in the unexplained infertility group were slightly older than the men in the fertile sperm group ($36{\pm}10$ years vs. $32{\pm}6$ years, p=0.051). No significant difference was observed between the two groups in semen analysis parameters ($p{\geq}0.05$). Men in the unexplained infertility group with normal semen parameters had a significantly higher DNA fragmentation index (DFI) than men in the fertile sperm group ($27.5%{\pm}7.0%$ vs. $14.1%{\pm}5.3%$, p<0.05). In the unexplained infertility group, 63% of the men had a DFI greater than 20%, compared to 4% in the fertile sperm group. In the unexplained infertility group, 15.2% of the subjects had a DFI greater than 30%, compared to 1% in the fertile sperm group. Conclusion: Our study showed that the SCSA may be a more reliable predictor of fertility potential than traditional semen analysis in cases of unexplained infertility.