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"The study on the dimethylformamid (DMF) optimization for maintenance sperm quality of the carp fish (Cyprrnus carpio L.) post-thawing has been performed. Sperm were collected by stripping method and were dissolved in the extender solution (DMF : Kurokura solution = 1 : 3). DMF concentrations were 0%, 5%, 10%, and 15%, respectively. The soluble sperm were kept on the 0.2mL of straw and were cryopreserved by the slow freezing method.Sperm analysis were carried out at the time of collection, post-equilibration, and post-thawing, respectively. Some parameters of the sperm quality observed were spermatozoa motility, viability, and abnormality. All data were analyzed by Analysis of Varians (ANOVA). DMF 10% were significantly (cc = 0.01) kept spermatozoa motility, viability, and abnormality relatively higher than DMF 0%, 5%, and 15%. These results suggest that DMF 10% is the optimize condition for maintenance sperm quality post-thawing."

"Platelet rich plasma (PRP) sebagai bahan suplemen medium kriopreservasi, mengandung plasma yang merupakan bagian dari darah dan mengandung banyak sekali albumin. Albumin diketahui sebagai CPA ekstraseluler alami yang bekerja dengan cara menstabilkan membran sel yang dapat terganggu akibat kriopreservasi. Bahan suplementasi medium kriopreservasi selama ini menggunakan bahan yang berasal dari hewan. Penggunaan bahan suplementasi dari hewan telah diketahui memiliki berbagai kendala seperti tersandung dengan komunitas perlindungan hewan dan juga dapat mencetuskan reaksi immunologi jika digunakan pada manusia. Karena itu, perlu dicari alternatif lain untuk bahan suplementasi medium kriopreservasi. Penelitian ini meneliti apakah PRP dapat digunakan sebagai alternatif FBS sebagai medium kriopreservasi sel punca asal tali pusat manusia dengan menilai viabilitas, morfologi dan proliferasi sel punca pasca kriopreservasi. Penelitian diawali dengan isolasi dan propagasi sel punca asal tali pusat manusia dari satu buah tali pusat manusia yang memenuhi kriteria dengan metode eksplan. Sel punca kemudian disubkultur sampai mencapai jumlah sel yang dibutuhkan untuk kriopreservasi. Kriopreservasi sel punca dilakukan dalam delapan protokol kriopreservasi dengan variasi bahan suplemen, konsentrasi bahan suplemen dan konsentrasi sel. Hasil penelitian menunjukkan tidak ada perbedaan antara FBS dan PRP dalam mempertahankan viabilitas dan morfologi sel bahkan PRP lebih baik ketika dilihat dari ukuran dan proliferasi pasca kriopreservasi. Sebagai kesimpulan, penelitian ini menunjukkan bahwa PRP dapat digunakan sebagai alternatif penggunaan FBS dalam medium kriopreservasi sel punca asal tali pusat manusia.

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Platelet rich plasma ( PRP ) as supplemental material cryopreservation medium , containing plasma which is part of the blood and contains a lot of albumin. Albumin is known as a natural extracellular CPA works by stabilizing cell membranes that can be disrupted by cryopreservation .One of the materials in cryopreservation medium that is used nowadays is derived from animals. Use of animal derived metarial has been known to pose various problems such as facing the animal protection community and also can trigger immunological reactions when used in humans. So it is necessary to find an alternative for animal derived material in cryopreservation medium. This study examined whether PRP could be used as an alternative to FBS as cryopreservation medium for human umbilical cord stem cells by assessing the viability, morphology and proliferation of stem cells after cryopreservation. The study began with the isolation and propagation of human umbilical cord stem cells from one human umbilical cord that meets the criteria using explant method. Stem cells then subcultured to achieve the required number of cells for cryopreservation. Cryopreservation of stem cells was done in eight cryopreservation protocols with various supplements, concentrations of the supplements, and cell concentrations. The results showed no difference between FBS and PRP in maintaining cell viability and morphology. PRP was even better when viewed from the size and proliferation of the cells after cryopreservation . In conclusion, this study shows that PRP can be used as an alternative to FBS in cryopreservation medium for human umbilical cord stem cells."

"Penelitian kriopreservasi spermatozoa ikan patin albino bertujuan untuk menganalisis ultrastruktur, fisiologi, dan molekuler spermatozoa ikan patin albino pasca kriopreservasi. Kriopreservasi dilakukan pada suhu -80°C selama 14 hari menggunakan kombinasi krioprotektan intraseluler yaitu metanol 10% dan krioprotektan ekstraseluler yaitu susu skim. Hasil ultrastruktur spermatozoa menunjukkan bahwa pada spermatozoa segar bagian membran sel kepala, mid piece, dan bagian flagel masih dalam kondisi utuh dan baik. Ultrastruktur spermatozoa pasca ekuilibrasi nampak ada perbesaran lebar dan panjang kepala spermatozoa dibandingkan spermatozoa segar, walaupun secara struktur masih tampak utuh. Ultrastruktur spermatozoa pasca pencairan tampak terjadi kerusakan membran bagian kepala dan flagel. Hasil pengukuran morfometri spermatozoa menunjukkan adanya peningkatan lebar kepala spermatozoa yaitu 1,59 µm pada spermatozoa segar menjadi 1,97 µm pada spermatozoa pasca ekuilibrasi dan 2,40 µm pada spermatozoa pasca pencairan. Demikian pula, terdapat perubahan panjang kepala spermatozoa yaitu 3,70 µm pada spermatozoa segar menjadi 3,81 µm pada spermatozoa pasca ekuilibrasi, dan 3,90 µm pada spermatozoa pasca pencairan. Analisis viabilitas spermatozoa didapatkan penurunan viabilitas spermatozoa pasca pencairan (61±2,30%) dibandingkan spermatozoa segar (92±0,58%) dan spermatozoa pasca ekuilibrasi (80±3,51%). Analisis fisiologi spermatozoa didapatkan penurunan fungsi mitokondria pada spermatozoa pasca ekuilibrasi (57±7%) dan spermatozoa pasca pencairan (42±3,2%) dibandingkan spermatozoa segar (98±2%). Analisis motilitas spermatozoa menunjukkan penurunan motilitas spermatozoa pasca ekuilibrasi (79±4,5%) dan spermatozoa pasca pencairan (30±3,2%) dibandingkan spermatozoa segar (87±1,5%). Penetasan telur pasca 24 jam fertilisasi pada perlakukan spermatozoa pasca ekuilibrasi didapatkan hasil lebih tinggi (64±17%) dibandingkan spermatozoa segar (38±4%), sedangkan spermatozoa pasca pencairan tidak ditemukan ada penetasan telur. Analisis molekular spermatozoa pada gen CO1 dan SOD2 didapatkan jumlah lesi gen SOD2 spermatozoa pasca ekuilibrasi yaitu 15,83 lesi / 10 kb dan spermatozoa pasca pencairan yaitu 17,14 lesi / 10 kb. Lesi gen CO1 pada spermatozoa pasca ekuilibrasi yaitu 9,24 lesi / 10 kb dan spermatozoa pasca pencairan yaitu 10,26 lesi / 10 kb. Sehingga disimpulkan kriopreservasi spermatozoa berpengaruh terhadap ultrastruktur, fisiologi, dan molekuler spermatozoa ikan patin albino.

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Research of cryopreservation on albino Pangasius catfish spermatozoa aims to analyze about ultrastructure, physiology, and molecular spermatozoa of albino Pangasius catfish post cryopreservation. Cryopreservation was carried out at -80°C for 14 days using a combination of intracellular cryoprotectants which is 10% methanol and extracellular cryoprotectant which is skim milk. The results of the spermatozoa ultrastructure showed that the cell membrane of the spermatozoa head, the midpiece, and the flagellum of fresh spermatozoa were still intact and good. The spermatozoa ultrastructure after post equilibration, shown enlargement of the head width and length compared to the fresh spermatozoa, although structurally were still intact. The ultrastructure of frozen-thawed spermatozoa, appeared a membrane damage at the head and flagellum. The results of spermatozoa morphometric measurements showed an increase at the head width of spermatozoa from 1.59 µm in fresh spermatozoa to 1.97 µm in post-equilibration spermatozoa and 2.40 µm in frozen-thawed spermatozoa. Similarly, there was an increase in the head length of spermatozoa, from 3.70 µm in fresh spermatozoa, to 3.81 µm in post-equilibration spermatozoa, and 3.90 µm in frozen-thawed spermatozoa. The viability analysis showed a decrease of frozen-thawed spermatozoa viability (61±2.30%) compared to fresh spermatozoa (92±0.58%) and post-equilibration spermatozoa (80±3.51%). The analysis physiology of spermatozoa showed a decrease in mitochondrial function in post equilibration spermatozoa (57±7%) and frozen-thawed spermatozoa (42±3.2%) compared to fresh spermatozoa (98±2%). The analysis of motility of spermatozoa showed a decrease in post equilibration spermatozoa (79±4.5%) and frozen-thawed spermatozoa (30±3.2%) compared to fresh spermatozoa (87±1.5%). Egg hatching after 24 hours of fertilization for the post-equilibration spermatozoa was higher (64±17%) than fresh spermatozoa (38±4%), whereas frozen-thawed spermatozoa were not hatched. The analysis of molecular on CO1 and SOD2 genes obtained the number of gene lesions in the spermatozoa SOD2 gene after equilibration were 15.83 lesions/10 kb and frozen-thawed were 17.14 lesions/10 kb. The CO1 gene lesions in post-equilibration spermatozoa were 9.24 lesions/10 kb, while the CO1 gene lesions in frozen-thawed spermatozoa were 10.26 lesions/10 kb. It can be concluded that there is an effect of cryopreservation on ultrastructure, physiology, and molecular in spermatozoa of albino Pangasius catfish."

"Pruatjan (Pimpinella pruatjan Molk) is an Indonesian endangered medicinal plant that included in Appendix I based on CITES. Therefore it is a highly protected species...."

"Telah dilakukan penelitian kriopreservasi spermatozoa ikan tawes, Barbonymus gonionotus (Bleeker, 1850) menggunakan berbagai konsentrasi susu skim dalam larutan pengencer yang dicampurkan dengan 5% metanol. Penelitian bertujuan mengetahui pengaruh campuran 5% metanol dengan 15% susu skim terhadap kualitas (motilitas, viabilitas, dan abnormalitas) spermatozoa ikan tawes satu hari pascakriopreservasi. Semen ikan tawes didapatkan melalui metode pengurutan (stripping). Semen diencerkan dalam larutan pengencer berupa larutan fish ringer yang ditambahkan dengan 5% metanol dan berbagai konsentrasi susu skim. Pengamatan kualitas spermatozoa dilakukan sebelum dan sesudah kriopreservasi. Hasil terbaik ditunjukkan oleh campuran 5% metanol dengan 20% susu skim dengan nilai persentase motilitas, viabilitas, dan abnormalitas secara berurutan sebesar 83,23 + 3,27%; 81,75 + 8,22%; dan 26,25 + 1,89%."

"Telah dilakukan penelitian untuk mengetahui pengaruh sari buah jeruk siam Citrus nobilis Lour. dalam pengencer tris kuning telur TKT terhadap kualitas spermatozoa domba garut Ovis aries L. 24 jam pascakriopreservasi. Semen ditampung dari lima pejantan domba garut satu kali dalam satu minggu menggunakan vagina buatan. Segera setelah dievaluasi, semen diencerkan dengan pengencer tris kuning telur yang mengandung sari buah jeruk 0 KK, 5 KP 1, 10 KP 2, 15 KP 3, dan 20 KP 4. Semen dikemas dalam straw 0,25 ml dengan konsentrasi akhir 50x106 spermatozoa/ml. Semen diekuilibrasi pada suhu 5oC selama dua jam kemudian dibekukan dan disimpan dalam tabung N2 cair. Semen dievaluasi terhadap parameter motilitas, viabilitas, integritas membran, integritas akrosom, dan abnormalitas. Hasil uji ANAVA satu arah menunjukkan terdapat perbedaan nyata antar kelompok perlakuan terhadap persentase motilitas, viabilitas, dan integritas membran p < 0,05. Hasil uji lanjut Duncan menunjukkan bahwa KP 2 memiliki perbedaan nyata terhadap semua kelompok perlakuan. Berdasarkan hasil tersebut, penambahan sari buah jeruk pada konsentrasi 10 KP 2 merupakan konsentrasi terbaik yang mampu memperkecil penurunan kualitas spermatozoa domba garut pascakriopreservasi berdasarkan persentase motilitas 58,68 0,68, viabilitas 59,73 6,47, dan integritas membran 54,87 5,58.

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The research aimed to find out the effect of siam orange juice in extender on garut ram spermatozoa quality 24 hours postcryopreseration. Semen was collected from five rams once a week using artificial vagina. Semen sample was diluted in tris egg yolk based extender containing 0 KK, 5 KP 1, 10 KP 2, 15 KP 3 and 20 KP 4 siam orange juice. Semen was packed in 0.25 ml straw with a final concentration of 50x106 spermatozoa ml. Sperm was equilibrated at 5 C for two hours then frozen and stored in a liquid nitrogen N2 tube. Sperm parameters including motility, viability, membrane integrity, acrosome integrity, and abnormality were assessed. One way ANAVA test results showed significant differences between treatment groups on the percentage of motility, viability, and membrane integrity."

"Telah dilakukan penelitian untuk mengetahui pengaruh berbagai konsentrasi sari buah nanas Ananas comosus L. Merr. dalam pengencer terhadap kualitas spermatozoa domba garut Ovis aries L. 24 jam pascakriopreservasi. Sampel semen ditampung dari lima ekor domba garut jantan satu kali dalam satu minggu menggunakan vagina buatan. Semen diencerkan dengan pengencer tris kuning telur yang mengandung sari buah nanas 0 KK, 5 KP 1, 10 KP 2, 15 KP 3, dan 20 KP 4. Semen dikemas dalam straw mini 0,25 ml dengan dosis 50 juta sel/ml. Semen diekuilibrasi pada suhu 5oC selama dua jam, kemudian dibekukan dan disimpan dalam nitrogen cair. Parameter yang dievaluasi adalah motilitas, viabilitas, integritas membran, integritas akrosom, dan abnormalitas. Hasil uji ANAVA satu faktor yang dilanjutkan dengan uji Duncan menunjukkan perbedaan nyata P < 0,05 antara KK dan KP 3 terhadap persentase motilitas dan integritas membran spermatozoa. Konsentrasi sari buah nanas 15 mampu memperkecil penurunan kualitas spermatozoa berdasarkan persentase motilitas 52,19 5,32 dan integritas membran spermatozoa 38,52 4,85.

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The research aimed to find out the effect of various concentrations of pineapple Ananas comosus L. Merr. juice in extender on garut ram Ovis aries L. 24 hours postcryopreservation. Semen samples were collected from five garut rams once a week using an artificial vagina. The samples were diluted in tris egg yolk extender with pineapple juice 0 KK, 5 KP 1, 10 KP 2, 15 KP 3, and 20 KP 4. The diluted semen samples were loaded into mini straws 0,25 ml with 50 million cell ml dosage. Samples were equilibrated at 5oC for two hours, then freezed and stored in liquid nitrogen. Parameters evaluated were motility, viability, membrane integrity, acrosome integrity, and abnormality. The one factor ANOVA and continued with Duncan test showed significant differences P 0.05 between KK and KP 3 on the percentage of spermatozoa motility and membrane integrity. Pineapple juice 15 was able to minimize the reduction of spermatozoa quality based on the percentage of spermatozoa motility 52.19 5.32 and membrane integrity 38.52 4.85."

"Penelitian mengenai pengaruh pemberian berbagai konsentrasi konsentrat kurma Phoenix dactylifera L. terhadap kualitas spermatozoa ikan koi Cyprinus carpio, Linnaeus 1759 48 jam pascakriopreservasi telah dilakukan. Penelitian tujuan untuk mengetahui pengaruh pemberian kombinasi metanol 10 dengan berbagai konsentrasi konsentrat kurma 0, 3, 5, 7, dan 9 terhadap motilitas, viabilitas, dan abnormalitas spermatozoa ikan koi 48 jam pascakriopreservasi. Semen ikan koi yang digunakan untuk kriopreservasi dikoleksi dengan metode pengurutan stripping yang kemudian dievaluasi secara makroskopis dan mikroskopis.Hasil evaluasi selanjutnya diuji dengan uji normalitas Shapiro-Wilk, uji homogenitas Levene, uji analisis variansi ANAVA faktor tunggal, dan uji perbandingan berganda Tukey. Hasil uji ANAVA faktor tunggal menunjukkan perbedaan nyata P < 0,05 terhadap persentase motilitas, viabilitas, dan abnormalitas spermatozoa ikan koi pascakriopreservasi. Metanol 10 dengan konsentrat kurma 5 merupakan kombinasi krioprotektan optimum karena menghasilkan nilai rata-rata tertinggi pada persentase motilitas 61,74 4,47 dan viabilitas 72,60 2,96, serta memberikan nilai rata-rata terendah pada persentase abnormalitas 22,00 3,16 spermatozoa ikan koi 48 jam pascakriopreservasi.

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Effects of various concentrations of dates palm juice Phoenix dactylifera L. to spermatozoa quality of koi fish Cyprinus carpio, Linnaeus 1758 48 hours post cryopreservation was investigated. The aim of this research is to determine combination effects of 10 methanol and various concentration of dates palm juice 0, 3, 5, 7, and 9 to koi fish spermatozoas motility, viability, and abnormality 48 hours post cryopreservation. Semen for cryopreservation was collected by stripping, then evaluated by macroscopic and microscopic.

The results of the evaluation were tested by Shapiro Wilk normality test, Levene homogeneity test, variants analysis ANAVA single factor test, and Tukey multiple comparison test. According to ANAVA, the percentage of motility, viability, and abnormality post cryopreservation was significantly different P 0,05. 10 methanol and 5 dates palm juice was the optimum concentration of cryoprotectant because it provides the highest average value in motility percentage 61,74 4,47 and viability percentage 72,60 2,96, and provides the lowest average value in abnormality percentage 22,00 3,16 spermatozoa of koi fish 48 hours post cryopreservation."

"Ikan kancra Tor soromerupakan ikan air tawar yang digemari oleh masyarakat untuk dikonsumsi dan digunakan untuk upacara adat. Sehingga permintaan konsumen terhadap ikan kancra meningkat. Hal tersebut menyebabkan terjadinya penangkapan ikan kancra yang berlebih di alam sehingga populasi ikan kancra menurun. Oleh karena itu, perlu dilakukan pelestarian ikan kancra dengan cara kriopreservasi spermatozoa. Penelitian menggunakan rancangan acak lengkap (RAL) dengan empat perlakuan dan enam ulangan. Empat perlakuan terdiri atas sari buah tomat 0% + DMSO 10% (SBt 0%); sari buah tomat 10% + DMSO 10% (SBt 10%); sari buah tomat 20% + DMSO 10% (SBt 20%); dan sari buah tomat 30% + DMSO 10% (SBt 30%). Tujuan dari penelitian ini adalah untuk mengetahui pemberian sari buah tomat Lycopersicon esculentumsebagai antioksidan alami terhadap spermatozoa ikan kancra 24 jam pascakriopreservasi. Parameter uji kualitas spermatozoa meliputi motilitas, viabilitas, abnormalitas, dan kemampuan fertilisasi. Data hasil penelitian yang diperoleh diuji dengan menggunakan uji Analisis Variansi (ANAVA) satu faktor. Hasil penelitian menunjukkan adanya perbedaan nyata (P < 0,05) pada nilai rata-rata persentase viabilitas dan kemampuan fertilisasi serta tidak adanya perbedaan nyata (P > 0,05) pada nilai rata-rata persentase motilitas dan abnormalitas. Hasil penelitian membuktikan bahwa penambahan konsentrasi 10% sari buah tomat dalam ekstender memberikan pengaruh yang cukup positif terhadap kualitas spermatozoa ikan kancra 24 jam pascakriopreservasi, yaitu dengan ditunjukkan nilai rerata persentase motilitas (32,57 ± 5,94%); viabilitas (14,5 ± 4,88%); abnormalitas (74,16 ± 2,13%); dan kemampuan fertilisasi (81,25 ± 6,07%).  Kancra fish Tor soro are freshwater fish that are favored by the community for consumption and use for traditional ceremonies. So that consumer demand for kancra fish increases. This causes the occurrence of over-fishing in the wild which causes the population of fish to decrease. Therefore, it is necessary to preserve kancra fish by cryopreservation of spermatozoa. The study used a completely randomized design with four treatments and six replications. Four treatments consisted of 0% tomato juice + 10% DMSO (0% SBt); 10% tomato juice + 10% DMSO (10% SBt); 20% tomato juice + 10% DMSO (20% SBt); and 30% tomato juice + 10% DMSO (SBt 30%). The purpose of this study was to determine the administration of tomato juice Lycopersicon esculentum as a natural antioxidant on spermatozoa of kancra fish 24 hours post cryopreservation. Spermatozoa quality test parameters include motility, viability, abnormality, and fertilization rates. The research data obtained were tested using the one-factor Variance Analysis (ANOVA) test. The results showed a significant difference (P < 0.05) in the average value of the percentage of viability and fertilization rates, and no significant difference (P > 0.05) in the average value of the percentage of motility and abnormality. The results of the study prove that the addition of 10% concentration of tomato juice in the extender has a quite positive influence on the quality of spermatozoa of kancra fish 24 hours post cryopreservation, that is indicated by the mean value of the percentage of motility (32.57 ± 5.94%); viability (14.5 ± 4.88%); abnormalities (74.16 ± 2.13%); and fertilization ability (81.25 ± 6.07%)."

"ABSTRAKKriopreservasi merupakan metode penyimpanan material genetik pada suhu rendah dalam jangka waktu tertentu. Kriopreservasi spermatozoa dapat digunakan sebagai usaha konservasi ex situ ikan kancra (Tor soro) yang populasinya semakin menurun. Pemilihan krioprotektan yang tepat dapat meminimalkan kerusakan akibat terbentuknya kristal es dan mempertahankan kualitas spermatozoa selama kriopreservasi. Gula merah sebagai krioprotektan alami dan metanol 10% memiliki potensi untuk kriopreservasi spermatozoa ikan kancra. Tujuan penelitian yaitu mendapatkan konsentrasi optimal dari berbagai konsentrasi gula merah dan metanol 10% terhadap motilitas, viabilitas, dan abnormalitas spermatozoa pascakriopreservasi ikan kancra, serta mengevaluasi persentase fertilitas spermatozoa pascakriopreservasi ikan kancra. Konsentrasi gula merah yang digunakan yaitu 0%, 5%, 10%, 15%, 20%, dan 25%. Rasio antara semen dan larutan pengencer yang digunakan yaitu 1:10. Ekuilibrasi dilakukan pada suhu 5 °C selama 10 menit. Pembekuan dilakukan pada suhu -10 °C selama 48 jam. Pencairan dilakukan pada suhu 40 °C selama 60 detik. Analisis data dilakukan dengan menggunakan uji ANAVA yang dilanjutkan dengan uji Tukey. Hasil penelitian menunjukkan bahwa krioprotektan gula merah memberikan pengaruh (P<0,05) pada kualitas spermatozoa pascakriopreservasi. Krioprotektan gula merah 15% dan metanol 10% menghasilkan persentase motilitas (81,85 ± 1,11%), viabilitas (83,75 ± 1,71%), fertilitas (89,75 ± 1,71%) tertinggi, serta abnormalitas terendah (14,50 ± 1,73%) pada spermatozoa pascakriopreservasi.

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ABSTRACTCryopreservation is a method of storing genetic material at low temperatures for a certain period of time. Spermatozoa cryopreservation can be used as an ex situ conservation method for kancra fish (Tor soro) whose population is declining. Selection of the right cryoprotectant can minimize damage caused by the formation of ice crystals and maintain the quality of spermatozoa during cryopreservation. Brown sugar as natural cryoprotectant and 10% methanol have potential for cryopreservation of spermatozoa. The research objective was to obtain the optimal concentration of various concentrations of brown sugar and 10% methanol on the motility, viability, and abnormalities of kancra fish spermatozoa after cryopreservation, and evaluate the fertility of kancra fish spermatozoa after cryopreservation. The concentration of brown sugar used were 0%, 5%, 10%, 15%, 20%, and 25%. The ratio between semen and diluent solution used was 1:10. Equilibration was carried out at 5 °C for 10 minutes. Freezing was carried out at -10 °C for 48 hours. Thawing was done at 40 °C for 60 seconds. Data was analyzed using ANOVA test followed by Tukey test. The results showed that the cryoprotectant brown sugar had an significant influence (P<0.05) on the quality of post-cryopreserved spermatozoa. 15% brown sugar together with 10% methanol can produce the highest percentage of motility (81.85 ± 1.11%), viability (83.75 ± 1.71%), fertility (89.75 ± 1.71%), and the lowest abnormality (14.50 ± 1.73%) in spermatozoa post-cryopreservation."