scholarly journals EFEK INTERAKSI MASA EKUILIBRASI DAN LAJU PENURUNAN SUHU TERHADAP PENINGKATAN KEUTUHAN MEMBRAN PLASMA SPERMA DOMBA PRIANGAN PASCA THAWING (The Interaction Effect of Equilibration Time and Freezing Rate toward the Membran Plasma Sperm Integrity of Priangan Ram Sperm Post thawing)

2020 ◽  
Vol 8 (2) ◽  
pp. 73-78
Author(s):  
Ken Ratu Gharizah Alhuur ◽  
Soeparna Soeparna ◽  
Rd. S. Darodjah

In general, the freezing process of semen will have a detrimental effect on the structure and function of spermatozoa. The critical point in the semen freezing process is when the temperature drops to freezing, but this can be overcome by applying the right equilibration period. This study aimed to find out the best the equilibration length, the freezing rate,  their interaction in the semen freezing process towards membrane sperm cell integrity of the frozen semen post thawing. This study used Priangan sheep’s semen and was carried out according to a completely randomized design with a 3 x 3 split-plot design arrangement. The main plot was equilibration length, i.e., 0.5,  1.5, and 2.5 hours. The subplot was the freezing rate, i.e., 7.5ºC/mins, 13.5ºC/mins, and 19.5ºC/mins. The result of the study showed that there was a significant interaction (P<0.05) between the equilibration length of 1.5 hours and the freezing rate of 13.5°C/mins on the motility and the membrane sperm cell integrity. In conclusion, the best sperm quality was obtained from the equilibration length of 1.5 hours and the freezing rate of 13.5°C/mins.

2015 ◽  
Vol 27 (1) ◽  
pp. 127
Author(s):  
C. P. Freitas-Dell'aqua ◽  
C. Ramires Neto ◽  
Y. F. R. Sancler-Silva ◽  
P. M. Papa ◽  
J. A. Dell'aqua ◽  
...  

Commercial freeze extenders have different composition and ratio of cryoprotectors; freezing and thawing protocols are different for each extender. The aim of this experiment was to observe the effect of thawing curve in stallion frozen semen with 2 commercial extenders. Two ejaculates from each of 9 stallions of different breeds (Quarter Horses and Mangalarga Marchador) were used. Semen was collected using an artificial vagina, and the ejaculate was divided into 2 groups following the manufacture's protocol: group 1 (INRA), in which the semen was diluted 1 : 1 with the extender INRA 96TM (IMV, Paillette Crista, France) and group 2 (BC), in which the semen was diluted (1 : 1) with the extender Botu-SemenTM (Botupharma, Brazil). The samples of the 2 groups were centrifuged at 600 × g for 10 min, the supernatant was discarded, and the pellet was resuspended with INRA FreezeTM (group INRA, IMV) and with BotucrioTM (group BC, Botupharma) at the concentration of sperm 100 × 106 sperm mL–1. After this, the semen was packaged in 0.5-mL straws. For each group the freezing process was carried out according to the manufacturer's instructions. The straws were thawed in a water bath with 3 different thawing curves: 37°C for 30 s (37/30), 46°C for 20 s (46/20), and 75°C for 7 s (75/7) before analysis. The aim of these rates is to keep the semen in 37°C post-thaw. The sperm kinetic analysis was performed by computerized method (CASA, HTM-IVOS, IMV, USA) and the analysis of plasma membrane integrity by flow cytometer (BD LSR Fortessa, Becton Dickinson, Mountain View, CA, USA). Data of sperm kinetic and of plasma membrane integrity were compared among the 3 thawing curves for one extender using analysis of variance. Differences were considered significant at a probability level of 5%. No differences were observed in total motility (%, BC 37/30 = 72.8 ± 14.4; BC 46/20 = 70.0 ± 14.2; BC 75/7 = 70.3 ± 12.0 v. INRA 37/30 = 57.2 ± 19.1; INRA 46/20 = 50.0 ± 21.9; BC 75/7 = 58.8 ± 20.8), progressive motility (%, BC 37/30 = 36.9 ± 8.2; BC 46/20 = 34.4 ± 10.5; BC 75/7 = 33.6 ± 7.8 v. INRA 37/30 = 25.3 ± 12.7; INRA 46/20 = 21.9 ± 13.9; BC 75/7 = 28.9 ± 14.8), rapid sperm (%, BC 37/30 = 59.7 ± 16.4; BC 46/20 = 56.8 ± 17.1; BC 75/7 = 58.1 ± 14.9 v. INRA 37/30 = 38.3 ± 20.9; INRA 46/20 = 35.3 ± 22.9; BC 75/7 = 44.4 ± 23.8), and plasma membrane integrity (%, BC 37/30 = 49.1 ± 14.8; BC 46/20 = 43.1 ± 13.1; BC 75/7 = 46.7 ± 11.8 v. INRA 37/30 = 32.2 ± 10.7; INRA 46/20 = 29.6 ± 10.1; BC 75/7 = 37.4 ± 9.1) among the 3 thawing curves for INRA and BC groups. In this study, we can conclude there is no influence of the 3 tested thawing curves in sperm quality for stallion frozen semen with INRA Freeze and Botucrio extenders.


2020 ◽  
Vol 32 (2) ◽  
pp. 187
Author(s):  
M. Felix ◽  
I. Ortiz ◽  
H. Resende ◽  
J. Brom-de-Luna ◽  
C. Love ◽  
...  

Equine semen used for intracytoplasmic sperm injection (ICSI) is typically frozen-thawed and may be of poor quality. To prepare sperm for ICSI, semen is typically centrifuged to remove freezing extender. However, centrifugation can cause damage to sperm, which is especially meaningful if sperm quality is already poor. We evaluated a method for selection of sperm without centrifugation, using a “swim-over” technique, and assessed the effect of pentoxifylline, a phosphodiesterase inhibitor that increases sperm motility in other species. To mimic poor-quality semen, we thawed frozen semen (1×) and re-froze it three additional times (4×). Aliquots (0.25 µL; 50,000 sperm) of 1× or 4× semen were placed at the bottom of the right leg of an “H,” made using 15µL of medium by tracing a template placed below a Petri dish. The medium used (Hanks’ balanced salt solution with 40mg mL BSA and added lactate and pyruvate) contained different concentrations of pentoxifylline (0, 0.5, 1, 2 or 4mgmL−1). One µL of medium was removed from the tip of the left arm of the H after 15 and 30min incubation, and the number of sperm were counted. In a second study, we evaluated the effect of pentoxifylline on sperm motility parameters using computer-assisted sperm motility analysis. After thawing, 1× and 4× semen was washed to remove freezing extender and resuspended in the same medium but with 7mgmL−1 bovine serum albumin (BSA), containing the different pentoxifylline concentrations. In Study 1, the number of collected sperm did not differ significantly for 1× sperm exposed to 0 to 4mgmL−1 pentoxifylline (means of 15 to 23 sperm at 15min, and 18 to 25 sperm at 30min). Similarly, in 4× frozen semen, there was no significant difference in number of collected sperm between 0mgmL−1 and 2 or 4mgmL−1 pentoxifylline concentrations (&lt;1 to 6 at 15 min; 5 to 6 at 30min). In Study 2, at 0min,% total motility was significantly higher in 1 and 2mgmL−1 pentoxifylline than in 0mgmL−1 for 1× sperm (47.8±1.7 and 49.3±1.9, vs. 32.1±3.9, respectively; P=0.018) and significantly higher for 1, 2, and 4mgmL−1 pentoxifylline than for 0mgmL−1 for 4× sperm (3.9±0.9, 5.7±0.4, and 8.2±0.5, vs. 1.2±0.4; P=0.0001). Similar results were found at 15 and 30min for 1×, and at 15min for 4×. Pentoxifylline at 1 to 4mgmL−1 significantly increased the percentage of progressive motility in 1× sperm at 30min (17.8±1.3, 21.8±2.7, and 20.3±1.2, vs. 10.0±0.4; P=0.002) and, at 4mgmL−1, increased the percentage of progressive motility in 4× sperm at 0min (1.43±0.1 vs. 0.2±0.1; P=0.005) and 15min (1.4±0.2 vs. 0.1±0.0; P=0.0001). Exposure of poor-quality semen to pentoxifylline at 4mgmL−1 improved total and progressive motility but did not increase the recovery of motile sperm in a swim-over collection preparation.


2019 ◽  
Vol 6 (1) ◽  
pp. 119
Author(s):  
Enike Dwi Kusumawati ◽  
Syam Rahadi ◽  
Sugeng Santoso ◽  
Dyah Lestari Yulianti

ABSTRAK Penelitian ini bertujuan untuk mengetahui pengaruh lama thawing yang berbeda pada suhu 250C untuk mendapatkan kualitas spermatozoa semen beku sapi ongole yang optimal. Penelitian dilakukan di Laboratorium Fakultas Peternakan Universitas Kanjuruhan Malang. Metode Penelitian yang digunakan adalah metode percobaan dengan menggunakan Rancangan Acak Lengkap (RAL) Faktorial. Setiap perlakuan thawing diberikan ulangan sebanyak 10 sampel semen beku sapi ongole. Hasil penelitian menunjukkan bahwa perlakuan dengan lama thawing 7, 15, 30 detik pada suhu 25oC memberikan perbedaan pengaruh yang sangat nyata (p<0.01) pada motilitas, viabilitas dan abnormalitas spermatozoa semen beku sapi ongole. Motilitas tertinggi diperoleh pada perlakuan P3 dengan rata-rata sebesar 40,8%, viabilitas tertinggi diperoleh pada perlakuan P3 dengan rata-rata sebesar 82,39%, sedangkan abnormalitas terendah diperoleh pada perlakuan P3 dengan rata-rata sebesar 11,95%. Berdasarkan penelitian ini maka dapat disimpulkan bahwa thawing pada suhu 25oC dengan lama waktu 30 detik memberikan kualitas spermatozoa yang paling baik sehingga disarankan untuk melakukan thawing pada suhu 25oC dengan lama waktu 30 detik. Kata kunci: abnormalias, mortalitas, ongole, thawing, viabilitas ABSTRACT This study aims to determine the effect of different thawing time at 250C to get the sperm quality of frozen semen ongole bull. Research conducted at the Laboratory of Animal Husbandry Faculty Kanjuruhan University of Malang. The method of study by using Completely Randomized Design (CRD) factorial. Any treatment given repeated thawing of frozen samples of 10 times. The study show that treatment with time thawing 7 (P1), 15 (P2), 30 (P3) seconds at 25oC gives a very significant difference (P<0.01) on motility, viability and abnormalities of ongole Bull sperm. The highest motility and viability were obtained at P3 (40,8%) and (82,39%), while the lowest abnormalities obtained on P3 (11,95%). Based on this research it can be concluded that thawing at 25oC with 30 seconds to give the best quality sperm that is recommended for thawing. Keywords: abnormality, motility, ongole, thawing, viabilit


2017 ◽  
Vol 38 (6) ◽  
pp. 3613
Author(s):  
Adriana Novaes dos Reis ◽  
Moysés Dos Santos Miranda ◽  
Lílian Kátia Ximenes Silva ◽  
Aluizio Otavio Almeida da Silva ◽  
José Silva de Sousa ◽  
...  

For artificial insemination, it is essential to use frozen semen, however the freezing process causes deleterious changes to the structure and integrity of sperm membranes that compromise the function of sperm. To avoid this cellular damage, extenders and suitable substrates must be used to recover the highest possible number of viable cells post-thaw. To this end, in the first experiment, we evaluated three different extenders: TES-TRIS, which is widely used for buffaloes; and an extender composed of powdered coconut water-based (ACP-112®) with or without milk (ACP-112®-milk) for buffalo semen freezing. In the second experiment, we evaluated the effect of Lippia origanoides oil extract on protecting buffalo sperm against cryoinjury arising from freezing semen. Semen was collected from ten buffalo bulls (10 ejaculates/bull) and diluted in TES-TRIS (control), ACP-112® or ACP-112®-Milk in the first experiment. In the second experiment, the samples were diluted in the diluent with the best results for sperm quality obtained in experiment I, and 2.5 ?g mL-1, 5 ?g mL-1 or 10 ?g mL-1 of the plant extract was added to treatments; and a control group containing only the diluent was also included. The fresh semen was analyzed for conventional features such as motility, concentration, morphology and viability. After thawing, the samples were evaluated again for motility, vigor and supra-vital staining, and then, were performed the of thermal-resistance test, hypoosmotic test and evaluated sperm membrane integrity with the fluorescent probes PI, FITC-PSA and JC-1 using flow cytometry. The data were submitted to ANOVA, and the results were compared by Tukey’s test at a significance of 5%. In the first experiment, the extender TES-TRIS showed better results for the various characteristics evaluated compared to ACP-112® and ACP-112®-Milk (P < 0.05), demonstrating greater protection of the buffalo sperm structures during cryopreservation. In the second experiment, the treatments with different concentrations of Lippia origanoides essential oil extract showed no differences among the assessed variables regarding the protection of sperm structures during cryopreservation (P > 0.05). Based on these data, we demonstrated the beneficial effects of TES-TRIS for post-thaw buffalo sperm quality; however, no protective effect was observed for buffalo sperm cryopreserved with the different tested concentrations of Lippia origanoides extract oil.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Luís Crisóstomo ◽  
Ivana Jarak ◽  
Luís P. Rato ◽  
João F. Raposo ◽  
Rachel L. Batterham ◽  
...  

AbstractThe consumption of energy-dense diets has contributed to an increase in the prevalence of obesity and its comorbidities worldwide. The adoption of unhealthy feeding habits often occurs at early age, prompting the early onset of metabolic disease with unknown consequences for reproductive function later in life. Recently, evidence has emerged regarding the intergenerational and transgenerational effects of high-fat diets (HFD) on sperm parameters and testicular metabolism. Hereby, we study the impact of high-fat feeding male mice (F0) on the testicular metabolome and function of their sons (F1) and grandsons (F2). Testicular content of metabolites related to insulin resistance, cell membrane remodeling, nutritional support and antioxidative stress (leucine, acetate, glycine, glutamine, inosine) were altered in sons and grandsons of mice fed with HFD, comparing to descendants of chow-fed mice. Sperm counts were lower in the grandsons of mice fed with HFD, even if transient. Sperm quality was correlated to testicular metabolite content in all generations. Principal Component Analysis of sperm parameters and testicular metabolites revealed an HFD-related phenotype, especially in the diet-challenged generation and their grandsons. Ancestral HFD, even if transient, causes transgenerational “inherited metabolic memory” in the testicular tissue, characterized by changes in testicular metabolome and function.


2020 ◽  
Vol 10 (1) ◽  
pp. 122
Author(s):  
Lilly-Ann Mohlkert ◽  
Jenny Hallberg ◽  
Olof Broberg ◽  
Gunnar Sjöberg ◽  
Annika Rydberg ◽  
...  

Preterm birth has been associated with altered cardiac phenotype in adults. Our aim was to test the hypothesis that children surviving extremely preterm birth have important structural or functional changes of the right heart or pulmonary circulation. We also examined relations between birth size, gestational age, neonatal diagnoses of bronchopulmonary dysplasia (BPD) and patent ductus arteriosus (PDA) with cardiac outcomes. We assessed a population-based cohort of children born in Sweden before 27 weeks of gestation with echocardiography at 6.5 years of age (n = 176). Each preterm child was matched to a healthy control child born at term. Children born preterm had significantly smaller right atria, right ventricles with smaller widths, higher relative wall thickness and higher estimated pulmonary vascular resistance (PVR) than controls. In preterm children, PVR and right ventricular myocardial performance index (RVmpi’) were significantly higher in those with a PDA as neonates than in those without PDA, but no such associations were found with BPD. In conclusion, children born extremely preterm exhibit higher estimated PVR, altered right heart structure and function compared with children born at term.


FEBS Journal ◽  
2021 ◽  
Author(s):  
Mengqi Dong ◽  
Ève Mallet Gauthier ◽  
Marilaine Fournier ◽  
Heather J. Melichar

Author(s):  
Patrizia Bisiacchi ◽  
Elisa Cainelli

AbstractAsymmetry characterizes the brain in both structure and function. Anatomical asymmetries explain only a fraction of functional variability in lateralization, with structural and functional asymmetries developing at different periods of life and in different ways. In this work, we perform a scoping review of the cerebral asymmetries in the first brain development phases. We included all English-written studies providing direct evidence of hemispheric asymmetries in full-term neonates, foetuses, and premature infants, both at term post-conception and before. The final analysis included 57 studies. The reviewed literature shows large variability in the used techniques and methodological procedures. Most structural studies investigated the temporal lobe, showing a temporal planum more pronounced on the left than on the right (although not all data agree), a morphological asymmetry already present from the 29th week of gestation. Other brain structures have been poorly investigated, and the results are even more discordant. Unlike data on structural asymmetries, functional data agree with each other, identifying a leftward dominance for speech stimuli and an overall dominance of the right hemisphere in all other functional conditions. This generalized dominance of the right hemisphere for all conditions (except linguistic stimuli) is in line with theories stating that the right hemisphere develops earlier and that its development is less subject to external influences because it sustains functions necessary to survive.


2016 ◽  
Vol 21 (4) ◽  
pp. 203-211 ◽  
Author(s):  
Lawrence A. Patterson ◽  
Samuel Berry

Purpose The purpose of this paper is to explore experiences of team culture, structure and function of an intensive support service (ISS) within the context of the recent service guidance “Building the Right Support” (NHS England, Local Government Association and Association of Directors of Adult Social Services, 2015). Reflections on the Hampshire and Southampton ISS set up in 2010 are discussed with a view to informing a debate about frameworks for ISS services nationally. Design/methodology/approach A reflective piece, drawing on experience and case examples. Findings This paper describes that a key function of an ISS is making individuals safe and this is significantly assisted by using shared team formulation, which can enable information and perspectives to be shared between and within teams as rapidly as possible. Further, a case is made for recognising the importance of inter-disciplinary practice, as the Southampton and Hampshire ISS has removed the “old fashioned” demarcations that led to individuals seeing a “procession” of different professionals from different disciplines. This relates to team structure, but importantly is about a culture of holding a shared identity based on positive behavioural support values, rather than a traditional uni-disciplinary perspective. Practical implications ISS models are being proposed by NHS England and this paper suggests some important practical aspects. Originality/value Limited literature exists examining the team culture within ISSs, which contributes to desired outcomes for service users. This paper opens a debate about structural and functional aspects of service delivery in this service model.


2015 ◽  
Vol 75 (3) ◽  
pp. 662-669 ◽  
Author(s):  
EG Sanches ◽  
IR Oliveira ◽  
PCS Serralheiro ◽  
VR Cerqueira

AbstractThis study aims developing and evaluate a protocol of semen cryopreservation of the lane snapper Lutjanus synagris. Firstly, sperm motility rate, motility time, density and spermatocrit were appraised to characterize the sperm quality of the lane snapper. The effect of three extenders with distinct ionic compositions and pH values combined with seven concentrations of cryoprotector dimethylsulfoxide (0; 2.5; 5.0; 7.5; 10.0; 12.5 e 15.0%), five cooling rates (110, 90, 60, 45 e 30°C –min), nine equilibration time (1; 2,5; 5; 10; 15; 20; 25; 30 e 60 minutes) e five dilutions ratio (1:1; 1:3; 1:6; 1:10 e 1:20) on the sperm motility rate and motility time were analyzed. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The higher sperm motility rate and motility time (P<0.05) was achieved by combining extender with pH 8.2 with 10% concentration of dimethylsulfoxide and cooling rate 60°C –min, 1 minute of equilibration time and 1:3 (v/v) dilution ratio. The use of cryopreserved sperm presented fertilization rates >60% validating the present protocol for lane snapper. The cryoconserved sperm of lane snapper is a viable alternative, being possible to maintain appropriate sperm viability.


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