Cryopreservation of Human Nasal Ciliated Epithelium

1996 ◽  
Vol 10 (5) ◽  
pp. 291-298 ◽  
Author(s):  
Bin Yang ◽  
Thomas V. McCaffrey ◽  
Eugene B. Kern
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Storage Time ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Slow Freezing ◽  
Freezing And Thawing ◽  
Ciliary Function ◽  
Nasal Cilia ◽  
Fast Freezing

The purpose of this study was to compare the effects on human nasal cilia of various freezing and thawing methods in order to determine a reliable method of cryopreservation. Ten samples each were preserved by a slow freezing and a fast freezing method. All samples were stored in liquid nitrogen (–196°C) for 1 week. The frozen samples were thawed by one of two methods: 1) rapid thawing—37°C water bath for 3–4 minutes; and 2) slow thawing—room temperature for 15 minutes. Prefreeze and postthaw ciliary beat frequency (CBF) was measured. The slow freezing and fast thawing method (SFFT) resulted in the best viability. An additional 10 samples preserved using the SFFT method stored at –70°C to –90°C did not retain ciliary function. Thirty ciliated samples preserved by the SFFT method were examined after freezing in liquid nitrogen (–196°C) for 1 week, 2 weeks, and 1 month. There was no significant decrease in CBF after cryopreservation at –196°C for 1 week or 2 weeks (P> 0.05). As the storage time increased to 1 month, postthaw CBF decreased 7.25 ± 0.87% when compared to the prefreeze CBF (P < 0.05). We conclude that human nasal cilia preserved by SFFT at –196°C retain activity for up to 1 month.

Rapid freezing of the mouse blastocyst: effects of cryoprotectants and of time and temperature of exposure to cryoprotectant before direct plunging into liquid nitrogen

1991 ◽  
Vol 3 (2) ◽  
pp. 175 ◽  
Author(s):  
R Li ◽  
A Trounson
Keyword(s):  
Liquid Nitrogen ◽  
Room Temperature ◽  
Rapid Freezing ◽  
High Concentration ◽  
Freezing And Thawing ◽  
Initial Exposure ◽  
Survival And Development ◽  
High Concentrations

This study investigates the effects of time and temperature of exposure to a high concentration (4.5 M) of dimethyl sulfoxide (DMSO), glycerol, 1,2-propanediol (PROH), or a mixture of DMSO and glycerol (DG) in a solution containing 0.25 M sucrose, on the survival and development of rapidly frozen mouse blastocysts. Embryos had significantly (P less than 0.01) higher rates of survival and development when exposed to cryoprotectant at 0 degree C compared with room temperature. The time of exposure to cryoprotectant at either 0 degree C or room temperature before being plunged into liquid nitrogen significantly (P less than 0.01) affected the survival and development of frozen-thawed embryos. Survival and development of blastocysts in vitro and in vivo was significantly (P less than 0.05) higher when exposed at 0 degree C for 10 min to DG, DMSO and glycerol than to PROH. It is concluded that, unlike early-cleavage stage embryos, blastocysts need to be equilibrated at a low temperature (0 degree C) with high concentrations of cryoprotectant before rapid freezing. Exposure of blastocysts to 4.5 M cryoprotectant and 0.25 M sucrose at room temperature either was toxic or else markedly reduced their viability after freezing and thawing, depending on the duration of the initial exposure.

scholarly journals Post-mortem glycolysis in ox skeletal muscle. Effect of pre-rigor freezing and thawing on the intermediary metabolism

1968 ◽  
Vol 109 (2) ◽  
pp. 197-202 ◽  
Author(s):  
R. K. Scopes ◽  
R. P. Newbold
Keyword(s):  
Carbon Dioxide ◽  
Creatine Phosphate ◽  
Slow Freezing ◽  
Hexose Monophosphate ◽  
Post Mortem ◽  
Solid Carbon ◽  
Freezing And Thawing ◽  
Solid Carbon Dioxide ◽  
Triose Phosphate ◽  
Fast Freezing

1. Ox sternomandibularis muscle was ‘slow-frozen’ by placing it in air at −22° or ‘fast-frozen’ by immersion in liquid air or acetone–solid carbon dioxide. In all cases muscles were frozen pre-rigor. Changes in length, pH and the concentrations of Pi, creatine phosphate, hexose monophosphate (glucose 1-phosphate+glucose 6-phosphate+fructose 6-phosphate), fructose diphosphate (fructose 1,6-diphosphate+½ triose phosphate), lactate, ATP, ADP, AMP and NAD+ during freezing and during subsequent thawing were determined. In addition some measurements were made of the changes in α-glycerophosphate, 3-phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate and pyruvate concentrations during slow freezing. 2. Appreciable shortening and marked changes in chemical composition took place during slow freezing but not during fast freezing. 3. During slow freezing the hexose monophosphate concentration fell and fructose 1,6-diphosphate and triose phosphate increased substantially. Increases also took place in 3-phosphoglycerate, 2-phosphoglycerate and phosphoenolpyruvate, but not in pyruvate. 4. On thawing, most of the chemical changes were similar to those in unfrozen muscle post mortem, but took place much more rapidly; loss of NAD+ was particularly rapid. Fast-frozen muscle metabolized at a faster rate on thawing than did slow-frozen muscle. 5. The overall changes in length during freezing and thawing were about the same in slow-frozen as in fast-frozen muscle.

scholarly journals Reference Ranges for Respiratory Ciliary Function for Chinese Children May Not Be Extrapolated From European Data

2020 ◽  
Author(s):  
So-Lun LEE ◽  
Christopher O'Callaghan ◽  
Yu-Lung Lau ◽  
Chun-Wai Davy Lee
Keyword(s):  
High Speed ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Chinese Children ◽  
Second Hand Smoke ◽  
Reference Ranges ◽  
Ciliary Function ◽  
Age Related ◽  
Beat Pattern ◽  
Healthy Chinese

Abstract Background:To aid in the diagnosis of Primary Ciliary Dyskinesia (PCD) and to evaluate the respiratory epithelium in respiratory disease, normal age related reference ranges are needed for ciliary beat frequency (CBF), beat pattern and ultrastructure. These reference data are not available for Asian or Chinese children. Our aim was to establish reference ranges for healthy Chinese children.Methods:Ciliated epithelial samples were obtained from 135 health Chinese children aged below 18 years by brushing the inferior nasal turbinate. CBF and beat pattern were analysedfrom high speed video recordings. Epithelial integrity and ciliary ultrastructure were assessed using transmission electronic microscopy. Results:The mean CBF from 135 children studied was 10.1 Hz (95% CI 9.8 to 10.4). Approximately 20% (ranged 18.0-24.2 %) of ciliated epithelial edges were found to have areas of dyskinetically beating cilia. Normal beat pattern was observed in ciliated epithelium from all subjects. We did not find any effect of exposure to second hand smoke on CBF in our subjects. Microtubular defects were found in 9.3% of all of the cilia counted in these children while other ciliary ultrastructural defects were found in less than 3%.Conclusions: We established the reference range for CBF, beat pattern and ultrastructure in healthy Chinese children. Using similar methodology, we found a lower overall mean CBF than previously obtained European values. This study highlights the need to establish normative data for ciliary function in different populations.

scholarly journals Ciliary function of the nose in patients with Osler`s disease and the effect of topically applied estrogens as a nose ointment

2011 ◽  
Vol 49 (4) ◽  
pp. 407-412
Author(s):  
J.U. Sommer ◽  
B.A. Stuck ◽  
C. Heiser ◽  
S.S. Kassner ◽  
K. Hormann ◽  
...  
Keyword(s):  
High Speed ◽  
Beat Frequency ◽  
Digital Camera ◽  
Ciliary Beat Frequency ◽  
Control Group ◽  
Mucus Clearance ◽  
Ciliary Function ◽  
The Mean ◽  
Positive Effect ◽  
Ciliary Beat

Background: In recent years, the positive effect of topically applied estriol nose ointment in the adjuvant therapy of Morbus-Rendu-Osler (HHT) has been proven. Due to the induced metaplasia, a complete destruction of the ciliated cells may be expected. However, data regarding the ciliary function of HHT patients with and without the use of topical estriol application are currently lacking. Methodology/principal: Ciliated samples were obtained by gently brushing the inferior nasal turbinate of 19 healthy volunteers and 15 patients with known HHT (8 of them regularly using 0.1% estriol nose ointment for 2 years (HHTwE) and 7 of them not using the ointment in the last 12 months (HHTwoE)). Analysis was done with an inverted phase contrast microscope connected to a high-speed digital camera. Recorded parameters were the visual integrity (VI) of the ciliary beat and its frequency (CBF) in Hz. Results: The VI index of all samples showed an undisrupted, even beating pattern with a difference between the three groups. The mean CBF in all HHT patients was reduced compared to the control group`s mean CBF. Within the HHT group itself, the mean CBF was reduced in the HHTwE group compared to the HHTwoE group. Conclusions: The ciliary beat frequency of HHT patients is impaired compared to the control group and even more so if the HHT patients topically apply estriol more than 6 months. An undisrupted beating pattern is found in the HHTwE group despite the fact that estrogens induce a transformation of the ciliated columnar into a keratinizing squamous epithelium. This data may justify the adjuvant application of estriol as a nose ointment in the treatment of epistaxis in HHT patients without the fear of damage to the nose`s mucus clearance.

84 COMPUTER-ASSISTED SPERM ANALYSIS OF FROZEN SPERM MOTILITY AFTER REPEATED EXPOSURES TO ROOM TEMPERATURE

2010 ◽  
Vol 22 (1) ◽  
pp. 200
Author(s):  
A. Alvaro Garcia Guerra ◽  
G. M. Brogliatti
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Room Temperature ◽  
Cell Damage ◽  
Beat Frequency ◽  
Computer Assisted ◽  
Initial Exposure ◽  
Frozen Semen ◽  
Casa System ◽  
Motility Parameters

The key factorin long-term cryopreservation is the very low temperature of liquid nitrogen. Several studies suggest temperatures should be maintained at -130°C or less to avoid cell damage. Damage due to initial exposure may not be overt; however, after repeated exposures a reduction in postthaw viability may become evident (Barth A 1991 Proc. 10th Annu. Conv. Am. ET Assoc, 20-26). The CASA system provides an opportunity to assess multiple motility characteristics on a semen sample objectively and with high repeatability. An experiment was designed to evaluate the effect that repeated exposure of frozen semen in 0.5-mL straws during 15 s to room temperature produces on motility characteristics assessed by CASA system. Groups were formed according to the number of exposures per straw; groups were as follows: 0, 3, 5, and 10 times of exposure during 15 s. Thirty-two ejaculates from different bulls (15 Angus, 3 Hereford, 8 Brangus, 3 others) were diluted using a chemically semi-defined media (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, Paillette Crista, France). Four frozen straws per bull were used, one for each group. Straws were exposed to a room temperature (15°C ± 1.28) and then placed back into liquid nitrogen. Semen thawing was conduced in a water bath at 37°C during 1 min. Motility characteristics were evaluated by the IVOS Sperm Analyzer (Hamilton Thorne Research). Two chambers of 20 μm depth and 5 fields per chamber were analyzed (30 frames/0.5 s for each field). Seven motility parameters were evaluated: % of motile sperm; % of progressive sperm; VAP (path velocity, μms-1); VCL (track speed, μm/s); ALH (lateral amplitude, μm); BCF (beat frequency, Hz); and LIN (linearity, %). The Kruskal Wallis test was used to compare variables among groups, and results are shown in Table 1. The average temperature inside the straw after 15 s of exposure was of -122.6°C. No difference (P > 0.05) was found among the groups for any of the 7 motility parameters. In conclusion, sperm motility seems not to be affected if straws are exposed up to 10 times during 15 s to room temperature. More research should be done to test higher room temperatures and pregnancy rates after AI. Table 1.CASA parameters of frozen sperm after different numbers of exposures at 15°C

scholarly journals The impact of zinc supplementation on the outcome of endoscopic sinus surgery and ciliary function of the nasal mucosa in children with chronic rhinosinusitis

2021 ◽  
pp. 73-82
Author(s):  
С.И. Алексеенко ◽  
А.В. Скальный ◽  
С.А. Карпищенко ◽  
С.А. Артюшкин ◽  
С.В. Барашкова ◽  
...  
Keyword(s):  
Chronic Rhinosinusitis ◽  
Nasal Mucosa ◽  
Endoscopic Sinus Surgery ◽  
Inductively Coupled Plasma ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Sinus Surgery ◽  
Ciliary Activity ◽  
Ciliary Function ◽  
The Impact

Несмотря на указания на роль цинка в функции реснитчатого эпителия, данные относительно влияния коррекции обмена цинка на цилиарную функцию у пациентов с хроническим риносинуситом отсутствуют. Целью настоящего исследования явилась оценка эффективности хирургического лечения, активности мукоцилиарного аппарата, а также местной воспалительной реакции слизистой оболочки носа в послеоперационном периоде у детей с хроническим риносинуситом на фоне применения цинка. Методы. Обследовано 192 ребенка с хроническим риносинуситом, в лечении которых применялась эндоскопическая риносинусохирургия. Из них 131 ребенок в послеоперационном периоде получал стандартные назначения, тогда как 61 ребенок дополнительно получал цинк в суточной дозе 10 мг в течение 90 дней. Оценка концентрации цинка в сыворотке крови осуществлялась методом ИСП-МС. Для определения функции цилий использована видеоцитоморфометрия. Определение эффективности лечения осуществлялось с использованием опросника SNOT-20, а также шкал Лунд-Кеннеди и Лунд-Маккей. Результаты. Проведение эндоскопической операции приводило к достоверному снижению значений шкал SNOT-20, Лунд-Кеннеди и Лунд-Маккей у пациентов вне зависимости от приема цинка. В то же время, пациенты, принимавшие цинк, характеризовались достоверно меньшим риском ревизионной хирургии. Цинк также оказывал значительное влияние на функционирование реснитчатого эпителия. В частности, к 12-му месяцу наблюдения количество клеток с подвижными цилиями, частота биения цилий, длина цилий и выживаемость клеток у пациентов, принимавших цинк, превышала соответствующие показатели в контрольной группе. Повышение уровня цинка в организме также сопровождалось многократным снижением количества инфильтрирующих слизистую оболочку нейтрофилов и лимфоцитов. Заключение. Предполагается, что цинк ускоряет репаративные процессы в слизистой оболочке носа и обладает противовоспалительным действием. Despite the presence of certain indications of the role of zinc in ciliated epithelium functioning, data on the potential effect of Zn supplementation in ciliary function in patients with chronic rhinosinusitis are absent. The aim of this study was to evaluate the efficiency of surgical treatment, ciliary activity, as well as local inflammation of nasal mucosa in children with chronic rhinosinusitis undergoing functional endoscopic surgery. Methods. 192 pediatric patients with chronic rhinosinusitis were examined. 131 patients received standard postoperative prescriptions, whereas 61 children received 10 mg Zn daily for 90 days. Serum Zn concentrations were measured by inductively coupled plasma mass spectrometry. Ciliary function was evaluated by video cytomorphometry. Surgery efficiency was assessed with a SNOT-20 questionnaire, as well as with Lund-Kennedy and Lund-Mackay scales. Results. Functional endoscopic sinus surgery resulted in a significant decrease in total SNOT-20, Lund-Kennedy, and Lund-Mackay scores independent of Zn supplementation. At the same time, Zn supplemented patients had a significantly lower risk of revision surgery. Zn also had a significant impact on ciliary function. Specifically, up to the 12th mo postoperatively, Zn supplementation resulted in higher number of cells with motile cilia, ciliary beat frequency, ciliary length, and cell viability as compared to controls. Increased Zn status was also associated with a manyfold decrease in mucosal neutrophil and lymphocyte infiltration. Conclusions. It is proposed that Zn enhances reparative processes in nasal mucosa and possesses anti-inflammatory activity.

91 MOTILITY (CASA) AND VIABILITY OF FROZEN SPERM AFTER DIFFERENT TIMES OF EXPOSURE AT 15°C

2010 ◽  
Vol 22 (1) ◽  
pp. 204
Author(s):  
A. Garcia Guerra ◽  
M. G. Lüssenhoff ◽  
G. M. Brogliatti
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Room Temperature ◽  
Bos Taurus ◽  
Cell Damage ◽  
Subjective Evaluation ◽  
Beat Frequency ◽  
Bos Indicus ◽  
Frozen Semen ◽  
Live Sperm

One of the key factors for successful long-term cryopreservation in liquid nitrogen is maintaining the samples at -130°C or lower at all times to avoid cell damage (Barth A 1991 Proc. 10th Annu. Conv. Am. ET Assoc., 20-26). Previous data reported that exposure of semen straw to ambient temperature for more than 15 s can raise the temperature above -130°C and could reduce sperm motility by subjective evaluation (Berndston et al. 1976 Proc. 6th NAAB Tech. Conf. Artif. Insem. Reprod., 51-60). The CASA system provides an opportunity to assess multiple motility characteristics on a semen sample objectively and with high repeatability. Two experiments were designed to evaluate the effect of exposing frozen semen in 0.5-mL straws for 15, 30, or 60 s to room temperature on motility characteristics assessed by CASA and viability parameters by vital stain, HOS test, and acrosome integrity. Thirty-three ejaculates from different bulls (88% British breeds) were used for CASA evaluation, and 12 ejaculates were from other bulls (7 Bos indicus and 4 Bos taurus) were used for viability evaluation. All ejaculates were diluted using a chemically semi-defined media (Andromed, Minitub, Germany) and frozen in an automatic freezer (Digicool, IMV, France). Five frozen straws per bull were used, one for each time of exposure and one as control (0 s = 0 time). Straws were exposed to room temperature (15°C ± 1.78) for different times and then placed back into liquid nitrogen. Semen thawing was conduced in a water bath at 37°C during 1 min. Motility characteristics were evaluated by the IVOS Sperm Analyzer (Hamilton Thorne Research). Two chambers of 20 μm depth and 5 fields per chamber were analyzed (30 frames/0.5 s for each field). Six motility parameters were evaluated: % of motile sperm; % of progressive sperm; VAP (path velocity, μm s-1); ALH (lateral amplitude, μm); BCF (beat frequency, Hz); and LIN (linearity, %). Viability characteristics were evaluated by % of live sperm (eosin-nigrosin); % positive to HOS test, and % of intact acrosome (Giemsa stain). A nonparametric AOV (Kruskal Wallis) test was used to compare variables among groups, and results are shown in Table 1. There was a reduction (P < 0.05) in the percentage of motile and progressive sperm when exposure to 15°C was longer than 30 s. The alive cells have similar motile characteristics as VAP, VCL, ALH, BCF, and LIN. The viability of spermatozoa was reduced (P < 0.05) when they were exposed to room temperature beyond 30 s. Also, a lower proportion of positive spermatozoa for HOS test was detected for exposures beyond 15 s. In conclusion, these results suggest sperm motility and viability would not be affected if straws are exposed up to 30 s to 15°C. Further study should be done regarding viability tests. Table 1.Motility and viability parameters of exposed frozen semen

Ciliary function and sinonasal mucosal cytology in pediatric patients with chronic rhinosinusitis during a year after functional endoscopic sinus surgery

2021 ◽  
Vol 0 (0) ◽  
pp. 0-0
Author(s):  
S. Alekseenko ◽  
S. Karpischenko ◽  
S. Artyushkin ◽  
S. Barashkova ◽  
I. Anikin
Keyword(s):  
Chronic Rhinosinusitis ◽  
Endoscopic Sinus Surgery ◽  
High Speed ◽  
Beat Frequency ◽  
Neutrophil Infiltration ◽  
Ciliary Beat Frequency ◽  
Substantial Improvement ◽  
Sinus Surgery ◽  
Cell Height ◽  
Ciliary Function

Background: The objective of the study is evaluation of ciliary function and mucosal cytology after endoscopic sinus surgery in children with chronic rhinosinusitis (CRS). Methodology: A total of 132 children with CRS who underwent endoscopic sinus surgery, as well as 15 healthy controls were involved in the study. In this follow-up study patients were examined preoperatively, as well as 3, 6, 9, and 12 months after endoscopic sinus surgery. Assessment of ciliary function and sinonasal mucosal cytology was performed using high-speed videomicroscopy. Lund-Kennedy, Lund-Mackay, and sinonasal outcome test 20 (SNOT20) scores were also evaluated. Results: Total SNOT-20, Lund-Mackay, and Lund-Kennedy values significantly decreased after sinus surgery. In contrast, ciliary function and mucosal cytology only tended to improve after 6 months. 9 months after surgery the number of ciliated cells, ciliary beat frequency, cell viability, and ciliary length were significantly higher than preoperatively. The most significant improvement of ciliary function and cell height was observed 12 months after operation, whereas epithelial dystrophy and neutrophil infiltration were significantly reduced. Conclusions: Substantial improvement was observed only in a year after surgery, whereas 0 to 3 months after the surgery ciliary function was severely impaired thus predisposing to recurrent sinusitis or other complications.

A pilot study exploring the impact of cardiac medications on ciliary beat frequency: possible implications for clinical management

2018 ◽  
Vol 29 (5) ◽  
pp. 565-571
Author(s):  
Rohit S. Loomba ◽  
Abhinav Bhushan ◽  
Adeleye J. Afolayan
Keyword(s):  
Intensive Care Unit ◽  
Intensive Care ◽  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Cardiac Intensive Care Unit ◽  
Ciliary Function ◽  
Cardiac Intensive Care ◽  
Ciliary Dyskinesia ◽  
Observation Period ◽  
Ciliary Beat

Abstract Background Cilia are involved in several physiologic processes, and at least a single primary cilium can be found on nearly every cell in the human body. Various factors, such as pH, temperature, exposure to medications and toxins can impact ciliary function as is manifested by changes in the ciliary beat frequency. Those with ciliary dyskinesia may also have congenital cardiac malformations and may require care in a cardiac intensive care unit. This study investigates the effect on the ciliary beat frequency of medications frequently used in a cardiac intensive care unit. Methods The ciliated epithelial cells were obtained via nasal swab from a relatively healthy individual. These cells were cultured for 24 h. Video microscopy was then employed to determine the ciliary beat frequency at baseline and then at 15, 30, 60 and 90 min after exposure to either normal saline (control) or one of several medications. The ciliary beat frequency at each time point was then compared to the ciliary beat frequency at the same time point in the control sample as well as the baseline value for that particular sample. Results Epinephrine increased the ciliary beat frequency compared to the baseline and the controls up to 30 min and then subsequently led to a significant decrease in ciliary beat frequency at 90 min. On the one hand, norepinephrine, dexmedetomidine, procainamide, propranolol and enalapril all decreased ciliary beat frequency significantly throughout the 90-min observation period. On the other hand, Milrinone significantly increased the ciliary beat frequency throughout the observation period, while heparin had no impact on ciliary beat frequency. Conclusions The medications frequently used in cardiac intensive care unit impact ciliary function, with most being ciliodepressant. Further investigation is needed to determine the clinical impacts and whether these effects are exaggerated in those with ciliary dyskinesia.

76 COMPUTER-ASSISTED SPERM ANALYSIS OF FROZEN SPERM MOTILITY AFTER DIFFERENT TIMES OF EXPOSURE AT 15°C

2008 ◽  
Vol 20 (1) ◽  
pp. 119
Author(s):  
A. Garcia Guerra ◽  
M. P. Etcheverry ◽  
D. Rodriguez ◽  
G. Larraburu ◽  
G. M. Brogliatti
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Room Temperature ◽  
Cell Damage ◽  
Semen Analysis ◽  
Beat Frequency ◽  
Computer Assisted ◽  
Frozen Semen ◽  
Significant Difference ◽  
Casa System

One of the key factors for successful long-term cryopreservation in liquid nitrogen is maintaining the samples at –130°C or lower at all times to avoid cell damage (Barth 1991 Proc. 10th Ann. Conv. Am. Embr. Transf. Assoc., 20–26). Previous data indicated that exposure of the semen straw to ambient temperature for more than 15 s can raise the temperature above –130°C and reduce sperm motility, as determined by subjective evaluation (Berndtson et al. 1976 Proc. 6th NAAB Tech. Conf. Artif. Insem. Reprod., 51–60). The computer-assisted semen analysis (CASA) system provides an opportunity to assess multiple motility characteristics on a semen sample objectively and with high repeatability. An experiment was designed to evaluate the effect of exposing frozen semen in 0.5-mL straws to room temperature for 15, 30, 60, or 120 s on motility characteristics assessed by CASA system. Twenty-eight ejaculates from different bulls (19 Angus, 7 Hereford, 1 Brangus, 1 Shorthorn) were diluted using a chemically semi-defined media (Andromed, Minitüb, Tiefenbach, Germany) and frozen in an automatic freezer (Digicool, IMV, Paillette Crista, France). Five frozen straws per bull were used, one for each time of exposure and one as control (0 s = 0 time). Straws were exposed to room temperature (15°C ± 0.78) for different times and then placed back into liquid nitrogen. Semen thawing was conducted in a water bath at 37°C for 1 min. Motility characteristics were evaluated by the IVOS SpermAnalyzer (Hamilton Thorne Research, Beverly, MA, USA). Two chambers of 20-μm depth and 5 fields per chamber were analyzed (30 frames/0.5 s for each field). Seven motility parameters were evaluated: motile sperm (%), progressive sperm (%), VAP (path velocity, μm s–1), VCL (track speed, μm s–1), ALH (lateral amplitude, μm), BCF (beat frequency, Hz), and LIN (Linearity, %). The Kruskal–Wallis test was used to compare variables among groups, and results are shown in Table 1. There is a significant difference (P < 0.05) in the % of motile and progressive sperm when time of exposure was increased. There was a drastic and significant reduction in the percentage of motile and progressive sperm when exposure to 15°C was longer than 30 s. The live cells had similar motile characteristics: VAP, VCL, ALH, BCF, and LIN. In conclusion, sperm motility would be affected if straws are exposed for more than 30 s. More research should be done to test higher room temperatures, detect viability effects, and determine pregnancy rates after AI. Table 1. CASA of frozen sperm motility characteristics at different times of exposure at 15°C This research was supported by Centro Genetico Bovino Eolia S.A.

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