Choice of staining technique affects the morphological assessment of epididymal feline sperm

https://doi.org/10.17221/132/2015-VETMEDCitation:Perez-Marin C.C., Jimenez E., Aguera E. (2016): Choice of staining technique affects the morphological assessment of epididymal feline sperm. Veterinarni Medicina, 61: 560-566.
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Morphological examination of spermatozoa remains a valuable tool for determining the quality of feline sperm for cryopreservation and/or for use in artificial insemination. Numerous staining techniques have been employed for feline sperm assessment, and several variations have also been reported. Forty-eight domestic cats were castrated and epididymal sperm samples were obtained. Sperm morphology was assessed using two staining methods, an air-dried stain (Giemsa-Wright) and a wet stain (eosin-nigrosin), to determine their relative abilities for estimation of the proportion of normal and abnormal spermatozoa. The effects of centrifugation and the cooling process on sperm morphology were also analysed. The mean percentage of morphologically normal spermatozoa was significantly lower in eosin-nigrosin-stained samples than in Giemsa-Wright-stained preparations (46.5% vs 53.4%; P < 0.01), both examined by conventional light microscopy. Results concerning head, midpiece and tail sperm abnormalities suggest that these methods differ in their capacity to identify sperm anomalies in cat. Although no significant differences were observed for secondary abnormalities, cytoplasmic droplets were significantly lower in Giemsa-Wright-stained samples (P < 0.01). Centrifugation or refrigeration of sperm samples did not affect the epididymal sperm morphology in evaluated samples. This study highlights the necessity of minimising morphological assessment variations in order to eliminate subsequent errors and to improve the reproducibility of sperm morphology assessment in different laboratories.

References:
Axnér E, Linde Forsberg C (2007): Sperm Morphology in the Domestic Cat, and its Relation with Fertility: A Retrospective Study. Reproduction in Domestic Animals, 42, 282-291  https://doi.org/10.1111/j.1439-0531.2007.00780.x
 
Axnér E., Ström B., Linde-Forsberg C. (1997): Sperm morphology is better in the second ejaculate than in the first in domestic cats electroejaculated twice during the same period of anesthesia. Theriogenology, 47, 929-934  https://doi.org/10.1016/S0093-691X(97)00048-4
 
Axnér E, Holst B.Strom, Linde-Forsberg C (1998): Morphology of spermatozoa in the cauda epididymidis before and after electroejaculation and a comparison with ejaculated spermatozoa in the domestic cat. Theriogenology, 50, 973-979  https://doi.org/10.1016/S0093-691X(98)00200-3
 
Axnér E., Linde-Forsberg C., Einarsson S. (1999): Morphology and motility of spermatozoa from different regions of the epididymal duct in the domestic cat. Theriogenology, 52, 767-778  https://doi.org/10.1016/S0093-691X(99)00170-3
 
Axnér Eva, Hermansson Ulrika, Linde-Forsberg Catharina (2004): The effect of Equex STM paste and sperm morphology on post-thaw survival of cat epididymal spermatozoa. Animal Reproduction Science, 84, 179-191  https://doi.org/10.1016/j.anireprosci.2003.11.003
 
Baran A, Bacinoglu S, Evecen M, Sahin BE, Alkan S, Demir K (2004): Freezing of cat semen in straws with different glycerol levels containing Tris extender. Turkish Journal of Veterinary and Animal Science 28, 545–552.
 
BILGILI S. F., RENDEN J. A., SEXTON K. J. (1985): The Influence of Staining Techniques and Examiners on Evaluation of the Morphology of Fowl Spermatozoa. Poultry Science, 64, 2358-2361  https://doi.org/10.3382/ps.0642358
 
Björndahl Lars, Söderlund Inger, Johansson Sofia, Mohammadieh Majid, Pourian Mohammad Reza, Kvist Ulrik (2004): Why the WHO Recommendations for Eosin-Nigrosin Staining Techniques for Human Sperm Vitality Assessment Must Change. Journal of Andrology, 25, 671-678  https://doi.org/10.1002/j.1939-4640.2004.tb02839.x
 
Blottner S, Jewgenow K (2007): Moderate Seasonality in Testis Function of Domestic Cat. Reproduction in Domestic Animals, 42, 536-540  https://doi.org/10.1111/j.1439-0531.2006.00817.x
 
Chatdarong K, Thuwanut P, Suksamai P, Patanatiradaj S, Sangwornrachasup A (2009): Survival of Frozen-Thawed Cat Spermatozoa Pre-Cooled in the Epididymides. Reproduction in Domestic Animals, 44, 377-380  https://doi.org/10.1111/j.1439-0531.2009.01412.x
 
Chatdarong K., Thuwanut P., Morrell J.M. (2010): Single-layer centrifugation through colloid selects improved quality of epididymal cat sperm. Theriogenology, 73, 1284-1292  https://doi.org/10.1016/j.theriogenology.2009.12.009
 
Cooper Trevor G., Yeung Ching-Hei (): Acquisition of volume regulatory response of sperm upon maturation in the epididymis and the role of the cytoplasmic droplet. Microscopy Research and Technique, 61, 28-38  https://doi.org/10.1002/jemt.10314
 
Glover T.E., Watson P.F. (1985): The effect of buffer osmolality on the survival of cat () spermatozoa at 5°C. Theriogenology, 24, 449-456  https://doi.org/10.1016/0093-691X(85)90051-2
 
Glover T.E., Watson P.F. (1987): The effects of egg yolk, the low density lipoprotein fraction of egg yolk, and three monosaccharides on the survival of cat (Felis catus) spermatozoa stored at 5°C. Animal Reproduction Science, 13, 229-237  https://doi.org/10.1016/0378-4320(87)90112-6
 
Goodrowe K.L., Hay M. (1993): Characteristics and zona binding ability of FRESH and cooled domestic cat epididymal spermatozoa. Theriogenology, 40, 967-975  https://doi.org/10.1016/0093-691X(93)90365-C
 
Howard JG, Brown JL, Bush M, Wildt DE (1990): Teratospermic and normospermic domestic cats: ejaculate traits, pituitary-gonadal hormones and improvement of spermatozoal motility and morphology after swim-up processing. Journal of Andrology 11, 204–215.
 
Jewgenow K., Neubauer K., Blottner S., Schon J., Wildt D. E., Pukazhenthi B. S. (): Reduced Germ Cell Apoptosis During Spermatogenesis in the Teratospermic Domestic Cat. Journal of Andrology, 30, 460-468  https://doi.org/10.2164/jandrol.108.006726
 
Johnson C, Jacobs J, Walker R (1991): Diagnosis and control of Brucella canis in kennel situations. Proceedings of the Annual Meeting of the Society for Theriogenology. 236–239.
 
Lengwinat T., Blottner S. (1994): In vitro fertilization of follicular oocytes of domestic cat using fresh and cryopreserved epididymal spermatozoa. Animal Reproduction Science, 35, 291-301  https://doi.org/10.1016/0378-4320(94)90044-2
 
MESCHEDE D., KECK C., ZANDER M., COOPER T. G., YEUNG C.-H., NIESCHLAG E. (1993): Influence of three different preparation techniques on the results of human sperm morphology analysis. International Journal of Andrology, 16, 362-369  https://doi.org/10.1111/j.1365-2605.1993.tb01363.x
 
Mota Paula C., Ramalho-Santos João (2006): Comparison between different markers for sperm quality in the cat: Diff-Quik as a simple optical technique to assess changes in the DNA of feline epididymal sperm. Theriogenology, 65, 1360-1375  https://doi.org/10.1016/j.theriogenology.2005.08.016
 
Palmer C.W., Persson Y., Söderquist L. (2013): Classification of the potential breeding ability of range beef bulls based on semen quality parameters in samples collected by transrectal massage—A comparison of the Swedish and Canadian systems. Animal Reproduction Science, 140, 124-130  https://doi.org/10.1016/j.anireprosci.2013.06.001
 
Parks J.E., Graham J.K. (1992): Effects of cryopreservation procedures on sperm membranes. Theriogenology, 38, 209-222  https://doi.org/10.1016/0093-691X(92)90231-F
 
Penfold L. M. (2003): Normospermic Versus Teratospermic Domestic Cat Sperm Chromatin Integrity Evaluated by Flow Cytometry and Intracytoplasmic Sperm Injection. Biology of Reproduction, 69, 1730-1735  https://doi.org/10.1095/biolreprod.103.016089
 
Platz CC, Seager SWJ (1978): Semen collection by electroejaculation in the domestic cat. Journal of the American Veterinary Medical Association 173, 1353–1355.
 
Pukazhenthi B. (1999): Sensitivity of Domestic Cat (Felis catus) Sperm from Normospermic versus Teratospermic Donors to Cold-Induced Acrosomal Damage. Biology of Reproduction, 61, 135-141  https://doi.org/10.1095/biolreprod61.1.135
 
Pukazhenthi Budhan, Noiles Esther, Pelican Katharine, Donoghue Annie, Wildt David, Howard JoGayle (2000): Osmotic Effects on Feline Spermatozoa from Normospermic versus Teratospermic Donors. Cryobiology, 40, 139-150  https://doi.org/10.1006/cryo.2000.2233
 
Pukazhenthi Budhan, Spindler Rebecca, Wildt David, Bush Lena May, Howard JoGayle (2002): Osmotic properties of spermatozoa from felids producing different proportions of pleiomorphisms: influence of adding and removing cryoprotectant. Cryobiology, 44, 288-300  https://doi.org/10.1016/S0011-2240(02)00035-4
 
Root Kustritz MV, Olson PN, Johnston SD, Root TK (1998): The effects of stains and investigators on assessment of morphology of canine spermatozoa. Journal of the American Animal Hospital Association, 34, 348-352  https://doi.org/10.5326/15473317-34-4-348
 
Schäfer S., Holzmann A. (2000): The use of transmigration and Spermac™ stain to evaluate epididymal cat spermatozoa. Animal Reproduction Science, 59, 201-211  https://doi.org/10.1016/S0378-4320(00)00073-7
 
Sprecher D.J., Coe P.H. (1996): Differences in bull spermiograms using eosin-nigrosin stain, feulgen stain, and phase contrast microscopy methods. Theriogenology, 45, 757-764  https://doi.org/10.1016/0093-691X(96)00005-2
 
Tebet J.M., Martins M.I.M., Chirinea V.H., Souza F.F., Campagnol D., Lopes M.D. (2006): Cryopreservation effects on domestic cat epididymal versus electroejaculated spermatozoa. Theriogenology, 66, 1629-1632  https://doi.org/10.1016/j.theriogenology.2006.02.013
 
Tittarelli Claudia, Savignone Cesar Augusto, Arnaudín Evangelina, Stornelli María Cecilia, Stornelli María Alejandra, de la Sota Rodolfo Luzbel (2006): Effect of storage media and storage time on survival of spermatozoa recovered from canine and feline epididymides. Theriogenology, 66, 1637-1640  https://doi.org/10.1016/j.theriogenology.2006.01.021
 
Villaverde A.I. Silva Balbin, Martins M.I. Mello, Castro V. Basto, Lopes M.D. (2006): Morphological and functional characteristics of chilled semen obtained from domestic feline epididymides (Felis catus). Theriogenology, 66, 1641-1644  https://doi.org/10.1016/j.theriogenology.2006.02.011
 
Zambelli Daniele, Cunto Marco (2006): Semen collection in cats: Techniques and analysis. Theriogenology, 66, 159-165  https://doi.org/10.1016/j.theriogenology.2006.01.054
 
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