Carluccio, A

Contri, A.

De Amicis, I

Pennelli, M

Ponzio, Patrizia

Robbe, D

Tosi, U

English

Institutional Research Information System University of Turin

Animal Reproduction Science 97 (2007) 186–190Short communicationSemen evaluation in the chamois of Abruzzi(Rupicapra pyrenaica ornata)Augusto Carluccio a,∗, Patrizia Ponzio b, Umberto Tosi a,Ippolito De Amicis a, Domenico Robbe a,Massimiliano Pennelli a, Alberto Contri aa Faculty of Veterinary Medicine, Department of Clinical Veterinary Science, University of Teramo,212,Viale F. Crispi, 64100 Teramo, Italyb Faculty of Veterinary Medicine, Department of Animal Pathology, University of Torino,44, Via L. Da Vinci, 10095 Grugliasco (To), ItalyReceived 27 October 2005; received in revised form 1 February 2006; accepted 7 February 2006Available online 15 March 2006AbstractThe chamois of Abruzzi (Rupicapra pyrenaica ornata) has been classified as endangered by the WorldUnion for Conservation.The objective of this study was to analyze seasonal differences in the characteristics of various malereproductive organs and in semen quality. The study was conduced during 2004 in the reserve of Lama deiPeligni (Italy) on three chamois males aged between 2 and 5 years. Males were captured during March–Maymonths and October–December months. Various testicular and scrotal measurements were taken and semenwas collected using an electroejaculator. Sperm motility pattern was evaluated using computer assisted spermanalyzer, membrane integrity using differential staining and morphology with phase contrast microscopy.Testicular size, sperm motility membrane integrity and the percent of morphological normal spermatozoawere greater during October–December.The authors concluded that sperm characteristics are strongly influenced by season and that semen col-lected in this period (breeding season) has suitable quality for manipulation and long-term preservation.© 2006 Elsevier B.V. All rights reserved.Keywords: Chamois; Electroejaculation; Semen evaluation; Season∗ Corresponding author. Tel.: +39 0861 266975; fax: +39 0861 266975.E-mail address: carluccio@unite.it (A. Carluccio).0378-4320/$ – see front matter © 2006 Elsevier B.V. All rights reserved.doi:10.1016/j.anireprosci.2006.02.005A. Carluccio et al. / Animal Reproduction Science 97 (2007) 186–190 1871. IntroductionSince 1973 the trade of the chamois of Abruzzi was allowed by CITES only for scientificresearch programs. This species is classified as “EN = Endangered” in the “Red Data Book” ofthe World Union for Conservation (IUCN).Scientific references on chamois concern only the ethological aspects, no data has been pub-lished on reproductive tract and sperm characteristics.Some authors described chamois reproductive season in October–December and the length ofgestation is about 23–27 weeks (Lovari, 1986; Lovari and Pellegrini, 1998).Because it is important to improve the knowledge on reproductive male characteristics forpreventing this species extinction, the objective of this study was to describe some testicular andsperm characteristics of chamois of Abruzzi and to find in which months semen quality wassuitable for manipulation and long-term preservation.2. Materials and methods2.1. ExtenderThe collected semen was diluted with Tris (hydroxymethyl-aminoethane)–citric acid–glucose(TCG) obtained according to Aboagla and Terada (2003) with Tris (375.0 mM, Sigma–AldrichS.r.l., Milan, Italy), citric acid (124.0 mM, Sigma–Aldrich S.r.l.) and glucose (41.0 mM,Sigma–Aldrich S.r.l.) and buffered to pH 7.0 and osmolarity 375 mOsm (Salomon and Ritar,1982).2.2. Animal captures and collectionThe research, was carried out during 2004, in the Reserve of Lama dei Peligni (Chieti, Italy),extended for about 1700 ha, nine of these limited by an enclosure. Eleven animals are housedin the reserve: three adult males aged between 2 and 5 years (object of the study), four adultfemales and four kids born in 2003. Males were captured in two different periods: March–May(MM period) and October–December (OD period). Captures were performed by tele-injection aspreviously described (Cuomo et al., 2003).Scrotal circumference and upper–inferior (UI), lateral–medial (LM) and cranial–caudal (CC)testicular diameters were measured. The seminal collection was obtained by electroejaculation andvolume was measured in a conical sterile glass tube graduated with 0.1 ml optical visible intervals.Then, semen was diluted 1:100 with TCG and placed in the Equitainer® (Hamilton ThorneBiosciences, Beverly, MA, USA). The evaluation was performed at the laboratory within 2 h.2.3. Semen evaluationSemen concentration was evaluated by haemocytometer after dilution (1:400) with 0.05%formal saline.Sperm motility was evaluated by computer assisted sperm analyzer (CASA) system IVOSversion 12.2 (Hamilton Thorne Biosciences) at regular intervals of 3, 10 and 34 h after collection.This device identified motile sperm by their brightness and mean size. Sperm kinetic characteristicsconsidered were motility (M), progressive motility (PM), and point-to-point curvilinear velocity(VCL).188 A. Carluccio et al. / Animal Reproduction Science 97 (2007) 186–190Table 1Means (±S.D.) of the scrotal and testicular measurements (cm) in March–May (MM) period and October–December(OD) period, respectively, considered non-breeding and breeding seasonMeasurement March–May (MM) period October–December (OD) periodRight testicle Left testicle Right testicle Left testicleUpper–inferior diameter 5.8 ± 0.4 b 5.3 ± 0.2 b 7.2 ± 0.3 a 6.2 ± 0.2 aLateral–medial diameter 3 ± 0.1 a 2.3 ± 0.2 a 3.5 ± 0.2 a 2.5 ± 0.2 aCranial–caudal diameter 4.7 ± 0.3 a 4.1 ± 0.1 b 5.2 ± 0.1 a 4.8 ± 0.2 aEpididymis tail 0.4 ± 0.2 a 0.4 ± 0.1 a 0.5 ± 0.1 a 0.5 ± 0.1 aScrotal circumference 14.3 ± 1.2 b 16.8 ± 0.5 aDifferent letters in the same column differ significantly (P≤ 0.05).Sperm plasmalemma integrity was referred to viability using SYBR-14 and propidium iodide(PI) (Live/Dead Sperm Viability Kit®, Molecular Probes, Eugene, OR, USA) as described byGarner and Johnson (1995). A semen aliquot (50l) was mixed with 5l of SYBR-14 (10M)and 2.5l of PI (120M) and incubated at 38 ◦C for 15 min. Slide was executed by placing5l of stained semen with 1l of glutaraldehide solution (3%) and evaluated with an epiflu-orescence microscope (BX 51, Olympus Italia, Segrate, Milan, Italy), at 1000× magnification.Green spermatozoa (SYBR-14) were considered live, these partially or completely red (PI) wereconsidered dead. The percentage of plasmalemma-intact spermatozoa was determined on at least200 spermatozoa.Percentage of morphologic alterations was determined by analysis of sperm with normal mor-phology on two different smears prepared by liquid sample preserved in formal saline buffer(0.05%) using a phase contrast microscopy (1000×) and at least 200 cells were observed.2.4. Statistical analysisScrotal circumference and testicular diameters in MM and OD periods were compared bypaired t-test and where considered significant with P≤ 0.05. Volume, concentration, M, PM,VCL, viability and morphologic alterations in March–May versus October–December periodswere compared in the three subjects at 3, 10 and 34 h by Wilcoxon matched—pairs signed-rankstest (t-test for non-paired value) and by paired t-test. The results were considered significant withP≤ 0.05.3. ResultsTesticular diameters in MM and OD periods are reported in Table 1. Every animal presented alight asymmetry between right and left testis. The scrotal circumference was significantly differentin MM than in OD periods, as like as UI diameters in both testicles and CC diameter in sx testicle(P≤ 0.05) while the other diameters were no significant (P≥ 0.05). During the MM period theoccipital gland, with a diameter of about 1 cm, was only palpable, while during the OD periodit was visible as a protuberance of about 3.5 cm in diameter. Semen color was yellowish duringMM and white and lactescent during OD.Sperm kinetic characteristics, viability and sperm morphologic alterations during MM andOD periods are reported in Table 2. The statistical analysis showed significant difference in MMA. Carluccio et al. / Animal Reproduction Science 97 (2007) 186–190 189Table 2Means (±S.D.) of seminal parameters calculated at 3, 10 and 34 h in March–May (MM) period and October–December(OD) period, respectively, considered non-breeding and breeding seasonParameters March–May (MM) period October–December (OD) periodTime post-collection (h) 3 10 34 3 10 34Volume (ml) 1.1 ± 0.2 0.6 ± 0.1Concentration (×109/ml) 1.9 ± 0.2 1.6 ± 0.1Total motility (%) 10.2 ± 1.5 3.2 ± 1.4 0 78.5 ± 3.2 72.1 ± 2.7 53.7 ± 3.5Progressive motility (%) 1.7 ± 0.4 0 0 69.6 ± 1.7 61.2 ± 2.1 43.9 ± 2.9VCL (m/s) 32.2 ± 1.1 15.5 ± 2.2 0 92.1 ± 13 83.2 ± 8.9 65.4 ± 7.4Viability (%) 3.5 ± 0.8 0 0 81.3 ± 1.8 73.4 ± 2.2 65.9 ± 1.5Morphologic alterations (%) 41.2 ± 1.3 42.9 ± 1.2 43.5 ± 1.4 3.2 ± 0.6 4.8 ± 0.8 5.2 ± 1.2versus OD period for volume (P≤ 0.05), M and viability (P≤ 0.01), PM, VCL and morphologicalterations (P≤ 0.001).4. DiscussionThe limitations of this research were the scant number of valued subjects (3), the large extensionof the protected area, the restricted number of captures (6) in comparison with the attempts.According to Karagiannidis et al. (2000) a significant relationship between testicular diameters,ejaculate volume and breeding season exists in buck. This correlation evidenced that the animalsin reproductive activity with repeated coupling showed a reduction in semen volume for ejaculate.Other semen parameters values reported in bibliography in small ruminants were: volume0.7–2 ml, concentration 2–5 × 109 spermatozoa/ml, motility 70–90% and a sperm abnormalitiespercentage 5–15% (Mann and Lutwac-Mann, 1981; Chemineau, 1986; Chemineau et al., 1991;Roca et al., 1992; Prado et al., 2002).Data reported in this study on chamois of Abruzzi semen characteristics were comparable.In fact, semen concentrations in chamois were 1.9 ± 0.2 and 1.6 ± 0.1 × 109 spermatozoa/ml,respectively, in MM and OD periods.Seminal quality parameters in chamois of Abruzzi during OD period showed a M of78.5 ± 3.2% and PM of 69.6 ± 1.7% after 2 h maintenance in Equitainer® which was pre-served at 44% after 34 h. The trend of viability and percentage of abnormal spermatozoawas related after 5 ◦C semen storage (56.1 ± 7.1–32.5 ± 4.8m/s, 45.9 ± 2.3–38.8 ± 2.1m/s,81.3 ± 1.8–65.9 ± 1.5% and 3.2 ± 0.6–5.2 ± 1.2%, respectively). In MM period the values showeda poor quality of semen, but the decrease was comparable with other studies (Table 2).Value of VCL in OD period (92.1 ± 13m/s) was significantly greater than in MM period(32.2 ± 1.1m/s). Because some studies reported that VCL was strongly correlated with IVFrates (Holt et al., 1994; Chan et al., 1990), it could be suitable a semen collection for long-termsemen preservation in October–December months.In chamois there seems to be a clear seasonal influence on both semen production and charac-teristics (Table 2) as it is also reported in some goat breeds (Karagiannidis et al., 2000). In goatsliving in temperate conditions, intense sexual activity was observed between October and April(Delgadillo et al., 1991), while in subtropical areas breeding season seems to start earlier than intemperate areas (Delgadillo et al., 1999). On the contrary, Karagiannidis et al. (2000) reported abreeding season in late summer and autumn with high quantity (volume and concentration) and190 A. Carluccio et al. / Animal Reproduction Science 97 (2007) 186–190quality (percentage of motile and progressive spermatozoa, percentage of abnormal spermatozoa).In the buck the breeding season in temperate areas seems to be different from the chamois. Apossible hypothesis could be related to the chamois different sensitivity to photoperiod or otherinfluences on melatonin release.The studied periods (March–May and October–December) could represent non-breeding andbreeding season, if compared with bibliography, but this observation must be confirmed with theanalysis of reproductive characteristics during whole year.5. ConclusionSemen characteristics of chamois of Abruzzi are highly influenced by season. The satisfactorysemen production to try cryopreservation programs was produced in October–December, but moreinvestigations are needed in order to identify the exact breeding period. Frozen semen could beutilized in artificial insemination, for genetic studies, to increase genotypic variability and resolvethe problem of rupicaprine species extinction.ReferencesAboagla, E.M.E., Terada, T., 2003. Threalose-enhanced fluidity of the goat sperm membrane and its protection duringfreezing. Biol. Reprod. 69, 1245–1250.Chan, S.Y., Tsoi, W.L., Leung, J., Ng, V., Lo, T., Wang, C., 1990. The accuracy of sperm concentration determination bythe automated Cellsoft semen analyzer before and after discontinuous Percoll gradient centrifugation. Andrologia 22,55–61.Chemineau, P., 1986. Sexual behaviour and gonadal activity during the year in the tropical Creole meat goat. II. Malemating behaviour, testis diameter, ejaculate characteristics and fertility. Reprod. Nutr. Dev. 26, 453–460.Chemineau, P., Cognie, Y., Guerin, V., Orgeur, P., Vallet, J.C., 1991. Factors involved in variation of reproductive charac-teristics. In: Training Manual of Artificial Insemination in Sheep and Goats. FAO, Rome, pp. 61–102.Cuomo, A., Gentile, L., Britti, D., Di Bartolomeo, F., Di Matteo, V., Fano, U., 2003. Romifidine–acepromazine–ketaminevs. medetomidine–ketamine in the Rupicapra pyrenaica ornata capture. In: Proceedings of the III InternationalSymposium on Wild Fauna, Ischiam, Italy, pp. 267–270.Delgadillo, J.A., Leboeuf, B., Chemineau, P., 1991. Decrease in seasonality of sexual behaviour and sperm production inbucksby exposure to short photoperiodic cycles. Theriogenology 36, 755–770.Delgadillo, J.A., Canedo, G.A., Chemineau, P., Guillaume, D., Malpaux, B., 1999. Evidence for an annual reproductiverhythm independent of food availability in male Creole goats in subtropical northern Mexico. Theriogenology 52,727–737.Garner, D.L., Johnson, L.A., 1995. Viability assessment of mammalian sperm using SYBR-14 and propidium iodide.Biol. Reprod. 53, 276–284.Holt, W., Watson, P., Curry, M., Holt, C., 1994. Reproducibility of computer-aimed analysis: comparison of five differentsystems used in a practical workshop. Fertil. Steril. 62, 1277–1282.Karagiannidis, A., Varsakely, S., Karatzas, G., 2000. Characteristics and seasonal variations in the semen of Alpine.Saanen and Damascus goat bucks born and raised in Greece. Theriogenology 53, 1285–1293.Lovari, S., 1986. Camoscio d’Abruzzo. In: Regione Abruzzo, Assessorato Urbanistica e Beni Ambientali, Ufficio Parchie Riserve Naturali, Fauna d’Abruzzo 1986, pp. 181–183.Lovari, S., Pellegrini, M., 1998. Camoscio Appenninico. In: Bulgarini, F., Calvario, E., Fraticelli, F., Petretti, F., Sarrocco,S. (Eds.), Libro Rosso degli animali d’Italia (vertebrati), vol. 102. WWF Italia, Rome, pp. 129–171.Mann, T., Lutwac-Mann, C., 1981. Male Reproductive Function and Semen. Springer-Verlag, New York, pp. 269–326.Prado, V., Orihuela, A., Lozano, S., Perez-Leon, I., 2002. Management of female stimulus during semen collection andits association with libido re-establishment and semen characteristics of goat. J. Anim. Sci. 80, 1520–1523.Roca, J., Martinez, E., Vasquez, J.M., Coy, P., 1992. Characteristics and seasonal variations in the semen of Murciano-Granadina goats in Mediterranean area. Anim. Reprod. Sci. 29, 255–262.Salomon, S., Ritar, A.J., 1982. Deep freezing of Angora goat semen: effects of diluent composition and method and rateof dilution on survival of spermatozoa. Aust. J. Biol. Sci. 35, 295–303.

Semen evaluation in the Chamois of Abruzzi (rupicapra pyrenaica ornata)

https://iris.unito.it/bitstream/2318/8204/1/semen%20evaluation%20in%20chamois%20abruzzi.pdf

Semen evaluation in the Chamois of Abruzzi (rupicapra pyrenaica ornata)

Abstract

Similar works

Full text

Available Versions

Institutional Research Information System University of Turin