Characterization of Red-Legged Partridge (Alectoris Rufa)Sperm: Seasonal Changes and Influence of Genetic Purity
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Characterization of red-legged partridge (Alectoris rufa)sperm: Seasonal changes and influence of genetic purity J. Santiago-Moreno,∗1,2 C. Casta˜no,∗ A. Toledano-D´ıaz,∗ M. C. Esteso,∗ A. L´opez-Sebasti´an,∗ N. Ga˜n´an,∗ M. J. Hierro,† F. Marchal,‡ J. L. Campo,§ and E. Blesbois∗∗ ∗Dpto. Reproducci´on Animal, INIA, Spain; †Consejer´ıa Medio Ambiente y Ordenaci´on del Territorio, Junta de Andaluc´ıa, Spain; ‡Agencia de Medio Ambiente y Agua de Andaluc´ıa, Junta de Andaluc´ıa, Spain; §Dpto. Mejora Gen´etica Animal, INIA, Spain; and ∗∗INRA 0085 UMR PRC INRA-CNRS-University Fran¸cois Rabelais-Haras Nationaux, 37380 Nouzilly, France Downloaded from https://academic.oup.com/ps/article-abstract/94/1/80/1596569 by guest on 20 March 2019 ABSTRACT The general decline in wild Iberian popu- the hybrids. Reproductive activity in both groups de- lations of the red-legged partridge (Alectoris rufa)has creased in June, to reach a minimum in August to De- been accompanied by an increase in game-farm facili- cember among the hybrids and in September to De- ties producing hybrids with chukar partridges (Alectoris cember among the pure birds. Spermatogenic activity chukar). Genetic introgression from chukar partridges is resumed in January in both groups. The sperm concen- thought to modify male red-legged partridge reproduc- tration produced by the pure birds was smaller than tive indicators. The aim of the present study was to that of the hybrids (P < 0.001), but the percentage of determine the effects of such genetic introgression on motile sperm was higher in the pure birds (P < 0.001). seasonal reproductive patterns by comparing the sperm The sperm of the hybrids showed greater straight-line and plasma testosterone concentrations of males from velocity (P < 0.05), linearity (P < 0.001), straightness pure red-legged and hybrid red-legged/chukar popula- (P < 0.001), sperm wobble (P < 0.05), and beat-cross tions. Semen was collected twice monthly over a 12-mo frequency values (P < 0.001). The length and area of period using a massage technique. Both types of bird the sperm head were smaller in the pure birds (P < showed a clear seasonal pattern of spermatogenic ac- 0.05). The seasonal plasma testosterone concentration tivity. The proportion of males ejaculating sperm was pattern followed a trend roughly parallel to the ejac- higher (P < 0.05) among the pure red-legged birds. The ulatory response. The present results suggest that ge- greatest sperm production was recorded in March to netic introgression influences the reproductive variables May among the pure birds and April to May among of the red-legged partridge. Key words: breeding activity, introgression, red-legged partridge, sperm morphometry, testosterone 2015 Poultry Science 94:80–87 http://dx.doi.org/10.3382/ps/peu020 INTRODUCTION cilities producing birds for release to maintain hunt- ing quotas. It is estimated that at least 3 to 4 mil- The red-legged partridge (Alectoris rufa) is a south- lion farm-bred red-legged partridges are released every western European game bird that inhabits farmland year in Spain (Garrido, 2002;S´anchez-Garc´ıa et al., and open stony areas. The greater part of its global 2009), but populations have failed to recover because population is found within Spain, where it breeds nat- these raised birds show reduced antipredator behaviour. urally. Despite its ecological (biodiversity, food chain) Their mortality rate is therefore high. The release of and economic (game resource) importance, wild popu- hybrids produced by crossing the red-legged partridge lations have declined sharply since the 1970s, the con- and the chukar partridge (Alectoris chukar), a Eurasian sequence of changes in agricultural practices and over- species, is threatening the genetic integrity of wild hunting (Blanco-Aguiar et al., 2003; Casas and Vi˜nuela, Spanish birds. Indeed, at least 63% of supposedly red- 2010;D´ıaz-Fern´andez et al., 2012). The general de- legged partridges now raised on game farms, and 30 cline of wild populations in the Iberian Peninsula has to 45% of birds living in the wild, possess chukar par- been accompanied by an increase in game-farm fa- tridge genes (Blanco-Aguiar et al., 2008; Casas et al., 2013). Although the release of hybrid partridges has C 2015 Poultry Science Association Inc. been forbidden under Spanish law, the competitiveness Received June 23, 2014. of markets has led to their continued, illegal, produc- Accepted September 16, 2014. tion. Indeed, A. rufa × A. chukar hybrids are preferred 1This work was funded by INIA grant RZ2012-00013-C02. 2Corresponding author: E-mail: [email protected]; Tel: +34 91347 4020; by game breeders because they are less sensitive to Fax: +34 91347 4014 stress, and their productivity is better under captive 80 SPERM PRODUCTION AND QUALITY IN RED-LEGGED PARTRIDGES 81 conditions (longer laying period and heavier body domly selected from the commercial farm; the results weight) (Potts, 1989). The preservation of the wild pop- revealed that 35% of sampled birds showed genetic in- ulation of red-legged partridges is also affected by newly trogression with A. chukar. emerging infectious diseases, the propagation of which All the birds were 2-y-old at the beginning of the ex- is facilitated by hybridization (Vi˜nuela et al., 2013; periment. They were housed in outdoor cages (90 by 82 Ag¨uero et al., 2011). by 60 cm) in groups of three birds under natural pho- It is here hypothesized that hybridization, and the ar- toperiod and temperature conditions at the El Enc´ın tificial selection of productive features under farms con- Research Station (Alcal´a de Henares, Spain, 40o31N]). ditions, might determine variations in sperm produc- Naturally occurring variations in day length at that lati- tion in red-legged partridges and perhaps even modify tude were from 10 h 17 min to 16 h 3 min light/d (winter the morphological and functional characteristics of their to summer solstices), including twilight. All birds were sperm. A. chukar is more promiscuous than A. rufa, fed (ad libitum) a commercial feed containing 16% CP, which is monogamous (Vidal and Colominas, 2007), and 2,700 kcal of ME/kg, 3.5% Ca, and 0.5% available P Downloaded from https://academic.oup.com/ps/article-abstract/94/1/80/1596569 by guest on 20 March 2019 it is well known that polyandry creates a background over the entire experimental period, which lasted 12 for sexual selection among males both before and after mo. Periodically, they were supplemented with amino copulation (sperm competition) that influences a num- acids and vitamins (Aminolid, Intervet Schering Plough ber of sperm variables (Pizzari et al., 2007;Parkerand Animal Health, Barcelona, Spain) via their water. The Pizzari, 2010; Collet et al., 2012), including the num- birds were weighed using a 1000 g Pesola (Pesola light ber of sperm produced, the percentage of sperm abnor- line, Oryx) precision scale (nearest 10 g) at the start of malities, sperm cell size (Birkhead, 2000, Kleven et al., the experiment. 2009), and swimming velocity variables (Kleven et al., 2009, Santiago-Moreno et al., 2014). Sperm Collection and Assessment In this study, the reproductive indicators of pure red- of Sperm Variables legged and hybrid red-legged/chukar partridges were compared over a period of one year to examine the po- Sperm was collected twice per month using the tential impact on the former species of chukar partridge massage technique of Burrows and Quinn (1937), as genetic introgression. adapted to this species. Two people were required for this. One caught the male and held him firmly over a MATERIALS AND METHODS table, with one hand holding the legs and the other im- mobilizing the body and wings. The other person stim- Animals ulated the bird, simultaneously stroking the back with the right hand and the abdomen with the left hand. The The studied birds were 19 pure red-legged partridges, bird was then “milked” by making the copulatory organ all adult males, from the Lugar Nuevo red-legged par- protrude by mild stimulation and then gripping its base tridge breeding facility (And´ujar, Spain, 38o16N; this with the thumb and index finger of the right hand. Se- facility belongs to the regional government of Andalusia men was recovered by capillarity using a microhaema- and maintains a genetically pure red-legged partridge tocrit tube (GmbH + Co. KG, Wertheim, Germany) population [D´avila, 2009]), and 22 adult male red- and immediately diluted 1:1 (v:v) at ambient tem- legged partridges showing genetic introgression from perature using a medium-containing sodium glutamate Alectoris chukar. These birds were obtained from a (1.92 g), glucose (0.8 g), magnesium acetate (4H2O; commercial farm (Sequera, Segovia, Spain, 41o22N), 0.08 g), potassium acetate (0.5 g), polyvinylpyrrolidone and genetic introgression by A. chukar is >35%. The (Mr 10,000; 0.3 g) and 100 mL H2O (pH 7.08, osmolal- genetic purity and A. chukar introgression shown by ity 343 mOsm kg−1; hereinafter referred to as Lake and these two populations were tested according to official Ravie medium) (Lake and Ravie, 1984). Diluted semen genetic analysis procedures recommended by the re- was immediately refrigerated at 5oC and transported to gional governments (Andalusian, Castilla y Le´on) (FE- the laboratory. All individual sperm samples were ex- DENCA, 2011). Genetic introgression was tested us- amined within 45 min of collection, and ejaculate vol- ing mitochondrial cytochrome b gene (mtDNA Cyt- ume, sperm concentration, motility, and morphometric b), 7 species-specific nuclear DNA microsatellites for variables were recorded. red-legged partridges (Aru1E45, Aru1E78, Aru1I68, Sperm volume was calculated by measuring the Aru1F138, Aru1V16, Aru1H15, and Aru1F32), and 7 length of the semen column in the microcapillary tube single nucleotide polymorphisms (SNPs) in mtDNA.