利用非损伤法分析鳄蜥粪便激素水平的变化
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摘要
利用粪便样品检测性腺类固醇激素代谢水平在很多物种中已得到有效应用,但是在爬行动物中相应的研究还较少。2011年5-10月对广东罗坑自然保护区的10只(5雄5雌)鳄蜥进行粪便采样,采用乙醇加热法提取粪便中类固醇激素,并采用酶联免疫法成功检测鳄蜥粪便中雌二醇、睾酮和孕酮的浓度。研究结果显示:雄性个体睾酮水平在6月份有明显升高,提示6月份可能是鳄蜥的主要发情期;在进入冬眠前的10月份,鳄蜥怀孕个体的孕酮水平明显升高,与5月份的孕酮水平差异显著(LSD:P<0.05);雌性怀孕和非怀孕个体雌二醇水平无季节性变化,因此认为雌二醇有可能在鳄蜥繁殖周期中并不起重要作用。
The noninvasive technique of gonadal steroid metabolite measurement in feces for evaluation of reproductive activity has been studied on many species,but few studies have been used in reptile species.In this study,fecal samples were collected from ten Shinisaurus crocodiluruses(five males and five females)in Luokeng Nature Reserve,Guangdong from May to October 2011.Ethanol-heating method was used to extract the hormone in feces,and enzyme immunoassays(EIA)were validated for the analysis of fecal samples from Chinese Crocodile Lizard for estradiol(E2),testosterone(T),and progesterone(P)and their metabolites.The progesterone concentration of female pregnant individual showed a high point in October(LSD:P<0.05),male testosterone levels have increased significantly in June.There were no significant differences in pregnant and non pregnant females estradiol levels in the breeding season,therefore,estradiol may not play an important role in S.crocodiluruses.
The noninvasive technique of gonadal steroid metabolite measurement in feces for evaluation of reproductive activity has been studied on many species,but few studies have been used in reptile species.In this study,fecal samples were collected from ten Shinisaurus crocodiluruses(five males and five females)in Luokeng Nature Reserve,Guangdong from May to October 2011.Ethanol-heating method was used to extract the hormone in feces,and enzyme immunoassays(EIA)were validated for the analysis of fecal samples from Chinese Crocodile Lizard for estradiol(E2),testosterone(T),and progesterone(P)and their metabolites.The progesterone concentration of female pregnant individual showed a high point in October(LSD:P<0.05),male testosterone levels have increased significantly in June.There were no significant differences in pregnant and non pregnant females estradiol levels in the breeding season,therefore,estradiol may not play an important role in S.crocodiluruses.
引文
[1]EDWARDS A,JONES S M.Changes in plasma progesterone,estrogen,and testosterone concentrations throughout the reproductive cycle in female viviparous blue-tongued skinks,Tilique nigrolutea(Scinidael),in Tasmania[J].General and Comparative Endocrinology,2001,122(3):260-269.
[2]BRASFIELD S M,TALENT L G,JANZ D M.Reproductive and thyroid hormone profiles in captive Western fence Lizards(Sceloporus occidentalis)after a period of brumation[J].Zoo Biology,2008,27(1):36-48.
[3]LASLEY B L,KIRKPATRICK J F.Monitoring ovarian function in captive and free-ranging wildlife by means of urinary and fecal steroids[J].Journal of Zoo and Wildlife Medicine,1991,22(1):23-31.
[4]SCHWARZENBERGER F,MSTL E,PALME R,et al.Faecal steroid analysis for non-invasive monitoring of reproductive status in farm,wild and zoo animals[J].Animal Reproduction Science,1996,42(1-4):515-526.
[5]TOUMA C,PALME R.Measuring fecal glucocorticoid metabolites in mammals and birds:the importance of validation[J].Annals of the New York Academy of Sciences,2005,1046:54-74.
[6]王慧平,高云芳,张新利,等.川金丝猴粪尿中类固醇性激素抽提方法比较[J].兽类学报,2005,25(3):297-301.
[7]李春,魏辅文,胡锦矗.雌性小熊猫粪样中雌二醇与孕酮水平的变化与繁殖启动的关系[J].动物学研究,2005,26(2):147-151.
[8]ALKINDI A Y A,MAHMOUD I Y,WOLLER M J,et al.Oviductal morphology in relation to hormonal levels in the snapping turtle,Chelydra serpentina[J].Tissue and Cell,2006,38(1):19-33.
[9]EDWARDS A,JONES S M.Changes in plasma testosterone,estrogen,and progesterone concentrations throughout the annual reproductive cycle in male viviparous blue-tongued skinks,Tiliqua nigrolutea,in Tasmania[J].Journal of Herpetology,2001,35(2):293-299.
[10]RITTENHOUSE C D,MILLSPAUGH J J,WASHBURN B E,et al.Effects of radiotransmitters on fecal glucocorticoid metabolite levels of three-toed box turtles in captivity[J].Wildlife Society Bulletin,2005,33(2):706-713.
[11]CASARES M.Untersuchungen zum Fortpflanzungsgeschehen bei Riesenschildkroeten(Geochelone elephantopus und G.gigantea)und Landschildkroeten(Testudo graeca und T.hermanni)anhand von Ultraschalldiagnostik und Steroidanalysen im Kot[J].Zoologische Garten,1995,12(1):65.
[12]ATKINS N,JONES S M,EDWARDS A.Fecal testosterone concentrations may not be useful for monitoring reproductive status in male blue-tongued Lizards(Tiliqua nigrolutea:Scincidae)[J].Journal of Herpetology,2002,36(1):106-109.
[13]KUMMROW M S,GILMAN C,MACKIE P,et al.Noninvasive analysis of fecal reproductive hormone metabolites in female veiled chameleons(Chamaeleo calyptratus)by enzyme immunoassay[J].Zoo Biology,2011,30(1):95-115.
[14]张玉霞.鳄蜥的卵胎生[J].大自然,1983(2):33.
[15]张玉霞.中国鳄蜥[M].北京:中国林业出版社,1991:22-25.
[16]刘晓燕.鳄蜥,大壁虎内耳的解剖学研究[J].解剖学研究,1999,21(1):4-7.
[17]万建军.鳄蜥(Shinisaurus crocodilurus Ahl)行为初步观察和遗传多样性的RADP分析[D].桂林:广西师范大学,2004:69-71.
[18]王振兴,武正军,于海,等.广东罗坑自然保护区鳄蜥的体温调节及静止代谢率的热依赖性[J].动物学报,2008,54(6):964-971.
[19]武正军,戴冬亮,宁加佳,等.广东罗坑自然保护区鳄蜥生境选择的季节性差异[J].生态学报,2012,32(15):4691-4699.
[20]于海,黄乘明,武正军,等.鳄蜥生活习性的观察[J].四川动物,2006,25(2):364-366.
[21]王振兴,武正军,陈亮,等.年龄和怀卵对鳄蜥选择体温及静止代谢率的影响[J].广西师范大学学报:自然科学版,2009,27(2):80-84.
[22]万建军.广东罗坑自然保护区瑶山鳄蜥昼间活动节律和领域行为研究[D].成都:四川大学,2009:75-77.
[23]张也,刘以祥.酶联免疫技术与食品安全快速检测[J].食品科学,2003,24(8):200-204.
[24]AMEY A P,WHITTIER J M.Seasonal patterns of plasma steroid hormones in males and females of the bearded dragon lizard,Pogona barbata[J].General and Comparative Endocrinology,2000,117(3):335-342.
[2]BRASFIELD S M,TALENT L G,JANZ D M.Reproductive and thyroid hormone profiles in captive Western fence Lizards(Sceloporus occidentalis)after a period of brumation[J].Zoo Biology,2008,27(1):36-48.
[3]LASLEY B L,KIRKPATRICK J F.Monitoring ovarian function in captive and free-ranging wildlife by means of urinary and fecal steroids[J].Journal of Zoo and Wildlife Medicine,1991,22(1):23-31.
[4]SCHWARZENBERGER F,MSTL E,PALME R,et al.Faecal steroid analysis for non-invasive monitoring of reproductive status in farm,wild and zoo animals[J].Animal Reproduction Science,1996,42(1-4):515-526.
[5]TOUMA C,PALME R.Measuring fecal glucocorticoid metabolites in mammals and birds:the importance of validation[J].Annals of the New York Academy of Sciences,2005,1046:54-74.
[6]王慧平,高云芳,张新利,等.川金丝猴粪尿中类固醇性激素抽提方法比较[J].兽类学报,2005,25(3):297-301.
[7]李春,魏辅文,胡锦矗.雌性小熊猫粪样中雌二醇与孕酮水平的变化与繁殖启动的关系[J].动物学研究,2005,26(2):147-151.
[8]ALKINDI A Y A,MAHMOUD I Y,WOLLER M J,et al.Oviductal morphology in relation to hormonal levels in the snapping turtle,Chelydra serpentina[J].Tissue and Cell,2006,38(1):19-33.
[9]EDWARDS A,JONES S M.Changes in plasma testosterone,estrogen,and progesterone concentrations throughout the annual reproductive cycle in male viviparous blue-tongued skinks,Tiliqua nigrolutea,in Tasmania[J].Journal of Herpetology,2001,35(2):293-299.
[10]RITTENHOUSE C D,MILLSPAUGH J J,WASHBURN B E,et al.Effects of radiotransmitters on fecal glucocorticoid metabolite levels of three-toed box turtles in captivity[J].Wildlife Society Bulletin,2005,33(2):706-713.
[11]CASARES M.Untersuchungen zum Fortpflanzungsgeschehen bei Riesenschildkroeten(Geochelone elephantopus und G.gigantea)und Landschildkroeten(Testudo graeca und T.hermanni)anhand von Ultraschalldiagnostik und Steroidanalysen im Kot[J].Zoologische Garten,1995,12(1):65.
[12]ATKINS N,JONES S M,EDWARDS A.Fecal testosterone concentrations may not be useful for monitoring reproductive status in male blue-tongued Lizards(Tiliqua nigrolutea:Scincidae)[J].Journal of Herpetology,2002,36(1):106-109.
[13]KUMMROW M S,GILMAN C,MACKIE P,et al.Noninvasive analysis of fecal reproductive hormone metabolites in female veiled chameleons(Chamaeleo calyptratus)by enzyme immunoassay[J].Zoo Biology,2011,30(1):95-115.
[14]张玉霞.鳄蜥的卵胎生[J].大自然,1983(2):33.
[15]张玉霞.中国鳄蜥[M].北京:中国林业出版社,1991:22-25.
[16]刘晓燕.鳄蜥,大壁虎内耳的解剖学研究[J].解剖学研究,1999,21(1):4-7.
[17]万建军.鳄蜥(Shinisaurus crocodilurus Ahl)行为初步观察和遗传多样性的RADP分析[D].桂林:广西师范大学,2004:69-71.
[18]王振兴,武正军,于海,等.广东罗坑自然保护区鳄蜥的体温调节及静止代谢率的热依赖性[J].动物学报,2008,54(6):964-971.
[19]武正军,戴冬亮,宁加佳,等.广东罗坑自然保护区鳄蜥生境选择的季节性差异[J].生态学报,2012,32(15):4691-4699.
[20]于海,黄乘明,武正军,等.鳄蜥生活习性的观察[J].四川动物,2006,25(2):364-366.
[21]王振兴,武正军,陈亮,等.年龄和怀卵对鳄蜥选择体温及静止代谢率的影响[J].广西师范大学学报:自然科学版,2009,27(2):80-84.
[22]万建军.广东罗坑自然保护区瑶山鳄蜥昼间活动节律和领域行为研究[D].成都:四川大学,2009:75-77.
[23]张也,刘以祥.酶联免疫技术与食品安全快速检测[J].食品科学,2003,24(8):200-204.
[24]AMEY A P,WHITTIER J M.Seasonal patterns of plasma steroid hormones in males and females of the bearded dragon lizard,Pogona barbata[J].General and Comparative Endocrinology,2000,117(3):335-342.