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Articles 19

Articles 19

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Published by brentlion
articles concerning the semen characteristics and mean testosterone of asiatic lions and indian tigers.
articles concerning the semen characteristics and mean testosterone of asiatic lions and indian tigers.

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Published by: brentlion on Mar 25, 2009
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09/30/2012

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Assessment of inbreeding depression in big cats: Testosterone levels and semen analysis
S. Shivaji* D. Jayaprakash and Suresh B. PatilCentre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, IndiaDubai Gynaecology and Fertility Centre PO Box No. 8729, DubaiStudies on Asiatic lions earlier suggested that they were highly inbred and have very low levels of geneticvariations. However, subsequent studies indicated higher degree of DNA polymorphism in Asiatic lions andIndian tigers and suggested that the low genetic variability was a characteristic feature of these species andwas not the consequence of intensive inbreeding. Therefore, the present study was undertaken to ascertainwhether inbreeding depression has set in these species. A total of 16 tigers and 7 lions from three Indian zooswere evaluated for inbreeding depression effects by analysis of semen samples with respect to spermatozoalnumber, percentage motile spermato- zoa, percentage morphologically abnormal sper- matozoa, ejaculatevolume and fertilizing ability of spermatozoa. Majority of the animals exhibited good spermatozoal number,high percentage of motile spermatozoa and low incidence of abnormal spermatozoa unlike inbred animals,thus implying that inbreeding depression has not yet affected these animals. The high fertilizing ability of thesemen samples and the high levels of serum testosterone further support the view that the Asiatic lions andIndian tigers are not completely inbred. THE Indian tiger (Panthera tigris tigris) and the Asiatic lion (Panthera leo persica) are today the mostcritically endangered animals. The decimation of these mega cats to a few thousand tigers and a few hundredlions could be attributed to habitat shrinkage, large-scale indiscriminate hunting for sport and pleasure anddue to poaching for their body parts which are in great demand in China, South East Asia, Middle East,Western Europe and USA1. Thus there is a need to conserve these mega cats and increase their numbers.Species survival is critically dependent on reproductive performance which in turn seems to correlate withgenetic heterozygosity2-7. Wildt et al.7 based on a comparative study of the Asiatic lions of Gir forest andseveral African lions (Panthera leo leo) established a direct correlation between the lack of genetic variabilityin the Asiatic lion and the high incidence of morphological abnormal spermatozoa and low levels of the malesteroid hormone testosterone8,9 and predicted7-9 that the Asiatic lion has suffered a population bottleneck followed by inbreeding. However, a recent study10 on the tigers and lions of India based on randomlyamplified polymorphic DNA (RAPD), microsatellite analysis of five repeat loci and multilocus fingerprintingindicated a higher degree of genetic heterozygosity than reported and they concluded that the low geneticvariability may be a characteristic feature of these species and not the result of intensive inbreeding. Thepresent study was, therefore, undertaken to evaluate the extent of inbreeding depression in the Indian tigersand the Asiatic lions based on semen characteristics such as spermatozoal number, percentage motilespermatozoa, incidence of abnormal spermatozoa, levels of serum testosterone and fertilizing ability of spermatozoa. Our analyses corroborate Singh et al's.10 observation in that the majority of the animals exhibitvery good semen parameters characteristic of proven fertile animals and do not seem to match that of inbredanimals under the grip of inbreeding depression as suggested by Wildt et al.7.
Materials and methods
AnimalsSemen and blood samples were collected from adult Indian tigers and Asiatic lions maintained in various zoosin India (Table 1). All the animals were reared in cages with an access to a crawl during the day time and astandard diet of beef (8 kg without bones) supplemented with adequate amount of calcium, was provided six
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days in a week. Water was always available to the animals.
Collection of semen
Tigers and lions were anesthetized prior to electroejaculation using a combination of ketamine HCl (2.22 to2.66 mg/kg body weight; Troy Labs, New South Wales, Australia) and xylazine (1.11 to 1.33 mg/kg bodyweight; Troy Labs, New South Wales, Australia) which were loaded into a projectile dart (Dis Inject, HelproHealth Products and Services, Delhi, India) and injected intramuscularly using a blow pipe or a dart pistol(Figure 1a). By this procedure, it took the animals about 20 to 30 min to be completely under the influence of the anaesthesia. Further, to maintain a surgical plane of anaesthesia till blood and semen samples werecollected, which normally took about 30 min, an additional injection of ketamine HCl (50 to 100 mg) wasgiven intravenously.The anesthetized animals were then shifted out from the cages onto an elevated platform and allowed to lie ontheir side. Prior to electroejaculation, the entire area surrounding the anus, testis and penis was washed withwater, dried with tissue paper and smeared with spirit which was allowed to evaporate. The prepuce and thepenis were also cleaned to avoid contamination of the ejaculate. Simultaneously a rectal probe (3.7 cm indiameter) was disinfected with Dettol (Reckitt and Colman of India Limited, Calcutta, India), lubricated withwhite petroleum jelly and gently inserted into the rectum of the animal such that the three longitudinalelectrodes of the probe faced the ventral side of the animal against the male accessory sex organs (Figure 1b). The probe was connected to a electrostimulator (Bioelectric Co., Washington, DC, USA) consisting of avoltmeter (0 to 45 V), current meter (0 to 4 amp), a control knob to set the desired volt and an on/off switchto deliver the electric pulses (Figure 1 b). In the field the electrostimulator was powered using an UPS(Kirolosker Electricals, Mysore, India).
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FIGURE 1
Just prior to stimulating the animal, the penis was extended beyond the prepuce and inserted into a 25 mlsterile specimen container (Figure 2 a). The stimulatory regime used for electroejaculation of tigers and lionsconsisted of a total of 80 stimuli between 2 and 5 V divided into 3 series. In series 1 the animal was stimulated30 times with 10 stimuli at 2, 3 and 4 V respectively; in series 2 also it was stimulated 30 times with 10 stimuliat 3, 4 and 5 V respectively and in series 3 the animal was stimulated 20 times with 10 stimuli at 4 and 5 Vrespectively. At times if the animal did not ejaculate even after the third series additional 10 stimuli weregiven at 5 and 6 V respectively. Each stimulus at the desired voltage was given for 2 s and was followed by abreak of 2 s till the next stimulus was given. The animals were rested for 3 to 5 min between each series of stimuli.
articles19.htmhttp://www.ias.ac.in/currsci/nov10/articles19.htm3 of 113/15/2009 2:47 PM

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