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PubMedDate not supplied

[Amur tiger: path to the third millennium (experience of development of the "Strategy of preservation of the Amur tiger in Russia")].

The development of a system of measures for conservation of the Amur tiger is summarized. In the middle of the 1990s, no marked reduction in the numbers of the Amur tiger took place in Russia; the animals were always present in most of the forest-covered area of the Primorskii and South Khababrovsk districts. However, new threats to the survival of the Amur tiger appeared. It is necessary to preserve the self-regulating population of the Amur tiger throughout its range. The key to solving this task is the identification of three functionally unequal zones in the southern Far East of Russia: a monolithic "tiger bastion" consisting of protected territories with different statuses, a zone of "stable compromise" (between the interests of tiger conservation and economic development of the territory), and a "tiger-free" zone, where the animals periodically migrate. The importance of Russian-Chinese cooperation in conservation of the Amur tiger has been stressed. A large international borderline reserve-cluster for this purpose has been proposed.

Animals↗
PubMed2005

Probable tiger-to-tiger transmission of avian influenza H5N1.

During the second outbreak of avian influenza H5N1 in Thailand, probable horizontal transmission among tigers was demonstrated in the tiger zoo. Sequencing and phylogenetic analysis of those viruses showed no differences from the first isolate obtained in January 2004. This finding has implications for influenza virus epidemiology and pathogenicity in mammals.

Animals↗
PubMedDate not supplied

Bit-mapped quantitative EEG analysis in a tiger (Tiger felis) with partial seizures: a case report.

We report electroencephalographic findings in an anesthetized 4-month-old female drug-naive tiger (Tiger felis) affected by partial seizures with secondary generalization. Both clinical signs and electroencephalographic abnormalities were consistent with a forebrain lesion. Recurring epileptiform activity was noted in the left frontal, central and temporal derivations upon visual inspection of the electroencephalogram (EEG). A quantified EEG, displayed on brain maps, showed the predominance of delta and theta slow rhythms. As regards the absolute power, a prevalence of left frontal-temporal activity was noted. An infectious or inflammatory condition was thought to be the most probable cause of the symptomatic epilepsy in our patient. Unfortunately, other differential diagnoses could not be ruled out.

Animals↗
PubMed2006

Developing a national tiger action plan for The Union of Myanmar.

A century ago, tigers were considered pests in Myanmar. Hunters claimed thousands, yet populations persisted. In the past century, because of habitat loss and prey depletion, coupled with the recent demand for traditional medicines, tiger populations have been reduced to a few hundred individuals. As a first step toward long-term planning for tigers, and to guide efforts to increase protected area coverage, the Myanmar government in 1998 initiated a project to develop a revised National Tiger Action Plan. Extensive surveys confirmed tigers in only 4 of 17 survey locations. Significant populations are thought to persist in the far North and far South of the country, where large, intact forests offer the potential for tiger recovery. With partnerships and collaborations, tiger populations can be protected in the short term (<5 years) by expanding protected areas and corridors, mobilizing enforcement staff to reduce poaching of tigers and prey, and amending existing wildlife legislation in accordance with international laws. Over the long term (5-20 years), recovery of Myanmar's tiger populations will depend on increasing support from local people, zoning tiger areas to reduce habitat loss and disturbance, and maintaining connectivity of existing national and transboundary forests. This article reviews the development of a new National Tiger Action Plan for the Union of Myanmar and discusses a blueprint for conservation measures aimed at saving tigers from extinction.

Animals↗
PubMed2004

Phylogeography and genetic ancestry of tigers (Panthera tigris).

Eight traditional subspecies of tiger (Panthera tigris),of which three recently became extinct, are commonly recognized on the basis of geographic isolation and morphological characteristics. To investigate the species' evolutionary history and to establish objective methods for subspecies recognition, voucher specimens of blood, skin, hair, and/or skin biopsies from 134 tigers with verified geographic origins or heritage across the whole distribution range were examined for three molecular markers: (1) 4.0 kb of mitochondrial DNA (mtDNA) sequence; (2) allele variation in the nuclear major histocompatibility complex class II DRB gene; and (3) composite nuclear microsatellite genotypes based on 30 loci. Relatively low genetic variation with mtDNA,DRB,and microsatellite loci was found, but significant population subdivision was nonetheless apparent among five living subspecies. In addition, a distinct partition of the Indochinese subspecies P. t. corbetti in to northern Indochinese and Malayan Peninsula populations was discovered. Population genetic structure would suggest recognition of six taxonomic units or subspecies: (1) Amur tiger P. t. altaica; (2) northern Indochinese tiger P. t. corbetti; (3) South China tiger P. t. amoyensis; (4) Malayan tiger P. t. jacksoni, named for the tiger conservationist Peter Jackson; (5) Sumatran tiger P. t. sumatrae; and (6) Bengal tiger P. t. tigris. The proposed South China tiger lineage is tentative due to limited sampling. The age of the most recent common ancestor for tiger mtDNA was estimated to be 72,000-108,000 y, relatively younger than some other Panthera species. A combination of population expansions, reduced gene flow, and genetic drift following the last genetic diminution, and the recent anthropogenic range contraction, have led to the distinct genetic partitions. These results provide an explicit basis for subspecies recognition and will lead to the improved management and conservation of these recently isolated but distinct geographic populations of tigers.

Alleles↗
PubMed2006

Genotyping faecal samples of Bengal tiger Panthera tigris tigris for population estimation: a pilot study.

BACKGROUND: Bengal tiger Panthera tigris tigris the National Animal of India, is an endangered species. Estimating populations for such species is the main objective for designing conservation measures and for evaluating those that are already in place. Due to the tiger's cryptic and secretive behaviour, it is not possible to enumerate and monitor its populations through direct observations; instead indirect methods have always been used for studying tigers in the wild. DNA methods based on non-invasive sampling have not been attempted so far for tiger population studies in India. We describe here a pilot study using DNA extracted from faecal samples of tigers for the purpose of population estimation. RESULTS: In this study, PCR primers were developed based on tiger-specific variations in the mitochondrial cytochrome b for reliably identifying tiger faecal samples from those of sympatric carnivores. Microsatellite markers were developed for the identification of individual tigers with a sibling Probability of Identity of 0.005 that can distinguish even closely related individuals with 99.9% certainty. The effectiveness of using field-collected tiger faecal samples for DNA analysis was evaluated by sampling, identification and subsequently genotyping samples from two protected areas in southern India. CONCLUSION: Our results demonstrate the feasibility of using tiger faecal matter as a potential source of DNA for population estimation of tigers in protected areas in India in addition to the methods currently in use.

Animals↗
PubMed2004

[Genetic diversity of microsatellite loci in captive Amur tigers].

The tiger is one of the most threatened wildlife species since the abundance and distribution of tiger have decreased dramatically in the last century. The wild Amur tiger (Panthera tigris altaica) only distributed in northeast China, the far east area of Russia and the north Korea and its size of wild population is about 450 in the world and 20 in China. Several hundred captive populations of Amur tigers are the main source to protect gene library of tiger and the source of recovering the wild populations. The Breeding Center for Felidae at Hengdaohezi and Haoerbin Tiger Park in Heilongjiang Province is the biggest captive breeding base in China. How to make clear the genetic pedigree and establish reasonable breeding system is the urgent issues. So we use the microsatellite DNA markers and non-invasive technology to research on the genetic diversity of captive Amur tiger in this study. Ten microsatellite loci (Fca005, Fca075, Fca094, Fca152, Fca161, Fca294, Pti002, Pti003, Pti007 and Pti010), highly variable nuclear markers, were studied their genetic diversity in 113 captive Amur tigers. The PCR amplified products of microsatellite loci were detected by non-denatured polyacrylamide gel electrophoresis. Allele numbers, allelic frequency, gene heterozygosity(H(e)), polymorphism information content(PIC) and effective number of allele(N(e)) were calculated. 41 alleles were found and their size were ranged from 110bp to 250bp in ten microsatellite loci, Fca152 had 6 alleles, Fca075, Fca094 and Fca294 had 5 alleles, Fca005 and Pti002 had 4 alleles and the others had 3 alleles in all tiger samples, respectively. The allelic frequencies were from 0.009 to 0.767; The He ranged from 0.385 to 0.707, and Fca294 and Pti010 locus had the highest and lowest value; the PIC were from 0.353 to 0.658, Fca294 and Pti010 locus had the highest and lowest value; and N(e) were from 1.626 to 3.409, Fca294 and Pti010 locus had the highest and lowest value, which showed the ten microsatellie loci had high or medium polymorphism in these Amur tigers and had high genetic diversity. At the same time, we only found even bases variability which showed the even bases repeat sequence (CA/GT) maybe the basic unit for length variability of microsatellite in all loci. In this study, the samples were made up of 75 hair specimens, 23 blood specimens and 15 tissue specimens, we obtained the genome DNA from hairs using the non-invasive DNA technology and demonstrated that DNA derived from hair samples is as good as that obtained from blood samples for the analysis of microsatellite polymorphism. These results imply that microsatellite DNA markers and non-invasive DNA technology can help study the genetic diversity of Amur tiger. This method could be used in the captive management of other endangered species.

Alleles↗
PubMed2006

Assessing tiger population dynamics using photographic capture-recapture sampling.

Although wide-ranging, elusive, large carnivore species, such as the tiger, are of scientific and conservation interest, rigorous inferences about their population dynamics are scarce because of methodological problems of sampling populations at the required spatial and temporal scales. We report the application of a rigorous, noninvasive method for assessing tiger population dynamics to test model-based predictions about population viability. We obtained photographic capture histories for 74 individual tigers during a nine-year study involving 5725 trap-nights of effort. These data were modeled under a likelihood-based, "robust design" capture-recapture analytic framework. We explicitly modeled and estimated ecological parameters such as time-specific abundance, density, survival, recruitment, temporary emigration, and transience, using models that incorporated effects of factors such as individual heterogeneity, trap-response, and time on probabilities of photo-capturing tigers. The model estimated a random temporary emigration parameter of gamma" = gamma' = 0.10 +/- 0.069 (values are estimated mean +/- SE). When scaled to an annual basis, tiger survival rates were estimated at S = 0.77 +/- 0.051, and the estimated probability that a newly caught animal was a transient was tau = 0.18 +/- 0.11. During the period when the sampled area was of constant size, the estimated population size N(t) varied from 17 +/- 1.7 to 31 +/- 2.1 tigers, with a geometric mean rate of annual population change estimated as lambda = 1.03 +/- 0.020, representing a 3% annual increase. The estimated recruitment of new animals, B(t), varied from 0 +/- 3.0 to 14 +/- 2.9 tigers. Population density estimates, D, ranged from 7.33 +/- 0.8 tigers/100 km2 to 21.73 +/- 1.7 tigers/100 km2 during the study. Thus, despite substantial annual losses and temporal variation in recruitment, the tiger density remained at relatively high levels in Nagarahole. Our results are consistent with the hypothesis that protected wild tiger populations can remain healthy despite heavy mortalities because of their inherently high reproductive potential. The ability to model the entire photographic capture history data set and incorporate reduced-parameter models led to estimates of mean annual population change that were sufficiently precise to be useful. This efficient, noninvasive sampling approach can be used to rigorously investigate the population dynamics of tigers and other elusive, rare, wide-ranging animal species in which individuals can be identified from photographs or other means.

Animal Identification Systems↗
PubMed2004

An extremely sensitive species-specific ARMs PCR test for the presence of tiger bone DNA.

The survival of the tiger (Panthera tigris) is seriously threatened by poaching to provide raw materials for Traditional Chinese Medicines (TCMs). Most highly prized are the tiger's bones, which are used in combination with other animal and plant derivatives in pills and plasters for the treatment of rheumatism and other ailments. Hundreds of patent remedies have been produced which claim to contain tiger bone, but proof of its presence is needed, if legislation prohibiting the trade in endangered species is to be enforced. A highly sensitive tiger-specific real-time PCR assay has been developed to address this problem. Using primers specific to the tiger mitochondrial cytochrome b gene, successful amplification has been reliably achieved from blood, hair and bone as well as from a range of TCMs spiked with 0.5% tiger bone. Although capable of detecting fewer than 10 substrate molecules, the seven varieties of TCM pills and plasters tested showed no detectable trace of tiger DNA before spiking. Furthermore, sequencing several "tiger bone" fragments seized from TCM shops has shown that they actually originated from cattle and pigs. The potential effects of traditional bone preparation methods, evidence that much lower concentrations are used than alleged on TCM packaging, and substitution of bones from other species all suggest a low likelihood of detecting tiger DNA in patent medicines. Despite this, the basic methods have been thoroughly proven and can be readily applied to derivatives from other CITES protected species providing a rapid and highly sensitive forensic test for species of origin. Potential applications to the monitoring of wild populations are demonstrated by the successful identification of shed hairs and faecal samples.

Animals↗
PubMed2005

Individualization of tiger by using microsatellites.

In investigating criminal cases of poaching and smuggling involving tigers (Panthera tigris), the number of tiger individuals involved is critical for determining the penalty. Morphological methodologies do not often work because tiger parts do not possess the distinctive characteristics of the individual. Microsatellite DNAs have been proved a reliable marker for the individualization of animals. Seven microsatellite loci derived from domestic cat (Felis catus) were selected to individualize tigers, namely F41, F42, F146, Fca304, Fca391, Fca441 and Fca453. A reference population containing 37 unrelated tigers were used to investigate alleles, allelic frequencies, genotypes and genotype frequencies of each locus. Consequently, the data was used to assess the validity of the combination of seven loci for tiger individualization. All loci were polymorphic and easy to amplify. Three out of the seven loci were significantly departure from the Hardy-Weinberg Equilibrium (P < 0.05). Cumulative discrimination power (DP) calculated with observed genotype frequencies was 0.99999789. Match probability of an individual in the reference population with a random individual in seven loci ranged from 7.34 x 10(-9) to 2.77 x 10(-5). This suggests that combining the seven microsatellite loci provides desirable power to individualize tigers. The combination of seven loci was applied to a case of tiger bone smuggling. Genotypes of all samples were identical in all seven loci, and the P(M) of the evidence samples in the seven loci hit 5.63 x 10(-7), provided evidence that the bones belong to a single tiger.

Animals↗
PubMed2002

An extremely sensitive species-specific ARMS PCR test for the presence of tiger bone DNA.

The survival of the tiger (Panthera tigris) is seriously threatened by poaching to provide raw materials for traditional Chinese medicines (TCMs). Most highly prized are the tiger's bones, which are used in combination with other animal and plant derivatives in pills and plasters for the treatment of rheumatism and other ailments. Hundreds of patent remedies have been produced which claim to contain tiger bone, but proof of its presence is needed if legislation prohibiting the trade in endangered species is to be enforced.A highly sensitive tiger-specific real-time PCR assay has been developed to address this problem. Using primers specific to the tiger mitochondrial cytochrome b gene, successful amplification has been reliably achieved from blood, hair and bone as well as from a range of TCMs spiked with 0.5% tiger bone. Although capable of detecting fewer than 10 substrate molecules, the seven varieties of TCM pills and plasters tested showed no detectable trace of tiger DNA before spiking. Furthermore, sequencing several "tiger bone" fragments seized from TCM shops has shown that they actually originated from cattle and pigs. The potential effects of traditional bone preparation methods, evidence that much lower concentrations are used than alleged on TCM packaging, and substitution of bones from other species all suggest a low likelihood of detecting tiger DNA in patent medicines. Despite this, the basic methods have been thoroughly proven and can be readily applied to derivatives from other Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) protected species, providing a rapid and highly sensitive forensic test for species of origin. Potential applications to the monitoring of wild populations are demonstrated by the successful identification of shed hairs and faecal samples.

Animals↗
PubMed1988

A comparative analysis of ejaculate and hormonal characteristics of the captive male cheetah, tiger, leopard, and puma.

Male cheetahs, tigers, leopards, and pumas maintained under the same conditions were anesthetized and 1) serially bled before, during, and after electroejaculation (EE); 2) serially bled only (AO); or 3) serially bled before and after receiving adrenocorticotropin hormone (ACTH). Ejaculates from leopards contained higher (p less than 0.05) sperm concentrations than cheetahs and pumas but lower (p less than 0.05) sperm motility ratings than all other species. Tigers produced a larger seminal volume and the greatest number of motile sperm/ejaculate (p less than 0.05). The percentage of morphologically abnormal spermatozoa was greater (p less than 0.05) in cheetahs (64.6%), leopards (79.5%), and pumas (73.5%) than in tigers (37.5%). The most prevalent spermatozoal deformities included a tightly coiled or bent flagellum, a deranged midpiece, or a residual cytoplasmic droplet. Mean baseline serum cortisol concentrations in leopards were 2- and 4-fold greater (p less than 0.05) than in tigers and cheetahs, respectively. Basal cortisol concentrations in pumas were similar to those of tigers, but irrespective of treatment increased 2-fold (p less than 0.01) during the bleeding period. An acute rise and fall in cortisol attributable to EE was observed only in cheetahs. In tigers and leopards, mean peak cortisol concentrations after ACTH were similar to maximal values observed after EE. However, peak cortisol levels in cheetahs and pumas after ACTH were greater (p less than 0.01) than the concentrations measured after EE, indicating that these manipulatory procedures were not eliciting a maximal adrenal response. In the EE groups, luteinizing hormone (LH) and testosterone levels in cheetahs were lower (p less than 0.05) than in other species, whereas levels of both hormones were comparable (p greater than 0.05) in tigers, leopards, and pumas. Elevated cortisol levels in cheetahs and pumas had no discernible effect on LH/testosterone patterns; however, the results were equivocal in tigers, and, among leopards, testosterone concentrations consistently declined over time. In this study, using a standardized approach, we identify different ejaculate and endocrine characteristics of captive cheetahs, tigers, leopards, and pumas. The data extend earlier observations and demonstrate that some, but not all, Felidae species ejaculate high numbers of pleiomorphic spermatozoa. However, inter-species differences in sperm integrity do not appear related to inter-species variations in cortisol, LH, or testosterone. The observation of continuously declining testosterone concentrations only in leopards after AO, EE, or ACTH treatment suggests that rising and/or elevated cortiso

Acinonyx↗
PubMed2004

Survey of transport environments of circus tigers (Panthera tigris).

The type of equipment used to transport circus tigers, environmental factors experienced during transport, and resultant body temperatures of tigers transported were surveyed during hot and cold weather conditions with six different circus tiger acts. Dataloggers recorded interior and exterior temperatures, relative humidity, and radiant heat at 5-min intervals during each trip. Microdataloggers fed to the tigers recorded body temperature and were recovered from eight Bengal (Panthera tigris tigris), Siberian (P. t. altaica), or Bengal-Sumatran (P. t. tigris-P. t. sumatrae) cross tigers from four different circuses. Three basic types of systems were used by circus acts to transport tigers: freestanding cages mounted on wheels that were winched or pushed into a semitrailer for transport, cages built into the trailer itself, and weather-resistant units transported on flatbed railcars or flatbed truck trailers. The highest temperature encountered inside a trailer was 37.3 degrees C in hot weather conditions, but overall, temperatures were usually between 21.1-26.7 degrees C. Temperature inside the trailers did not appear to be affected by movement and did not generally exceed ambient temperatures, indicating adequate insulation and passive ventilation. During cold weather trips, the lowest temperature inside the trailers was -1.1 degrees C, occurring during an overnight stop. Interior temperatures during cold weather transport stayed 2-6 degrees C warmer than ambient temperatures. The body temperatures of the tigers were unaffected by extreme temperatures. The only changes observed in body temperature were increases of 1-2 degrees C caused by activity and excitement associated with loading in several groups of tigers, regardless of whether it was hot or cold weather. Whenever measured, carbon monoxide and ammonia were below the detectable concentrations of 10 and 1 ppm. respectively. Overall, transport did not appear to have any adverse effects on the tigers' ability to thermoregulate.

Animals↗
PubMed2005

Rapid and repeated origin of insular gigantism and dwarfism in Australian tiger snakes.

It is a well-known phenomenon that islands can support populations of gigantic or dwarf forms of mainland conspecifics, but the variety of explanatory hypotheses for this phenomenon have been difficult to disentangle. The highly venomous Australian tiger snakes (genus Notechis) represent a well-known and extreme example of insular body size variation. They are of special interest because there are multiple populations of dwarfs and giants and the age of the islands and thus the age of the tiger snake populations are known from detailed sea level studies. Most are 5000-7000 years old and all are less than 10,000 years old. Here we discriminate between two competing hypotheses with a molecular phylogeography dataset comprising approximately 4800 bp of mtDNA and demonstrate that populations of island dwarfs and giants have evolved five times independently. In each case the closest relatives of the giant or dwarf populations are mainland tiger snakes, and in four of the five cases, the closest relatives are also the most geographically proximate mainland tiger snakes. Moreover, these body size shifts have evolved extremely rapidly and this is reflected in the genetic divergence between island body size variants and mainland snakes. Within south eastern Australia, where populations of island giants, populations of island dwarfs, and mainland tiger snakes all occur, the maximum genetic divergence is only 0.38%. Dwarf tiger snakes are restricted to prey items that are much smaller than the prey items of mainland tiger snakes and giant tiger snakes are restricted to seasonally available prey items that are up three times larger than the prey items of mainland tiger snakes. We support the hypotheses that these body size shifts are due to strong selection imposed by the size of available prey items, rather than shared evolutionary history, and our results are consistent with the notion that adaptive plasticity also has played an important role in body size shifts. We suggest that plasticity displayed early on in the occupation of these new islands provided the flexibility necessary as the island's available prey items became more depauperate, but once the size range of available prey items was reduced, strong natural selection followed by genetic assimilation worked to optimize snake body size. The rate of body size divergence in haldanes is similar for dwarfs (h(g) = 0.0010) and giants (h(g) = 0.0020-0.0025) and is in line with other studies of rapid evolution. Our data provide strong evidence for rapid and repeated morphological divergence in the wild due to similar selective pressures acting in different directions.

Adaptation, Physiological↗
PubMed2006

Natural and gonadotropin-induced ovarian activity in tigers (Panthera tigris) assessed by fecal steroid analyses.

Fecal samples were collected from female tigers (n = 17) to compare endocrine patterns associated with natural ovarian activity with those after chorionic gonadotropin ovulation induction and artificial insemination (AI). Baseline fecal estradiol concentrations were 65.77 +/- 3.61 ng/g with estrual peaks of 167.39 +/- 9.92 ng/g and an anovulatory cycle length of 17.96 +/- 0.70 days. Peak fecal estradiol was higher when females were housed with a male for breeding (262.30 +/- 41.43 vs. 165.30 +/- 3.67 ng/g; P < 0.05). The majority of animals showed some seasonal differences in fecal estradiol however, patterns were inconsistent. Fecal progestagens increased only after breeding confirming tigers are primarily induced ovulators. The non-pregnant luteal phase was 34.50 +/- 1.85 days in duration. In pregnant tigers, fecal progestagens remained elevated for 108 days until parturition and the diagnosis of pregnancy was possible based on the elevated fecal progestagens after 35 days of gestation. Tigers were administered equine chorionic gonadotropin (eCG) to stimulate follicular growth and human chorionic gonadotropin (hCG) to induce ovulation prior to AI [200 IU eCG/100 IU hCG (n = 5); 400 IU eCG/200 IU hCG (n = 2); 500 IU eCG/100 IU hCG (n = 2); 1000 IU eCG/750 IU hCG (n = 11); 1000 IU eCG/1000 IU hCG (n = 4)]. None of the tigers subjected to AI became pregnant (n = 9). Fecal endocrine patterns in gonadotropin-stimulated tigers were considerably different from those observed in naturally bred tigers. In particular, fecal estradiol concentrations were higher than those observed during natural estrus and remained elevated for longer periods of time in tigers administered the higher doses of gonadotropins typically used in conjunction with AI in this species. These abnormal endocrine patterns may help explain the poor success rate of AI in this species.

Animals↗
PubMed2007

Cell cycle analysis and interspecies nuclear transfer of in vitro cultured skin fibroblasts of the Siberian tiger (Panthera tigris Altaica).

The present study was conducted to examine the effect of cell culture conditions, antioxidants, protease inhibitors (PI), and different levels of dimethylsulfoxide (DMSO) for the promotion of synchronization of different cell cycles of Siberian tiger skin fibroblasts. We also compared the ability of somatic cell nuclei of the Siberian tiger in pig cytoplasts and to support early development after reconstruction. Cell cycle synchronization between nuclear donor and recipient cells is considered to be one of the most crucial factors for successful cloning. Five experiments were performed each with a one-way completely randomized design involving three replicates of all treatments. Least significant difference (LSD) was used to determine variation among treatment groups. Experiment I focused in the effects of cycling, serum starved and fully confluent stages of Siberian tiger cells on different cell cycles. In Experiment II, the effects of different antioxidants like beta-Mercaptoethanol (beta-ME, 10 microM), cysteine (2 mM), and glutathione (2 mM) were examined after cells were fully confluent without serum starvation for 4 hr. In Experiment III, three PI, namely 6-dimethylaminopurine (6-DMAP, 2 mM), cycloheximide (7.5 microg/ml) and cytochalasin B (7.5 microg/ml) were used in the sane manner as in Experiment II. In Experiment IV, different levels of DMSO at 0%, 0.5%, 1.0%, and 2.5% were tested on different cell cycle stages of Siberian tiger examined by Flowcytometry (FACS). In Experiment I, 67.2% of the Siberian tiger skin fibroblasts reached the G0/G1 stage (2C DNA content) in fully confluent conditions which was more than the cycling (49.8%) and serum starved (SS) medium (65.5%; P < 0.05). Among the chemically treated group, glutathione (72.6%) and cycloheximide (71.3%) had little bit better results for the synchronization of G0 + G1 phases than serum starved and fully confluent. After nuclear transfer we did not see any significant differences on the development of tiger-porcine reconstructed embryos at cycling, SS and fully confluent. Data indicate that prolonged culture of cells in the absence of serum as well as using different chemicals for this experiment does not imply a shift in the percentage of cells that enter G0/G1 and that confluency is sufficient to induce quiescence. This finding can be beneficial in nuclear transfer programs in Siberian tiger, because there are negative effects, such as apoptosis associated with serum starvation.

Animals↗