Feature Article MUKESH THAKUR & RUHINA JAVED Frozen Zoos! DespiteDespite therethere beingbeing numerousnumerous genegene repositoriesrepositories inin thethe country,country, therethere isis nono initiativeinitiative asas yetyet toto developdevelop aa nationalnational genegene bankbank exclusivelyexclusively forfor thethe preservationpreservation andand utilizationutilization ofof wildwild animalanimal germplasm.germplasm. GenomeGenome ResourceResource BankBank (GRB)(GRB) oror FrozenFrozen ZooZoo wouldwould provideprovide significantsignificant benefitsbenefits forfor wildlifewildlife managersmanagers asas itit holdsholds particularparticular promisepromise forfor endangeredendangered species.species. Artificial insemination (AI) is the process by which sperm is placed into the reproductive tract of a female for the purpose of impregnating the female by using means other than sexual intercourse. It has been practiced in the livestock industry for many years. It may be similarly practiced in T’S not a zoo that you would particularly enjoy visiting. wildlife management. Semen can be collected at one site Unlike the conventional zoo where antics of animals in and shipped anywhere in the world. captivity keep you entertained, in a Frozen Zoo you Embryo Transfer (ET) is another captive breeding tool are more likely to come across tanks containing semen currently used. It requires two genetically related species. Iand embryos of wild animals frozen in liquid nitrogen. One species is super-ovulated, inseminated, and the Sometime in the future, when an animal in the wild is fertilized eggs are collected. The fertilized eggs are implanted threatened with extinction, it could be reproduced thanks into surrogate mothers. The offspring carry the genetic to the gene material safely stored in the Frozen Zoo. makeup of the two original parents, not the surrogates. Wildlife reproduction is a new and upcoming trend in This science provides many opportunities to captive conservation of wildlife. Practiced widely in several breeding programmes. Animals no longer have to be countries, it is not yet frequently applied in India. There are moved to accommodate natural mating. Conservation several new methods available today in wildlife biologists can work to counter the effects of genetic drift. reproduction including Cross-fostering, Artificial Even if an animal dies, gametes stay viable up to 24 hours, incubation, Artificial insemination, and Embryo transfer. allowing the possibility to re-introduce diversity back into All these methods go a long way in boosting genetic wild populations. variability, increasing population size, and improving reproductive success. Genome Resource Bank (GRB) or Frozen Cross fostering is a technique of inselective breeding, where Zoo would provide significant benefits for offspring are removed from their biological parents at birth and raised by surrogates. This can also occasionally occur wildlife managers. It has particular promise in nature. The surrogate can be a close relative of the species and does not have to be the same species. for endangered species. 44 SCIENCE REPORTER, May 2010 Feature Article There are species that desperately need help from captive situations, as we can’t save them in the wild. But captive breeding is not always easy. Many animals are disoriented by living in the wrong climate or social system, or being squeezed into small “habitats” in the presence of humans. Scientists and keepers try to help animals breed naturally. But often, researchers do not even know how normal mating takes place. Therefore, AI and IVF might be helpful in such cases. Actually, Gene Bank is not a new concept for India. These have been earlier only used for domestic animals, cultivated agriculture crops and for microbes. Some of the prominent examples are: National Gene Bank Facility for conservation and preservation of domestic animal germplasm at the National Frozen Gene Bank Bureau of Animal Genetic Resources, Karnal, Haryana with The Frozen Zoo is a collection of animal genes in the form the responsibility of evaluating, certifying and conserving of frozen semen and embryos. In practical terms this is a the rich and varied germplasm resources available in the collection of sperm-holding straws stored in liquid nitrogen country and whose genetic base is shrinking fast. Thus, the tanks. The first so-called “frozen zoo” was created in 1975 establishment of the National Bureau of Animal Genetic by Kurt Benirschke, a visionary physician who switched Resources/National Institute of Animal Genetics in 1984 from his human practice to work with endangered species was the culmination of sustained efforts made by leading at the San Diego Zoo, studying gene pools and genetic geneticists in the country over the years. diseases made prevalent by inbreeding. National Genomic Resources Repository was established Modern technology allows the indefinite preservation in the premises of the National Bureau of Plant Genetic of genetic material through the freezing of semen and Resources, New Delhi for exchange, quarantine, collection, embryos (unfortunately, ova do not freeze very well). The conservation, evaluation and the systematic documentation implication of being able to freeze semen and embryos is of plant genetic resources. NBPGR act as an institutional that the species can be kept forever, even when they are framework for methodical and centralized efforts to collect, extinct, as their genes would be preserved. When the threat generate, conserve and distribute genomic resources for to the preserved species has been controlled, implantation agricultural research. and development of an embryo can then be carried out. On the research front it allows much to be learned about species-specific reproductive physiology and will help to 1. A researcher at the Cincinnati Zoo removes blood from a biopsy of a develop the technology required for related species not yet domestic cat’s follicles, the egg-nurturing structures in the ovary. endangered. Thus when the time comes to make 2. After the blood is extracted, the eggs in the sample are easier to conservation efforts for one of these species, the technology detect. would already be available. 3. A researcher opens up one of the chilled canisters containing sperm Today, in zoos around the world, these deep-freeze samples and frozen embryos. tanks act as a high-tech Ark, housing cells from the planet’s most endangered animals, including chimps, cheetahs, 4. Samples stored in liquid nitrogen cooled to -374 degrees Fahrenheit pandas, California condors and hundreds more. As a result, remain viable indefinitely. They are often used successfully after two long-dead animals are now becoming parents from the to five years in storage. grave, with their precious DNA still circulating in the gene pool. The real goal is the management of the eroding gene pools of endangered species. left: This ocelot kitten was conceived in the lab using sperm and a stored egg SCIENCE REPORTER, May 2010 45 Feature Article The San Diego Zoo where the first frozen zoo was created (above), Animals at LaCones (above right); and Samples preserved in a Frozen Zoo (below). Microbial Type Culture Collection and Gene Bank (MTCC), a national facility, was established in 1986, funded Only good jointly by the Department of Biotechnology (DBT) and the quality Council of Scientific and Industrial Research (CSIR), Government of India. The MTCC is a modern facility sperm housed at the Institute of Microbial Technology (IMTECH), Chandigarh. The main objectives of this national facility samples are to act as a depository, to supply authentic microbial cultures and to provide related services to scientists fulfilling all working in research institutions, universities and the industries. Presently, the MTCC has five sections, the Actinomycetes, Bacteria, Fungi, Yeasts and Plasmids essential collectively holding over nine thousand cultures. The MTCC scientists are actively involved in research parameters programmes relating to microbial diversity, ecology and taxonomy using both classical and molecular approaches. are banked. Laboratory for the Conservation of Endangered Species (LaCONES) at CCMB, Hyderabad aims to conserve endangered animals of India by using modern techniques Frozen Zoo—Great Opportunity of DNA fingerprinting, genetic polymorphism analysis and Genes from wild-caught endangered animals are valuable, assisted reproductive techniques such as artificial as they are needed to increase the founder population of insemination, in vitro fertilization, cryo-banking of DNA, breeding groups. Domestic cattle are constantly being cells and tissues and eventually somatic cell cloning. improved genetically by means of artificial breeding. The Scientists at Centre for Cellular and Molecular Biology advantage of speeding up the genetic progress in a breeding (CCMB) have been successful in the first artificial programme can similarly be used for exotic species. The insemination of a spotted deer at the Nehru Zoological Park frozen zoo could provide the valuable material to produce in Hyderabad. genetically superior stock so that natural breeding in one A group of scientists at LaCONES under the leadership herd or colony could be reproduced. Furthermore, the of Dr. S. Shivaji, has also been working towards developing production of genetically healthier animals will assisted reproductive technologies to successfully induce automatically reduce inbreeding problems caused by loss pregnancy in endangered animals by artificial