The study on social impact of mini zoos and deer parks in the three regions of the country namely, North, south and east were completed by A.E.D.R., JNU New Delhi; Indian Institute of Management, Bangalore, and Xavier Institute of Management, Bhubaneswar. The study revealed that :

2.    Molecular Characterization of Wild Animals by DNA Fingerprinting for their better Management in Indian Zoological Gardens.

The Asiatic lions and Indian tigers are not as inbred as previously reported by S.J. O' Brien and group from USA and do not suffer from inbreeding depression. The Asiatic lions exhibit a moderate genetic variability of about 26%. The Indian tigers also exhibit similar levels of genetic variation. It is our hypothesis that low levels of genetic variability is a characteristic of these species. The immune locus data reveals that the Asiatic lions reveal abudant immune diversity. Analysis of reproductive parameters by Dr. Shivaji from C.C.M.B. also does not reveal any significant differences in the incidence of sperm abnormalities and circulating testosterone levels and compared to other wild animals. These data collated provided evidence that Asiatic lions and Indian tigers do not suffer from inbreeding depression. The analysis of 50 to 125 years old tiger skin does not reveal any significant differences as compared to the modern population. This also supports our hypothesis that low genetic variability is not due to any recent bottlenecks but has been an inherent feature of these species.

The graphical representation shows that Asiatic lions Madhuri (1256), Chandra (1188), Hemlatha (1237) and Hadara from the Sakkarbaug zoo possess high amounts of genetic variation. These animals can especially be used for further breeding to improve the genetic diversity of the Asiatic lion population.

Using micro satellite and mitochondrial sequence analysis, we can confirm that the two Asiatic lions in the Bhubneswar zoo are pure Asiatic lions. All lions from the Sakkarbaug zoo are also pure Asiatic lions as designated by the International studbook. The lions in all other zoos were found to be hybrids.

The micro satellite analysis at loci Fca 77 and Fca 126 is able to differentiate pure Asiatic lions from hybrid lions. The Asiatic lions do not exhibit any polymorphism at these loci, while hybrids exhibits extensive variability. This variability could be due to the contribution of African alleles into the population. The mitochondrial D loop sequence analysis also confirms this point. Asiatic lions do not exhibit any variation, while the hybrids show extensive variability

The Indian tigers possess 29% genetic variation. This level of variability is comparable to any other free ranging carnivore. Using micro satellites, detection of hybrids between Siberian and Indian tigers was also achieved.

A rapid and simplified method has been developed to extract DNA for molecular analysis from fecal samples. Fecal samples are ideal for molecular analysis because they are non-invasive and complications with tranquilizing animals for blood collection are avoided. Using this technique a large scale study of wild animals is now possible. Routine monitoring of populations for genetic variation will also become significantly easier.

The Project on Cryopreservation of spermatozoa in wild animals is the first of its kind in India and it has yielded promising results that can be used in efficient management of captive animals, particularly the fields. The significant achievements of the project are as follows:

Protocols have been established for anaesthetics and electro-ejaculating a wide range of animals such as lions, tigers, leopards, hyenas, jackals, bears, dears and monkeys.

Repeated anesthesia or electro-ejaculation did not cause harm to the animals.

TALP and HF-10 media were found to be the most suitable for washing and processing of sperm for motility analysis and fertilising ability studies.

TEST-yolk buffer was most suitable for cryopreserving the sperm at the site of collection.

Zone free hamster oocytes could be used to test the penetration ability of the sperms both neat and freeze-thawed) of the wild animals.

CASA could be utilised for fast and accurate diagnosis of the quality of the spermatozoa of the wild animals.

Finding based on spermatology and hormonal analysis, indicate that inbreeding depretion vis-a-vis usefulness of the animals for captive breeding is not a matter of serious concern among the captive tigers, lions and leopards. These findings are in agreement with the study carried out by Dr. Singh and his colleagues on the genetic heterogeneity of the mega cates.

In an effort to understand the seroprevalance of viral infections and their impact on the Asiatic lions, a seroepizootiological study of cannine distemper virus(CDV), feline pervo virus(FPV), feline immunodeficiency virus(FIV) and feline leukaemia virus(FeLV) was done in asiatic lions, hybrid lions and sympatric leopard and domestic carnivores in zoological Park in Ahmedabad, Baroda, Junagarh< Rajkot and the Gir National Park and Sanctuary and Devalia, Safari park. fifty Lions, including 13 hybrid lions, 24 leopards, 30 domestic dogs were tested by agar gel immunodiffusion tests and dot- immunobinding assays for the serological evidence of above viruses. Antibodies to CDV were detected in 94.59%(35/37) of Asiatic lions. 76.92%(10/13) of the hybrid lions. 91.66% (22/24) of the leopards. 70% (21/30) of the domestic cats and 66.83%(131/196) of the domestic dogs. High seropositivity to CDV without overt symptoms of clinical disease suggested the possibility of field to field transmission. 100% seroprevalence to FPV antibodies was observed in both the domestic and exotic fields. Absence of clinical feline panleukopedia-like symptoms, suggested that FPV appeared to manifest itself as an in apparent infection in these domestic and non-domestic fields. 80% of lions. 62.5% of leopards and 80% of domestic cats sampled had high FPV antibody liters more than 1:160, suggesting repeated infection with an endemic parvo like virus. There were no detectable levels of antibodies to FIV or FeLV antigens in the lions, Leopards and domestic cats. The study found in captive non-domestic felids, a high prevalence to CDV and FPV, two viruses , known to have caused large scale mortalities in captive and free-living non-domestic felids worldwide. It is suggested that movement, translocation or re-introduction of these seropositive felids may be associated with disease risks and hence movement and translocation of these felids must be done after subjecting theme to standard quarantine and disease screening protocols. Vaccination may be considered using killed or other suitable viral vaccines, which have been proved to be safe, effective and efficacious in endangered felids.

Although our country is endowed with a tremendous wealth of mega diversity in plant and animals wildlife, their very existence is at stake owing to destruction of forest following industrialization, agriculture activities as well as poaching of wildlife for reasons of sport and money. The problem becomes confounded with the population growth as more forest area is brought under cultivation. Thus, the dwindling of forests and destruction of habitats has led to fragmentation of wildlife and inbreeding of animals. This results in reduction in their biodiversity, leading to sterility and extinction. It has been reported that approximately one hundred species in the world become extinct every day. Extinction threatens 11% of birds, 25% of mammals and 34% of fish species. Given current trends, many rare or endangered vertebrate species will soon be lost despite efforts to maintain biodiversity via habitat and wildlife conservation. There is, therefore, an urgent need to arrest this phenomenon and work out strategies to tackle these problems by using innovative techniques.

The Centre for Cellular and Molecular Biology (CCMB), Hyderabad is setting up a laboratory for Conservation of Endangered Species (La-CONES) at Attapur near Nehru Zoological park, Hyderabad, In partnership with the department of Environment and Forests, A.P. and the Nehru Zoological Park. Piloted by Dr. Lalji Singh, Director and Dr. S. Shivaji, Dy. Director, CCMB, this project has culminated as a result of the initiative taken by the department of Biotechnology of the Ministry of Science and Technology and the Central Zoo Authority of India of the Ministry of Environment and Forests, Govt. of India, recognizing the need for preservation of genetic diversity and thus conservation of wildlife in the country. The new laboratory (LaCONES) being set up has been designed as a national programme to tackle this issue by biotechnological intervention for the conservation as well as propagation of endangered species.

This unique state-of the art laboratory is being set up at an estimated total cost of Rs 94.8 million (the estimated cost for the for the first phase being Rs. 57.5 million). The Government of AP has allotted five acres of land at Attapur. LaCONES with the technical know-how and expertise would trouble-shoot events related to the biology of the conservation of endangered species. Its objective include:

Monitoring of genetic variation by modern techniques, such as DNA fingerprinting, So as to establish the degree of genetic variation in the species and to establish identify family lineage which would contribute positively towards planned in-house breeding strategies and probably wildlife parks.

Establishment of gene banks by Cryopreservation of semen, eggs and embryos of endangered species for use in future in under the conditions of inbreeding partners behaviourally incompatible.

Analysis of semen source to establish a database to identify fertile males and select the best for breeding.

Monitoring and establishment of the ovulatory cycle of females to facilitate artificial insemination.

Development of assisted reproduction technologies such as intrauterine insemination, in Vitro Fertilization, intra-cytoplasm sperm injection, embryo transfer, etc. So as to facilitate propagation of species under circumstances which do not support normal mating and conception.

Establishment of cell banks of endangered species to be used for cloning under conditions when all other conventional methods of assisted reproductive technologies fail.

The resurrection of extinct and several endangered species by cloning the frozen genetic material of the extinct species and oocytes of a closely related species to be used as a surrogate mother.

The CCMB scientists have also standardized many protocols for assisted reproduction such as semen collection by electro ejaculation, semen cryopreservation, semen profile evaluation in vitro fertilization and radioimmunoassay of steroids, and hope to extend their expertise to achieve pregnancy by intrauterine insemination and embryo transfer, so as to overcome difficulties owing to failure of normal reproductive performance in wild animals. The scientists have already developed a semen cryobank of tigers, lions and leopards and hope to create egg, embryo and cell banks for future use. A Mobile Van with appropriate facilities including Cryopreservation facility, sonography, computerized sperm motility analyzer. etc. has already been procured to enable the scientists reach the habitats for collecting samples and to attend on the animals for immidiate aid when necessary.