India’s wealth of biodiversity is increasingly under threat with the rampant destruction of forests. Without doubt, the consequences of this are far reaching. If governments were to choose between humans and biodiversity, it would but obviously be the former. We are aware that with rising population pressures, the destruction of forests cannot be stopped in totality, but it can most certainly be slowed down.
Destruction of forest area has resulted in fragmentation of wildlife habitats. Survival of primary predator species such as the lions, tigers and leopards is doubly threatened as the animals not only need a diverse but a large habitat for sustaining a viable population.
At the crux of dwindling wildlife population are two issues. One, the poaching of animals by traders for economic exploitation and the other – inbreeding leading to a reduction in genetic diversity and possible future extinction. Nationally both are of equal concern; however, at present the focus is on poaching. Virtually no steps have been taken to address the latter. It is high time for the country to engage in an effective and innovative action plan.
Considering the importance of genetic diversity, a project to make use of biotechnological intervention for the conservation of endangered animals (lions, tigers etc.) was conceived. A proposal was submitted in 1998, to Department of Biotechnology (DBT), Government of India and the Central Zoo Authority of India (CZA), New Delhi towards the said project.
Today this facility is fully functional. For the genetic programme, the genetic diversity of lions, tigers and other endangered animals needs to be determined and those genotypes selected which show a high degree of diversity. Indian zoological parks can play a pivotal role as laboratories for scientific breeding of such species. Animals with breeding problems could be assisted by artificial insemination or IVF – a technique which is equivalent to the test-tube babies in humans.
The cloning technology would be established to the extent that it would be used only for those rare species where the number of surviving animals is extremely small. Such scientifically bred animals could be nurtured in a twilight zone with minimum human intervention in order to maintain their wild characteristics. They could be released in a forest only when the number of animals falls below a critical level.
Sperm, egg and cell banks would help in producing specific animals if and when the necessity arises. For example, the Indian cheetah is now extinct in the country. Even if a single animal could have been found or had cheetah cell lines been available it would have been possible to resurrect the species. Unfortunately, in India we have lost the cheetah forever. However, a few Indian cheetahs are believed to be surviving in Iran. CZA has already procured a few cheetahs and these animals could be the source for cloning.
The LaCONES project is indeed unique in its conception, organisation, involvement and participation. It is perhaps for the first time that a number of organisations, Council of Scientific and Industrial Research (CSIR), Centre for Cellular and Molecular Biology (CCMB), DBT, CZA and the Government of Andhra Pradesh, have come together to jointly undertake a programme of this magnitude. The scope of the present project may be widened even further in terms of the spectrum of technologies, which are expected to be developed during the course of work.
LaCONES has developed species-specific microsatellite markers for lion, tiger and leopard and established that these animals are genetically polymorphic and not inbred. The Project has also put in place non invasive methods for DNA isolation from scat and hair samples and developed a universal primer for forensic use to identify animal species. It has identified the Himalayan wolf as a new species – Canis himalayensis and demonstrated that the Ganges River dolphin is probably a ‘living fossil’. Moreover, LaCONES Project has established that the Olive Ridleys are ancestors of all the other known ridleys.
Interestingly the Project has achieved pregnancy in the black buck, cheetal deer and blue rock pigeon by assisted reproduction. A molecular DNA based method has been developed for sexing of birds and for the first time in the world, the Laboratory standardised a method for the collection of semen from the white backed vulture. The Project has also fascinatingly demonstrated that decline of the Indian white backed vulture in areas free of diclofenac could be due to malaria.
Ongoing research activities
LaCONES is at present engaged in a variety of new studies and experiments. Foremost among them are the monitoring of genetic variation by microsatellite and mitochondrial gene analysis in endangered animals of India and estimation of the number of wild tigers in reserves by DNA profiling of faecal samples. The lions in the Gir forest are also being estimated using the faecal samples. As part of a global effort to DNA barcode all life forms LaCONES is barcoding the birds of India. DNA based diagnosis of common viral and bacterial diseases of wild animals is also in progress. Non invasive methods of sex steroid analysis using faecal samples of the big cats and ungulates is being carried out to establish the fertility status and pregnancy.
Harmony and balance in nature between the biotic and abiotic compartments is best achieved by habitat protection, which in turn supports the flora and fauna. Man’s selfishness due to urbanisation, industrialisation and poaching nullifies the benefits of habitat protection. LaCONES is a unique attempt to protect the endangered animals of India using a combination of genetics, assisted reproductive technologies and cryobanking.
Objectives of the LaCONES
Monitoring of genetic variation: Modern techniques such as DNA fingerprinting may be used to monitor genetic variation
Establishment of gene banks: Cryo-preservation of semen, eggs and embryos of endangered species may be undertaken.
Semen analysis: Semen quality may be studied for selecting animals for breeding.
Determination of the time of ovulation: Successful intra-uterine insemination may be achieved once the time is determined.
Artificial insemination: Standardisation of the technique for use in wild animals is needed – it is already well established for domestic animals.
In vitro fertilization (IVF) and embryo transfer: The process involves fusion of spermatozoon with oocyte carried out in vitro and the transfer of the resulting embryo to the true or surrogate mother.
Establishment of cell bank: The cells preserved under appropriate conditions to be used in future for various purposes including cloning.
Cloning: The technique may be developed only for those species which are extremely rare.