July 2000 - June 2002
Claudio Ciofi, Ph.D.

Research Progress Report

Gaylord Donnelley Environmental Fellowship
January 2001

Research in Molecular Ecology

Submitted by: Dr. Claudio Ciofi 
Yale Faculty Sponsors:  Dr. Gisella Caccone and Professor Jeffrey Powell, ECOSAVE Conservation Genetics Laboratory

Dr. Claudio Ciofi arrived at Yale on July 1, 2000 as a Gaylord Donnelley Environmental Fellow.  Dr. Ciofi received his Ph.D. in Conservation Biology from the University of Kent in England in 1998.  His dissertation was on the spectacular Komodo dragon and is one of the finest studies done on the Komodo dragon in the last 20 years.  He conducted arduous fieldwork in an effort to understand these lizards, including tagging them and determining range size.  He also collected blood from about 150 animals and conducted microsatellite DNA studies.  Finally, he analyzed the genetic data using sophisticated techniques that allowed him, among other things, to draw conclusions about the relative isolation among the remaining populations.  Thus he has demonstrated all the skills required in modern conservation biology: fieldwork, ecological studies, laboratory genetic studies, and sophisticated computer-based analysis.  In addition, he has been working with and training local Indonesians, not only to give them skills, but also to raise their appreciation for these endangered animals with the hopes of affecting public policy. 

After his arrival at Yale University as a Donnelley Post-Doctoral Fellow, he has been involved in a number of very productive research and educational activities, both at Yale and abroad, all part of the two main wildlife conservation projects that he has been involved in.  The giant Galápagos tortoise and the Komodo dragon projects are well-established studies where molecular genetic techniques are employed to better define the distinctiveness of natural populations, to obtain information on demographic parameters, and to provide recommendations on the amount of effort that should be devoted to conservation.  This is a fast evolving discipline, which is being integrated with ecological and demographic studies, to formulate appropriate management strategies for endangered species.

In the first three months of his appointment, he has participated in scientific expeditions in Ecuador and Indonesia to collect samples for genetic analysis from giant tortoises and Komodo dragons respectively.  The Ecuadorian expedition was carried out in the Galápagos archipelago, about 1,500 km off the coast of Ecuador in the Pacific Ocean.  The fieldwork was conducted on Isabela, the largest and geologically youngest of a set of 15 volcanic islands.  The study, organized at Yale, included four researchers of different nationalities.  Its purpose was to collect blood samples of giant tortoises inhabiting the slopes of two volcanoes on the north part of the island.  The group identified, measured, sexed, and sampled 93 tortoises of up to 200 kg in weight.  Samples were collected from the brachial vein located on one of the forelimbs.  During the work, Dr. Ciofi had the opportunity of training two Ecuadorian undergraduate students in the collection and storage of reptile blood.

The Indonesian expedition was conducted at Komodo National Park in the southeast part of the country.  The goal of the study was to collect blood samples from adult females once the location of their nests was identified.  Subsequent collection of samples from the hatchlings will allow a DNA fingerprinting analysis to assess single or multiple paternity.  This will help to clarify the mating system of the species.  Dr. Ciofi collected blood samples from five females.   As for his previous studies on the Komodo dragon, specimens were caught in baited traps (300x50x50 cm).  Young females were first restrained, and 500 micro liters of blood were collected from the caudal vein using a 5 ml syringe and a spinal needle (0.70x90 mm), while large specimens were kept in the trap and bled by nail-clipping.

Whole genomic DNA was extracted from blood samples soon after fieldwork.  Genetic analysis is now conducted by assessing allelic differences among individuals at nine microsatellite loci.  Microsatellites are short, tandemly repeated simple sequences of nuclear DNA.  Genetic differences between individuals are determined by identifying variation in the number of repeats.   Microsatellite loci are first amplified by polymerase chain reaction (a technique to obtain sizable specific DNA sequences from minute amounts of whole DNA), and individual genotypes are then obtained.  Differences in allele sizes between individuals as little as one DNA base pair can be detected, allowing the maximum degree of resolution.  Yale undergraduate students are currently working on the above techniques under Dr. Ciofi’s supervision.  During their training period, they will become proficient in DNA extraction and analysis methods and will gain knowledge of how the data obtained can be used to design and implement wildlife management plans.

The Komodo dragon study is part of a wider project Dr. Ciofi set up during his Ph.D. work.   At Yale, he has had the opportunity to continue and expand the work he initiated using genetics to obtain quantitative information on population divergence, reproductive biology, and demography of the species.

In addition, he has submitted a number of applications for funding to US Institutions to support his ongoing research.  In January 2001, three grants have been awarded by private Zoological foundations, and additional support has been confirmed by the Smithsonian Institution, Washington D.C.  These funds will allow Dr. Ciofi to continue his studies in the laboratory and in the field, and will also afford him the opportunity to continue his collaboration with Indonesian universities and support undergraduate short-term field studies on the biology and management of the Komodo dragon.