Biology Reference
In-Depth Information
and threatened species in the United States are regulated by the Fish and Wildlife
Service of the Department of Interior and permits must be obtained to collect,
possess, or transport any species on the List of Endangered and Threatened
Wildlife. Furthermore, the Convention on International Trade in Endangered
Species of Wild Fauna and Flora (CITES) requires collecting permits and restricts
collection or importation of any species on the international list. For example,
sampling DNA from the hind wing of the endangered butterfly
Neonympha
mitchellii mitchellii
was permitted only after
Hamm et al. (2010)
showed that
they could remove small amounts of hind wing (2-3 mm
2
) from two other butter-
fly species without having a “significant impact on the behavior or survival.”
Most countries limit importation of live organisms that could be harm-
ful to crops, livestock or humans. In the United States, for example, the Plant
Protection and Quarantine (PPQ) branch of the U.S. Department of Agriculture,
Animal and Plant Health Inspection Service (USDA-APHIS) must be contacted
before importing and transporting live plants, arthropod natural enemies,
arthropod plant pests, plant pathogens, and vectors of plant or animal disease.
The Office of Biosafety of the Centers for Disease Control regulates the impor-
tation of agents of human disease or vectors that could harbor these agents.
Shipping live insects by mail or by shipping services also is restricted; many will
not accept live insects. Shipment of insects in alcohol or in dry ice also is limited
due to concerns about airline safety.
13.5 Molecular Ecological Methods
Proteins, nuclear DNA, mitochondrial DNA, and DNA from symbionts such as
Wolbachia
can be analyzed to resolve ecological problems; each has advantages
and disadvantages. The ease of study, amount of variation that can be detected,
and cost differs with each target. Analysis of genetic variation at the individual,
population or species level can be achieved with both single- and multiple-copy
DNA sequences. Mitochondrial (mt) DNA analysis provides sufficient variation
for studies of individuals, populations, or species, depending upon the region of
the mitochondrion analyzed. Because the genomes of many arthropod species
have been, or soon will be, completely sequenced, many more genes soon will
become available for analysis in ecological studies.
Potential techniques include allozyme electrophoresis, restriction-fragment-
length polymorphisms (RFLPs) of mtDNA or nuclear DNA, DNA fingerprinting
by analysis of microsatellites, RAPD-PCR, heteroduplex analysis (HDA), amplified-
fragment-length polymorphisms (AFLP-PCR), sequencing of both nuclear and
mtDNA, allele-specific PCR by using standard or high-fidelity PCR protocols or
quantitative PCR, and microarray analysis. These methods were described in
