Selina Patel

Conserved primers for DNA barcoding historical and modern samples from New Zealand and Antarctic birds

Our ability to DNA barcode the birds of the world is based on the effective amplification and sequencing of a 648 base pair (bp) region of the mitochondrial cytochrome c oxidase (COI or cox1) gene. For many geographic regions the large numbers of vouchered specimens necessary for the construction of a DNA barcoding database have already been collected and are available in museums and other institutions. However, many of these specimens are old (>20 years) and are stored as either fixed study skins or dried skeletons. DNA extracted from such historical samples is typically degraded and, generally, only short DNA fragments can be recovered from such specimens making the recovery of the barcoding region as a single fragment difficult. We report two sets of conserved primers that allow the amplification of the entire DNA barcoding region in either three or five overlapping fragments. These primer sets allow the recovery of DNA barcodes from valuable historical specimens that in many cases are unique in that they are unable or unlikely to be collected again. We also report three new primers that in combination allow the effective amplification from modern samples of the entire DNA barcoding region as a single DNA fragment for 17 orders of Southern Hemisphere birds.

The world’s rarest whale

Summary The vast expanses of the South Pacific Ocean have, until recently, concealed the identity of the worlds rarest whale, the spade-toothed beaked whale ( Mesoplodon traversii ). Based on the scarcity of records and the total absence of previous sightings, this species is the least known species of whale and one of the worlds rarest living mammals. Two individuals of this species, previously known from only two skull fragments and a mandible, were recently discovered beach-cast in New Zealand. Although initially misidentified, we have used DNA analysis to reveal their true identity. We provide the first morphological description and images of this enigmatic species. This study highlights the importance of DNA typing and reference collections for the identification of rare species.

Bucking the trend: genetic analysis reveals high diversity, large population size and low differentiation in a deep ocean cetacean

Understanding the genetic structure of a population is essential to its conservation and management. We report the level of genetic diversity and determine the population structure of a cryptic deep ocean cetacean, the Gray’s beaked whale (Mesoplodon grayi). We analysed 530 bp of mitochondrial control region and 12 microsatellite loci from 94 individuals stranded around New Zealand and Australia. The samples cover a large area of the species distribution (~6000 km) and were collected over a 22-year period. We show high genetic diversity (h=0.933–0.987, π=0.763–0.996% and Rs=4.22–4.37, He=0.624–0.675), and, in contrast to other cetaceans, we found a complete lack of genetic structure in both maternally and biparentally inherited markers. The oceanic habitats around New Zealand are diverse with extremely deep waters, seamounts and submarine canyons that are suitable for Gray’s beaked whales and their prey. We propose that the abundance of this rich habitat has promoted genetic homogeneity in this species. Furthermore, it has been suggested that the lack of beaked whale sightings is the result of their low abundance, but this is in contrast to our estimates of female effective population size based on mitochondrial data. In conclusion, the high diversity and lack of genetic structure can be explained by a historically large population size, in combination with no known exploitation, few apparent behavioural barriers and abundant habitat.

High coverage of the complete mitochondrial genome of the rare Gray’s beaked whale (Mesoplodon grayi) using Illumina next generation sequencing

Abstract Using an Illumina platform, we shot-gun sequenced the complete mitochondrial genome of Gray’s beaked whale (Mesoplodon grayi) to an average coverage of 152X. We performed a de novo assembly using SOAPdenovo2 and determined the total mitogenome length to be 16,347 bp. The nucleotide composition was asymmetric (33.3% A, 24.6% C, 12.6% G, 29.5% T) with an overall GC content of 37.2%. The gene organization was similar to that of other cetaceans with 13 protein-coding genes, 2 rRNAs (12S and 16S), 22 predicted tRNAs and 1 control region or D-loop. We found no evidence of heteroplasmy or nuclear copies of mitochondrial DNA in this individual. Beaked whales within the genus Mesoplodon are rarely seen at sea and their basic biology is poorly understood. These data will contribute to resolving the phylogeography and population ecology of this speciose group.

Genetic structure of the grey side-gilled sea slug (Pleurobranchaea maculata) in coastal waters of New Zealand

Pleurobranchaea maculata is a rarely studied species of the Heterobranchia found throughout the south and western Pacific–and recently recorded in Argentina–whose population genetic structure is unknown. Interest in the species was sparked in New Zealand following a series of dog deaths caused by ingestions of slugs containing high levels of the neurotoxin tetrodotoxin. Here we describe the genetic structure and demographic history of P. maculata populations from five principle locations in New Zealand based on extensive analyses of 12 microsatellite loci and the COI and CytB regions of mitochondrial DNA (mtDNA). Microsatellite data showed significant differentiation between northern and southern populations with population structure being associated with previously described regional variations in tetrodotoxin concentrations. However, mtDNA sequence data did not support such structure, revealing a star-shaped haplotype network with estimates of expansion time suggesting a population expansion in the Pleistocene era. Inclusion of publicly available mtDNA sequence sea slugs from Argentina did not alter the star-shaped network. We interpret our data as indicative of a single founding population that fragmented following geographical changes that brought about the present day north-south divide in New Zealand waters. Lack of evidence of cryptic species supports data indicating that differences in toxicity of individuals among regions are a consequence of differences in diet.

Post-mortem examinations of New Zealand birds. 2. Long-tailed cuckoos (Eudynamys taitensis, Aves: Cuculinae)*

ABSTRACT Little is known about the biology of long-tailed cuckoos (Eudynamys taitensis). In this study, 79 dead cuckoos, mostly from the wider Auckland region, New Zealand, were examined and dissected to shed light on the sex ratio, morphology, breeding season and diet. There were no statistically significant sexual differences in the means for weight or body measurements. However, immature birds had significantly shorter bill, wing and tail lengths than adults. In adults, gonads were enlarged (for breeding) from October to January, while all cuckoos identified (from plumage) as immatures had small gonads. Of 888 food items identified from 62 gizzards, 94% were insects. The main foods were cicadas and shield-bugs (Hemiptera, 48% of food items), stick-insects (Phasmatodea, 19%) and wētās and katydids (Orthoptera, 13%). Small vertebrates (lizards and birds’ eggs and nestlings) were a minor dietary element (1% of food items; 13% of stomachs). Cicadas, stick-insects and praying mantids (large insects abundant in late summer) made up 57% of the immature diet; in adults these three categories made up just 13% of food items. Immatures leave New Zealand on migration several months after adults, and this delay in late summer and autumn coincides with an opportunity to exploit a seasonal abundance of large insects.

Complete mitochondrial genome of the green-lipped mussel, Perna canaliculus (Mollusca: Mytiloidea), from long nanopore sequencing reads

Abstract We describe here the first complete genome assembly of the New Zealand green-lipped mussel, Perna canaliculus, mitochondrion. The assembly was performed de novo from a mix of long nanopore sequencing reads and short sequencing reads. The genome is 16,005 bp long. Comparison to other Mytiloidea mitochondrial genomes indicates important gene rearrangements in this family.

Genetic Kinship Analyses Reveal That Gray’s Beaked Whales Strand in Unrelated Groups

Some marine mammals are so rarely seen that their life history and social structure remain a mystery. Around New Zealand, Grays beaked whales (Mesoplodon grayi) are almost never seen alive, yet they are a commonly stranded species. Grays are unique among the beaked whales in that they frequently strand in groups, providing an opportunity to investigate their social organization. We examined group composition and genetic kinship in 113 Grays beaked whales with samples collected over a 20-year period. Fifty-six individuals stranded in 19 groups (2 or more individuals), and 57 whales stranded individually. Mitochondrial control region haplotypes and microsatellite genotypes (16 loci) were obtained for 103 whales. We estimated pairwise relatedness between all pairs of individuals and average relatedness within, and between, groups. We identified 6 mother-calf pairs and 2 half-siblings, including 2 whales in different strandings 17 years and 1500 km apart. Surprisingly, none of the adults stranding together were related suggesting that groups are not formed through the retention of kin. These data suggest that both sexes may disperse from their mothers, and groups consisting of unrelated subadults are common. We also found no instances of paternity within the groups. Our results provide the first insights into dispersal, social organization, and the mating system in this rarely sighted species. Why whales strand is still unknown but, in Grays beaked whales, the dead can tell us much about the living.