Development of polymorphic microsatellite markers for the sorghum plant bug, Stenotus rubrovittatus (Heteroptera: Miridae)

Nine polymorphic microsatellite DNA markers from the sorghum plant bug, Stenotus rubrovittatus, were isolated and characterized. These markers were used to analyse 22 individuals from a single field population. The number of alleles at these nine loci ranged from two to 28 (mean = 11.4) and heterozygosity ranged from 0.27 to 0.86 (mean = 0.58). Stenotus rubrovittatus has shown rapid population growth in the decades since the first report in the 1980s of serious damage to a rice crop. These microsatellite markers will be of value for studying both the population genetics and population dynamics of S. rubrovittatus.     

Ten di- and trinucleotide microsatellite loci in the Caribbean spiny lobster, Panulirus argus, for studies of regional population connectivity

We describe 10 microsatellite loci for Panulirus argus (Caribbean spiny lobster). The number of alleles at each locus ranged from four to 39 (mean = 21.8) in 89 juvenile specimens collected at two different times at a recruitment site in south Florida. Levels of expected and observed heterozygosities ranged from 0.48 to 0.96 (mean = 0.83) and from 0.32 to 0.98 (mean = 0.71), respectively. Significant departures from Hardy–Weinberg equilibrium were observed at two loci. There was no evidence of genotypic disequilibrium for any pair of loci. Overall, the loci were well resolved, highly polymorphic and independently segregating, confirming their utility for population genetic studies.     

Development of 15 polymorphic microsatellite markers in the Atlantic tarpon (Megalops atlanticus) for capture-recapture studies

Fifteen polymorphic microsatellite DNA loci for Atlantic tarpon, Megalops atlanticus, were isolated by using PIMA, a polymerase chain reaction‐based technique. The number of alleles at each locus ranged from two to 24 (mean = 7.7) in 65 specimens from Tampa Bay, Florida. Observed and expected heterozygosities ranged from 0.27 to 0.92 (mean = 0.60) and from 0.28 to 0.95 (mean = 0.62), respectively. Genotypes at one locus deviated significantly from Hardy–Weinberg equilibrium. In exact tests for genotypic disequilibrium, there was no evidence of associations between any pair of loci. Overall, loci were well resolved and highly polymorphic, confirming their suitability for DNA fingerprinting applications and other genetic studies.     

PERMANENT GENETIC RESOURCES: Eighteen new polymorphic microsatellite markers for the endangered Florida manatee, Trichechus manatus latirostris

Here we describe 18 polymorphic microsatellite loci for Trichechus manatus latirostris (Florida manatee), isolated using a polymerase chain reaction‐based technique. The number of alleles at each locus ranged from two to four (mean = 2.5) in specimens from southwest (n = 58) and northeast (n = 58) Florida. Expected and observed heterozygosities ranged from 0.11 to 0.67 (mean = 0.35) and from 0.02 to 0.78 (mean = 0.34), respectively. Departures from Hardy–Weinberg equilibrium occurred at two loci. There was no evidence of genotypic disequilibrium for any pair of loci. For individual identification, mean random‐mating and θ‐corrected match probabilities were 9.36 × 10^?7 and 1.95 × 10^?6, respectively.     

New microsatellite loci for the endangered scalloped hammerhead shark, Sphyrna lewini

We isolated 15 microsatellite markers for the scalloped hammerhead shark, Sphyrna lewini. Loci were tested on 80 specimens of S. lewini from four Eastern Pacific samples. The number of alleles per locus ranged from 6 to 31 (mean = 14). Observed and expected levels of heterozygosity per locus ranged from 0.39 to 0.91 (mean = 0.70) and from 0.54 to 0.90 (mean = 0.76), respectively. No pairs of loci were in gametic disequilibrium after Bonferroni correction of α. One locus showed significantly lower heterozygosity than expected under Hardy–Weinberg proportions in two populations, possibly caused by null alleles.     

PERMANENT GENETIC RESOURCES: Twelve polymorphic microsatellite markers for the bonefish, Albula vulpes and two congeners

Twelve polymorphic microsatellite loci were isolated for the bonefish, Albula vulpes using a polymerase chain reaction‐based procedure. The number of alleles ranged from two to 23 (mean = 8.8) in 37 specimens from south Florida. Observed and expected heterozygosities ranged from 0.07 to 0.77 (mean = 0.42) and from 0.07 to 0.84 (mean = 0.48), respectively. There were no significant departures from Hardy–Weinberg equilibrium and no evidence of genotypic disequilibrium between any pair of loci. In a cross‐amplification test, all markers yielded appropriately sized alleles for specimens of the provisional Albula sp. B and 11 of the 12 loci amplified for those of Albula glossodonta.     

Isolation and characterization of microsatellite markers from the acacia-ant Crematogaster mimosae

We describe 10 microsatellite loci developed from Crematogaster mimosae, an ant species that nests mutualistically in Acacia drepanolobium trees in east Africa. Polymorphism ranged from 4 to 16 alleles per locus (mean = 7.3). Observed and expected heterozygosities ranged from 0.485 to 0.813 (mean 0.626), and from 0.502 to 0.894 (mean 0.674), respectively. These markers will foster studies of the population structure, colony structure, and reproductive strategies of these ants.     

Isoenzyme Variation in Melipona rufiventris (Hymenoptera: Apidae, Meliponina) in Minas Gerais State, Brazil

The stingless bee Melipona rufiventris is an important pollinator in several Brazilian ecosystems. Originally widely distributed in Minas Gerais (MG) state, this species is becoming very rare. Therefore this species was included in the endangered species list of MG. We used isoenzyme data for a better understanding of the genetic structure of several M. rufiventris colonies. Samples of 35 colonies were collected from 12 localities and evaluated by nine enzymatic systems, which yielded 17 loci. M. rufiventris genetic variation was found to be low, typical of an endangered species. The proportion of polymorphic loci was 5.88% in both ecosystems. Only Est-4 was polymorphic in colonies from the Forest and Mdh-1 in colonies from the Cerrado. The expected heterozygosity ranged from 0.0068 in the Cerrado to 0.0078 in the Forest. Despite this, enzyme electrophoretic analyses provided a good idea of the diversity between samples from Cerrado and Forest which reinforce the existence of two different forms of M. rufiventris in MG, one present in the Cerrado and the other in Forest. This information is of great importance for the conservation of, M. rufiventris in MG.     

Isolation and characterization of 23 microsatellite loci in the yellow tang, Zebrasoma flavescens (Pisces: Acanthuridae)

Twenty‐three microsatellites were isolated from the yellow tang (Zebrasoma flavescens), an ecologically and commercially important reef fish. Genetic diversity was assessed in 90 adults collected from Honokohau, Hawaii. The number of alleles per locus varied from four to 29 (mean = 13.8) and observed and expected heterozygosities ranged from 0.15 to 0.94 (mean = 0.70) and from 0.29 to 0.93 (mean = 0.81), respectively. Eight loci exhibited significant departure from Hardy–Weinberg equilibrium due to the presence of null alleles. Exact tests showed no evidence of genotypic disequilibrium between loci. Overall, loci were well resolved, easy to score and highly polymorphic.     

Isolation and characterization of 10 highly polymorphic di- and trinucleotide microsatellite markers in the mayfly Ameletus inopinatus (Ephemeroptera: Siphlonuridae)

We describe the isolation of ten polymorphic microsatellite loci from the mayfly Ameletus inopinatus. Loci had di‐ or trinucleotide repeat motifs and were highly variable with three to 17 alleles (mean = 7.15). Observed heterozygosity ranged from 0.143 to 0.905. One locus (Ami_202) showed significant deviation from Hardy–Weinberg equilibrium in one population, but no evidence for null alleles. One locus (Ami_73) was significantly linked with three other loci. The remaining nine loci should prove highly informative for population genetic studies.     

Intra- and interspecific nestmate recognition inMelipona workers (Hymenoptera: Apidae)

The possible significance of nestmate recognition in prevention of robbing and parasitism in three species of stingless bees was assessed. Nestmate discrimination abilities vary among them; Melipona quadrifasciataworkers attacked 74% of nonnestmate conspecifics that were encountered, while M. scutellarisand M. rufiventriswere less discriminating, attacking only 14 and 60% of non-nestmates, respectively. In tests of interspecific interactions, M. quadrifasciataand M. scutellariswere the least mutually tolerant of all species pairs tested. Tests with Apis melliferashowed a high degree of intolerance by two of the three Meliponaspecies.     

Distribution and movement of juvenile and sub‐adult Chinese sturgeon (Acipenser sinensis Gray, 1835) in the Three Gorges Reservoir and the adjacent upstream free‐flowing Yangtze River section: a re‐introduction trial

The aim of this 2008–2012 study was to prove the hypothesis that the Three Gorges Reservoir (TGR) and the upper free‐flowing section can offer a habitat for restocking the Chinese sturgeon, Acipenser sinensis. Thus, 498 cultured A. sinensis (including 28 sub‐adults tagged with ultrasonic transmitters) were released into the TGR and the adjacent free‐flowing section of the Yangtze River. The distribution, movement and growth of the released fish were collected by acoustic telemetry and bycatch information. Results indicated that only three sub‐adults migrated upstream to the free‐flowing section where they remained for the three‐year observation (limited by the longevity of the transmitter battery). However, most sub‐adults migrated downstream after release, with a migration distance of 15–877 km as well as a tracking duration from 19 to 6063 h. The downstream traveling speed in the free‐flowing section (mean = 3.70 km h^?1, range = 1.51 – 8.25 km h^?1) was significantly faster than that in the reservoir section (mean = 1.05 km h^?1, range = 0.12 – 1.82 km h^?1; p < 0.05). Only three sub‐adults migrated to within 3 km of Three Gorges Dam (TGD) and stayed in the reservoir. Sub‐adults stayed obviously deeper within the reservoir (mean = 11.47 m) than in the free‐flowing section (mean = 3.63 m; p < 0.05). Juveniles were recaptured from 45 – 810 km downstream their release sites with a mean traveling speed of 1.05 km h^?1 (range = 0.13 – 1.82 km h^?1), lower than that of the sub‐adults released in the free‐flowing section. Twelve percent of juveniles released in one of the tributaries of the Yangtze River found the entrance and migrated downstream to the reservoir within the first 30 days. Average body weight of the recaptured juveniles increased by 10.3% (range = 2.3% – 15.3%) and some crustaceans (e.g. river shrimp) were found in the stomach and gut of two juveniles. The results demonstrate that the TGR and its adjacent upstream free‐flowing Yangtze River section can serve as reintroduction sites to support a certain quantity of Chinese sturgeon to remain and mature there.     

Isolation and characterization of microsatellite markers from three species of swallows in the genus Tachycineta: T. albilinea, T. bicolor and T. leucorrhoa

We describe 30 microsatellite loci developed from three species of swallows in the genus Tachycineta: T. bicolor (tree swallow), T. albilinea (mangrove swallow), and T. leucorrhoa (white‐rumped swallow). These commonly studied birds nest in secondary cavities and are distributed from Alaska to Argentina. Primer pairs were designed for each species individually and tested for cross‐amplification in 40–48 individuals of all three species. Polymorphism ranged from 5 to 65 alleles per locus (mean = 19.1). These markers will allow comparative studies of extra‐pair paternity rates among members of the genus as well as the assessment of population structure.     

Characterization of nine microsatellite loci in the sea star Astropecten aranciacus and cross-species amplification for related taxa

So far, only few microsatellite markers have been developed and extensively tested for echinoderms. To study the population genetic structure of the sea star Astropecten aranciacus, we developed primers for nine polymorphic microsatellite loci and tested them on two populations from Faro in Portugal (N = 25) and from La Herradura in Spain (N = 20). Within populations, allele numbers varied from four to 20, while expected and observed heterozygosities ranged from 0.593 to 0.936 and from 0.222 to 0.900, respectively. Additional cross‐species amplifications were polymorphic at some loci, indicating their potential usefulness for related taxa.     

Similarities and differences in 13C and 15N stable isotope ratios in two non‐lethal tissue types from shovelnose sturgeon Scaphirhynchus platorynchus (Rafinesque, 1820)

We tested the hypothesis that δ¹³C and δ¹⁵N signatures of pectoral spines would provide measures of δ¹³C and δ¹⁵N similar to those obtained from fin clips for adult shovelnose sturgeon Scaphirhynchus platorynchus. Thirty‐two shovelnose sturgeon (fork length [FL] = 500–724 mm) were sampled from the lower Mississippi River, USA on 23 February 2013. Isotopic relationships between the two tissue types were analyzed using mixed model analysis of covariance. Tissue types differed significantly for both δ¹³C (P < 0.01; spine: mean = ?23.83, SD = 0.62; fin clip: mean = ?25.74, SD = 0.97) and δ¹⁵N (P = 0.01; spine: mean = 17.01, SD = 0.51; fin clip: mean = 17.19, SD = 0.62). Neither FL nor the FL × tissue type interaction had significant (P > 0.05) effects on δ¹³C. Fin clip δ¹³C values were highly variable and weakly correlated (r = 0.16, P = 0.40) with those from pectoral spines. We found a significant FL‐tissue type interaction for δ¹⁵N, reflecting increasing δ¹⁵N with FL for spines and decreasing δ¹⁵N with FL for fin clips. These results indicate that spines are not a substitute for fin clip tissue for measuring δ¹³C and δ¹⁵N for shovelnose sturgeon in the lower Mississippi River, but the two tissues have different turnover rates they may provide complementary information for assessing trophic position at different time scales.     

Isolation and characterization of 11 polymorphic trinucleotide microsatellite markers in the stonefly Arcynopteryx compacta (Plecoptera: Perlodidae)

We describe the isolation of 11 polymorphic trinucleotide microsatellite loci from the stonefly Arcynopteryx compacta. Loci were highly variable with 3 to 14 alleles (mean = 6.45). Observed heterozygosity ranged from 0 to 0.867. Seven loci showed significant deviation from Hardy–Weinberg equilibrium across both populations. There was no evidence for null alleles, and thus, Hardy–Weinberg departures could have resulted from genetic structure between populations or subpopulations. No linkage between loci was found. The 11 loci should prove highly informative for population genetic studies.     

Extracting DNA from museum bird eggs, and whole genome amplification of archive DNA

We present a comprehensive protocol for extracting DNA from egg membranes and other internal debris recovered from the interior of blown museum bird eggs. A variety of commercially available DNA extraction methods were found to be applicable. DNA sequencing of polymerase chain reaction (PCR) products for a 176‐bp fragment of mitochondrial DNA was successful for most egg samples (> 78%) even though the amount of DNA extracted (mean = 14.71 ± 4.55 ng/µL) was significantly less than that obtained for bird skin samples (mean = 67.88 ± 4.77 ng/µL). For PCR and sequencing of snipe (Gallinago) DNA, we provide eight new primers for the ‘DNA barcode’ region of COI mtDNA. In various combinations, the primers target a range of PCR products sized from 72 bp to the full ‘barcode’ of 751 bp. Not all possible combinations were tested with archive snipe DNA, but we found a significantly better success rate of PCR amplification for a shorter 176‐bp target compared with a larger 288‐bp fragment (67% vs. 39%). Finally, we explored the feasibility of whole genome amplification (WGA) for extending the use of archive DNA in PCR and sequencing applications. Of two WGA approaches, a PCR‐based method was found to be able to amplify whole genomic DNA from archive skins and eggs from museum bird collections. After WGA, significantly more archive egg samples produced visible PCR products on agarose (56.9% before WGA vs. 79.0% after WGA). However, overall sequencing success did not improve significantly (78.8% compared with 83.0%).     

Isolation and characterization of ten microsatellite loci in stingless bee Trigona spinipes (Apidae: Meliponini)

Stingless bees are the most abundant pollinators of Brazilian tropical flora. Trigona spinipes has some of the largest colonies of any stingless bee species found in several types of environment. This work describes the isolation and characterization of microsatellite loci for this species. A microsatellite‐enriched genomic library was constructed and ten primer pairs were designed for T. spinipes. The primers were tested in 20 unrelated individuals. The mean number of alleles was 8.10 and mean observed and expected heterozygosity were 0.655 and 0.680, respectively. Primers were also tested in cross‐species amplification and five loci were successfully amplified in Trigona chanchamayoensis, Trigona hyalinata, Tetragonisca angustula, Partamona mulata and Frieseomelitta varia. The microsatellite primers described herein will be useful for evaluating genetic variability and gaining a better understanding of the population structure of T. spinipes as well as other species of stingless bees.     

Spatial and Temporal Patterns of Frugivorous Hornbill Movements in Central Africa and their Implications for Rain Forest Conservation

Tropical forest conservation and restoration require an understanding of the movements and habitat preferences of important seed dispersers. With forests now being altered at an unprecedented rate, avian frugivores are becoming increasingly vital for forest regeneration. Seed movement, however, is highly dependent on the behavioral characteristics of their dispersers. Here, we examined the movements, habitat preferences, and range sizes of two African frugivores: the Black‐casqued (Ceratogymna atrata) and the White‐thighed (Bycanistes albotibialis) Hornbill, in the lowland rain forests of southern Cameroon. Using satellite transmitters, we tracked eight hornbills for 3 yr to characterize their movements and relate them to environmental landscape features. Hornbill movements differed significantly, with B. albotibialis ranging over larger areas (mean = 20,274 ha) than C. atrata (mean = 5604 ha), and females of both species covering over 15 times the area of males. Evidence suggests that movements are irruptive during particular periods, perhaps driven by low resource availability. In addition, hornbills often returned to the same localities within a year, although movements were not characterized as migratory. Both species displayed significant differences in habitat preference, with B. albotibialis utilizing disturbed habitat more frequently than C. atrata (t = ?22.04, P = 2.2 × 10^?16). Major roads were found to act as barriers for C. atrata, but not for B. albotibialis. The ability of both hornbill species to move large distances suggests hornbills will play a vital role in the maintenance and regeneration of rain forests in Central Africa as forest fragmentation increases and terrestrial vertebrates decline in numbers.     

Age,growth, mortality and sex ratio of the inshore population of the edible crab,Cancer pagurus (Linnaeus 1758) in South Wales (UK)

Age, growth, and mortality of the edible crab, Cancer pagurus, were determined for the native population in South Wales (UK). Sampling was carried out on a monthly basis between February 2001 and September 2002. Carapace width ranged between 10.4 and 163 mm. Based on the carapace width frequency distribution, the Swansea and Gower population was composed mainly of males belonging to the first and second age‐class (1 and 2), and of females belonging to the third and fourth age‐class (3 and 4). Sex ratio was 1.126 ± 0.27 in favour of males. Carapace width frequency distributions and weight‐at‐age data were used to estimate the von Bertalanffy growth equation parameters. For the population as a whole, these were: L∞ = 199 mm, W∞ = 1179.56 g, K = 0.24 year^?1, t₀ = ?0.1004 years. The overall carapace width–weight relationship was: W = 0.38(CW^2.69). Analysis of covariance indicated a significant difference in the carapace width–weight relationship between males and females in the study area. Total mortality Z and natural mortality M rates for combined sexes were 1.245 year^?1 and 0.567 year^?1, respectively. The exploitation ratio E was estimated to be 54.43%.