DNA extraction from museum specimens of parasitic Hymenoptera.

At the same time that molecular researchers are improving techniques to extract DNA from museum specimens, this increased demand for access to museum specimens has created tension between the need to preserve specimens for maintaining collections and morphological research and the desire to conduct molecular analyses. To address these concerns, we examined the suitability of non-invasive DNA extraction techniques on three species of parasitic Hymenoptera (Braconidae), and test the effects of body size (parasitoid species), age (time since collection), and DNA concentration from each extract on the probability of amplifying meaningful fragments of two commonly used genetic loci. We found that age was a significant factor for determining the probability of success for sequencing both 28S and COI fragments. While the size of the braconid parasitoids significantly affected the total amount of extracted DNA, neither size nor DNA concentration were significant factors for the amplification of either gene region. We also tested several primer combinations of various lengths, but were unable to amplify fragments longer than ~150 base pairs. These short fragments of 28S and COI were however sufficient for species identification, and for the discovery of within species genetic variation

Microsphere Spray System for Wound Coverage

Spinal fusion is used to treat diseases or disorders of the spine by fusing together two or more vertebrae. Two associated risks with spinal fusion are infection and blood loss. Administration of tranexamic acid is used to prevent blood loss, and transfusions are given following blood loss. Surgical site infections are prevented with vancomycin powder spread into the surgical wound, while established infections are treated by debridement and delivery of antibiotics for 4 to 6 weeks. The present research explored an alternate method to prevent and treat blood loss or infection in spinal fusion. Poly(lactic-co-glycolic acid) (PLGA) microspheres was used to encapsulate vancomycin for 42 days to treat infection. Vancomycin encapsulated in gelatin microspheres had a controlled release of 7 days to prevent infection. Tranexamic acid was dissolved into phosphate-buffered saline or carboxymethylcellulose to provide a release of 6 hours to prevent blood loss after surgery. The microspheres and tranexamic acid were delivered to a target region using a water based spray system. The spray system demonstrated the delivery and distribution of drugs to a target region. The microsphere spray system is capable of spraying drugs onto a target region to prevent or treat blood loss and infection over time

CALL BROADCASTING AND AUTOMATED RECORDERS AS TOOLS FOR ANURAN SURVEYS IN A SUBARCTIC TUNDRA LANDSCAPE

Relatively little is known about population ecology of anurans in arctic and subarctic tundra regions, in part because it is difficult to survey anurans in these landscapes. Anuran survey protocols developed for temperate regions have limited applicability in arctic and subarctic tundra landscapes, which may lack roads and vehicle access, and experience variable and inclement weather during short anuran breeding seasons. To evaluate approaches to address some of the limitations of surveying anurans in tundra landscapes, we assessed the effectiveness of using breeding call broadcasts to increase detection of Boreal Chorus Frogs (Pseudacris maculata) and Wood Frogs (Lithobates sylvaticus) near Cape Churchill, Manitoba, Canada. We also evaluated how counts of anurans derived from automated audio recorders compared with those obtained simultaneously by observers. We detected on average 0.4 additional Wood Frogs per survey when we broadcasted calls (x = 0.82, SD = 1.38), an increase of > 40% compared to surveys without broadcasts (x = 1.24, SD = 1.51; Wilcoxon test; Z = 2.73, P = 0.006). In contrast, broadcasting Boreal Chorus Frog calls did not increase the number of chorus frog detections (Wilcoxon test; Z < 0.001, P > 0.90). Detections of Wood Frogs in a 100-m radius were lower via automated recorders (x = 0.60, SD = 0.87 SD) than by observers during simultaneous surveys (x = 0.96, SD = 1.27 Z = 2.07, P = 0.038), but those of Boreal Chorus Frogs were not different (x = 1.72, SD = 1.31;x = 1.44, SD = 1.5; Z = 1.55, P > 0.121). Our results suggest that broadcasting calls can increase detection of Wood Frogs, and that automated recorders are useful in detecting both Wood Frogs and Boreal Chorus Fogs in arctic and subarctic tundra landscapes

A population-based survey on family intentions and fertility awareness in women and men in the United Kingdom and Denmark

Background: Across several European countries family formation is increasingly postponed. The aims of the study were to investigate the desire for family building and fertility awareness in the UK and Denmark.Methods: A population-based internet survey was used among women (n?=?1,000) and men (n?=?237) from the UK (40%) and Denmark (60%). Data covered socio-demographics, family formation, and awareness of female age-related fertility. Data analysis used descriptive statistics and logistic regression analysis for studying associations between low fertility awareness and desired family formation.Results: The majority of all participants desired two or three children. Two-thirds of the childless participants desired a first child at 30+ years, and one-fifth of the women and one-third of the men desired a last child at age 40. Overall, 83% of women and 73% of men were aware that female fertility starts to decline around 25–30 years. Men had significantly lower fertility awareness. Women who underestimated the impact of age on female fertility were significantly more likely to have a desire or attempted their first child at a higher age.Conclusion: Even though the majority were aware of the age-related decrease in female fertility, most desired having children at an age when female fertility has declined. Women who were not sufficiently aware of the impact of advanced age were significantly more likely to have their first child at a higher age. There is a need for developing educational programs for women and men in order to increase the population’s knowledge of fertility and risk factors for infertility

iMSAT: a novel approach to the development of microsatellite loci using barcoded Illumina libraries

BACKGROUND: Illumina sequencing with its high number of reads and low per base pair cost is an attractive technology for development of molecular resources for non-model organisms. While many software packages have been developed to identify short tandem repeats (STRs) from next-generation sequencing data, these methods do not inform the investigator as to whether or not candidate loci are polymorphic in their target populations. RESULTS: We provide a python program iMSAT that uses the polymorphism data obtained from mapping individual Illumina sequence reads onto a reference genome to identify polymorphic STRs. Using this approach, we identified 9,119 candidate polymorphic STRs for use with the parasitoid wasp Trioxys pallidus and 2,378 candidate polymorphic STRs for use with the aphid Chromaphis juglandicola. For both organisms we selected 20 candidate tri-nucleotide STRs for validation. Using fluorescent-labeled oligonucleotide primers, we genotyped 91 female T. pallidus collected in nine localities and 46 female C. juglandicola collected in 4 localities and found 15 of the examined markers to be polymorphic for T. pallidus and 12 of the examined markers to be polymorphic for C. juglandicola. CONCLUSIONS: We present a novel approach that uses standard Illumina barcoding primers and a single Illumina HiSeq run to target polymorphic STR fragments to develop and test STR markers. We validate this approach using the parasitoid wasp T. pallidus and its aphid host C. juglandicola. This approach, which would also be compatible with 454 Sequencing, allowed us to quickly identify markers with known variability. Accordingly, our method constitutes a significant improvement over existing STR identification software packages. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-858) contains supplementary material, which is available to authorized users