Reversal of motility loss in bongo antelope (Tragelaphus eurycerus isaaci) spermatozoa contaminated with hyposmotic urine during electroejaculation

Semen collected by a combination of ampullary (rectal) massage and electroejaculation of a bongo bull was incidentally contaminated with urine (1:3.7). At 1.5h post-collection, progressive motility was 0% but some spermatozoa had intermittently twitching tails. Subsequent dilution with media and processing improved the progressive motility (up to 50%) and intact membranes (up to 71%) of spermatozoa. After thawing, the respective values were 35 and 70%. The osmolarity and pH of the contaminated supernatant was 151 mOsm and 7.45, respectively. Initial progressive motility in a non-contaminated portion of semen collected during the same procedure was 80%, and, after thawing, 60 and 90%, of the spermatozoa showed progressive motility and intact membranes, respectively. In conclusion, urine-contaminated bongo spermatozoa can regain progressive motility after dilution with isosmotic solutions and survive cryopreservation.     

Genome scale assessment of a species translocation program

Despite increased use of species translocations, controversy remains regarding the efficacy and efficiency of the strategy in obtaining conservation goals. Much of this controversy results from vague program objectives, unclear definitions of success, and lack of follow-up monitoring. We used the translocation program initiated by Zoo Knoxville for the federally threatened Bog Turtle (Glyptemys muhlenbergii) as a case study to demonstrate how genomic assessments not only assess the success of program objectives, but also allow managers to quickly obtain baseline data from which program objectives and explicit definitions of ‘success’ can be determined. Here we used 7030 SNP markers derived from RADseq data to confirm the premise that different source populations are genetically differentiated. Then we tested whether the release population has enhanced genetic diversity, as expected from a deliberate admixture. Although the release population had greater diversity than any source population, variation was lower than expected from modeling admixture with equal source contribution. Our results support the premise that genetic diversity can be maximized by including representatives from as many natural populations as possible. But failure to achieve the expected level of diversity could result from nonrandom success of founders from different sources or unrecorded bias in the implementation of the release program. Many existing and future translocation programs would benefit from genetic assessment similar to that conducted here with Bog Turtles.     

Blastocyst development after intergeneric nuclear transfer of mountain bongo antelope somatic cells into bovine oocytes

Intergeneric embryos were constructed by nuclear transfer using Mountain Bongo antelope somatic cells fused with enucleated bovine oocytes and their subsequent development in vitro was investigated. After two to six passages, starved or non-starved skin fibroblast cells were used as donor nuclei. In vitro matured bovine oocytes were enucleated by squeezing the first polar body and surrounding cytoplasm through a slit in the zona pellucida. After injection of a somatic cell into the perivitelline space, couplets were fused electrically and activated chemically, then subjected to different embryo culture treatments. Serum starvation had no effect on the frequency of cleavage to two cells or on development to the blastocyst stage in either sequential hamster embryo culture medium (HECM)-6/TCM-199 + serum or HECM-9/TC-199 + serum, or modified synthetic oviduct fluid (mSOF) culture medium. When couplets from non-starved donor nuclei were cultured, the frequency of cleavage (66 +/- 8% vs. 44 +/- 5%), development to >/=9 cells (46 +/- 6% vs. 24 +/- 4%), and formation of blastocysts (24 +/- 5% vs. 11 +/- 2%) were all significantly higher (p < 0.05) in the HECM-6 medium than in mSOF medium. In conclusion, bovine oocytes can support blastocyst development after intergeneric fusion with bongo fibroblasts. This technique could potentially be used as an alternative to using scarce bongo oocytes in attempts to propagate these endangered animals.     

Meiotic chromosome pairing is promoted by telomere-led chromosome movements independent of bouquet formation

Chromosome pairing in meiotic prophase is a prerequisite for the high fidelity of chromosome segregation that haploidizes the genome prior to gamete formation. In the budding yeast Saccharomyces cerevisiae, as in most multicellular eukaryotes, homologous pairing at the cytological level reflects the contemporaneous search for homology at the molecular level, where DNA double-strand broken ends find and interact with templates for repair on homologous chromosomes. Synapsis (synaptonemal complex formation) stabilizes pairing and supports DNA repair. The bouquet stage, where telomeres have formed a transient single cluster early in meiotic prophase, and telomere-promoted rapid meiotic prophase chromosome movements (RPMs) are prominent temporal correlates of pairing and synapsis. The bouquet has long been thought to contribute to the kinetics of pairing, but the individual roles of bouquet and RPMs are difficult to assess because of common dependencies. For example, in budding yeast RPMs and bouquet both require the broadly conserved SUN protein Mps3 as well as Ndj1 and Csm4, which link telomeres to the cytoskeleton through the intact nuclear envelope. We find that mutants in these genes provide a graded series of RPM activity: wild-type>mps3-dCC>mps3-dAR>ndj1Δ>mps3-dNT = csm4Δ. Pairing rates are directly correlated with RPM activity even though only wild-type forms a bouquet, suggesting that RPMs promote homologous pairing directly while the bouquet plays at most a minor role in Saccharomyces cerevisiae. A new collision trap assay demonstrates that RPMs generate homologous and heterologous chromosome collisions in or before the earliest stages of prophase, suggesting that RPMs contribute to pairing by stirring the nuclear contents to aid the recombination-mediated homology search.     

Cryobanking of viable biomaterials: implementation of new strategies for conservation purposes

Cryobanking, the freezing of biological specimens to maintain their integrity for a variety of anticipated and unanticipated uses, offers unique opportunities to advance the basic knowledge of biological systems and their evolution. Notably, cryobanking provides a crucial opportunity to support conservation efforts for endangered species. Historically, cryobanking has been developed mostly in response to human economic and medical needs — these needs must now be extended to biodiversity conservation. Reproduction technologies utilizing cryobanked gametes, embryos and somatic cells are already vital components of endangered species recovery efforts. Advances in modern biological research (e.g. stem cell research, genomics and proteomics) are already drawing heavily on cryobanked specimens, and future needs are anticipated to be immense. The challenges of developing and applying cryobanking for a broader diversity of species were addressed at an international conference held at Trier University (Germany) in June 2008. However, the magnitude of the potential benefits of cryobanking stood in stark contrast to the lack of substantial resources available for this area of strategic interest for biological science — and society at large. The meeting at Trier established a foundation for a strong global incentive to cryobank threatened species. The establishment of an Amphibian Ark cryobanking programme offers the first opportunity for global cooperation to achieve the cryobanking of the threatened species from an entire vertebrate class.     

Association between parasite load and orange,but not blue,male nuptial colouration in Etheostoma caeruleum

The hypothesis that males possessing more vivid nuptial colouration are of better quality with regard to individual measures of health such as parasite infection was investigated by taking spectral measurements of orange and blue bars in the nuptial males of a sexually dichromatic stream fish rainbow darter Etheostoma caeruleum to determine if male colouration in this species varied with parasite load in the form of black spot disease. The yellow chroma of orange bars, i.e. the relative contribution of wavelengths in the range of 550–625 nm to total brightness, was the only spectral measurement significantly associated with parasite counts. These results are discussed in the context of sexual selection and the potential of orange bars in E. caeruleum to serve as honest indicators of quality to potential mates.     

Cerapeplus gen.n. and the classification of micropeplids (Coleoptera: Micropeplidae)

Cerapeplus siamensis gen.n., sp.n. from Thailand is described and illustrated. The new genus is characterized by filament-based lamellar secretions arising from the lateral body margin, a feature observed for the first time in Coleoptera. Possible autapomorphies for the micropeplid genera are discussed. It is concluded that Cerapeplus and Kalissus are monophyletic; Micropeplus and/Or Peplomicrus are probably polyphyletic but both genera together form a monophyletic group. A number of characters support the family status of the micropeplids recently proposed by some authors. A revised family diagnosis is provided.     

Flow microfluorometric analysis of living spermatozoa stained with Hoechst 33342

Bovine spermatozoa were stained with Hoechst 33342. The fluorescence distribution of stained spermatozoa was complex. Non-motile spermatozoa displayed a higher fluorescence than did motile spermatozoa. The fluorescence profile of the motile spermatozoa was bimodal. Sort and reanalysis, and orientation experiments suggested that there are two distinct populations of motile spermatozoa.