Low reproductive success of leatherback turtles, Dermochelys coriacea, is due to high embryonic mortality

We examined the mechanism responsible for low reproductive success in leatherback turtles (Dermochelys coriacea) at Playa Grande, Costa Rica: low egg fertilization versus high rates of embryonic death. Leatherbacks at this beach had a high rate of fertility (=93.3%±2.5%, n=819). We incubated 10 eggs from every clutch encountered of 19 females during 3 months of the 1998-1999 nesting season. Fertility rate of some females decreased during the nesting season, but overall was high. Detection of fertility was difficult using standard methods because fertility rates cannot be determined accurately from nests excavated after hatching because of egg decomposition. Removal and incubation of eggs from nests provided a better estimate. Embryonic death, particularly in the beginning of incubation before embryos are visible to the unaided eye, was the cause of low hatching success in this population. Hatching success increased with increasing fertility and differed between females, with some mothers having 71-81% success and others 23-32%. Embryonic death and not low egg fertility drives poor recruitment at Playa Grande. Improved conservation of this species at Playa Grande will require a better understanding of the mechanism behind embryonic death.     

Bioactive cinchona alkaloids from Remijia peruviana

Three known Cinchona alkaloids of the quinine type, quinine (1), cupreine (2), cinchonine (3), and the possible artifact cinchonine-HCl (3-HCl), along with two new ones, acetylcupreine (4) and N-ethylquinine (5), have been isolated from the bark of Remijia peruviana (Rubiaceae). Their stereochemical structures were established by high resolution NMR spectroscopy. Alkaloids 2-4 had antifeedant effects on Leptinotarsa decemlineata with varying potencies. Compound 4 was cytotoxic to both insect Sf9 and mammalian CHO cells after 48 h of incubation, while 3-HCl had stronger and selective cytotoxicity to Sf9. Quinine 1 had a moderate to low effect on Trypanosoma cruzi. Tumoral cells were also affected by these alkaloids, with 4 and 3-HCl being the most cytotoxic to all the cell lines tested. Overall, the 8R, 9S configurations, as in 3 and 3-HCl, as well as the C-6'acetylated alkaloid 4, with an 8S, 9R configuration, showed stronger biological effects.     

Oxidative dimers produced from protocatechuic and gallic esters in the DPPH radical scavenging reaction

DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging reactions of protocatechuic and gallic acids, and their methyl esters, have been investigated by NMR. In acetone, methyl protocatechuate was gradually converted to a Diels-Alder adduct of two molecules of the intermediate quinone in the reaction with DPPH radical, whereas methyl gallate rapidly gave a symmetrical dimer via a putative quinone precursor. Both dimers are rather unstable and their structures have been deduced by in situ NMR measurements of the reaction mixtures. Gallic acid also gave a corresponding symmetrical dimer in the same reaction as methyl gallate, although protocatechuquinone produced from protocatechuic acid did not yield a Diels-Alder adduct, unlike its methyl ester. Interestingly, these dimer formations were not observed in methanol solution.     

Serological characterization of the new genotype E of small ruminant lentivirus in roccaverano goat flocks

Maedi visna virus (MVV) and caprine arthritis encephalitis virus (CAEV) are a heterogeneous group of infectious agents affecting sheep and goats. Due to their natural cross-species infection they are referred to as small-ruminant lentiviruses (SRLV). Recently a new genetic cluster, highly divergent from MVV and CAEV was identified in the north-west part of Italy. A panel of genotype E specific antigens was developed and evaluated in flocks infected with B1 and E strains. The results clearly indicate that a strain specific antigen is required to correctly identify animals infected with different genotypes.     

Preliminary study on homing,site fidelity,and diel movement of black rockfish <Emphasis Type="Italic">Sebastes inermis</Emphasis> measured by acoustic telemetry

Several species of fish exhibit strong homing ability and distinct site fidelity to their original habitats and spawning sites. We have studied the homing ability, site fidelity, and diel movement patterns of three nocturnal black rockfish Sebastes inermis. The patterns were studied by radio-linked acoustic positioning system (VRAP) and fixed monitoring receivers (VR2) in Maizuru Bay, Japan from June to July 2004. All tagged fish were displaced by 80–120 m, but they returned to their respective captured sites at dusk, regardless of the time of release. The fish tagged with a depth sensor transmitter displayed active vertical movement just after the release, and subsequently swam near the bottom depth around the captured site. After homing, they displayed small home ranges [average 815 ± 265 m² (SD)] around a single rocky area with one or two core areas (average 64 ± 30 m²). One core area was utilized during the daylight and dark periods, whereas the other was used primarily during the dark period. The depth-tagged fish primarily used the depth of the rock crevice during the daylight, and it spent its dark period between the surface and the deeper bottom depth.     

Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.     

Isolation of chicken taste buds for real‐time Ca2+ imaging

We isolated chicken taste buds and used a real‐time Ca²⁺ imaging technique to investigate the functions of the taste cells. With RT‐PCR, we found that isolated chicken taste bud‐like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle‐shaped and approximately 61–75 μm wide and 88–98 μm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca²⁺ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L‐glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5′‐monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca²⁺ imaging can be informative.