Ontogenetic diet shift in the June sucker Chasmistes liorus (Cypriniformes,Catostomidae) in the early juvenile stage

Kreitzer JD, Belk MC, Gonzalez DB, Tuckfield RC, Shiozawa DK, Rasmussen JE. Ontogenetic diet shift in the June sucker Chasmistes liorus (Cypriniformes, Catostomidae) in the early juvenile stage.
Ecology of Freshwater Fish 2010: 19: 433–438. © 2010 John Wiley & Sons A/S Abstract – Ontogenetic diet shifts are common in fishes and often occur during early life stages. The larval and early juvenile period is critical in the life cycle of the endangered June sucker, Chasmistes liorus (Teleostei: Catostomidae). High larval and juvenile mortality leads to low recruitment to the breeding population and hence a declining natural population. To understand diet composition and dynamics in June sucker at early life stages, diet was quantified and compared to available food items in the natural environment during the early juvenile stage. Rotifers (Brachionus sp.) were the primary diet item at week 10, but by week 12 a small cyclopoid copepod (Microcyclops rubellus) became predominant. Availability of diet items varied little across the experimental period. The increase in size of young suckers may explain this rapid dietary shift, but there are some inconsistencies with the size selection argument. This diet shift represents an important nutritional change that should be considered in development of diets for young June sucker and in assessing suitability of nursery habitats.     

Characterization and ontogenetic development of digestive enzymes in Pacific bluefin tuna Thunnus orientalis larvae

The major digestive enzymes in Pacific bluefin tuna Thunnus orientalis larvae were characterized, and the physiological characteristics of the enzymes during early ontogeny were clarified using biochemical and molecular approaches. The maximum activity of trypsin (Try), chymotrypsin (Ct) and amylase (Amy) was observed at pH 6–11, 8–11 and 6–9, respectively. Maximum activity of Try, Ct and Amy occurred at 50 °C, that of lipase (Lip) was at 60 °C and that of pepsin (Pep) was at 40–50 °C. These pH and thermal profiles were similar to those for other fish species but differed from those previously reported for adult bluefin tuna. Enzyme activity for all enzymes assayed was found to decrease at high temperatures (Try, Ct, Amy and Pep: 50 °C; Lip: 40 °C), which is similar to findings for other fish species with one marked exception—increased Try activity was observed at 40 °C. Lip activity appeared to be dependent on bile salts under our assay conditions, resulting in a significant increase in activity in the presence of bile salts. Ontogenetic changes in pancreatic digestive enzymes showed similar gene expression patterns to those of other fish species, whereas marked temporal increases in enzyme activities were observed at 10–12 days post hatching (dph), coinciding with previously reported timing of the development of the pyloric caeca in bluefin tuna larvae. However, complete development of digestive function was indicated by the high pep gene expression from 19 dph, which contradicts the profile of Pep activity and previously reported development timing of the gastric gland. These findings contribute to the general knowledge of bluefin tuna larval digestive system development.     

Diversity of human copy number variation and multicopy genes

Copy number variants affect both disease and normal phenotypic variation, but those lying within heavily duplicated, highly identical sequence have been difficult to assay. By analyzing short-read mapping depth for 159 human genomes, we demonstrated accurate estimation of absolute copy number for duplications as small as 1.9 kilobase pairs, ranging from 0 to 48 copies. We identified 4.1 million "singly unique nucleotide" positions informative in distinguishing specific copies and used them to genotype the copy and content of specific paralogs within highly duplicated gene families. These data identify human-specific expansions in genes associated with brain development, reveal extensive population genetic diversity, and detect signatures consistent with gene conversion in the human species. Our approach makes ~1000 genes accessible to genetic studies of disease association.     

Establishment of ES cells from inbred strain mice by dual inhibition (2i)

A number of mouse ES cells from inbred strains have been established to date, but efficiency varies across the different strains. The 129 strain mouse is efficient to establish, whereas C57BL/6 and BALB/c strains are not. It is possible that their genetic backgrounds account for the difference in their ability to establish ES cell lines. In this study, we attempted to establish C57BL/6J and BALB/c Cr ES cells by dual inhibition (2i) using two inhibitors (PD0325901 and CHIR99021) of extracellular signal regulated-kinase (ERK) and glycogen synthase kinase-3 (GSK-3), which promote ES cell differentiation. The results revealed that the establishment efficiencies of C57BL/6J and BALB/c Cr ES cells were remarkably increased by 2i. These ES cells stably expressed pluripotent markers and generated high-contribution chimeras with germline transmission. Furthermore, we generated germline chimeras from C57BL/6J ES cells through the method of gene modification. These findings indicate that 2i is a powerful tool for establishing C57BL/6J and BALB/c Cr ES cells with the ability to generate germline chimeras.     

Sero-epidemiological survey for toxoplasmosis in wild New World monkeys (Cebus spp.; Alouatta caraya) at the Paraná river basin, Paraná State, Brazil

In this study, we captured 60 wild New World monkeys (Cebus spp.; Alouatta caraya) at the Paraná river basin, Paraná State, Brazil, and modified agglutination test (MAT) was performed to evaluate anti-Toxoplasma gondii antibodies. Prevalence was 30.2% (13/43) in Cebus spp. (capuchin monkeys) and 17.6% (3/17) for A. caraya (black and golden howler monkeys). MAT showed antibody titers of 16 (15/16) and 64 (1/16). Herein, we have observed an odds ratio (OR)=4.67 (1.060.05). The present work is the first report on serum occurrence of anti-T. gondii antibodies in wild capuchin monkeys and in wild black and golden howler monkeys.     

Numerical simulation on effect of irrigation conditions on water temperature distribution in a paddy field

Water management methods regulate water temperature in paddy fields, which affects rice growth and the environment. To understand the effect of irrigation conditions on water temperature in a paddy field, water temperature distribution under 42 different irrigation models including the use of ICT water management, which enables remote and automatic irrigation, was simulated using a physical model of heat balance. The following results were obtained: (1) Irrigation water temperature had a more significant effect on paddy water temperature close to the inlet. As the distance from the inlet increased, the water temperature converged to an equilibrium, which was determined by meteorological conditions and changes in water depth. (2) Increasing the irrigation rate with higher irrigation water amount increased the extent and magnitude of the effects of the irrigation water temperature. (3) When total irrigation water amount was the same, increasing the irrigation rate decreased the time-averaged temperature gradient effect over time across the paddy field. (4) Irrigation during the lowest and highest paddy water temperatures effectively decreased and increased the equilibrium water temperature, respectively. The results indicate that irrigation management can be used to alter and control water temperature in paddy fields, and showed the potential of ICT water management in enhancing the effect of water management in paddy fields. Our results demonstrated that a numerical simulation using a physical model for water temperature distribution is useful for revealing effective water management techniques under various irrigation methods and meteorological conditions.

     

In vitro production of viable eggs from isolated mouse primary follicles by successive culture

To produce viable eggs from single primary follicles in vitro, primary follicles containing oocytes (average 39.0 ± 0.2 µm in diameter) were isolated from the ovaries of 1-week-old mice, and cultured in combination with culture membranes for the first 8 days up to the secondary follicle stage, followed by the next 12 days to the later stages. After culture with a combination of first and second culture membranes using high and low adhesion characteristics, the average oocyte diameters of the surviving follicles increased by almost two-fold in all four groups. Further, the oocyte maturation rate was the highest (74.1%) in the culture group with low adhesion with collagenase and high adhesion. In this culture group, when the O₂ concentration was changed from 20% in the first culture to 5% in the second culture, the cleavage rate increased to 47.5%, which was comparable to the level of the in vivo control (34.6%). Finally, 39 embryos at the 2- to 8-cell stages were transferred into the oviducts of three pseudopregnant females, and eight live pups (20.5%) were obtained. Of the eight pups, six survived for at least six months and were fertile. The present study shows successive in vitro cultures of single isolated primary follicles for the production of viable eggs. We believe that this culture system, with a combination of culture membranes under controlled O₂ conditions, is applicable to other mammalian species, including humans.     

Identification and in vitro cytotoxicity of ochratoxin A degradation products formed during coffee roasting

The mycotoxin ochratoxin A is degraded by up to 90% during coffee roasting. In order to investigate this degradation, model heating experiments with ochratoxin A were carried out, and the reaction products were analyzed by HPLC-DAD and HPLC-MS/MS. Two ochratoxin A degradation products were identified, and their structure and absolute configuration were determined. As degradation reactions, the isomerization to 14-(R)-ochratoxin A and the decarboxylation to 14-decarboxy-ochratoxin A were identified. Subsequently, an analytical method for the determination of these compounds in roasted coffee was developed. Quantification was carried out by HPLC-MS/MS and the use of stable isotope dilution analysis. By using this method for the analysis of 15 coffee samples from the German market, it could be shown that, during coffee roasting, the ochratoxin A diastereomer 14-(R)-ochratoxin A was formed in amounts of up to 25.6% relative to ochratoxin A. The decarboxylation product was formed only in traces. For toxicity evaluations, first preliminary cell culture assays were performed with the two new substances. Both degradation products exhibited higher IC50 values and caused apoptotic effects with higher concentrations than ochratoxin A in cultured human kidney epithelial cells. Thus, these cell culture data suggest that the degradation products are less cytotoxic than ochratoxin A.