Assessment of antioxidant activity of cane brown sugars by ABTS and DPPH radical scavenging assays: determination of their polyphenolic and volatile constituents

Seven cane brown sugars (four from La Réunion, two from Mauritius, and one from France) were investigated for their polyphenol content and volatile composition in relation to their free radical scavenging capacity determined by ABTS and DPPH assays. The thin layer coated on the sugar crystal was extracted by Soxhlet extractor with dichloromethane. The volatile compounds of brown sugars were studied by GC-MS, and 43 compounds were identified. The total phenolic content of brown sugars was determined according to the Folin-Ciocalteu method. Phenolic compounds were quantified in the brown sugar extracts by LC-UV-ESI-MS. Brown sugar aqueous solutions exhibited weak free radical scavenging activity in the DPPH assay and higher antioxidant activity in the ABTS assay at relatively high concentration. The brown sugar extracts showed interesting free radical scavenging properties despite the low concentration of phenolic and volatile compounds. Sugar is a common foodstuff traditionally used for its sweetening properties, which might be accompanied by antioxidant properties arising from molecules (polyphenols, Maillard products) other than sucrose of the cane brown sugars.     

Isolation and characterization of an extracellular antimicrobial protein from Aspergillus oryzae

A 17 kDa antimicrobial protein was isolated from growth medium containing the filamentous fungus Aspergillus oryzae by extracting the supernatants from the culture media, ion exchange chromatography on CM-sepharose, and C18 reverse-phase high-performance liquid chromatography. This antimicrobial protein, which we considered to be an extracellular antimicrobial protein from A. oryzae (exAP-AO17), possessed antimicrobial activity but lacked hemolytic activity. The exAP-AO17 protein strongly inhibited pathogenic microbial strains, including pathogenic fungi, Fusarium moniliform var. subglutinans and Colletotrichum coccodes, and showed antibacterial activity against bacteria, including E. coli O157 and Staphylococcus aureus. To confirm that the protein acts as a regulation factor for extracellular secretion, we examined growth under varying conditions of N sources, C sources, ions, ambient pH, and stress. Various culture conditions were found to induce characteristic changes in the expression of protein synthesis as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Highly basic polypeptides were regulated by suppressing the ambient pH under acidic conditions and strongly induced under alkaline conditions, thus confirming that pH regulation is physiologically relevant. The expression of exAP-AO17 was upregulated by heat shock upon growth in the presence of NaCl. Automated Edman degradation showed that the N-terminal sequence of exAP-AO17 was NH 2-GLPGPAGAVGFAGKDQNM-. ExAP-AO17 showed partial sequence homology with a collagen belonging to the animal source. These results suggest that exAP-AO17 is an excellent candidate as a lead compound for the development of novel oral or other types of anti-infective agents.     

Rapid selection of catechin-rich tea trees (Camellia sinensis) by a colorimetric method

A rapid and efficient colorimetric method based on the use of Fast Blue B-salt (FBB) was established to select catechin-rich tea trees (Camellia sinensis L.). The catechin levels measured by the colorimetric method under optimized reaction conditions correlated closely with estimations by high-performance liquid chromatography (HPLC) analysis. The FBB colorimetric method was successfully used to classify 160 tea trees on the basis of their catechin contents into rich and poor lines. HPLC analysis of the FBB-selected tea tree extracts showed them to contain (−)-epigallocatechin 186 mg/g in tea tree line HR-29, (−)-epicatechin 43.7 mg/g in HR-82, (−)-epigallocatechin gallate 4.32 mg/g in HR-29, and (−)-epicatechin gallate 0.22 mg/g in HR-52. Classification of tea trees from the Hadong region into catechin-rich and -poor trees was independent of the growing season. Thus the FBB colorimetric method could find application as a reliable tool in screening and selection of tea trees on the basis of their catechin content.     

Diversity and origins of giant guitarfish and wedgefish products in Singapore

1. Giant guitarfishes (Glaucostegidae) and wedgefishes (Rhinidae) are some of the most threatened marine taxa in the world, with 15 of the 16 known species exhibiting global population declines and categorized as Critically Endangered according to the International Union for Conservation of Nature (IUCN) Red List of Threatened Species. The recent inclusion of all species in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) necessitates more rigorous enforcement by regulatory authorities.
2. Challenges in regulating the trade of giant guitarfish and wedgefish products due to difficulties in visual identification of processed products and labelling issues impede enforcement. The aim of this study is to characterize the diversity and origins of associated traded products that were commercially available in Singapore, one of the world's top importers and re-exporters of shark and ray products.
3. A total of 176 samples of elasmobranch products were obtained between June and December 2019 from fishery ports and various retailers in Singapore. By applying cytochrome c oxidase subunit I gene barcoding, 31 elasmobranch species were detected, with 55% of the species considered threatened (Critically Endangered, Endangered, or Vulnerable) based on the IUCN Red List and 35% of species listed in CITES Appendix II. Four species of giant guitarfishes and wedgefishes were commercially available to consumers in fresh forms of whole fish, fillet, and fin, as well as dried and cooked meats.
4. DNA barcoding has proven to be an effective tool for identifying elasmobranch products that are impossible to recognize visually and would aid enforcement of CITES trade regulations. This work underscores the urgent need to step up enforcement of marine wildlife regulations and draw public attention to the elasmobranch trade.

     

Co‐culture of Buffalo Preantral Follicles with Different Somatic Cells

The effect of co‐culture of buffalo preantral follicles (PFs) with different somatic cells, i.e, cumulus, granulosa, ovarian mesenchymal and oviductal epithelial cells was studied. Large PFs (250–450 μm) were isolated by microdissecting the trypsin (1%) digested ovarian cortical slices. Cumulus cells were isolated by repeated pipetting of oocytes, granulosa cells were isolated by aspirating from punctured PFs and ovarian mesenchymal cells were isolated from ovarian cortex by scraping the cortical slices and passing through 20 μm filter. Preantral follicles were cultured in standard culture medium without somatic cells or co‐cultured with cumulus cells, granulosa cells, ovarian mesenchymal cells and oviductal epithelial cells for 80 days. The growth rate (μm/day) of the PFs was monitored by measuring follicular diameter on day 0, 30, 60 and 80 days of culture. The viability of PFs was evaluated by trypan blue staining. The results indicated that PFs co‐cultured with cumulus, granulosa and ovarian mesenchymal cells had a better development and survivality compared with control and those co‐culture with oviductal epithelial cells. Maximum growth and survivality of PFs were achieved when cultured with cumulus cells. It is concluded that inclusion of somatic cells in PF culture media had beneficial effect on the growth of PFs and cumulus cells supported maximum growth and survivality of PFs in vitro of all somatic cells tested.     

Developmental capacity of reconstituted mouse embryos: influences of nucleus and cytoplasm sources.

This study was undertaken to examine the developmental capacity of reconstituted mouse embryos, and the influences of nucleus and cytoplasm on the development of these embryos following reciprocal pronuclear transplantation between in vitro 2-cell blocked and nonblocked embryos. Karyoplast containing pronuclei was transferred into the perivitelline space of the enucleated zygote and fused to cytoplasm with electrofusion. Maximum fusion rate was obtained when a field strength of 1.5 kV/cm was used. The fusion rates were high (86.2 +/- 3.2 to 90.6 +/- 2.0%) regardless of the strains of donor nucleus and recipient cytoplasm. Developmental rates of reconstituted embryos to the blastocyst stage, which were similar to that of the F1 (C57BL/6J x CBA) control were high when F1 embryos were used as the cytoplasm recipients (88.8 +/- 1.5 and 91.9 +/- 2.0%). When ICR embryos were used as the recipient cytoplasm, developmental rates were significantly reduced (71.5 +/- 2.9 and 54.1 +/- 3.2%), and affected by the source of nucleus. There were no significant differences in the cell number of embryos that developed to blastocysts and in the developmental rates to live young among the embryos reconstituted with different nuclei and cytoplasm, and the ICR control. The results of this study show that the development of reconstituted embryos is hardly affected by nuclear transplantation and electrofusion procedures. It is indicated that the recipient cytoplasm, rather than the donor nucleus, has the greater influence on the in vitro development of the reconstituted embryos to the blastocyst stage.     

Effect of twin birth calvings on milk production, reproductive performance, and survival of lactating cows

OBJECTIVE: To evaluate effect of twin birth calvings on milk production, reproductive performance, and survival of lactating cows. DESIGN: Retrospective observational cohort study. ANIMALS: 33,868 cows from 20 farms. PROCEDURES: Data on age at calving for primiparous cows and mature equivalent milk yield for multiparous cows, assistance at calving, stillbirths, twin births, gestation duration, pregnancy at the end of the data collection period, and culling-death for all cows were extracted from farm computer records and used for statistical analysis. RESULTS: Prevalence of twin parturitions was 1.3% (159/12,050) and 6.5% (1,410/21,818) for primiparous and multiparous cows, respectively. Primiparous and multiparous cows with singletons produced more milk than cows with live twins or at least 1 dead twin (primiparous, 33.1 vs 31.9 vs 31.2; multiparous, 36.5 vs 35.7 vs 35.0). Multiparous cows with dead twins produced less milk than cows with live twins. Compared with dams with singleton birth, cows with twins were 0.78 times as likely to conceive and 1.42 times as likely to die or be culled. Cows with dead twins also had increased time to conception, compared with live twins. CONCLUSIONS AND CLINICAL RELEVANCE: Twin birth was associated with decreased survival, milk production, and reproductive performance. Having at least 1 dead twin was even more detrimental than having live twins and resulted in decreased milk production and reproductive performance of lactating cows.