Impact of Endocrine‐Disrupting Chemicals on Reproductive Function in Zebrafish (Danio rerio)

The prevalence of endocrine‐disrupting chemicals (EDCs) in the aquatic environment has been associated with the wide detection of alterations in the development and physiology of vertebrates. Zebrafish, as a model species, has been extensively used in toxicological research. In this review, we focus on recent published evidence of the harmful effects of EDCs on reproductive function in zebrafish, including skewed sex ratio, immature gonads, diminished sexual behaviour, decreased sperm count, reduced spawning and fertilization. These impairments mostly result from disruption to sex‐steroid hormones induced by endocrine disruptors. We also discuss other effects of exposure to EDCs. In EDC exposure research, despite incomplete assessments of altered gonad histopathology and sexual behaviour, these present potential effective biomarkers or pathways for evaluating the reproductive function in zebrafish on EDC exposure. To date, the pernicious effects of some EDCs on the reproductive performance in laboratory zebrafish are well understood; however, similar alterations remain for further determination in wild‐type fish and more kinds of EDCs. More studies should be performed under established scientific regulatory criteria to investigate the impact of EDCs on reproduction in zebrafish. Moreover, further research is required to explain the definite mechanism of sexual differentiation, which helps in understanding the shift of sexual phenotype with EDC exposure.     

Status of giant clam resources around Okinawa‐jima Island,Ryukyu Archipelago,Japan

1. Giant clams provide and support valuable functions to coral reefs, as well as represent a sustainable resource for traditional fisheries throughout the Indo‐Pacific region. The Ryukyu Archipelago (southern Japan) is known to be the northern latitudinal limits of giant clam distribution, but there is only limited information in the literature regarding species diversity, status, and distribution in this region.
2. In this study, we report findings from a rapid survey in 2016, the first of its kind for the Ryukyu Islands, to determine species distribution and abundance of giant clams (tridacnines) around Okinawa‐jima Island.
3. Results indicate the presence of four species with an overall density of 5.03 per 100 m², from most to least abundant: Tridacna crocea, Tridacna maxima, Tridacna squamosa, and Tridacna noae. The previously reported species Tridacna gigas and Hippopus hippopus were both absent from the surveys. The densities and distributions of tridacnines varied among species and sites, which are likely attributable to efforts in replenishing and protecting stocks of selected species.
4. The most abundant species, T. crocea, is an important fishery species in Okinawa, and has been widely cultured and restocked to augment depleted populations. In comparison, restocking efforts for less popular species, such as T. squamosa, have been limited, and their current sizes and numbers suggest recruitment constraints.
5. Given the importance of the giant clam fishery in this region and the current declines of various species (except T. crocea), further regulations should focus on the protection of larger and mature clams that function as broodstock necessary to maintain spawning and natural recruitment.

     

Distribution and habitat characteristics of three sympatric cetacean species in the coastal waters of Matang,Perak, Peninsular Malaysia

1. Coastal cetaceans in Southeast Asia are poorly studied and are particularly vulnerable to anthropogenic threats, especially in intensive fishing grounds.
2. To investigate the distribution and habitat characteristics of cetaceans in the productive coastal waters of Matang, Perak, Malaysia, boat‐based line transect surveys were conducted between 2013 and 2016.
3. The Irrawaddy dolphin (Orcaella brevirostris) was most frequently encountered at 3.87 sightings per 100 km, followed by the Indo‐Pacific finless porpoise (Neophocaena phocaenoides) at 1.72 sightings per 100 km, and the Indo‐Pacific humpback dolphin (Sousa chinensis) at 0.66 sightings per 100 km.
4. The mean group size was largest for humpback dolphins (8.4 individuals), followed by Irrawaddy dolphins (6.4 individuals), and finless porpoises (2.8 individuals).
5. Humpback dolphins exhibited a clustered distribution concentrated mainly in shallow estuarine waters (<10 m deep and <5 km from river mouths), whereas Irrawaddy dolphins were more widely distributed in farther coastal waters (<15 m deep and <15 km from river mouths), and finless porpoises were mostly found farthest from the shore in coastal waters (10–25 m deep and >15 km from river mouths).
6. The spatial distribution of the three cetaceans overlapped minimally, and this is likely to reflect the distribution of preferred prey resources, species interactions, and their differential responses to anthropogenic activities and species dominance.
7. The results from our study serve as baseline information for future research, conservation, and habitat management of these vulnerable and endangered coastal cetaceans. Conservation actions are recommended for the Matang area.

     

Conservation of freshwater wildlife in Hong Kong: A genetic perspective

1. Freshwater fauna is highly threatened owing to various human disturbances and a lack of natural connectivity among populations. Immediate conservation actions are vital to save freshwater wildlife from population decline and extinction. Given the paucity of resources, conservation prioritization is essential to guide resource allocation for effective and practical management; however, the incorporation of quantitative ranking approaches in population genetic studies is highly limited.
2. This study scrutinized the genetic diversity and examined the population structure of three invertebrates (the atyid shrimp Caridina cantonensis, the pachychilid snail Sulcospira hainanensis, the palaemonid shrimp Macrobrachium hainanense) and five fishes (the Chinese barb Barbodes semifasciolatus, the paradise fish Marcopodus opercularis, the Hong Kong paradise fish Macropodus hongkongensis, the Beijiang thick‐lipped barb Acrossocheilus beijiangensis, and the rosy bitterling Rhodeus ocellatus) in Hong Kong, based on mitochondrial cytochrome c oxidase subunit I gene and control region, respectively. In general, the three fish species of conservation concern (M. hongkongensis, A. beijiangensis, and R. ocellatus) showed low to no genetic diversity while the other species showed high genetic diversity and strong population differentiation.
3. This study also investigated the use of Heightened Evolutionary Distinctiveness (HED) estimation on quantitatively prioritizing the 95 populations for conservation, including three previously studied species (the white cheek goby Rhinogobius duospilus, the sucker‐belly loach Pseudogastromyzon myersi, and the striped loach Schistura fasciolata). Only five sites had relatively high normalized HED values (>0.70) for all taxa occurring there, and thus are of relatively higher conservation priority. Many sites harboured evolutionarily distinct populations for only one to three species; therefore, a general plan for all freshwater inhabitants cannot be formulated.
4. This comprehensive comparative study highlights the importance of genetic attributes in conservation decision making and sheds light on plausible conservation management in Hong Kong and other highly developed areas with significant freshwater biodiversity.

     

Schisandrin B protects against solar irradiation-induced oxidative injury in BJ human fibroblasts

The effects of schisandrin B (Sch B) and its analogs on solar irradiation-induced oxidative injury were examined in BJ human fibroblasts. Sch B and schisandrin C (Sch C) increased cellular reduced glutathione (GSH) level and protected against solar irradiation-induced oxidative injury. The photoprotection was paralleled by decreases in the elastases-type protease activity and matrix-metalloproteinases-1 expression in solar-irradiated fibroblasts. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production. The results suggest that by virtue of its pro-oxidant action and the subsequent glutathione antioxidant response, Sch B or Sch C may offer the prospect of preventing skin photo-aging.     

Application of Two-Level Full Factorial Design for the Extraction of Fucoxanthin and Antioxidant Activities from Sargassum siliquosum and Sargassum polycystum

Two-level full factorial design was employed to identify the extraction parameters that can improve the derivation of fucoxanthin content (FC), total carotenoid content (TCC), and antioxidant from two brown seaweeds, Sargassum siliquosum (SS) and Sargassum polycystum (SP). These parameters included temperature (A: 4–45°C), time (B: 30–1,440 min), and solvent-to-solid ratio (C: 10–50 ml/g). Antioxidant activities were determined as trolox equivalent antioxidant capacity (TEAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, and ferric-reducing antioxidant power (FRAP). Results showed that all three factors were significant (p < 0.05) in providing higher FC in both species. These factors were also significant in obtaining higher TCC in SS; whereas in SP, TCC was only affected by solvent-to-solid ratio. Only temperature was found to contribute significantly to a higher TEAC in both species. However, none of the factors improved DPPH for SS, except temperature and time for SP. For SS, only time was significant in obtaining higher FRAP; whereas temperature and time were significant for SP. Hence, results indicate that a simple modification in the extraction temperature, time, and solvent-to-solid ratio will be able to improve the derivation of fucoxanthin, carotenoids, and antioxidant activities.     

Cloning of a novel phytase from an anaerobic rumen bacterium,Mitsuokella jalaludinii,and its expression in Escherichia coli

The full length phytase gene of Mitsuokella jalaludinii was successfully cloned and was found to be 1 047 bp in length, with 348 amino acids, and was designated as PHY7 phytase gene. A comparison of the sequence of PHY7 phytase gene of M. jalaludinii with various microbial phytase gene sequences showed that it was not similar to those from other bacteria except Selenomonas ruminatium, thus suggesting that they may both express a new class of phytase. The PHY7 phytase gene was subsequently subcloned into bacterial expression vector, p ET32 a, for expression in Escherichia coli strain Rosetta-gami. Expression of the recombinant phytase gene was optimised and characterised. The recombinant phytase was estimated to be approximately 55 k Da by SDS-PAGE analysis. The recombinant phytase exhibited optimum activity at 55°C, p H 4.5 and showed good p H stability from p H 3.5 to 5.5(78% relative activity). Metal ions such as Ca2+, Mg2+, and K+ were found to exert significant stimulatory effect on the recombinant phytase activity while Cu2+, Fe3+, and Zn2+ greatly inhibited the enzyme activity. The recombinant phytase showed moderate resistance to trypsin proteolysis, but susceptible to pepsin proteolysis. The results of the study showed that several characteristics of recombinant phytase were slightly different from the native enzyme. Unfavourable characteristics such as reduced p H stability and metal ion effects should be taken into consideration during feed enzyme formulation.     

Schisandrin B co-treatment ameliorates the impairment on mitochondrial antioxidant status in various tissues of long-term ethanol treated rats

The effect of schisandrin B (Sch B) on long-term ethanol-induced oxidative stress in various rat tissues was investigated. Long-term ethanol treatment increased reactive oxygen metabolites (ROM) level in plasma. The ethanol-induced oxidative stress was assessed by mitochondrial glutathione and α-tocopherol levels, antioxidant enzyme activities, malondialdehyde (mtMDA) production and heat shock protein (Hsp) 25/70 levels. Liver was most susceptible to oxidative stress with a significant increase in mtMDA production. Long-term Sch B treatment enhanced mitochondrial antioxidant status in a tissue non-specific manner. Sch B co-treatment ameliorated the alterations in plasma ROM levels, mtMDA production and Hsp 25/70 expression in rat tissues.     

High-efficiency regnerationin vitro from potato petioles with intact leaflets

The shoot/plantlet regenerationin vitro of seven potato (Solanum tuberosum L.) cultivars from petioles with intact leaflets was assessed using six treatment combinations-a basal medium with or without silver thiosul-phate (STS) or thidiazuron (TDZ) at two concentrations (2 or 0.5 mg/l) of the indoleacetic acid (IAA). The basal medium consisted of Murashige and Skoog (MS) salts and vitamins supplemented with 3 mg/1 6-benzylaminopurine, and 1 mg/1 gibberellic acid, 30 g/l sucrose, and 7.0 g/l PHYTOAGAR. Two full sets repeats and one partial set repeat of independent experiments were conducted and all produced similar results. Silver thiosulphate decreased the regeneration frequency and number of shoots per callus. No significant changes were observed with thidiazuron. Regeneration rates of (100% ) with up to 20 shoots/plantlets per callus were achieved at 2 mg/1 IAA with cultivars Désirée, Kennebec, Niska, and Lenape. These cultivars still showed high regeneration rates (87%–98% ) on media with 0.5 mg/1 IAA, and good regeneration rates were also achieved by the other three cultivars (48%, 94%, and 50% for Chieftain, Russet Burbank, and Shepody, respectively). Even with the single medium protocol (0.5% IAA without thiosulphate or thidiazuron), Désirée, Lenape, and Niska exhibited a regeneration rate of 98%. The use of petiole-with-leaflet explants could be ideal for the regeneration step of potato genetic transformation protocol because of their high regeneration efficiency and their small cut surface area forAgrobacterium elimination after co-incubation.