Newly-hatched Epinephelus coioides larvae were stocked in five 5-ton tanks at an initial density of 25,000 ind/tank. Copepod nauplii were propagated in four of these tanks by inoculating various densi ties (20 to 80 ind/1) of mixed copepodids of Acartia tsuensis, Pseudodiaptomus spp., and Oithona sp. three days before stocking larvae. Rotifers were added in these tanks on Day 7 at an initial density of 5,000 ind/1. Larvae in the remaining tank were fed rotifers (only) starting Day 2 at 5,000 ind/l. The feeding incidence, gut content, growth, and survival of larvae were better in tanks with higher density of copepodids (60-80 ind/l). These indices were lowest in larvae given rotifers only. Total n-3 HUFA of copepods was 2 to 3 times higher than rotifers. High percentages of 22:6n-3 (DHA) were detected in the fatty acid composition of Pseudodiaptomus (13%) and Acartia (24%) with DHA/EPA (20:5n-3) values of 1.4 and 2.6, respectively. By providing nauplii of copepods at the early feeding stage, an average survival of 3.4% at harvest (Day 36) was obtained in a pilot scale grouper seed production trial in three 10-ton tanks.

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Cellular incorporation of Cd involves multiple transport systems for other metals such as Fe, Zn, Mn, and Ca. Metal transporters including divalent metal transporter 1, Zrt/Irt-related protein (ZIP) 8, and ZIP14, and certain types of voltage-dependent Ca channels have been shown to be involved in cellular Cd uptake. However, tissue- or cell-specific roles of these metal transporters in the accumulation and toxicity of Cd remains unclear. In the present study, we compared the sensitivity to and accumulation of Cd, Mn, and Zn among four types of rat cell lines. Rat basophilic leukemia RBL-2H3 cells showed the highest sensitivity to Cd and Mn due to the highest accumulation of Cd and Mn among the four cell lines. The high accumulation of Cd and Mn was caused by high uptake rates of Cd and Mn. Since relatively high expression of ZIP8 and ZIP14 was found in RBL-2H3 cells, siRNAs of ZIP8 and ZIP14 were transfected into RBL-2H3 cells. The knockdown of ZIP8, but not of ZIP14, significantly reduced the uptake rates of Cd and Mn in RBL-2H3 cells, especially in the presence of bicarbonate. These results suggest that the high expression of ZIP8, which is known to have affinities for both Cd and Mn, resulted in high accumulation of Cd and Mn, leading to high sensitivity to these metals in RBL-2H3 cells. Thus, RBL-2H3 cells may serve as a good model for clarifying the mechanisms of Cd and Mn transport via ZIP8.

Pfs230, surface protein of gametocyte/gamete of the human malaria parasite, Plasmodium falciparum, is a prime candidate of malaria transmission-blocking vaccine. P. vivax has an ortholog of Pfs230 (Pvs230), however, there has been no study in any aspects on Pvs230 to date. To investigate whether Pvs230 can be a vivax malaria transmission-blocking vaccine, we performed evolutionary and population genetic analysis of the Pvs230 gene (pvs230: PVX_003905). Our analysis of Pvs230 and its orthologs in seven Plasmodium species revealed two distinctive parts: an interspecies variable part (IVP) containing species-specific oligopeptide repeats at the N-terminus and a 7.5 kb interspecies conserved part (ICP) containing 14 cysteine-rich domains. Pvs230 was closely related to its orthologs, Pks230 and Pcys230, in monkey malaria parasites. Analysis of 113 pvs230 sequences obtained from worldwide, showed that nucleotide diversity is remarkably low in the non-repeat 8-kb region of pvs230 (θπ = 0.00118) with 77 polymorphic nucleotide sites, 40 of which resulting in amino acid replacements. A signature of purifying selection but not of balancing selection was seen on pvs230. Functional and/or structural constraints may limit the level of polymorphism in pvs230. The observed limited polymorphism in pvs230 should ground for utilization of Pvs230 as an effective transmission-blocking vaccine.

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