Molecular characterization of coral sulfate transporter homolog that is up-regulated by the presence of symbiotic algae

Reef-building corals are in symbiosis with the dinoflagellates Symbiodinium spp. In our previous study, the expression of two mRNAs (AtSym-01 and 02) was up-regulated by the presence of Symbiodinium cells (strain PL-TS-1) in juveniles of the reef-building coral Acropora tenuis. In this study, the AtSym-01 mRNA was found to encode a cnidarian ortholog of the vertebrate SLC26A11 sulfate transporter. The AtSym-01 and human SLC26A11 proteins exhibited 46% identity over 542 amino acids. Real-time polymerase chain reaction analysis showed that the expression level of the AtSym-01 mRNA was also increased by the presence of Symbiodinium strain CCMP2467 cells. Immunohistochemical analysis was performed using polyclonal antibodies against the AtSym-01 protein, in order to study the distribution of the protein in A. tenuis tissues. Cell-specific immunoreaction was observed in diverse tissues in juvenile and adult specimens. Notable immunoreaction was observed in mucocytes (mucus cells) in the outer epithelium of juveniles, and gastrodermal cells located between the coelenteron and skeleton of the adult colony. These observations suggest the possibility that the AtSym-01 protein is involved in uptake of sulfate ion for synthesis of sulfated macromolecules that are contained in the mucus and organic matrix of the calcified skeleton.     

Development of novel intracytoplasmic sperm injection and somatic cell nuclear transfer techniques for animal reproduction

Animal biotechnology has made new biological experiments possible and new discoveries are being made on an almost daily basis. Among these discoveries is a method for directly injecting a spermatozoa or somatic nucleus into an oocyte that has brought a revolution in the world of micromanipulators. Experiments that were unfeasible until now have become possible, and normal offspring can be generated from infertile cells, such as using dead sperm or a dead frozen body. In this review, I will introduce the progress of animal reproductive biotechnology, including our current research.     

Full-term development of offspring using round spermatids produced ectopically from fetal male germ cells

The continuous production of mammalian sperm is maintained by the proliferation and differentiation of spermatogonial stem cells, which originate from primordial germ cells in the early embryo. Previously, we reported that the transplantation of fetal male gonadal tissue into the recipient testis was effective obtaining functional sperm. This transplantation technique is a promising new approach for the preservation of testicular function in a mutant animal with embryonic lethality. In the present study, we examined whether spermatogenesis from fetal male germ cells is induced under ectopic conditions in male and female recipients. Nine to 10 weeks after the transplantation of male gonads prepared from embryos at 12.5 or 16.5 days post gestation, male germ cell differentiation occurred under the skin of male and female recipient nude mice. Histological analyses revealed that grafted gonads contained haploid germ cells such as round or elongated spermatids. Furthermore, we succeeded in obtaining normal progeny by injecting the ectopically produced round spermatids into the cytoplasm of oocytes, even when the male germ cells had been generated in female recipients. These results indicate that the transplantation of fetal male gonads under the skin of recipient mice is a useful technique for obtaining functional male gametes.     

In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus and other vibrios

Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.     

Effect of fluorescent mercury light irradiation on in vitro and in vivo development of mouse oocytes after parthenogenetic activation or sperm microinjection

The detection of specific cellular components using fluorescent agents such as green fluorescent protein (GFP), red fluorescent protein or Hoechst dyes provides a powerful tool for studying cell biology. However, specimens must be exposed to high-intensity light, which might cause cellular damage. Here, we exposed mouse metaphase stage (M) II oocytes to fluorescent mercury vapor light at three wavelengths (539 nm, 488 nm and 341 nm) to determine the maximum exposure time that would avoid damage. When oocytes were activated parthenogenetically after exposure to these wavelengths for more than 20 min, 5 min or 4 sec, respectively, the percentages of dead oocytes after activation increased, and none of the surviving embryos developed to blastocysts. However, embryos fertilized by intracytoplasmic sperm injection (ICSI) were more tolerant to light damage, even though the quality of blastocysts, judged by cell number and cell allocation to the inner cell mass and trophectoderm measured by immunostaining for Oct4 and Cdx2, was reduced as exposure times increased. Live, healthy offspring were obtained when these exposed embryos were transferred into recipient pseudopregnant females at the 2-cell stage. In addition, MII oocytes collected from GFP-expressing transgenic mice after 5 min of irradiation with 488-nm light were also able to develop to full term following ICSI. Thus, we determined the safe period of exposure to several wavelengths for oocyte manipulation or observation that would permit subsequent development.     

Ammonia volatilization from Vietnamese acid sulfate paddy soil following application of digested slurry from biogas digester

Paddy and Water Environment - In some areas of Vietnam, digested slurry from biogas digesters is discharged into canals without wastewater treatment. In order to reduce environmental pollution,...     

Effects of carbamate, BPMC, on the central cholinergic functions and behavior of mice

O-sec-butylphenyl methylcarbamate (BPMC), an anticholinesterase carbamate, was injected once (100 mg/kg, s.c.) or repeatedly (50 mg/kg/day for 10 days) into mice. Animals were examined for their behavior and for parameters of cholinergic activity in the forebrain. Mice that received only a single injection exhibited reduced ambulation, hypothermia, and impairment of rotarod performance for up to 3 hr after a single injection. BPMC increased levels of acetylcholine (ACh) in the forebrain for up to 6 hr, and decreased acetylcholinesterase (AChE) activity for up to 24 hr. Both high-affinity choline uptake (HACU) and binding of [3H]quinuclidinyl benzilate (QNB) were reduced 20 min after a single injection without any effect on choline acetyltransferase (ChAT) activity. In behavioral tests conducted 10 min prior to the daily injections, rotarod performance and ambulation were slightly impaired for a few days before and after cessation of injection. Repeated treatment decreased HACU and binding of [3H]QNB for 24 hr after the final injection without any changes in levels of ACh content, AChE activity and ChAT activity. BPMC may reversibly impair cholinergic functions through effects not only on AChE activity but also on HACU and binding of [3H]QNB.     

New analogs of pochonicine,a potent β-N-acetylglucosaminidase inhibitor from fungus Pochonia suchlasporia var. suchlasporia TAMA 87

Three novel analogs of pochonicine (1) were isolated from a solid fermentation culture of the fungal strain Pochonia suchlasporia var. suchlasporia TAMA 87, and their structures were elucidated as 7-deoxypochonicine (2), 6-deoxypochonicine (3), and 6,7-dideoxypochonicine (4). These analogs were found to possess the same stereochemistry as pochonicine. Comparison of β-N-acetylglucosaminidase (GlcNAcase) inhibitory activity between these analogs and pochonicine suggested that the C-6 hydroxy group of pochonicine was essential to its potent GlcNAcase inhibitory activity and that the C-7 hydroxy group also contributed to the activity, but to a lesser extent than the C-6 hydroxy group.     

Construction of a high-density SSR marker-based linkage map of zoysiagrass (<Emphasis Type="Italic">Zoysia japonica</Emphasis> Steud.)

A consensus genetic linkage map with 447 SSR markers was constructed for zoysiagrass (Zoysia japonica Steud.), using 86 F₁ individuals from the cross ‘Muroran 2’ × ‘Tawarayama Kita 1’. The consensus map identified 22 linkage groups and had a total length of 2,009.9 cM, with an average map density of 4.8 cM. When compared with a previous AFLP-SSR linkage map, the SSR markers from each linkage group mapped to similar positions in both maps. Eight pairs of linkage groups from the AFLP-SSR map were joined into eight new groups in the current map. This zoysiagrass consensus map contained 35 SSR markers exhibiting high homology with rice genomic sequences from known chromosomal locations. This allowed synteny to be identified between Zoysiagrass linkage groups 2, 3, 9, 19 and rice chromosomes 3, 12, 2, 7 respectively. These results provide important comparative genomics information and the new map is now available for quantitative trait locus analysis, marker-assisted selection and breeding for important traits in zoysiagrass.     

Evaluation of the inhibitory activities of the extracts of Indonesian traditional medicinal plants against Plasmodium falciparum and Babesia gibsoni

Twenty-four kinds of water extracts derived from 22 plants that are traditionally used for the treatment of malaria on Java Island, Indonesia, were screened for their antibabesial and antimalarial activities. Among the extracts, 8 extracts displayed strong antimalarial activity, with an inhibition range from 89.6 to 100%, and 15 showed strong antibabesial activity, with an inhibition range from 84.2 to 98.1%. The extracts of Achillea millefolium, Baeckea frutenscens, Brucea javanica, Curcuma xanthorrhiza, Strychnos lucida and Swietenia macrophylla showed both strong antibabesial and antimalarial activities. The antimalarial activities paralleled the antibabesial activities, but the converse was not true.