Extractives ofJuniperus chinensis L. I: Isolation of podophyllotoxin and yatein from the leaves ofJ. chinensis

Two compounds, yatein and podophyllotoxin, were isolated from the chloroform-soluble fraction in the methanolic extractives of byakushin (Juniperus chinensis L.) leaves for the first time.This report was presented at the 47th annual meeting of the Japan Wood Research Society, Kochi, 1997     

Faster growth before metamorphosis leads to a higher risk of pseudoalbinism in juveniles of the starry flounder <Emphasis Type="Italic">Platichthys stellatus</Emphasis>, as suggested by otolith back-calculation

The mechanism underlying malformation during flatfish metamorphosis is not clear. Here, the relationship between growth before metamorphosis and the occurrence of malformation after metamorphosis was examined in individuals reared in a single tank. Otoliths of starry flounder Platichthys stellatus larvae were labeled with alizarin complexone before metamorphosis [12 days after hatching (DAH)]. The positive linear regression between the lapillus otolith diameter and body length at 12 DAH enabled the estimation of individual larval size after completion of metamorphosis. At 12 DAH, the average body length of the typical pseudoalbino juveniles was significantly greater (5.37 mm) than that of the normal juveniles (5.13 mm) (P < 0.01, n = 100). In addition, of the larvae that were estimated to measure less than 4.8 mm at 12 DAH, approximately 80% metamorphosed into normal juveniles, whereas more than 60% of the larvae that measured more than 5.4 mm became pseudoalbino juveniles. These results indicate that the larvae that grow rapidly during the first 12 days are more likely to become pseudoalbino individuals after metamorphosis.     

Precocious sex change and spermatogenesis in the underyearling Malabar grouper Epinephelus malabaricus by androgen treatment

The Malabar grouper Epinephelus malabaricus is an important candidate species for commercial aquaculture in tropical and subtropical areas. In nature, this species requires more than 10 years to change sex from female to male and have active spermatogenic tissues in the testis. Thus, it is essential to find a means of producing sperm for seed production. This is the first report of artificial sex change in underyearling E. malabaricus . Female E. malabaricus with immature ovaries at 144 days post-hatch (DPH) were fed a diet with 17α-methyltestosterone (MT) at 50 μg g⁻¹ diet for 6 months. Sex change occurred in most of the treated fish, which had testis with all stages of spermatogenic germ cells including spermatozoa. In contrast, most of the control fish had immature ovaries. These results, which reveal that germ cells in the underyearling grouper have the ability to produce spermatozoa in response to exogenous androgen, demonstrate that sex change can be artificially induced during ovarian development.     

Social capital accumulation through public policy systems implementing paddy irrigation and rural development projects

In Japan, a developed country in Asia, the Land Improvement Act systems have worked as a measure of social capital accumulation as well as an adjective law in implementing irrigation projects since 1949. This paper illustrates the status and problems in applying the concept of social capital to agricultural and rural development policy systems in developing countries, EU and Japan. The results obtained are summarized as follows: (1) it is desirable that governments formulate public policies that are appropriate for correcting inefficiencies in resource distribution for accumulating social capital, so that the governments play a significant role in developing and supporting social capital; (2) the effect of policies in the areas, where government-supported land improvement projects have been implemented, has been improved, subsequent to which the efficient use of the national budget allocated for the projects has been improved, while the projects significantly contribute to the national land conservation and social stability by continuously promoting minimum social capital accumulation nationwide. In addition, Japan and countries in the Asian monsoon region, which hold the five elements in common that constitute the characteristics of rice field irrigation in humid climates, should take over the advantages of the land improvement project system while learning from the success achieved by the LEADER+ programme in Europe, where society precedes Asian countries concerning the problems of decline in the birth rate, and expansion of an aging society.

Determination of sex and gonadal maturity in the honeycomb grouper, <Emphasis Type="Italic">Epinephelus merra</Emphasis>, through biopsy

Determination of the sex and gonadal maturity of fish is necessary for successful commercial aquaculture operations. Sex identification is problematic in grouper fish, because they change sex at certain ages or sizes. Here we represent an accurate and reliable method for sex determination of the live grouper, using 5–10 mg gonadal tissue samples, harvested using non-lethal gonadal biopsy. Sex and sexual maturity of biopsied individuals were determined using standard histological method. All biopsied fish survived and there were no serious infections resulting from the surgery. Therefore, this biopsy technique is a simple and cost-effective tool for grouper fishery management.     

Change in antimicrobial resistance pattern in Salmonella enterica serovar Typhimurium isolates detected in a beef cattle farm

Multidrug-resistant Salmonella enterica serovar Typhimurium (S. Typhimurium) isolates with four different antimicrobial resistance patterns obtained from a beef cattle farm were characterized to determine their clonality. Macrorestriction analysis of genomic DNA revealed that these four isolates are closely related to each other and can be classified as a newly emerged pulsed-field gel electrophoresis type among cattle: cluster VII. Three of the four isolates showed resistance to extended-spectrum cephalosporins (ESCs), and this resistance was mediated by AmpC β-lactamase encoded by the bla(CMY-2) gene in a 190-kbp IncA/C plasmid. Results of restriction analysis and IncA/C backbone PCR suggest that the three 190-kbp plasmids are identical and that a 70-kbp IncA/C plasmid of the ESC-susceptible isolate is derived from the 190-kbp plasmid by a deletion event. Three isolates harboured a virulence-resistance plasmid (165 or 180 kbp), and restriction analysis revealed that these plasmids were identical or closely related to each other. These results suggest that the four S. Typhimurium cluster VII isolates originate from a common ancestor that probably invaded the farm prior to the salmonellosis outbreak. Antimicrobial resistance patterns may not necessarily reflect the relationships of the isolates.     

Prevalence and mechanism of antimicrobial resistance in Staphylococcus aureus isolates from diseased cattle, swine and chickens in Japan

Antimicrobial administration is essential for the control and treatment of diseases in animals, but the emergence and prevalence of antimicrobial-resistant Staphylococcus aureus is a significant concern during animal production. Here we investigated the antimicrobial susceptibility of S. aureus from diseased food-producing animals and molecularly characterized the methicillin-resistant and fluoroquinolone-resistant isolates. A total of 290 S. aureus isolates obtained from cattle (n=246), swine (n=16), and chickens (n=28) between 2003 and 2009 were examined for antimicrobial susceptibility against 9 antimicrobials using an agar dilution method. Resistance to penicillin (PC) was most frequently found (24.8%), followed by oxytetracycline (OTC, 10.0%), dihydrostreptomycin (4.1%), erythromycin (EM, 3.1%), enrofloxacin (ERFX, 2.1%), and kanamycin (1.7%). The PC resistance rate was significantly higher in swine than in cattle (P<0.01) and chickens (P<0.01). The resistance rates to OTC, EM and ERFX were significantly higher in swine and chickens than in cattle (P<0.05). Methicillin-resistant S. aureus (MRSA) was recovered from milk derived from a cow with mastitis in 2003; sequence type 8, SCCmec type IV and spa type t024. In the six ERFX-resistant strains isolated after 2003, amino acid substitutions in ParC with/without GyrA were detected. As the prevalence of MRSA and FQ-resistant S. aureus in the animals should be noticed, continuous monitoring is necessary to control resistance to clinically important antimicrobials in S. aureus from food-producing animals.     

Growth suppression of human cancer cells by polyphenolics from sweetpotato (Ipomoea batatas L.) leaves

Sweetpotato leaves (Ipomoea batatas L.) contain a high content of polyphenolics that consist of caffeic acid, chlorogenic acid, 3,4-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, 4,5-di-O-caffeoylquinic acid, and 3,4,5-tri-O-caffeoylquinic acid. We investigated the suppression of the proliferation of selected human cancer cells by phenolic compounds isolated from sweetpotato leaf. The human cancer cells used in this research included a stomach cancer (Kato III), a colon cancer (DLD-1), and a promyelocytic leukemia cell (HL-60). Caffeic acid and di- and tricaffeoylquinic acids dose-dependently depressed cancer cell proliferation, and the difference in sensitivity between caffeoylquinic acid derivatives and each kind of cancer cell was observed. Specifically, 3,4,5-tri-O-caffeoylquinic acid effectively depressed the growth of three kinds of cancer cells, and caffeic acid had an exceptionally higher effect against HL-60 cells than other di- and tricaffeoylquinic acids. In attempting to clarify the mechanism of growth suppression with the addition of the apoptotic inhibitor N-ethylmaleimide, we observed that the nuclear granulation in 3,4,5-tri-O-caffeoylquinic acid-treated HL-60 cells suggested apoptosis induction. This effect was confirmed by DNA fragmentation, an increase of caspase-3 activity, and expression of c-Jun. Growth suppression of HL-60 cells by 3,4,5-tri-O-caffeoylquinic acid was determined to be the result of apoptotic death of the cells. These results indicate that 3,4,5-tri-O-caffeoylquinic acid may have potential for cancer prevention.