Mass balance of anti-influenza drugs discharged into the Yodo River system,Japan, under an influenza outbreak

In February 2011, at the peak of an influenza outbreak, we performed a comprehensive analysis of the mass balances of four anti-influenza drugs—oseltamivir (OS), oseltamivir carboxylate (OC), amantadine (AMN), and zanamivir (ZAN)—in the urban area of the Yodo River system. This area includes three main river catchments (the Katsura, Uji, and Kidzu Rivers) and is home to 12 million people, about 10% of Japan’s population. Water was sampled at six main rivers and 13 tributary sites and eight sewage treatment plants (STPs). We concluded that the STP effluents were the major sources of the anti-influenza drug load in the Yodo River system (68–94% of total mass fluxes). Extended measurement throughout the Yodo River system further showed only small fluctuations of the ratio of OS to OC from 0.2 to 0.3, suggesting that OS and its metabolite are environmentally stable. The results also clearly showed the importance of reducing the levels of anti-influenza drugs in the water environment by reducing their emission at STPs.     

Development of extraction method of pharmaceuticals and their occurrences found in Japanese wastewater treatment plants

In this study, occurrence of 66 PPCPs (pharmaceuticals and personal care products) in liquid and solid phases of sewage sludge was elucidated. The extraction methods for the PPCPs from sludge were newly developed employing Pressurized Liquid Extraction (PLE) and Ultrasonic Solvent Extraction (USE). As an appropriate method, PLE using water (pH2), PLE using methanol (pH4), and USE using mixture of methanol and water (1/9,v/v, pH11) was found most effective because total recovery of most of the PPCPs indicated 40 to 130%. The developed extraction method with previously developed method for liquid phase analysis was applied to field survey at wastewater treatment plants (WWTPs) in Japan. 56 compounds were detected from the primary sludge and 61 compounds were detected from the excess sludge. The concentration was ranged between several ng/g and several μg/g. Solid-water distribution coefficient (Log K_d) ranged between 0.9 L/kg (Caffeine) and 3.7 L/kg (Levofloxacin) for primary sludge and between 1.4 L/kg (Sulpirid) and 4.3 L/kg (Mefenamic acid) for excess sludge.     

Fluorinated alkyl compounds including long chain carboxylic acids in wild bird livers from Japan

A wide range of fluorinated alkyl compounds (FACs) has been reported in wildlife in various locations in the world. However, such information regarding Japanese wildlife is rarely found. In the present study, we investigated the occurrence of 21 FACs, including perfluorinated alkyl sulfonates (PFASs), perfluorinated carboxylates (PFCAs), and fluorotelomer acids, in the livers of 10 wild bird species from two regions in northern Japan. To avoid interferences, FACs were quantified by a recently developed method using acetonitrile and solid-phase extraction followed by an ion exchange HPLC column separation. Apart from perfluorooctane sulfonate (PFOS), which was found at the highest levels of all the compounds detected, several long chain perfluorinated carboxylates (PFCAs) from C8 to C16, particularly perfluorotetradecanoic acid (PFTeDA) and perfluorohexadecanoic acid (PFHxDA), were detected for the first time. Additionally, 7:3 FTCA, a fluorotelomer acid, was also detected in most swan livers from Miyagi prefecture and all the birds from Tochigi prefecture. However, none of the sulfonamides and unsaturated telomer acids were detected in any species. Swans seem to be the least exposed wild birds to FACs among the investigated birds, signifying that feeding habits may reflect FAC accumulation in wild birds. The highest total concentration of detected FACs was 405 ng g⁻¹ wet wt., which was found in a Japanese sparrowhawk, indicating that the top predatory wild birds can accumulate several long chain carboxylic acids. However, the current FAC concentrations found in livers may suggest that these compounds alone would not cause a severe toxic effect in these species.     

Bioremediation of cadmium contaminated soil using symbiosis between leguminous plant and recombinant rhizobia with the MTL4 and the PCS genes

Cadmium contamination in rice grains is one of the important issues in Asian countries. We have developed a novel bio-remediation system based on the symbiosis between leguminous plant and genetically engineered rhizobia. We designed two types of recombinant rhizobia, carrying two genes, synthetic tetrameric metallothionein (MTL4) and cDNA encoding phytochelatin synthase from Arabidopsis thaliana (AtPCS). The MTL4 and AtPCS genes were transferred to Mesorhizobium huakuii subsp. rengei B3, which can infect and form nodules on Chinese milk vetch, Astragalus sinicus. The two genes were fused to the nolB or nifH promoter, which generated nodule specific expression of these genes in strain B3. The two recombinant strains, B3(pMPnolBMTL4nifHPCS) and B3::nifHMTL4(pMPnifHPCS), showed 25 and 12-fold increase in Cd concentration, in the free-living cells, respectively. When these recombinant strains established the symbiotic relationship with A. sinicus, the symbionts increased Cd accumulation in nodules by two-fold in hydroponic culture. The expression of the both MTL4 and AtPCS genes showed additive effect on cadmium accumulation in nodules. We also applied these recombinant bacteria to rice paddy soil polluted with Cd (1mgkg(-1) dry weight soil). The accumulation of Cd increased not only in nodules but also in the roots of A. sinicus infected by the recombinant rhizobia. The accumulation of Cd in the plant roots infected by B3(pMPnolBMTL4nifHPCS) achieved three-fold than that by the wild-type B3. After two months of cultivation of the symbiont, a maximum of 9% of Cd in paddy soil was removed. Thus, the symbiosis will be useful in phytoremediation for heavy metals.