Integration of chemical and toxicological tools to assess the bioavailability of copper derived from different copper-based fungicides in soil.

Because the extensive use of Cu-based fungicides, the accumulation of Cu in agricultural soil has been widely reported. However, little information is known about the bioavailability of Cu deriving from different fungicides in soil. This paper investigated both the distribution behaviors of Cu from two commonly used fungicides (Bordeaux mixture and copper oxychloride) during the aging process and the toxicological effects of Cu on earthworms. Copper nitrate was selected as a comparison during the aging process. The distribution process of exogenous Cu into different soil fractions involved an initial rapid retention (the first 8 weeks) and a following slow continuous retention. Moreover, Cu mainly moved from exchangeable and carbonate fractions to Fe-Mn oxides-combined fraction during the aging process. The Elovich model fit well with the available Cu aging process, and the transformation rate was in the order of Cu(NO3)2 > Bordeaux mixture > copper oxychloride. On the other hand, the biological responses of earthworms showed that catalase activities and malondialdehyde contents of the copper oxychloride treated earthworms were significantly higher than those of Bordeaux mixture treated earthworms. Also, body Cu loads of earthworms from different Cu compounds spiked soils were in the following order: copper oxychloride > Bordeaux mixture. Thus, the bioavailability of Cu from copper oxychloride in soil was significantly higher than that of Bordeaux mixture, and different Cu compounds should be taken into consideration when studying the bioavailability of Cu-based fungicides in the soil.

Morphological and enzymatical observations in Oncomelania hupensis after molluscicide treatment: implication for future molluscicide development.

A preparation of niclosamide named 50 % wettable powder of niclosamide ethanolamine salt (WPN), the only chemical molluscicide available in China, has been widely used for Oncomelania hupensis control over the past 20 years, but its molluscicidal mechanism has not been elucidated yet. Recently, a derivative of niclosamide, the salt of quinoid-2',5-dichloro-4'-nitro-salicylanilide (Liu Dai Shui Yang An, LDS), has been proven to have equivalent molluscicidal effects as WPN but with lower cost and significantly lower toxicity to fish than WPN. In our previous study, gene expression profiling of O. hupensis showed significantly effects after these two molluscicides had been applied. This study was designed to use morphological and enzymological analyses to further elucidate the mechanism by which these molluscicides cause snail death. After WPN or LDS treatment, the number of mitochondria of O. hupensis was reduced and their cristae appeared unclear, heterochromatin gathered to be polarized, ribosome numbers of the rough endoplasmic reticulums (rERs) decreased, myofilaments in muscle cells became disordered and loose, and cytoplasm in some liver cells was concentrated. Damage of cell structures and organelles suggested inhibited movement ability and effects on liver and energy metabolism following treatment. In parallel, activities of enzymes related with carbohydrate metabolism were inhibited except lactate dehydrogenase (LDH) increased in muscle tissue, and activities of enzymes related with stress response increased followed by decreasing to lower levels than those of the H2O-treated group. This shift of carbohydrate metabolism patterns led to insufficient energy supply and lactic acid accumulation, and variations of nitric oxide synthase (NOS), alanine aminotransferase (ALT), and superoxide dismutase (SOD) during process of molluscicide treatment suggested a stress response of snail to the molluscicides at early stages and later fatal damage in liver and nervous system. In general, effects of WPN and LDS were similar although LDS-treated snails showed more serious damage in the liver and a stronger inhibition of enzymes related with aerobic respiration and stress response. This was consistent with the transcriptome profile obtained previously. However, considering enzyme activities at post-transcriptional and protein levels, comprehensive identification and annotation of potential enzyme-related genes and regulation pattern would be necessary to provide great benefit for understanding of potential mechanism of these molluscicides and even for future molluscicide development.

[Preparation and Molluscicidal Effect of Chlorosalicylicamide Sustained-release Granules].

Objective: To prepare the 5% and 10% chlorosalicylicamide（quinoid-2', 5-dichloro-4'-nitrosalicylanilide from niclosamide） sustained-release granules （LDS-SRG） and evaluate their molluscicidal effect. Methods: The 5% and 10% LDS-SRG were prepared with screened carriers, surfactants, adhesives, defoamers and lubricants. Their bulk density, water content, repose angle, critical relative humidity, thermal stability and release rate were determined. Spraying method was used to test the molluscicidal effect of LDS-SRG at 1.6 g/m2. Meanwhile, 50% wettable powder of niclosamide ethanolamine salt (WPN) was applied as the positive control at 1.0 g/m2, and dechlorinated water was used as the blank control. The mortality of snails was calculated on days 3, 7 and 14 after administration. Results: The 5% and 10% LDS-SRG were red brown in color, showed good fluidity, and had bulk density of 0.655 g/ml and 0.594 g/ml, moisture content of 1.15% and 1.28%, repose angle of 39.8° and 39.7°, and critical relative humidity of 64.98% and 61.63%, respectively. Moreover, both showed good thermal stability. The release curve was stable for both 5% and 10% LDS-SRG during day 1 to day 9, and faster release for 5% LDS-SRG than for 10% LDS-SRG. The burst release occurred on days 10 and 15, and the steady release occurred from days 14 and 20 for 5% and 10% LDS-SRG respectively. The snail mortality on day 7 after 5% LDS-SRG 1.6 g/m2 administration and on day 14 after 10% LDS-SRG 1.6 g/m2 administration was both higher than 95%, and higher than that of the 50% WPN 1.0 g/m2 control (P<0.05). Conclusion: The 5% and 10% LDS-SRG show sustained-release potential and satisfactory molluscicidal effect by spraying, reaching the evaluation standard for molluscicidal agents.

[The Molluscicidal Effect of Chlorosalicylicamide Sustained-release Granules on Oncomelania hupensis].

Objective: To evaluate the molluscicidal effect of the chlorosalicylicamide sustained-release granules (LDS-SRG) on Oncomelania hupensis. Methods: Seven effective concentrations or dosages of LDS-SRG, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2 and 6.4 mg/L （for immersion test） or g/m2（for spraying test）, were prepared from the original 5% and 10% concentrations or dosages in the laboratory. In the immersion test, each concentration of LDS-SRG was incubated with 3 packs of snails（30 snails in each pack）, and each pack was taken for snail counting at 24, 48 and 72 h respectively. In the spraying test, each dosage of LDS-SRG was applied to 200 snails, and the snail mortality was calculated in 50 randmoly collected snails on days 3 and 7, and in the whole on day 14 after administration. In the field immersion test, LDS-SRG at concentrations of 0.4, 0.8 and 1.6 g/m3 was incubated with 6 packs of snails （30 snails in each pack）, and each 2 packs were taken at 24, 48, and 72 h to calculate the snail mortality. In the field spraying test, 0.8, 1.6 and 3.2 g/m2 LDS-SRG was sprayed in 3 snail-positive ditches （～100 m2）, and 10 boxes of snails were selected in each ditch on days 3, 7 and 14 to calculate the snail mortality. The 50% wettable powder of niclosamide ethanolamine salt （WPN） with effective concentrations or dosages of 1.0 mg/L （or g/m2 and g/m3） was used as the positive control. Fresh water served as the blank control. Results: In the labratory immersion test using the original concentration of 5%, both 0.1-6.4 mg/L LDS-SRG for 72 h and 1.6-6.4 mg/L LDS-SRG for 48 h caused 100% mortality; and the concentration lethal to 50% （LC50） at 24, 48 and 72 h was 0.70, 0.01 and 0.01 mg/L respectively. When using the original concentration of 10%, both 0.1-6.4 mg/L LDS-SRG for 72 h and 0.2-6.4 mg/L LDS-SRG for 48 h caused 100% mortality; and the LC50 at 24, 48 and 72 h was 0.15, 0.01 and 0.01 mg/L respectively. The labratory spraying test showed that 7-day administration of 1.6 and 6.4 g/m2 LDS-SRG as well as 14-day administration of 3.2 and 6.4 g/m2 LDS-SRG prepared from 5% dosage, resulted in a snail mortality>95%, with the LD50 on days 3, 7 and 14 being 0.06, 0.16, and 0.18 g/m2; 14-day administration of 1.6 g/m2 LDS-SRG as well as 7-day administration of 6.4 g/m2 LDS-SRG prepared from 10% dosage, resulted in a snail mortality>95%, with the LD50 on days 3, 7 and 14 being 3.29, 0.75, and 0.16 g/m2. The mortality by various dosages of LDS-SRG prepared from 5% dosage was significantly higher than that of the control group (P<0.05). In the field immersion test, the snail mortality by 1.6 g/m3 LDS-SRG prepared from 5% and 10% concentrations for 72 h was 96.43% and 98.21% respectively (P>0.05 versus the control group). In the field spraying test, the snail mortality by 3.2 g/m2 LDS-SRG prepared from 5% dosage for 3, 7 and 14 days was 93.99%, 91.18% and 86.48% respectively, and that from 10% dosage was 94.95%, 93.50% and 85.43%, all significantly higher than that of the control group （82.83%, 72.38% and 48.38%）（P<0.05）; the snail mortality by 0.8 g/m2 LDS-SRG prepared from 5% dosage for 14 days（66.51%） and that by 1.6 g/m2 LDS-SRG prepared from 5% dosage for 3 days（84.61%） were both significantly higher than that by 10% LDS-SRG（20.13% and 43.06%） （P<0.05）. Conclusion: The 5% and 10% LDS-SRG used separately in the immersion test and the spraying test both meet the requirements of the national standard of Efficacy Test Methods and Evaluation of Molluscicide for Pesticide Registration.

Splice variants of the P2X7 receptor reveal differential agonist dependence and functional coupling with pannexin-1.

P2X7 receptors function as ATP-gated cation channels but also interact with other proteins as part of a larger signalling complex to mediate a variety of downstream responses that are dependent upon the cell type in which they are expressed. Receptor-mediated membrane permeabilization to large molecules precedes the induction of cell death, but remains poorly understood. The mechanisms that underlie differential sensitivity to NAD are also unknown. By studying alternative variants of the mouse P2X7 receptor we show that sensitivity to NAD is mediated through the P2X7k variant, which has a much more restricted distribution than the P2X7a receptor, but is expressed in T lymphocytes. The altered N-terminus and TM1 of the P2X7k receptor enhances the stability of the active state of this variant compared with P2X7a, thereby increasing the efficacy of NAD-dependent ADP ribosylation as measured by ethidium uptake, a rise in intracellular Ca(2+) and the activation of inward currents. Co-expression of P2X7k and P2X7a receptors reduced NAD sensitivity. P2X7k-receptor-mediated ethidium uptake was also triggered by much lower BzATP concentrations and was insensitive to the P451L single nucleotide polymorphism. P2X7k-receptor-mediated ethidium uptake occurred independently of pannexin-1 suggesting a pathway intrinsic to the receptor. Only for the P2X7aL451 receptor could we resolve a component of dye uptake dependent upon pannexin-1. Signalling occurred downstream of the activation of caspases rather than involving direct cross talk between the channels. However, an in situ proximity assay showed close association between P2X7 receptors and pannexin-1, which would facilitate ATP efflux through pannexin-1 acting in an autocrine manner.

Metabolic profiles of Oncomelania hupensis after molluscicidal treatment: Carbohydrate metabolism targeted and energy deficiency.

Oncomelania hupensis is the intermediate host of Schistosoma japonicum, one of the Schistosoma species that can cause human schistosomiasis. Molluscicidal treatment remains the primary means to control snail. Niclosamide is the only molluscicide recommended by the World Health Organization, and it has been used throughout schistosomiasis-endemic areas in China for almost 30 years. In our previous studies on transcriptomics, morphology, and enzymology of snails after molluscicidal treatment, two effective molluscicides were used, 50% wettable powder of niclosamide ethanolamine salt (WPN) and a new molluscicide derived from niclosamide, the salt of quinoid-2', 5-dichloro-4'-nitro-salicylanilide (LDS, simplified for Liu Dai Shui Yang An). Genes involved in cell structure mintenance, inhibition of neurohumoral transmission, and energy metabolism showed significant differential expression after molluscicide treatments. Damages in the structure of liver and muscle cells were accompanied by inhibited activities of enzymes related to carbohydrate metabolism and energy supply. This study was designed to clarify the dynamic metabolic process by metabonomics, together with the previous transcriptomic and enzymological profiles, to identify potential metabolite markers and metabolism pathways that related to the toxic mechanism of the molluscicide. In total, 56 metabolites were identified for O. hupensis, and 75% of these metabolites consisted of amino acids and derivatives, organic acids, and nucleic acid components. The concentration of glucose, maltose, succinate, choline, and alanine changed significantly after molluscicide treatments. These changes in metabolites mainly occurred in the process of carbohydrate metabolism, energy metabolism, and amino acid metabolism, primarily related to glycolysis/gluconeogenesis, oxidative phosphorylation, and transamination by KEGG pathway identification. Most of the identified pathways were also related to those differentially expressed unigenes and observed enzymes from our previous studies. Inhibited aerobic respiration and oxidative phosphorylation, and energy deficiency were implied further to be the leading causes of the final death of snails after molluscicide treatments. The hypothesised mathematical model in this study identified the rational hysteresis to explain the inconsistency of responses of unigenes, enzymes, and metabolites to molluscicide treatments. This study contributes to the comprehensive understanding of the molluscicidal mechanism in the metabolic process and this could assist in improving existing molluscicide formulations or development of new molluscicides.

Effect of floods on the transmission of schistosomiasis in the Yangtze River valley, People's Republic of China.

The aim of the present study was to assess the effect of floods on the transmission of schistosomiasis in the Yangtze River valley, People's Republic of China. Retrospective analyses of malacologic, clinical and epidemiologic data, covering a 22-year period, were carried out to elucidate the dispersal patterns of intermediate host snails (Oncomelania hupensis), and acute and chronic infections with Schistosoma japonicum in humans in relation to floods. Potential O. hupensis habitats in years with floods were 2.6-2.7 times larger than in years when water levels were normal. Both the density and infection rate of O. hupensis dropped in the first two years after a flood, but significantly increased in the third year. The number of acute cases with schistosomiasis japonica was markedly higher in years characterized by floods; on average, 2.8 times more cases were observed when compared to years that the Yangtze River had normal water levels. In view of our findings, emergency responses are warranted as soon as possible after the occurrence of a flood in order to avoid or mitigate the reemergence and spread of human schistosomiasis in the People's Republic of China.

Purification of eutrophic water containing chlorpyrifos by aquatic plants and its effects on planktonic bacteria.

In this study, the removal of nutrients and chlorpyrifos as well as shifts of planktonic bacterial communities in constructed microcosms were investigated to evaluate the influence of Phragmites australis, Nymphaea alba, and Myriophyllum verticillatum, and their combination, on the restoration of eutrophic water containing chlorpyrifos. Plant-treated groups showed a higher pollutant removal rate than did no-remediation controls, indicating that treatment with plants is effective at remediation of eutrophic water containing chlorpyrifos. Different plants showed different performance on the remediation of eutrophic water, e.g., P. australis manifested stronger capacity for removal of sediment chlorpyrifos. This finding indicated that an appropriate plant combination is needed to deal with complex wastewater. During the treatments, the planktonic bacterial communities were influenced by the concentrations of nutrients and pollutants. The changes of composition of bacterial communities indicated a strong correlation between the bacterial communities and the concentrations of pollutants. The plants also influenced the planktonic bacterial communities, especially at the early phase of treatments. For example, P. australis increased the abundance of Limnohabitans and Nevskia significantly and decreased the abundance of Devosia, Luteolibacter, Methylibium, and Caulobacter significantly. The abundance of Hydrocarboniphaga significantly increased in N. alba-treated microcosms, whereas in M. verticillatum-treated microcosms, the abundance of Limnohabitans and Bdellovibrio significantly increased. Our results suggest that the planktonic bacterial communities may be altered during phytoremediation, and the functions of the affected bacteria should be concerned.

Epidemiology of schistosomiasis in the People's Republic of China, 2004.

Results from the third nationwide cluster sampling survey on the epidemiology of schistosomiasis in the People's Republic of China, conducted by the Ministry of Health in 2004, are presented. A stratified cluster random sampling technique was used, and 239 villages were selected in 7 provinces where Schistosoma japonicum remains endemic. A total of 250,987 residents 6-65 years of age were included in the survey. Estimated prevalence rates in the provinces of Hunan, Hubei, Jiangxi, Anhui, Yunnan, Sichuan, and Jiangsu were 4.2%, 3.8%, 3.1%, 2.2%, 1.7%, 0.9%, and 0.3%, respectively. The highest prevalence rates were in the lake and marshland region (3.8%) and the lowest rates were in the plain region with waterway networks (0.06%). Extrapolation to all residents in schistosome-endemic areas indicated 726,112 infections. This indicates a reduction of 16.1% compared with a nationwide survey conducted in 1995. However, human infection rates increased by 3.9% in settings where transmission is ongoing.

[Study on molluscicidal effect of chlorosalicylicamide].

OBJECTIVE: To observe the molluscicidal effect of chlorosalicylicamide on Oncomelania hupensis and toxicity on fishes. METHODS: Different concentrations of chlorosalicylicamide were prepared, and immersion and spraying methods were used to test its molluscicidal effect and toxicity to fishes. Niclosamide ethanolamine salt was used as control. RESULTS: Under water temperature of 28 C with the dose of chlorosalicylicamide at 0.25 mg/L and 0.4 mg/L, the mortality rate of snails by immersion was 97.8% and 100% respectively after 48 h, and the LC50 was 0.0674 mg/L. While for niclosamide at the same doses, the mortality rate of snails was 22.2% and 66.7% respectively, and the LC50 was 0.397 6 mg/L. With the concentration of 0.4 mg/L for 72 h, the fish death rate in chlorosalicylicamide and niclosamide was 50.0% and 100% respectively. CONCLUSION: Chlorosalicylicamide shows a molluscicidal effect on Oncomelania snails and a lower toxicity to fishes than that of niclosamide.

[Evaluation on schistosomiasis control effect of the intervention measures adapted to the ecological environment changes in Jiang Han Plain due to establishment of the Three Gorges dam].

OBJECTIVE: To evaluate the effect of the intervention measures for schistosomiasis control adapted to the ecological environment changes in Jiang Han plain caused by the establishment of the Three Gorges Dam. METHODS: Four villages in Qianjiang City were selected to implement paddy-upland rotation, crawfish-paddy alternation, water control and soil improvement, and adjusting agricultural structure to rebuild the waterlogging low yielding land and to change the snail habitat environment respectively. The snail habitat area, mean density of living snails and prevalence of schistosomiasis in human and cattle were compared with those of the control villages. Miracidia hatching methods were used to examine the prevalence in human and cattle. RESULTS: In the four experimental villages, the snail-ridden area decreased by 100%, 51.35%, 62.16% and 87.88% respectively; mean density of living snails decreased by 100%, 69.41%, 52.30% and 75.77%, with a t value of 9.37, 4.91, 2.31 and 9.16, I'<0.01. Human prevalence of schistosomiasis in 2005 in village with crawfish-paddy alteration decreased significantly than control (chi2=39.84, I'<0.01); decreased by 73.10% in village with water control and soil improvement in 1990 than in 1987 (chi2=236.10, P<0.01). CONCLUSION: Implementation of the four intervention measures reaches a remarkable benefit in reforming snail habitat and protecting environment, which can be recommended to the inner embankment type endemic regions.

Transmission control of schistosomiasis japonica: implementation and evaluation of different snail control interventions.

The great progress made in the control of schistosomiasis japonica in China is to some extent explained by successful intermediate host snail control, in particular with environmental management commencing some 50 years ago. By 1995, interruption of Schistosoma japonicum transmission had been achieved in five of the 12 schistosome-endemic provinces while endemic areas in the remaining provinces had been reduced significantly, and snail habitats had decreased by 74%. In this paper, we review the role and approaches towards control and give an account of compounds toxic to snails. Mid- and long-term national plans for schistosomiasis control are discussed, including integrated snail control approaches as an important measure. It is anticipated that implementation of these plans will accelerate snail control, which in turn will consolidate and mutually reinforce other control measures, most notably praziquantel-based chemotherapy. The ultimate aim is to eliminate schistosomiasis as a public health problem in China.

Spirodela polyrhiza stimulates the growth of its endophytes but differentially increases their fenpropathrin-degradation capabilities.

In situ remediation of organic contaminants via physical, chemical, and biological approaches is a practical technique for cleansing contaminated water and soil. In the present study, we showed that the three bacterial strains Pseudomonas sp. E1, Klebsiella terrigena E42, and Pseudomonas sp. E46, which can infect and colonize the aquatic plant Spirodela polyrhiza, utilize fenpropathrin as the sole carbon source for growth. S. polyrhiza helped enhance fenpropathrin degradation by E46 by 17.5%, only slightly improved fenpropathrin degradation by E42, and had no effect on strain E1. The application of plant exudates and extracts from fenpropathrin-unexposed/induced plants stimulated bacterial growth of the three strains, but resulted in differential fenpropathrin degradation, suggesting that not all plants and their endophytic bacteria are suitable for coupling phytoremediation and microbial-remediation. Moreover, addition of soil sediments to a microcosm not only stimulated the growth of strain E46 but also increased the rate of fenpropathrin degradation.

De Novo transcriptome analysis of Oncomelania hupensis after molluscicide treatment by next-generation sequencing: implications for biology and future snail interventions.

The freshwater snail Oncomelania hupensis is the only intermediate host of Schistosoma japonicum, which causes schistosomiasis. This disease is endemic in the Far East, especially in mainland China. Because niclosamide is the only molluscicide recommended by the World Health Organization, 50% wettable powder of niclosamide ethanolamine salt (WPN), the only chemical molluscicide available in China, has been widely used as the main snail control method for over two decades. Recently, a novel molluscicide derived from niclosamide, the salt of quinoid-2',5-dichloro-4'-nitro-salicylanilide (Liu Dai Shui Yang An, LDS), has been developed and proven to have the same molluscicidal effect as WPN, with lower cost and significantly lower toxicity to fish than WPN. The mechanism by which these molluscicides cause snail death is not known. Here, we report the next-generation transcriptome sequencing of O. hupensis; 145,008,667 clean reads were generated and assembled into 254,286 unigenes. Using GO and KEGG databases, 14,860 unigenes were assigned GO annotations and 4,686 unigenes were mapped to 250 KEGG pathways. Many sequences involved in key processes associated with biological regulation and innate immunity have been identified. After the snails were exposed to LDS and WPN, 254 unigenes showed significant differential expression. These genes were shown to be involved in cell structure defects and the inhibition of neurohumoral transmission and energy metabolism, which may cause snail death. Gene expression patterns differed after exposure to LDS and WPN, and these differences must be elucidated by the identification and annotation of these unknown unigenes. We believe that this first large-scale transcriptome dataset for O. hupensis will provide an opportunity for the in-depth analysis of this biomedically important freshwater snail at the molecular level and accelerate studies of the O. hupensis genome. The data elucidating the molluscicidal mechanism will be of great benefit in future snail control efforts.

[Acute toxicity of LDS, a molluscicide, to non-target organisms].

OBJECTIVE: To evaluate the acute toxicity of 10% LDS, a new molluscicide, to non-target organisms. METHODS: Based on "Chemical pesticide environmental safety test evaluation standard", an acute toxicity test was carried out with Coturnix coturnix japonica (quail), Apis mellifera L (bee), Bombyx mori (silkworm), and Brachydonio rerio (zebra fish) , and the skin stimulus test was also performed with guinea pig. RESULTS: The quails had no toxic symptoms while the maximum poisoning concentration of LDS was 200 mg/kg (no toxicity). LC50 of bees was 2.68 x 10³ mg/L (low toxicity). After 96 hours, no silkworms died in each group of different concentrations of LDS while the most concentration was 6.00 x 10² mg/kg, but there were some toxic symptoms such as inappetence and inactive in the high concentration group as compared to the blank control group (low toxic). LC50 (96 h) of zebra fish was 6.16 mg/L (medium toxicity). CONCLUSIONS: LDS has no toxicity to Coturnix coturnix japonica, low toxicity to Apis mellifera L and Bombyx mori, and medium toxicity to Brachydonio rerio. Compared with niclosamide ethanolamine salt, the toxicity to the fish is lower, and therefore, it is more suitable for the field application.

[Evaluation on molluscicidal effect of chlorosalicylicamide on Oncomelania hupensis in schistosomiaisis endemic areas of eight provinces in China].

OBJECTIVE: To evaluate the effects of a novel molluscicide, the salt quinoid-2', 5-dichloro-4'-nitrosalicylanilide from niclosamide (LDS), with 10% wettable powder, in main schistosomiasis epidemic areas of China, including Hunan, Jiangxi, Hubei, Anhui, Jiangsu, Sichuan, Yunnan and Zhejiang Province. METHODS: In the immersion test, 6 effective concentrations of 10% LDS were tested respectively: 0.1, 0.2, 0.4, 0.6, 0.8, 1.0 g/m3 in the field; at the same time, 50% wettable powder of niclosamide ethanolamine salt (WPN) with effective concentrations of 1.0 g/m was used as the molluscicide control, and the fresh water as the blank control, then the mortality rates of 0. hupensis snails were recorded at 24 h, 48 h and 72 h after the immersion. In the spraying test and powder-spraying test, 5 effective dosages of 10% LDS were tested respectively: 0.2, 0.4, 0.6, 0.8, 1.0 g/m2, while 50% WPN 1.0 g/m2 was used as the molluscicide control, and the fresh water as the blank control in the field for 1 d, 3 d and 7 d, then the mortality rates of O. hupensis snails were recorded at 1 d, 3 d and 7 d after the spraying and powder-spraying. RESULTS: The snail mortality rates of LDS using the immersion test for 72 h were more than 95% in the field of eight provinces (0.1 g/m in Sichuan, Jiangxi, Jiangsu and Zhejiang provinces, 0.2 g/m3 in Yunnan, Hunan and Hubei provinces, and 0.4 g/min Anhui Province); the snail mortality rates of LDS using the spraying test for 7 d were more than 85% (0.2 g/m2 in Hunan, Hubei, Jiangxi and Zhejiang provinces, 0.4 g/m2 in Sichuan and Anhui provinces, 0.6 g/m2 in Yunnan and Jiangsu provinces). The snail mortality rates of LDS the powder-spraying test for 7 d were more than 85% (0.6 g/m2 in Yunnan, Sichuan, Hubei, Jiangxi, Anhui, Jiangsu and Zhejiang provinces). According to the standards of "Efficacy test methods and evaluation of molluscicide for pesticide registration (NY/T 1617-2008)", LDS is a qualified molluscicide. CONCLUSIONS: LDS has good molluscicidal effects through the immersion, spraying and powder-spraying test in the fields. It is suitable for a variety of environments to control O. hupensis snails of schistosomiasis endemic areas in China. The recommended dosages of LDS are 0.1-0.2 g/m3 by the immersion method, 0.2-0.4 g/m2 by the spraying method, and 0.4-0.6 g/m2 by the powder-spraying method in the fields.

Relationship between the transmission of schistosomiasis japonica and the construction of the Three Gorge Reservoir.

OBJECTIVE: To study the relationship between the changes of environment and the transmission of Schistosomiasis japonica after the construction of the Three Gorge Reservoir. METHODS: On the basis of the predictive data on changes of water level and sediment in the middle and lower reaches of the Yangtze River after the dam construction provided by the Yangtze River Committee on Water Conservancy, corresponding data were collected and analyzed. In order to understand the effects of changes of water level and sediment on Oncomelania hupensis, the intermediate host of Schistosoma japonicum, field surveys and ecologically intimated experiments were done. The prevalence status and factors related to transmission were investigated by epidemiology, medical geography and public health. RESULTS: Positive impact on control of schistosomiasis is that floods in the Yangtze River will occur less and will decrease the dispersal of snails and the chances of infection for humans and animals as well. Snail habitats' beach will be reduced in the flushed area along the Yangtze River. Exploitation of beach and elimination of snails will benefit from the beach's uplift in flushed area. The floods ahead of time in autumn will interfere with the development of snails. Negative impact is that the flushed beaches and migratory settlements at certain altitude may become snail habitats. So the reservoir area will be a potential transmission area of schistosomiasis due to dispersal of infectious resources and Oncomelania snails. The uprising of the water body level may cause the expending of the snail habitats in some places of Jianghan plain during the transition of spring to summer. The destruction of the embankments of the Dongting Lake will be put off by its delayed flush, which causes the beach in a situation of growing reed-grass or grass for a long time that benefits the reproduction of the snails. The chances for schistosome infection to humans and animals will be on the increase, because the water regression occurs ahead of time in the Dongting Lake beach in autumn. The distribution of snails will not be affected by the changes of water level in the lower reaches of the Yangtze River after the dam is built, but the flushed section in Anhui in the recent 80 years are suitable for the snails' development. The environmental changes due to dam construction will not obviously affect the development of snails and the transmission of schistosomiasis in the Poyang Lake area. CONCLUSION: The construction of the huge dam and the formation of the Three Gorge Reservoir on the Yangtze River may have both positive and negative impact on schistosomiasis transmission and control. Great effort must be made to decrease the negative impact to avoid or, at least, to decrease S. japonicum transmission.

A Bayesian approach to estimate the prevalence of Schistosomiasis japonica infection in the Hubei Province Lake Regions, China.

A Bayesian inference model was introduced to estimate community prevalence of Schistosomiasis japonica infection based on the data of a large-scale survey of Schistosomiasis japonica in the lake region in Hubei Province. A multistage cluster random sampling approach was applied to the endemic villages in the lake regions of Hubei Province in 2011. IHA test and Kato-Katz test were applied for the detection of the S. japonica infection in the sampled population. Expert knowledge on sensitivities and specificities of IHA test and Kato-Katz test were collected based on a two-round interview. Prevalence of S. japonica infection was estimated by a Bayesian hierarchical model in two different situations. In Situation 1, Bayesian estimation used both IHA test data and Kato-Katz test data to estimate the prevalence of S. japonica. In Situation 2, only IHA test data was used for Bayesian estimation. Finally 14 cities and 46 villages from the lake regions of Hubei Province including 50,980 residents were sampled. Sensitivity and specificity for IHA test ranged from 80% to 90% and 70% to 80%, respectively. For the Kato-Katz test, sensitivity and specificity were from 20% to 70% and 90% to 100%, respectively. Similar estimated prevalence was obtained in the two situations. Estimated prevalence among sampled villages was almost below 13% in both situations and varied from 0.95% to 12.26% when only using data from the IHA test. The study indicated that it is feasible to apply IHA test only combining with Bayesian method to estimate the prevalence of S. japonica infection in large-scale surveys.

Assessing the effect of an integrated control strategy for schistosomiasis japonica emphasizing bovines in a marshland area of Hubei Province, China: a cluster randomized trial.

INTRODUCTION: More than 80% of schistosomiasis patients in China live in the lake and marshland regions. The purpose of our study is to assess the effect of a comprehensive strategy to control transmission of Schistosoma japonicum in marshland regions. METHODOLOGY/PRINCIPAL FINDINGS: In a cluster randomized controlled trial, we implemented an integrated control strategy in twelve villages from 2009 through 2011 in Gong'an County, Hubei Province. The routine interventions included praziquantel chemotherapy and controlling snails, and were implemented in all villages. New interventions, mainly consisting of building fences to limit the grazing area for bovines, building safe pastures for grazing, improving the residents' health conditions and facilities, were only implemented in six intervention villages. Results showed that the rate of S. japonicum infection in humans, bovines, snails, cow dung and mice in the intervention group decreased from 3.41% in 2008 to 0.81% in 2011, 3.3% to none, 11 of 6,219 to none, 3.9% to none and 31.7% to 1.7%, respectively (P<0.001 for all comparisons). In contrast, there were no statistically significant reductions of S. japonicum infection in humans, bovines and snails from 2008 to 2011 in the control group (P>0.05 for all comparisons). Moreover, a generalized linear model showed that there was a higher infection risk in humans in the control group than in the intervention group (OR = 1.250, P = 0.001) and an overall significant downward trend in infection risk during the study period. CONCLUSIONS/SIGNIFICANCE: The integrated control strategy, designed to reduce the role of bovines and humans as sources of S. japonicum infection, was highly effective in controlling the transmission of S. japonicum in marshland regions in China. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR-PRC-12002405.

Expression, assembly and function of novel C-terminal truncated variants of the mouse P2X7 receptor: re-evaluation of P2X7 knockouts.

BACKGROUND AND PURPOSE: Splice variants of P2X7 receptor transcripts contribute to the diversity of receptor-mediated responses. Here, we investigated expression and function of C-terminal truncated (ΔC) variants of the mP2X7 receptor, which are predicted to escape inactivation in one strain of P2X7(-/-) mice (Pfizer KO). EXPERIMENTAL APPROACH: Expression in wild-type (WT) and Pfizer KO tissue was investigated by reverse transcription (RT)-PCR and Western blot analysis. ΔC variants were also cloned and expressed in HEK293 cells to investigate their assembly, trafficking and function. KEY RESULTS: RT-PCR indicates expression of a ΔC splice variant in brain, salivary gland (SG) and spleen from WT and Pfizer KO mice. An additional ΔC hybrid transcript, containing sequences of P2X7 upstream of exon 12, part of exon 13 followed in-frame by the sequence of the vector used to disrupt the P2X7 gene, was also identified in the KO mice. By blue native (BN) PAGE analysis and the use of cross linking reagents followed by SDS-PAGE, P2X7 trimers, dimers and monomers were detected in the spleen and SG of Pfizer KO mice. The molecular mass was reduced compared with P2X7 in WT mice tissue, consistent with a ΔC variant. When expressed in HEK293 cells the ΔC variants were inefficiently trafficked to the cell surface and agonist-evoked whole cell currents were small. Co-expressed with P2X7A, the ΔC splice variant acted in a dominant negative fashion to inhibit function. CONCLUSIONS AND IMPLICATIONS: Pfizer KO mice are not null for P2X7 receptor expression but express ΔC variants with reduced function.