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Schistosomiasis is a fatal zoonotic parasitic disease that also threatens human health. The main pathological features of schistosomiasis are granulomatous inflammation and subsequent liver fibrosis, which is a complex, chronic, and progressive disease. Extracellular vesicles (EVs) derived from schistosome eggs are broadly involved in host-parasite communication and act as important contributors to schistosome-induced liver fibrosis. However, it remains unclear whether substances secreted by the EVs of Schistosoma japonicum, a long-term parasitic "partner" in the hepatic portal vein of the host, also participate in liver fibrosis. Here, we report that EVs derived from S. japonicum worms attenuated liver fibrosis by delivering sja-let-7 into hepatic stellate cells (HSCs). Mechanistically, activation of HSCs was reduced by targeting collagen type I alpha 2 chain (Col1α2) and downregulation of the TGF-β/Smad signaling pathway both in vivo and in vitro. Overall, these results contribute to further understanding of the molecular mechanisms underlying host-parasite interactions and identified the sja-let-7/Col1α2/TGF-β/Smad axis as a potential target for treatment of schistosomiasis-related liver fibrosis. Author summary: Schistosomiasis is a neglected parasitic disease that affects over 250 million people in tropical and subtropical regions worldwide. As common pathogenic species, the eggs of Schistosoma japonicum and S. mansoni cause hepatic schistosomiasis, which can lead to granulomatous inflammation and subsequent liver fibrosis. Hence, elucidation of the mechanisms underlying liver fibrosis and host-parasite crosstalk is essential to inhibiting the progression of schistosomiasis. Accumulating evidence supports the pivotal role of schistosome egg-derived extracellular vesicles (EVs) in the onset and progression of schistosomiasis. However, the potential involvement of schistosome worm-derived EVs in the activation and modulation of hepatic stellate cells (HSCs) in hepatic schistosomiasis remains largely unknown. The results of this study demonstrate that the release of sja-let-7 by S. japonicum worm-derived EVs reduced activation of HSCs by targeting Col1α2 and further attenuated the progression of liver fibrosis by mediating the TGF-β/Smad signaling pathway. These findings provide evidence for the application of worm-derived substances and the sja-let-7/Col1α2/TGF-β/Smad axis as a novel therapeutic target for treatment of liver fibrosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Liver fibrosis (LF) is a common pathologic feature of multiple liver diseases. However, there is currently no effective treatment to slow the progression of LF. In this study, sja-let-7 from Schistosoma japonicum was shown to regulate host immune responses and attenuate the progression of carbon tetrachloride-induced LF via the Col1α2 and TGF-β/Smad signaling pathway, thereby providing references for the application of worm-derived molecules for the treatment of LF. Liver fibrosis (LF) is a chronic progressive disease with no definitive treatment. The aim of this study was to assess helminth-derived molecules as potential therapeutic targets to prevent or reverse LF. A mouse model of carbon tetrachloride (CCL4)-induced LF was established and sja-let-7 was overexpressed by treatment with a miRNA agomir once per week. After four weeks, serum biochemistry, hepatic hydroxyproline content measurements, liver histology, mRNA expression profiling of fibrotic markers, the dual-luciferase reporter assay, and fluorescence in situ hybridization (FISH) were performed. Administration of the sja-let-7 agomir markedly ameliorated hepatosplenomegaly and reduced the liver hydroxyproline content. Liver histological analysis showed significant reductions in collagen deposition in the sja-let-7 agomir-treated mice. Additionally, the mRNA levels of both pro-fibrotic markers and pro-inflammatory cytokines were diminished after treatment. Furthermore, the dual-luciferase reporter assay and FISH identified the α2 chain of collagen type 1 (Col1α2) as the direct target of sja-let-7. Accordingly, the progression of LF was attenuated by targeting Col1α2 and the TGF-β/Smad signaling pathway. [ABSTRACT FROM AUTHOR]

Simple Summary: Molecular diagnostic methods based on nucleic acid detection possess more advantages in high sensitivity and specificity, and low cross-reactivity with other pathogens. We assessed the detection efficacy of a real-time fluorescent quantitative PCR assay for schistosomiasis japonica in mice, before and after treatment with praziquantel. The sensitivity of the method was 99.3% (152/153, 95% CI: 96.41–99.98%) and its specificity was 100% (77/77, 95% CI: 95.32–100%). The results showed that the method exhibited good sensitivity and specificity, and its sensitivity correlated with the infection intensity in mice. After the oral administration of praziquantel, mice infected with 10 cercariae or 40 cercariae were all Schistosoma japonicum-negative via this method 6 weeks after treatment. This method had advantages over a soluble-egg-antigen-based enzyme-linked immunosorbent assay, and possessed better potential utility for evaluating the treatment efficacy of praziquantel in schistosome-infected mice. The prevalence of schistosomiasis japonica in China is now characterized by a low epidemic rate and low-intensity infections. Some diagnostic methods with high sensitivity and specificity are urgently needed to better monitor this disease in the current situation. In this study, the detection efficacy of a real-time fluorescent quantitative PCR (qPCR) assay was assessed for schistosomiasis japonica in mice, and before and after treatment with praziquantel (PZQ). Our results showed that the sensitivity of the qPCR was 99.3% (152/153, 95% CI: 96.41–99.98%) and its specificity was 100% (77/77, 95% CI: 95.32–100%) in mice infected with different numbers of Schistosoma japonicum. After the oral administration of PZQ, mice infected with 10 cercariae or 40 cercariae were all Schistosoma japonicum-negative 6 weeks after treatment. However, the negativity rates on a soluble egg antigen (SEA)-based enzyme-linked immunosorbent assay (ELISA) were only 34.8% (8/23, 10 cercariae group) and 6.7% (1/15, 40 cercariae group) at the sixth week after PZQ treatment. These results demonstrated that the qPCR method had good sensitivity and specificity, and suggested that its sensitivity correlated with the infection intensity in mice. Moreover, this method had better potential utility for evaluating the treatment efficacy of PZQ in schistosome-infected mice than SEA-based ELISA. [ABSTRACT FROM AUTHOR]

Background: N-acetyltransferase 13 (NAT13) is a probable catalytic component of the ARD1A-NARG1 complex possessing alpha (N-terminal) acetyltransferase activity. Results: In this study, a full-length complementary DNA (cDNA) encoding Schistosoma japonicum NAT13 (SjNAT13) was isolated from schistosome cDNAs. The 621 bp open reading frame of SjNAT13 encodes a polypeptide of 206 amino acids. Real-time PCR analysis revealed SjNAT13 expression in all tested developmental stages. Transcript levels were highest in cercariae and 21-day-old worms, and higher in male adult worms than female adult worms. The rSjNAT13 protein induced high levels of anti-rSjNAT13 IgG antibodies. In two independent immunoprotection trials, rSjNAT13 induced 24.23% and 24.47% reductions in the numbers of eggs in liver. RNA interference (RNAi) results showed that small interfering RNA (siRNA) Sj-514 significantly reduced SjNAT13 transcript levels in worms and decreased egg production in vitro. Conclusions: Thus, rSjNAT13 might play an important role in the development and reproduction of schistosomes. [ABSTRACT FROM AUTHOR]

Background: N-acetyltransferase 13 (NAT13) is a probable catalytic component of the ARD1A-NARG1 complex possessing alpha (N-terminal) acetyltransferase activity. Results: In this study, a full-length complementary DNA (cDNA) encoding Schistosoma japonicum NAT13 (SjNAT13) was isolated from schistosome cDNAs. The 621 bp open reading frame of SjNAT13 encodes a polypeptide of 206 amino acids. Real-time PCR analysis revealed SjNAT13 expression in all tested developmental stages. Transcript levels were highest in cercariae and 21-day-old worms, and higher in male adult worms than female adult worms. The rSjNAT13 protein induced high levels of anti-rSjNAT13 IgG antibodies. In two independent immunoprotection trials, rSjNAT13 induced 24.23% and 24.47% reductions in the numbers of eggs in liver. RNA interference (RNAi) results showed that small interfering RNA (siRNA) Sj-514 significantly reduced SjNAT13 transcript levels in worms and decreased egg production in vitro. Conclusions: Thus, rSjNAT13 might play an important role in the development and reproduction of schistosomes. [ABSTRACT FROM AUTHOR]

Background: Schistosomiasis japonica is an infectious disease caused by Schistosoma japonicum that seriously endangers human health. Domestic animals have important roles in disease transmission and goats are considered a primary reservoir host and source of infection. The prevalence and intensity of schistosomiasis infections have significantly decreased in China, and a more sensitive, specific detection method is urgently needed. The aim of this study was to develop a real-time PCR assay for accurate detection of S. japonicum infection in goats. Methods: A real-time PCR method for detecting schistosomiasis japonica in goats was developed by amplification of a specific S. japonicum DNA fragment, and validated using a total of 94 negative and 159 positive plasma and serum samples collected in our previous study of S. japonicum infection. Both plasma and serum samples were evaluated by real-time PCR and enzyme-linked immunosorbent assay (ELISA). In addition, 120 goat plasma samples from an S. japonicum-endemic area (Wangjiang) and 33 from a non-endemic region (Weihai) were collected and evaluated using our method. Results: The sensitivity and specificity of the real-time PCR for detecting infected samples were 98.74% (157/159, 95% CI: 95.53–99.85%) and 100% (94/94, 95% CI: 96.15–100%), respectively. For the ELISA, sensitivity and specificity were 98.11% (156/159, 95% CI: 94.59–99.61%) and 90.43% (85/94, 95% CI: 82.60–95.53%), respectively. Further, we found positivity rates for S. japonicum infection in Wangjiang and Weihai of 8.33% (10/120, 95% CI: 4.07–14.79%) and 0% (0/33, 95% CI: 0–10.58%), respectively. Conclusions: The results of this study indicate that our real-time PCR method exhibits higher sensitivity and specificity than ELISA and is a useful method for detection of S. japonicum infection in goats. [ABSTRACT FROM AUTHOR]

• The Lwr gene of Schistosoma japonicum was used for bioinformatical analysis. • SjLwr protein was greatest in internal tissues and the surface layer on day 14 in female worms. • SjLwr functions in the growth, development, and reproduction of S. japonicum. The lesswright (lwr) gene and its products are essential molecules in mitosis, DNA repair, and embryo formation in many eukaryotes. In this study, immunohistochemical analysis revealed that the Lwr protein was located in the internal tissues and the surface layer of the adult Schistosoma japonicum (Sj) worms. The mRNA expression levels of SjLwr at different points were evaluated by quantitative real-time RT-PCR. The expression of SjLwr peaked at 14 days and then decreased thereafter. SjLwr expression was relatively more stable in male worms than in female worms. The functions of SjLwr were explored by siRNA-based gene silencing with a simple soaking method. The results showed that knockdown of the SjLwr gene impaired the growth and development of S. japonicum in mice, as well as survival, morphology, reproductive capacity, and egg vitality. These observations imply that SjLwr presents a novel target for the development of immuno- and/or small molecule-based therapeutics for the control and treatment of schistosome infections. [ABSTRACT FROM AUTHOR]

Highlights • We confirmed that buffalos received twice pre-infection have developed a resistance against S. japonicum reinfection. • There was a significant reduction in worm burden, worm length and egg counts in buffalos received twice pre-infection. • After reinfection levels of IL-4, IL-10, IFN-g, and specific IgG were higher. • Resistance against S. japonicum reinfection was mainly due to acquired immunity. • These findings may aid in the future vaccine design for water buffalos. Abstract Schistosomiasis is a zoonotic parasitic disease threatening tens of millions people and farm animals. Water buffalos are a major reservoir for schistosomiasis and a control target. Epidemiological surveys suggest that buffalos can develop resistance against Schistosoma japonicum reinfection. In the present paper, relative to control animals, we report an over 97% worm burden reduction after two rounds of infection with S. japonicum and treatment with Praziquantel (PZQ). Relative to control animals, shorter length of female worms, and lower egg counts (over 87.7% reduction rates) were observed in reinfected buffalos. We also found that the reinfected buffalos had significantly higher levels of IL-4, IL-10, and IFN-γ, 4–9 weeks after the secondary infection, and a significantly higher level of specific IgG antibodies before infection. Our results confirmed that after infection buffalos develop resistance against S. japonicum reinfection, and that this resistance is mainly due to acquired immunity. These findings may aid in the future vaccine design for water buffalos. [ABSTRACT FROM AUTHOR]

• PZQ-pretreated mice could resist infection after PZQ is completely eliminated. • The effective dose was 300 mg/kg twice orally given or one injection at 200 mg/kg. • The protection period of PZQ injection was 18 days. • PZQ could induce physiological changes of mice. Praziquantel (PZQ) is the first line drug for the treatment of schistosomiasis. Several studies have confirmed that PZQ regulates host immunity, and we have recently found that pretreatment with PZQ enhances resistance against Schistosoma japonicum infection in buffaloes. We speculate that PZQ induces physiological changes in mice that prevent S. japonicum infection. To test this hypothesis and provide a practical measure to prevent S. japonicum infection, we determined the effective dose (the minimum dose), protection period and onset time of protection by comparing the worm burden, female worm burden and egg burden in PZQ-pretreated mice and blank control mice. Morphological differences between parasites were observed by measuring the total worm length, oral sucker, ventral sucker and ovary. The levels of cytokines, nitrogen monoxide (NO), 5-hydroxytryptamine (5-HT) and specific antibodies were measured using kits or soluble worm antigens. Hematological indicators on day 0 were analyzed in mice that received PZQ on days -15, -18, -19, -20, -21 and -22. The PZQ concentrations in plasma and blood cells were monitored using high performance liquid chromatography (HPLC). The effective dose was found to be two oral administrations (interval of 24 h) at 300 mg/kg body weight (BW) or one injection at 200 mg/kg BW, and the protection period of PZQ injection was 18 days. The optimal preventive effect was observed at two days post-administration, with a >92% worm reduction rate and significant worm reduction until 21 days after administration. Adult worms from PZQ-pretreated mice were runtish showing a shorter length, smaller organs and fewer eggs in the uteri of females. Detection of cytokines, NO, 5-HT and hematological indicators showed that PZQ induced immune-physiological changes, including higher levels of NO, IFN-γ and IL-2, and a lower level of TGF-β. No significant difference in the anti- S. japonicum specific antibody levels was observed. The PZQ concentrations in plasma and blood cells 8 and 15 days post-administration were lower than the detection limit. Our results confirmed that pretreatment with PZQ promotes the protection of mice against S. japonicum infection within 18 days. Although we observed some immune-physiological changes in the PZQ-pretreated mice, the exact mechanisms involved in the preventive effect require further study. [ABSTRACT FROM AUTHOR]

Domestic animals infected with Schistosoma japonicum are a major source of infection and play an important role in transmission to humans. A key strategy for the elimination of schistosomiasis is to control the sources of infection. In the present study, we identified a candidate diagnostic antigen-encoding gene, SjMRP1, the putative multidrug resistance protein 1 gene, by screening a cDNA phage display library from 44-day-old S. japonicum worms using IgGs from goat, cattle, and buffalo infected with S. japonicum . We cloned and expressed the fragment of SjMRP1 and subsequently evaluated the diagnostic potential of the recombinant protein rSjMRP1. In the enzyme-linked immunosorbent assay of rSjMRP1 (rSjMRP1-ELISA), the sensitivity in goat, cattle, and buffalo was 95.6% (86/90), 100% (22/22), and 90% (81/90), respectively, and the specificity was 100% (30/30) in goat and cattle and 96.67% (29/30) in buffalo. These results were not significantly different from soluble egg antigen (SEA)-ELISA results. Notably, rSjMRP1-ELISA has no cross reaction with Haemonchus contortus , a most common nematode seen in goat and bovine in China, in 13 infected goats, and with Orientobilhazia turkestanica , which is genetically under Schistosoma , in 36 infected goats; whereas SEA-ELISA showed false positive rate of 15.38% and 83.33% in the two respective animal groups. The results obtained here suggest that rSjMRP1 may be used for diagnosis of S. japonicum infection of domestic animals. [ABSTRACT FROM AUTHOR]

• SjAAP protein was mainly distributed in tegument and parenchyma of schistosomes. • Purified recombinant SjAAP protein has enzymatic activity in vitro. • The level of expression in 42-day-old male worms was significantly higher than that in females. • There was no significant immunoprotection effect in vaccination test. The tegument of schistosomes is the interface between the worm and the host environment. Some molecules distributed on the tegument participate in host–parasite interactions. Aspartyl aminopeptidase (AAP), identified on the tegument of Schistosoma japonicum (S. japonicum), facilitate protein turnover by acting in concert with other aminopeptidases. In this study, the gene encoding S. japonicum aspartyl aminopeptidase (SjAAP) was cloned, expressed and characterized. Quantitative real-time PCR analysis showed that SjAAP was expressed in all studied developmental stages. The transcript level was higher in 8, 14, 21, and 28 days old worms than the other detected stages. Moreover, the level of expression in 42-day-old male worms was significantly higher than that in females. The recombinant SjAAP (rSjAAP) was expressed as both supernatant and inclusion bodies in Escherichia coli BL21 cells. The enzymatic activity of rSjAAP was 4.45 U/mg. The Km and Vmax values for H-Asp-pNA hydrolysis were discovered to be 5.93 mM and 0.018 mM·min–1. Immunofluorescence analysis revealed that SjAAP is primarily distributed on the tegument and parenchyma of schistosomes. Western blot showed that rSjAAP possessed good immunogenicity. Although specific antibodies were produced in BALB/c mice vaccinated with rSjAAP emulsified with ISA 206 adjuvant, no significant reduction of worm burden and number of eggs in the liver was observed. Therefore, rSjAAP may not be suitable to act as a potential vaccine candidate against schistosomiasis japonica in mice. However, this study provides some foundation for further exploration of the biological function of this molecule. [ABSTRACT FROM AUTHOR]

Wnt signaling is a key pathway involving the regulation of cell development and growth in metazoa. An analysis of Wnt signaling in Schistosoma japonicum might provide information regarding the molecular mechanisms underlying parasite development, which might be useful for vaccine screening and identification of pharmaceutical targets. The SjWnt5 gene, a member of the Wnt gene family, contained an 1149-bp open reading frame that encoded a 382-aa protein. Analysis of the SjWnt5 amino acid sequence revealed a domain that was conserved among members of the Wnt protein family. Expression of SjWnt5 was observed at all of the developmental stages in definitive hosts, and the highest level of SjWnt5 messenger RNA (mRNA) was detected at the schistosomula stage. Higher levels of SjWnt5 mRNA and protein were observed in mature male worms, compared with those in mature females. SjWnt5 mRNA was expressed at higher levels in maldeveloped worms from nonpermissive host or single-sex infection than in normal worms from permissive host and mixed-sex infection. The immunohistochemical analysis showed that SjWnt5 protein was expressed in the subtegumental musculature and acetabulum musculature of schistosomulum and adult worms, suggesting that SjWnt5 may play a role in regulation of parasite muscle development. Furthermore, SjWnt5 was found prominently expressed in the testes of the male and the ovary as well as the vitellarium of the female, suggesting that SjWnt5 may involve in the development of the reproductive organs of both sexes. [ABSTRACT FROM AUTHOR]

Water buffalo are less susceptible to Schistosoma japonicum infection than yellow cattle. The factors that affect such differences in susceptibility remain unknown. A Bos taurus genome-wide gene chip was used to analyze gene expression profiles in the peripheral blood of water buffalo and yellow cattle pre- and post-infection with S. japonicum. This study showed that most of the identified differentially expressed genes(DEGs) between water buffalo and yellow cattle pre- and post-infection were involved in immune-related processes, and the expression level of immune genes was lower in water buffalo. The unique DEGs (390) in yellow cattle were mainly associated with inflammation pathways, while the unique DEGs (2,114) in water buffalo were mainly associated with immune-related factors. The 83 common DEGs may be the essential response genes during S. japonicum infection, the highest two gene ontology (GO) functions were associated with the regulation of fibrinolysis. The pathway enrichment analysis showed that the DEGs constituted similar immune-related pathways pre- and post-infection between the two hosts. This first analysis of the transcriptional profiles of natural hosts has enabled us to gain new insights into the mechanisms that govern their susceptibility or resistance to S. japonicum infections. [ABSTRACT FROM AUTHOR]

Inhibitor of apoptosis proteins (IAP) play an important role in the regulation of apoptotic processes and are defined by the presence of baculoviral IAP repeat (BIR) domains. Here, we characterized a cDNA fragment (SjBIRP) synthesized from the RNA of Schistosoma japonicum, which was found to contain the BIR domain, suggesting that it could encode a potential IAP. Real-time PCR analyses indicated that SjBIRP transcription was detected at several stages of the schistosome's lifecycle, with increased levels present in schistosomula (7 days). In addition, the SjBIRP was highly expressed in adult females as compared to adult males. A functional assay showed that SjBIRP could inhibit caspase3/7 activity in both HeLa cells and schistosome lysates. Furthermore, SjBIRP expression profiles varied between different hosts of S. japonicum. Taken together, our preliminary studies suggest that SjBIRP may play a functional role in the regulation of apoptosis in schistosomes, and that it could be a potential drug target for schistosomiasis control. [ABSTRACT FROM AUTHOR]

Schistosomiasis is a worldwide parasitic disease, and while it can be successfully treated with chemotherapy, this does not prevent reinfection with the parasite. Adenovirus vectors have been widely used for vaccine delivery, and a vaccination approach has the potential to prevent infection with Schistosoma. Here, we developed a recombinant adenoviral vector that expresses Schistosoma japonicum inhibitor apoptosis protein (Ad-SjIAP) and assessed its immunoprotective functions against schistosomiasis in mice. Murine immune responses following vaccination were investigated using enzyme-linked immunosorbent assays (ELISA), lymphocyte proliferation, and cytokine assays. The protective immunity in mice was evaluated by challenging with S. japonicum cercariae. Our results indicated that immunization with the Ad-SjIAP in mice induced a strong serum IgG response against IAP including IgG1, IgG2a, and IgG2b. In addition, lymphocyte proliferation experiments showed that mice treated with Ad-SjIAP significantly increased the lymphocyte response upon stimulation with recombinant Schistosoma japonicum inhibitor apoptosis protein (rSjIAP). Moreover, cytokine assays indicated that vaccination of Ad-SjIAP significantly increased the production of interferon (IFN)-γ and IL-2 as compared to the corresponding control group. Furthermore, following the challenge with S. japonicum cercariae, the vaccine conferred moderate protection, with an average rate of 37.95 % for worm reduction and 31.7 % for egg reduction. Taken together, our preliminarily results suggested that schistosoma IAP may be a potential vaccine against S. japonicum and that adenoviral vectors may serve as an alternative delivery vehicle for schistosome vaccine development. [ABSTRACT FROM AUTHOR]

Lethal giant larvae (Lgl) are an evolutionarily conserved tumor suppressor present in fungi and animals. It plays an essential role in establishing apical-basal cell polarity, cell proliferation, differentiation, and tissue organization. Here, we report the presence of Lgl gene in the blood fluke Schistosoma japonicum (SjLgl) (GenBank: KF246684). SjLgl protein was mainly distributed in the unique surface tegument structure by immunofluorescence microscopic staining. Using a simple soaking method, a short interfering RNA (siRNA)-based RNA interference approach knocked down the expression of SjLgl in schistosomula in vitro by up to 89.0 %. Moreover, tail vein injection of SjLgl-siRNA into the infected mice reduced SjLgl mRNA levels in vivo by 48.6-85.3 %, depending on the duration of treatments. SjLgl-specific siRNA treatment during the infection in mice significantly altered the surface structure of adult worm, featured by the disappearance or significant reduction of sharp spines on the inner all of oral and ventral suckers. The siRNA also reduced the hatching rates in eggs produced by treated mice by up to 85.3 %. These observations implied that Lgl plays an important role in the development of tegument in schistosomes, and may be explored as a novel target for developing immuno- and/or small molecule-based therapeutics to control and treat the infections caused by schistosome and other flatworms. [ABSTRACT FROM AUTHOR]

Background: Water buffalo and goats are natural hosts for S. japonicum in endemic areas of China. The susceptibility of these two hosts to schistosome infection is different, as water buffalo are less conducive to S. japonicum growth and development. To identify genes that may affect schistosome development and survival, we compared gene expression profiles of schistosomes derived from these two natural hosts using high-throughput microarray technology. Results: The worm recovery rate was lower and the length and width of worms from water buffalo were smaller compared to those from goats following S. japonicum infection for 7 weeks. Besides obvious morphological difference between the schistosomes derived from the two hosts, differences were also observed by scanning and transmission electron microscopy. Microarray analysis showed differentially expressed gene patterns for parasites from the two hosts, which revealed that genes related to lipid and nucleotide metabolism, as well as protein folding, sorting, and degradation were upregulated, while others associated with signal transduction, endocrine function, development, immune function, endocytosis, and amino acid/carbohydrate/glycan metabolism were downregulated in schistosomes from water buffalo. KEGG pathway analysis deduced that the differentially expressed genes mainly involved lipid metabolism, the MAPK and ErbB signaling pathways, progesterone-mediated oocyte maturation, dorso-ventral axis formation, reproduction, and endocytosis, etc. Conclusion: The microarray gene analysis in schistosomes derived from water buffalo and goats provide a useful platform to disclose differences determining S. japonicum host compatibility to better understand the interplay between natural hosts and parasites, and identify schistosome target genes associated with susceptibility to screen vaccine candidates. [ABSTRACT FROM AUTHOR]

Type V collagen is a component of non-cartilaginous tissues and is important in the determination of fibril structure and matrix organization, although its functions are still poorly understood. In this report, RNA interference (RNAi) approaches were used to investigate the effects of knockdown of the schistosome type V collagen ( SjColV) gene. In this study, three different short interfering (si) RNAs targeting different regions of the gene were designed to suppress the expression of SjColV in Schistosoma japonicum using a soaking method. By establishing controls for measuring off-target RNAi effects, we found that different siRNA sequences had different levels of effectiveness. Although all the siRNAs tested reduced SjColV transcript levels, the S1 siRNA consistently reduced SjColV expression to >99 % of the control. In the following experiments, S1 siRNA was adapted to inhibit SjColV expression, and the silencing effects were detected by real-time PCR and Western blot. The spawning and egg hatching of parasites were calculated, while the worms' morphology was taken by scanning electron microscopy. The results show that silencing the expression of SjColV significantly affects the spawning and egg hatching of S. japonicum, and it also affects the worms' morphology. [ABSTRACT FROM AUTHOR]

Schistosomiasis is an important global public health problem, as millions of people are at risk of acquiring this infection. An ideal method for sustainable control of schistosomiasis is using a vaccine alone or in combination with drugs. In the present study, we cloned the SjGALE gene and generated the expression product in E. coli. The expression level of SjGALE during different developmental stages of S. japonicum was evaluated by real-time RT-PCR and western blotting. Immunolocalization indicated that the protein was mainly located on the tegument of the parasite. Infection of rSjGALE-immunized mice demonstrated a 34% and 49% reduction of the mean worm burden and liver egg burden, respectively, in two independent experiments, indicating immune protection. The liver egg count from each female adult worm was significantly reduced by 63% in the two trials. The cytokine profile and IgG isotype analysis demonstrated the induction of a Th1 immune profile in response to immunization with this protein, further suggesting protection against infection. In conclusion, these findings indicated that SjGALE is a potential vaccine against S. japonicum. [ABSTRACT FROM AUTHOR]

The reed vole, Microtus fortis, is the only known mammalian host in which schistosomes of Schistosoma japonicum are unable to mature and cause significant pathogenesis. However, little is known about how Schistosoma japonicum maturation (and, therefore, the development of schistosomiasis) is prevented in M. fortis. In the present study, the ultrastructure of 10 days post infection schistosomula from BALB/c mice and M. fortis were first compared using scanning electron microscopy and transmission electron microscopy. Electron microscopic investigations showed growth retardation and ultrastructural differences in the tegument and sub-tegumental tissues as well as in the parenchymal cells of schistosomula from M. fortis compared with those in BALB/c mice. Then, microarray analysis revealed significant differential expression between the schistosomula from the two rodents, with 3,293 down-regulated (by $2-fold) and 71 up-regulated ($2 fold) genes in schistosomula from the former. The up-regulated genes included a proliferation-related gene encoding granulin (Grn) and tropomyosin. Genes that were down-regulated in schistosomula from M. fortis included apoptosisinhibited genes encoding a baculoviral IAP repeat-containing protein (SjIAP) and cytokine-induced apoptosis inhibitor (SjCIAP), genes encoding molecules involved in insulin metabolism, long-chain fatty acid metabolism, signal transduction, the transforming growth factor (TGF) pathway, the Wnt pathway and in development. TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) and PI/Annexin V-FITC assays, caspase 3/7 activity analysis, and flow cytometry revealed that the percentages of early apoptotic and late apoptotic and/or necrotic cells, as well as the level of caspase activity, in schistosomula from M. fortis were all significantly higher than in those from BALB/c mice. [ABSTRACT FROM AUTHOR]

Abstract: The development of an effective vaccine against the schistosome is thought to be the most desirable means to control schistosomiasis, even though there is an effective means of chemotherapy with praziquantel. A full-length cDNA encoding the Schistosoma japonicum proteasome subunit alpha type 5 protein (SjPSMA5) was first isolated from 18-day-schistosomulum cDNAs. The cDNA had an open reading frame (ORF) of 747bp and encoded 248 amino acids. Real-time quantitative RT-PCR analysis revealed that SjPSMA5 is up-regulated in 18-day and 32-day schistosomes, and the level of expression in male is around fourfold higher than that in female worms at 42days. The SjPSMA5 was subcloned into pET28a(+) and expressed as inclusion bodies in Escherichia coli BL21 (DE3) cells. Western blotting showed that the recombinant SjPSMA5 (rSjPSMA5) was immunogenic. After immunization of BALB/c mice with rSjPSMA5, reductions of 23.29% and 35.24% were obtained in the numbers of worms and eggs in the liver, respectively. The levels of specific IgG antibodies and cells were significantly higher (P< 0.01) in the group vaccinated with rSjPSMA5 combined with Seppic 206 adjuvant than in the other groups, as detected by enzyme linked immunosorbent assay (ELISA) and flow cytometry. The study suggested that rSjPSMA5 induced partial immunoprotection against S. japonicum in BALB/c mice, and it could be a potential vaccine candidate against schistosomiasis. [ABSTRACT FROM AUTHOR]

• The diagnostic method for schistosomiasis control in China is introduced. • Many different diagnostic methods in S. japonicum were analyzed and compared. • New diagnostic strategies need to be studied in the future. Schistosomiasis is an acute and chronic parasitic disease caused by blood flukes (trematode worms) of the genus Schistosoma. Schistosoma japonicum (S. japonicum) infection has decreased significantly in prevalence and intensity of infection in China. However, this disease still remains a serious public health problem in some endemic areas of the Philippines and Indonesia. Thus, more accurate and sensitive methods are much needed for further control of this disease. Here, we review the research progress in techniques for the diagnosis of S. japonicum infection. [ABSTRACT FROM AUTHOR]

In schistosomiasis, eggs produced by bisexual infected mature female schistosome worms (FMS) are the main cause of pathological damage to the host and the dissemination of the disease. Single-sex infected female worms (FSS) cannot completely develop to sexual maturity or produce normal eggs. In this study, isobaric tags for relative and absolute quantitation (iTRAQ)-coupled LC-MS/MS was used to explore the proteome of FSS and FMS of Schistosoma japonicum. A total of 1477 differentially expressed proteins (fold change >1.2, P <.05) between FSS and FMS were identified. Bioinformatics analysis indicated that FMS expressed more proteins related to biosynthetic processes, such as eggshell synthesis, ribosomal synthesis, protein folding, cellular detoxification, and metabolic processes such as protein metabolism and glucose metabolism, whereas more proteins related to locomotion and oxidative phosphorylation were expressed in FSS. Our identification and analysis of differentially expressed proteins between FMS and FSS provides new insights to elucidate the molecular biological mechanisms of female worm sexual maturation and reproduction. Female Schistosome worms must maintain constant pairing contact with male worms for differentiation of their reproductive organs. Mature female worms can produce infectious eggs, cause serious pathological damage to the host and the dissemination of the disease. Unpaired female worms remain small and sexually immature; they do not spawn normally. In this study, iTRAQ-coupled LC-MS/MS was used to explore the whole proteome of single-sex infected female worms (FSS) and bisexual infected mature female worms (FMS) of Schistosoma japonicum. 1477 differentially expressed proteins (DEPs) between FSS and FMS were identified and analyzed. Further research on DEPs' functions in schistosome sexual maturation and reproductive development might provide theoretical bases to explore female maturation and spawning. Unlabelled Image • The morphology of 25d single-sex infected and bisexual infected female worms were visualized by light microscopy. • The proteome of single-sex infected and bisexual infected female worms were explored by iTRAQ-coupled LC-MS/MS approach. • 1477 DEPs between single-sex infected and bisexual infected female worms were identified and analysed. [ABSTRACT FROM AUTHOR]

Background: Schistosoma japonicum (S. japonicum) is an important zoonotic parasite that is prevalent in China and parts of Southeast Asia. Water buffaloes are an important reservoir and the main transmission sources of S. japonicum. However, self-curing and resistance to re-infection have been observed in water buffaloes. Results: In this study, we compared the morphometry and differences in transcriptional expression of adult S. japonicum worms recovered from primary-infected and re-infected water buffaloes using Illumina RNA-sequencing (RNA-Seq) technology. Results of morphometry analysis revealed that adult S. japonicum worms recovered from re-infected water buffaloes were runtish with smaller organs. The ventral length of male worms was shorter in re-infected buffaloes (328 ± 13 vs 273 ± 8 µm, P < 0.05), and in female worms the oral sucker length (44 ± 3 vs 33 ± 5 µm, P < 0.05), ovary length (578 ± 23 vs 297 ± 27 µm, P < 0.05) and width (150 ± 8 vs 104 ± 9 µm, P < 0.05) were shorter, with fewer eggs in the uteri (41 ± 2 vs 12 ± 1, P < 0.05). Of 13,605 identified genes, 112 were differentially expressed, including 51 upregulated and 61 downregulated genes, in worms from re-infected compared with primary-infected water buffaloes. Gene ontology (GO) enrichment analysis revealed that GO terms such as "oxidation-reduction process", "calcium-dependent phospholipid binding", "lipid binding" and "calcium ion binding" were significantly enriched in downregulated genes, whereas GO terms related to metabolism and biosynthesis were significantly enriched in upregulated genes. The results revealed that the downregulation of some important genes might contribute to a reduction in worm numbers and maldevelopment of surviving worms in re-infected water buffaloes. Furthermore, upregulation of genes related to metabolic processes and biosynthesis might be a compensatory mechanism of worms in disadvantageous environments. Conclusions: To our knowledge, our results present the first large-scale transcriptional expression study identifying the differences between adult S. japonicum worms from primary-infected and re-infected water buffaloes, and particularly emphasize differential expression that may affect the survival and growth of worms in re-infected water buffalo. This will provide new insight into screening for anti-schistosome targets and vaccine candidates. [ABSTRACT FROM AUTHOR]

Background: Yellow cattle and water buffalo are important natural reservoir hosts and the main transmission sources of Schistosoma japonicum in endemic areas of China. The worms from the two hosts have marked differences in general worm morphology and ultrastructure, gene transcription and protein expression profiles. Results: To investigate microRNAs (miRNAs) involved in the regulation of schistosome development and survival, we compared miRNA expression profiles of adult schistosomes derived from yellow cattle and water buffalo by using high-throughput sequencing with Illumina Hiseq Xten. Schistosoma japonicum from water buffalo and yellow cattle yielded 63.78 million and 63.21 million reads, respectively, of which nearly 50% and 49% could be mapped to selected miRNAs in miRbase. A total of 206 miRNAs were identified, namely 79 previously annotated miRNAs of S. japonicum and 127 miRNAs that matched with the S. japonicum genome and were highly similar to the annotated miRNAs from other organisms. Among the 79 miRNAs, five (sja-miR-124-3p, sja-miR-219-5p, sja-miR-2e-3p, sja-miR-7-3p and sja-miR-3490) were significantly upregulated in the schistosomes from water buffalo compared with those from yellow cattle. A total of 268 potential target genes were predicted for these five differentially expressed miRNAs. Eleven differentially expressed targets were confirmed by qRT-PCR among 15 tested targets, one of which was further validated through dual-luciferase reporter assay. Among the 127 'possible' S. japonicum miRNAs, ten were significantly differentially expressed in the schistosomes from these two hosts. Conclusions: These results highlight the important roles of miRNAs in regulating the development and survival of schistosomes in water buffalo and yellow cattle and facilitate understanding of the miRNA regulatory mechanisms in schistosomes derived from different susceptible hosts. [ABSTRACT FROM AUTHOR]