Mulberry leaf extract (MLE), an active substance extracted from mulberry leaves, is known to have a positive effect on several physiological functions. The current study examined the effects of dietary MLE in feed at concentrations 0, 3.0, 6.0, 9.0, 12.0 and 15.0 g/kg on the growth performance and gastrointestinal and hepatic functions of Andrias davidianus for 12 weeks. Results indicated that the final body weight, weight gain rate (WGR), specific growth rate and feed intake (FI) of A. davidianus increased with the increase in the dietary MLE up to 9.0 g/kg and declined thereafter, while the feed conversion ratio (FCR) exhibited an opposite trend. Meanwhile, A. davidianus with a dietary intake of 9.0 g/kg MLE showed higher levels of crude protein in muscles and lower levels of moisture and crude lipid levels in the liver and muscles when compared with the control. In addition, dietary MLE increased the density and length of the villi and decreased the cavity rate in the foregut, enhancing the activities of carbonic anhydrase, H+-K+-ATPase, pepsin, intestinal trypsin, lipase and Na+-K+-ATPase (p < .05) in the stomach. Furthermore, dietary MLE increased the intestinal and hepatic superoxide dismutase activities and total antioxidative capacities but decreased their malondialdehyde contents in A. davidianus. Dietary MLE also significantly increased the immune parameters, and the plasma total protein, albumin and immunoglobulin M contents but significantly decreased the aspartate aminotr-ansferase, alanine aminotransferase and diamine oxidase activities, and the total bilirubin, direct bilirubin, cholesterol, triglyceride and endotoxin contents (p < .05). In conclusion, a quadratic regression analysis of WGR and FCR indicated that the optimum level of MLE for A. davidianus was between 8.21 and 8.30 g/kg of the diet.

Schistosomes are causative agents of human schistosomiasis, which is endemic in tropical and subtropical areas of the world. Adult schistosomes can survive in their final hosts for several decades, and they have evolved various strategies to overcome the host immune response. Consequently, understanding the mechanisms that regulate parasitic cell survival will open avenues for developing novel strategies against schistosomiasis. Our previous study suggested that an inhibitor of apoptosis protein in Schistosoma japonicum (SjIAP) may play important roles in parasitic survival and development. Here, we demonstrated that SjIAP can negatively regulate cellular apoptosis in S. japonicum by suppressing caspase activity. Immunohistochemistry analysis indicated that SjIAP ubiquitously expressed within the worm body including the tegument. Silencing of SjIAP expression via small interfering RNA led to destruction of the tegument integrity in schistosomes. We further used co-immunoprecipitation to identify interaction partners of SjIAP and revealed the tegument protein SjTeg-20 as a putative interacting partner of SjIAP. The interaction between SjIAP and SjTeg-20 was confirmed by a yeast two-hybrid (Y2H) assay. Moreover, results of a TUNEL assay, RNA interference, scanning and transmission electron microscopy, caspase assays, transcript profiling, and protein localization of both interacting molecules provided first evidence for an essential role of SjIAP and SjTeg-20 to maintain the structural integrity of the tegument by negatively regulating apoptosis. Taken together, our findings suggest that the cooperative activities of SjIAP and SjTeg-20 belong to the strategic inventory of S. japonicum ensuring survival in the hostile environment within the vasculature of the final host. Schistosomiasis is a worldwide public health concern particularly in developing countries. The causative agents, schistosomes, can survive within the vascular system of their final hosts for several decades despite facing the host’s immune response. Therefore, elucidating the mechanism of cell survival will contribute to the understanding of host-parasite interaction and may lead to the identification of suitable targets for developing novel strategies against schistosomiasis. Inhibitor of apoptosis proteins are highly conserved proteins functioning as endogenous inhibitors of apoptotic cell death. Here, we demonstrated that an inhibitor of apoptosis protein of Schistosoma japonicum (SjIAP) governs the integrity of the tegument of schistosomes by inhibiting cellular apoptosis of the parasite. Further studies revealed that SjTeg-20, an S. japonicum tegumental protein, cooperates with SjIAP to inhibit apoptosis in schistosomes. Our findings provide new insights into the role of SjIAP and SjTeg-20 in maintaining the integrity of the worm tegument by negatively regulating apoptosis.