Introduction: Anopheles stephensi is the main malaria vector in Southeast Asia. Recently, plant-sourced larvicides are attracting great interests. Methods: The essential oil was extracted from the leaf of Cinnamomum camphora (L.), and a bioassay was conducted to determine the larvicidal efficacy. The chemical composition of the essential oil was determined by GC-MS analysis. Results: The oil showed strong, dose-dependent larvicidal activities. The onset of larvicidal efficiency was rapid. The LC50 and LC95 were determined as 0.146% and 1.057% at 1 h, 0.031% and 0.237% at 12 h, 0.026% and 0.128% at 24 h, respectively. The oil contains 32 compounds. Conclusions: The essential oil of C. camphora leaf has an excellent larvicidal potential for the control of A. stephensi.
Paragonimiasis skrjabini is a kind of zoonosis and prevalent in 16 provinces in China, such as Chongqing, Fujian, Sichuan, and Yunnan. However, sensitive and efficient diagnostic methods for the infection are limited. In order to provide a more convenient and simple method for serologic diagnosis, the recombinant P. skrjabini cysteine protease (PsCP) was expressed, purified, and then used to develop an indirect enzyme-linked immunosorbent assay (ELISA) for detecting anti-PsCP antibodies in human. Given the positive/negative cutoff value as 0.606, the maximum dilution of human sera in which anti-PsCP antibodies could be detected was 1:12,800. In addition, the coefficients of variation (CVs) of inter-assay and intra-assay experiments were both below 10 %. Furthermore, the sensitivity of the PsCP-based ELISA was 95.5 %, and the indirect ELISA displays no cross-reactivity with human antisera against Echinococcus granulosus, Taenia solium, Schistosoma japonicum, and Trichinella spiralis, either. In conclusion, recombinant PsCP was readily produced and used to establish a simple PsCP-based ELISA that provided a highly specific and sensitive method for analysis of clinical samples. Besides, the method can also probably be used to diagnose P. skrjabini infection in animals.
Malaria is still the most widespread parasitic disease and causes the most infections globally. Owing to improvements in sanitary conditions and various intervention measures, including the use of antimalarial drugs, the malaria epidemic in many regions of the world has improved significantly in the past 10 years. However, people living in certain underdeveloped areas are still under threat. Even in some well-controlled areas, the decline in malaria infection rates has stagnated or the rates have rebounded because of the emergence and spread of drug-resistant malaria parasites. Thus, new malaria control methods must be developed. As the spread of the Plasmodium parasite is dependent on the part of its life cycle that occurs in mosquitoes, to eliminate the possibility of malaria infections, transmission-blocking strategies against the mosquito stage should be the first choice. In fact, after the gametocyte enters the mosquito body, it undergoes a series of transformation processes over a short period, thus providing numerous potential blocking targets. Many research groups have carried out studies based on targeting the blocking of transmission during the mosquito phase and have achieved excellent results. Meanwhile, the direct killing of mosquitoes could also significantly reduce the probability of malaria infections. Microorganisms that display complex interactions with Plasmodium, such as Wolbachia and gut flora, have shown observable transmission-blocking potential. These could be used as a biological control strategy and play an important part in blocking the transmission of malaria.
Mosquitoes are important vectors of many infectious diseases. Bacillus sphaericus (Bs) is an ideal larvicide and has attracted more and more attention, recently. However, the fundamental research of its application is very limited, especially on the subsequent impact of Bs exposure on mosquito's fecundity and resistance emergence. Through bioassay, LC50 and LC95 of Bs in killing Anopheles dirus larvae were determined as 9.793 ± 1.878 IU/L and 62.4 ± 6.438 IU/L at 48 h posttreatment, 7.128 ± 0.913 IU/L and 34.385 ± 12.547 IU/L at 72 h post treatment, respectively. After being treated with a sub-lethal dose of Bs, gravidity, oviposition, hatch, pupation, and eclosion of the surviving mosquitoes were counted and analyzed to elucidate the subsequent effects of Bs exposure on the reproductive capacity of A. dirus. The result interestingly showed that the exposure of Bs significantly reduced the oviposition ability of the surviving A. dirus, without effect on egg formation/gravidity, hatch, pupation, and eclosion. The surviving mosquitoes were also maintained routinely for generations to test the sustained effect of Bs exposure on the fecundity of the offsprings. After conventional breeding for generations, the capacity of egg laying totally recovered. To explore the rules of resistance development, bioassays were performed after treatment twice with a sub-lethal dose of Bs on two continuous generations of A. dirus larvae. The killing efficacies between the Bs treated group and control group were compared. The results showed that LC50 and LC95 increased by 4.35- and 7.37-folds after treatment with the sub-lethal dose of Bs on two consecutive generations, respectively. The results indicated that A. dirus was sensitive to Bs, which could reduce oviposition of the surviving A. dirus. The subsequent effect might help to further decrease the mosquito population. However, a sub-lethal dose of Bs exposure could easily cause resistance development. Our study provides a dose standard and reference for the rational use of Bs, which will be helpful for mosquito control.
Background: Vector control with Bacillus sphaericus (Bs) is an effective way to block the transmission of malaria. However, in practical application of Bs agents, a sublethal dose effect was often caused by insufficient dosing, and it is little known whether the Bs exposure would affect the surviving mosquitoes' vector capacity to malaria. Methods: A sublethal dose of the Bs 2362 strain was administrated to the early fourth-instar larvae of Anopheles dirus to simulate shortage use of Bs in field circumstance. To determine vector competence, mosquitoes were dissected and the oocysts in the midguts were examined on day 9-11 post-infection with Plasmodium yoelii. Meanwhile, a SYBR quantitative PCR assay was conducted to examine the transcriptional level of the key immune molecules of mosquitoes, and RNA interference was utilized to validate the role of key immune effector molecule TEP1. Results: The sublethal dose of Bs treatment significantly reduced susceptibility of An. dirus to P. yoelii, with the decrease of P. yoelii infection intensity and rate. Although there existed a melanization response of adult An. dirus following challenge with P. yoelii, it was not involved in the decrease of vector competence as no significant difference of melanization rates and densities between the control and Bs groups was found. Further studies showed that Bs treatment significantly increased TEP1 expression in the fourth-instar larvae (L4), pupae (Pu), 48 h post-infection (hpi) and 72 hpi (P < 0.001). Further, gene-silencing of TEP1 resulted in disappearance of the Bs impact on vector competence of An. dirus to P. yoelii. Moreover, the transcriptional level of PGRP-LC and Rel2 were significantly elevated by Bs treatment with decreased expression of the negative regulator Caspar at 48 hpi, which implied that the Imd signaling pathway was upregulated by Bs exposure. Conclusions: Bs exposure can reduce the vector competence of An. dirus to malaria parasites through upregulating Imd signaling pathway and enhancing the expression of TEP1. The data could not only help us to understand the impact and mechanism of Bs exposure on Anopheles' vector competence to malaria but also provide us with novel clues for wiping out malaria using vector control.
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