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According to the WHO there are more than 700,000 deaths every year involving vector-borne diseases such as malaria, dengue, Chagas disease, yellow fever and Japanese encephalitis. Aedes aegypti, the principal vector of the dengue virus, is of great concern in various parts of the world, especially in tropical and subtropical countries. Vector control through insecticide application is one of the best ways to control the disease’s transmission. Thus, insecticide resistance in Ae. aegypti poses a significant threat to public health worldwide. The use of plant natural product-based insecticides that are less harmful to the environment, and without known resistance development, constitutes an alternative to chemical insecticides. Given this, a methanolic extract from Vitex ovata plants was prepared and tested for its larvicidal effect against Ae. aegypti. A susceptibility test on Ae. aegypti larvae was conducted using the standard WHO method. Results showed that the methanolic extract of V. ovata had larvicidal activity against Ae. aegypti with LC50 values of 2114 mg/L, and achieved 84% mortality with the highest concentration at 10,000 mg/L. This study showed that the crude extract of V. ovata bioactive molecules could be potentially developed as biolarvicides for Aedes mosquito vector control. This study recommends future research on using different solvents in the isolation of active ingredients from V. ovata, identification of phytochemicals with larvicidal properties, a toxicity study and lastly, an evaluation of the effectiveness of controlling Aedes in small-scale field trials for environmentally safe botanical insecticide intervention.

Metarhizium anisopliae (Metchnikoff) Sorokin, a pathogenic fungus to insects, infects the subterranean termite, Coptotermes curvignathus Holmgren, a devastating pest of plantation trees in the tropics. Electron microscopy and proteomics were used to investigate the infection and developmental process of M. anisopliae in C. curvignathus. Fungal infection was initiated by germ tube penetration through the host’s cuticle as observed at 6 h post-inoculation (PI), after which it elongated into the host’s integumental tissue. The colonization process continued as seen from dissemination of blastospores in the hemocoel at 96 h PI. At this time point, the emergent mycelia had mummified the host and forty-eight hours later, new conidia were dispersed on the termites’ body surface. Meanwhile, hyphal bodies were observed in abundance in the intercellular space in the host’s body. The proteomes of the pathogen and host were isolated separately using inoculated termite samples withdrawn at each PI-time point and analyzed in two-dimensional electrophoresis (2-DE) gels. Proteins expressed in termites showed evidence of being related to cell regulation and the immune response, while those expressed in M. anisopliae, to transportation and fungal virulence. This study provides new information on the interaction between termites and its entomopathogen, with potential utilization for developing future biopesticide to control the termite population.

Simple Summary The use of Metarhizium anisopliae as a biological control of insect pests has been experimented in the laboratory as well as in field trials. This includes against the termite Coptotermes curvignathus, however the results have varying degrees of success. One reason could be due to the lack of detailed knowledge on the molecular pathogenesis of M. anisopliae. In the current study, the conidial suspension of M. anisopliae isolate PR1 was first inoculated on the C. curvignathus, after which the pathogenesis was examined using two different approaches: electron microscopy and protein expression. At the initiation stage, the progression observed and documented including adhesion, germination, and penetration of the fungus on the cuticle within 24 h after inoculation. Later, this was followed by colonization and spreading of the fungus at the cellular level. Proteomics of C. curvignathus witnessed the expression of proteins related to cell regulation and defense, while in M. anisopliae, protein related to transport and fungal virulence were expressed throughout the infection. These findings offer relevant knowledge for use in the development of M. anisopliae as a prospective biological control agent for termites in the future. Abstract Metarhizium anisopliae (Metchnikoff) Sorokin, a pathogenic fungus to insects, infects the subterranean termite, Coptotermes curvignathus Holmgren, a devastating pest of plantation trees in the tropics. Electron microscopy and proteomics were used to investigate the infection and developmental process of M. anisopliae in C. curvignathus. Fungal infection was initiated by germ tube penetration through the host’s cuticle as observed at 6 h post-inoculation (PI), after which it elongated into the host’s integumental tissue. The colonization process continued as seen from dissemination of blastospores in the hemocoel at 96 h PI. At this time point, the emergent mycelia had mummified the host and forty-eight hours later, new conidia were dispersed on the termites’ body surface. Meanwhile, hyphal bodies were observed in abundance in the intercellular space in the host’s body. The proteomes of the pathogen and host were isolated separately using inoculated termite samples withdrawn at each PI-time point and analyzed in two-dimensional electrophoresis (2-DE) gels. Proteins expressed in termites showed evidence of being related to cell regulation and the immune response, while those expressed in M. anisopliae, to transportation and fungal virulence. This study provides new information on the interaction between termites and its entomopathogen, with potential utilization for developing future biopesticide to control the termite population.

Trypanosoma (Megatrypanum) theileri is a non-pathogenic or weakly pathogenic parasite of domestic cattle that is cyclically transmitted by blood-sucking insects, mainly tabanid flies. It has been reported in several countries like Brazil, Venezuela, Japan, Taiwan, Thailand, Vietnam, and the Philippines. Although the ruminant industry is actively expanded in Malaysia, T. theileri and T. theileri-like trypanosomes have never been reported from Malaysia. The low pathogenicity of this species might be the main reason for overlooking T. theileri in this country. This paper describes an unforeseen finding of T. theileri from the outbreak of T. evansi in the state of Kelantan, Malaysia. This is the first time T. theileri reported in Malaysia, and also the first time T. theileri is reported in equid. Clinical signs compatible with infection by blood protozoa were observed; however, it was uncertain whether they were due to T. theileri infection. The detection of T. theileri from the blood sample and Tabanus sp. were confirmed through molecular analysis with PCR and DNA sequencing. In the present study, T. theileri from one horse and one Tabanus sp. were clustered with sequences of the previously described phylogenetic lineages from Japan, Chad and Brazil cattle. Even though this species is claimed to be host-specific with ruminant host restriction, the finding from this study suggested that T. theileri can infect equine whilst other isolates are known to infect ruminant species only. It is suspected there were two genotypes of T. theileri circulating in at least two districts of Kelantan. Thus, further study on multiple DNA regions should be conducted to determine the strains of detected T. theileri in Malaysia. Its impact on the horse and cattle industry should also be revised.

The termite, Coptotermes curvignathus has been reported in Malaysia and it is responsible for the destruction of various timberbased products, plantation and orchard trees. Controlling this pest population using biological pesticide agents, such as entomopathogenic fungi are highly desirable when compared to chemical pesticides because the latter often leads to many environmental concerns. In this study, we isolated Metarhizium anisopliae from four different locations in Peninsular Malaysia and tested their pathogenicity on C. curvignathus. Another six isolates from a previous work were also included. Conidia in the form of suspension at 1x10 7 conidia/mL were applied on worker termites and observed for their mortality within a 12dayperiod. In addition, fungal progression rate was calculated based on the number of days it took for the mycelia to emerge and the conidia to form on the infected termites. Among the ten fungal isolates tested, PR1 yielded the highest mortality (97%) and the shortest median lethal time (LT50= 1.5 days), while TFFH3 and PKLG isolates had the highest rate in mycelia (88%) and conidia (80%) formation, respectively, and they both caused mortality of 93% and LT50 within2 days. PR1’s performance in infecting C. curvignathus evidently could augment its potential use as a microbial pesticide for a sustainable and environment friendly management of termites