RNAi-mediated mortality of Culex quinquefasciatus using two delivery methods of potential field application.

With increasing reports of resistance to traditional insecticides, there is a need for innovative ways for mosquito control. RNAi is a sequence-specific molecular biology technique for gene silencing through degradation of mRNA and prevention of protein translation. Some genes are essential for insect life and their silencing can lead to insect morbidity and/or mortality. Searching for lethal genes in Culex quinquefasciatus, we found dynamin, ROP, HMGR and JHAMT to be lethal targets for RNAi in initial screening through larval soaking in dsRNA solution. Two delivery methods, chitosan nanoparticles and genetically modified yeast cells, were used in this study and proved effective in inducing high larval mortality and low adult emergence. Adult emergence after chitosan nanoparticles/dsRNA treatment was 12.67% ± 1.76 (HMGR), 17.33% ± 1.76 (dynamin), 18.67% ± 0.67 (ROP), and 35.33% ± 0.67 (JHAMT). Genetically modified yeast increased mortalities as adult emergence was 8.33% ± 1.67 (HMGR), 13.33% ± 3.33 (dynamin), and 10% ± 2.89 (JHAMT and ROP). Chitosan nanoparticles retained 75% of its biological activity whereas yeast cells retained >95% of their activities after 7 days of incubation in water. In conclusion, our results showed that these four genes are good targets for C. quinquefasciatus control using RNAi packaged in either chitosan nanoparticles or genetically modified yeast cells.

RNAi-Mediated Screening of Selected Target Genes Against Culex quinquefasciatus (Diptera: Culicidae).

Culex quinquefasciatus, a member of the Culex pipiens complex, is widespread in Saudi Arabia and other parts of the world. It is a vector for lymphatic filariasis, Rift Valley fever, and West Nile virus. Studies have shown the deleterious effect of RNA interference (RNAi)-mediated knockdown of various lethal genes in model and agricultural pest insects. RNAi was proposed as a tool for mosquito control with a focus on Aedes aegypti and Anopheles gambiae. In this study, we examined the effect of RNAi of selected target genes on both larval mortality and adult emergence of Cx. quinquefasciatus through two delivery methods: soaking and nanoparticles. Ten candidate genes were selected for RNAi based on their known lethal effect in other insects. Disruption of three genes, chitin synthase-1, inhibitor of apoptosis 1, and vacuolar adenosine triphosphatase, resulted in the highest mortality among the selected genes using the two treatment methods. Silencing the other seven genes resulted in a medium to low mortality in both assays. These three genes are also active against a wide range of insects and could be used for RNAi-based mosquito control in the future.

Delivery Methods for RNAi in Mosquito Larvae.

Mosquito-transmitted diseases pose a threat for a great portion of the world population. Chemical insecticides are the main tool for mosquito control. Heavy dependence on chemicals created several problems such as resistance development in many mosquito species, environmental effects, and human health issues. Other tools for mosquito control were developed and used in some parts of the world. Ribonucleic acid interference (RNAi) is a reverse genetic mechanism that was recently introduced as a new tool for pest control. Regarding mosquito, RNAi was used to study gene function and to discover genes that can be used as targets for control purposes. Several delivery methods are used to induce RNAi in mosquito larvae. Some methods such as injection and soaking are used routinely in RNAi research but have no application in the field. Other methods such as nanoparticles and microbes have some characteristics that make them good candidates for field application. In this report, we will focus on delivery methods for RNAi in mosquito larvae and will give examples for each method.

Assessment and an updated list of the mosquitoes of Saudi Arabia.

BACKGROUND: Mosquito-borne pathogens are important causes of diseases in the Kingdom of Saudi Arabia. Knowledge of the mosquito fauna is needed for the appropriate control of the vectors that transmit the pathogens and prevent the diseases they cause. An important first step is to have an up-to-date list of the species known to be present in the country. Original occurrence records were obtained from published literature and critically scrutinized to compile a list of the mosquito species that occur within the borders of the Kingdom. RESULTS: Fifty-one species have been recorded in the Kingdom; however, the occurrence of two of these species is unlikely. Thus, the mosquito fauna of the Kingdom comprises 49 species that include 18 anophelines and 31 culicines. Published records are provided for each species. Problematic records based on misidentifications and inappropriate sources are discussed and annotated for clarity. CONCLUSION: Integrated morphological and molecular methods of identification are needed to refine the list of species and accurately document their distributions in the Kingdom.

A new species of the genus Amyrsidea (Phthiraptera: Amblycera: Menoponidae) parasitizing domestic chickens in Saudi Arabia.

Amyrsidea Ewing, 1927 is a genus of chewing lice comprising 52 species, divided into five subgenera: Amyrsidea sensu stricto, Argimenopon Eichler, 1947, Cracimenopon Carriker, 1954, Desumenopon Carriker, 1954 and Numidimenopon Scharf & Price, 1977 (see Price et al. 2003: 86), which parasitise a wide range of hosts belonging to the avian order Galliformes (see Price et al. 2003: 321). Species of Amyrsidea can be distinguished from species of Menacanthus Neumann, 1912-the other menoponid genus with species parasitic on the same hosts-by lacking ventral spinous processes in the head (Scharf & Price 1977: 815). Revisions of all subgenera and species of Amyrsidea, including keys for their identification, were published by Scharf & Price (1977, 1983) and Scharf & Emerson (1983, 1984).

New host/parasite record for very rare chewing louse Cuculotogaster arabicus (Clay, 1938) (Phthiraptera: Ischnocera) on endemic mountain partridges of Arabian Peninsula.

The endemic avifauna of Arabia is unique and characteristic through the whole region. Little is known about these birds and their ectoparasites. The Arabian partridge Alectoris melanocephala (Rüppell, 1835) and Philby's partridge Alectoris philbyi Lowe, 1934 are two endemic species which are distributed through the Sarawat Mountains in Saudi Arabia and Yemen. Captive breeding population of these birds were examined for chewing lice at the National Wildlife Research Center (NWRC) near Ta'if. Only one rare species of louse, Cuculotogaster arabicus (Clay 1938), was found to infest these birds. The occurrence of such species is considered a first record of the genus Cuculotogaster from Saudi Arabia, and the association of C. arabicus with Philby's partridge is considered a new host/parasite association. The clear diagnostic characters, high definition photos, and drawing of male genitalia are available through this paper.

Little known chewing lice (Phthiraptera) infesting crab plover Dromas ardeola Paykull, 1805 (Charadriiformes: Dromadidae) from the Red Sea.

Scanty information is available for many species of chewing lice of marine birds. Through this work we investigated one of most characteristic marine bird for chewing lice. Seven individuals of crab plovers Dromas ardeola Paykull, 1805 were trapped using standard mist nets on Humr Island in Farasan Archipelago, Saudi Arabia. Two species of chewing lice were found to infest these birds: Actornithophilus ardeolae Timmermann, 1954 of suborder Amblycera and Quadraceps brunneus (Nitzsch in Giebel, 1866) of suborder Ischnocera. Diagnostic characters, data of specimens examined, high definition photos and host distribution map are provided through this paper.

New records of species of Saemundssonia (Insecta: Phthiraptera: Philopteridae) infesting breeding terns in the Arabian Peninsula, with notes on their phylogeny and ecology.

Six species of terns, which breed on the Arabian Peninsula, were examined for head chewing lice of the genus Saemundssonia in four different islands around the coasts of Saudi Arabia, both in the Red Sea and in the Arabian Gulf. Four louse species were collected: Saemundssonia laticaudata, Saemundssonia melanocephalus, Saemundssonia meridiana and Saemundssonia sternae, of which three are recorded for the first time from this region. Also, we record three new host-louse associations for the world-Saemundssonia laticaudata and Saemundssonia sternae from white-cheeked terns and Saemundssonia melanocephalus from Saunders's terns-including a host-switch event of Saemundssonia laticaudata on white-cheeked terns in the Karan Island population. Gene bank data for the COI gene from seven species of Saemundssonia that infest marine birds were used to propose evolutionary trees using two different statistical methods: maximum parsimony (MP) and neighbour joining (NJ). The result indicated that the tree which was produced by NJ is likely to be more accurate as it appeared more compatible with hosts' phylogeny. The trees indicate relationships between tern Saemundssonia and congeneric species from other marine birds, especially from gulls. An ANOVA was also conducted to test the mean parasite load for each tern species studied, and results indicate that there is a relation between louse loads and colonization behaviour of the hosts. Data from lice examined and illustrations of lice and their hosts are also included.

Myiasis in sheep farms in Riyadh Region, Saudi Arabia.

During a survey for myiasis in Riyadh Region, 3712 sheep were examined. 73 animals (2%) were infested with different dipterous larvae. Out of the 115 larvae recovered, 100 larvae (87%) were Chrysomya bezziana, 10 (8.7%) were C. albiceps, and 5 (4.3%) were Wohlfahrtia nuba. The prevalence rate of larval myiasis among young sheep was 60%, and 40% among adults. The myiasis incidences were highest during Mar-May (60%) and Sept-Nov (31.5%) where temperature and R.H. are optimum. In the dry hot season (Jun-August) and cold season (Dec-Feb) infestation incidences were low (5%&1.5% respectively). Temperature and R.H. are conducive in controlling myiasis in other regions of the Kingdom.

Acaricidal effects of cardiac glycosides, azadirachtin and neem oil against the camel tick, Hyalomma dromedarii (Acari: Ixodidae).

The cardiac glycoside, digitoxin, from Digitalis purpurea L (Scrophulariaceae), a cardiac glycosidal (cardenolide) extract from Calotropis procera (Ait) R Br (Asclepiadaceae), azadirachtin and neem oil from Azadirachta indica A Juss (Meliaceae) were tested for their effects against larvae and adult stages of the camel tick, Hyalomma dromedarii Koch (Acari: Ixodidae). The contact LC50 values of the first three materials against adults were 4.08, 9.63 and >40.7 microg cm(-2), respectively, whereas the dipping LC50 values of the four materials were 409.9, 1096, >5000 and >5000 mg litre(-1), respectively. Contact and dipping LC50 values of the extract and azadirachtin against larvae were 6.16, >20.3 microg cm(-2) and 587.7 and >2500 mg litre(-1), respectively. Azadirachtin had no effects on egg production or feeding of adults up to 5000 mg litre(-1); however at 2500 mg litre(-1), it caused significant reduction in feeding activity of larve, prolonged the period for moulting to nymphal stage, and caused 60% reduction in moultability. Results of the two cardiac glycoside materials are comparable with those of several commercial acaricides. The risks and benefits associated with the use of cardiac glycosides are considered.