Identification of Wolbachia new strains from Aedes aegypti mosquitoes, the vector of dengue fever in Jeddah Province.

Wolbachia are endosymbiotic bacteria found within many arthropods, including insects. A variety of benefits are provided by these bacteria to human and insect hosts, including protection from viruses and parasites and the ability to kill males. In this study, Wolbachia was identified in Aedes aegypti present in Jeddah, Saudi Arabia. A population of mosquitoes was collected from eight different areas, processed, and tested for Wolbachia using 16 S rRNA specific to Wolbachia bacteria and Wolbachia surface protein (wsp) under optimized PCR conditions. In five ecologically diverse sites to determine Wolbachia prevalence, we identified eleven diverse novel resident Wolbachia strains within Ae. Aegypti for the first time in Jeddah, Saudi Arabia. Future studies to evaluate the possible use of Wolbachia as a control agent in Aedes sp. in Saudi Arabia are necessary. Wolbachia prevalence rates and strain characterization through Sanger sequencing with multilocus sequence typing (MLST) and phylogenetic analysis revealed significant diversity. In developing biocontrol strategies, it is beneficial to consider the implications of resident Wolbachia strains.

Resistance development to bioinsecticides in Aedes aegypti (Culicidae: Diptera), the vector of dengue fever in Saudi Arabia.

A laboratory strain of Aedes aegypti (L) was subjected repeatedly to larval selection pressure with two bacterial insecticides, spinosad (Saccharopolyspora spinosa) and bacilod (Bacillus thuringiensis israelensis). The results indicated that the mosquito Ae. aegypti acquired low resistance to spinosad and bacilod by about 3.1 and 2.4-fold, respectively, due to selection pressure for fifteen successive generations. The slope values of the selected strains were increased gradually from one generation to the next, indicating moderate homogeneity between individuals in their response to the test bio-insecticide. Moreover, larval selection with current bacterial bioinsecticides prolonged the time required to digest a blood meal. It showed an evident decrease in the reproductive potential of adult mosquitoes surviving selected larvae.

Synthesis and characterization of different complexes derived from Schiff base and evaluation as a potential anticancer, antimicrobial, and insecticide agent.

The condensation of (1H-benzimidazole-2-yl) methanamine, with 2-hydroxy naphthaldehyde lead to Schiff base ligand (H2L) (1). This was later reacted with metal salts (ZnCl2, CrCl3·6H2O, and MnCl2·4H2O) to afford the corresponding metal complexes. Biological activity findings indicate that the metal complexes have promising activity against Escherichia coli and Bacillus subtilis and modest activity against Aspergillus niger. The in vitro anticancer activities of Zn (II), Cr (III), and Mn (II) complexes were investigated and the best results were observed with Mn (II) complex as the most potent cytotoxic agent toward human cell lines colorectal adenocarcinoma HCT 116, hepatocellular carcinoma HepG2 and breast adenocarcinoma MCF-7 with 0.7, 1.1 and 6.7 µg of inhibitory concentration IC50 values respectively. Consequently, the Mn (II) complex and ligand were docked inside the energetic site of ERK2 and exhibited favorable energy for binding. The investigation of biological tests towards mosquito larvae indicates that Cr (III) and Mn (II) complexes manifest strong toxicity against Aedes aegypti larvae with 3.458 and 4.764 ppm values of lethal concentration LC50, respectively.

Biological comparative study between Wolbachia-infected Aedes aegypti mosquito and Wolbachia-uninfected strain, Jeddah city, Saudi Arabia.

In this study, samples of Wolbachia-infected Aedes aegypti mosquitoes were collected from Al-Safa district in Jeddah city, Saudi Arabia. The presence of Wolbachia bacteria in mosquitoes was confirmed by PCR technique and they were reared and propagated in the laboratory. Comparative studies were conducted between Wolbachia-infected A. Aegypti and the Wolbachia-uninfected laboratory strain in terms of their ability to withstand drought, resist two types of insecticides and the activities of pesticide detoxification enzymes. The Wolbachia-infected A. aegypti strain proved less able to withstand the drought period, as the egg-hatching rate of the Wolbachia-uninfected strain was greater than that of the Wolbachia-infected strain after one, two and three months of dry periods. Compared to the Wolbachia-uninfected strain, the Wolbachia-infected strain demonstrated a relatively greater resistance to tested pesticides, namely Baton 100EC and Fendure 25EC which may be attributed to the higher levels of the detoxification enzymes glutathione-S-transferase and catalase and the lower levels of esterase and acetylcholine esterase.

Molecular analysis of knockdown resistance (kdr) mutations in the voltage-gated sodium channel gene of Aedes aegypti populations from Saudi Arabia.

BACKGROUND: The Aedes aegypti mosquito is the primary vector for dengue, chikungunya, yellow fever and Zika viruses worldwide. The first record of Ae. aegypti in southwestern Saudi Arabia was in 1956. However, the first outbreak and cases of dengue fever were reported in 1994, and cases have increased in recent years. Vector control for Ae. aegypti mainly uses pyrethroid insecticides in outdoor and indoor space spraying. The constant use of pyrethroids has exerted intense selection pressure for developing target-site mutations in the voltage-gated sodium channel (vgsc) gene in Ae. Aegypti against pyrethroids-mutations that have led to knockdown resistance (kdr). METHODS: Aedes aegypti field populations from five regions (Jazan, Sahil, Makkah, Jeddah and Madinah) of southwestern Saudi Arabia were genotyped for known kdr mutations in domains IIS6 and IIIS6 of the vgsc gene using polymerase chain reaction (PCR) amplification and sequencing. We estimated the frequency of kdr mutations and genotypes from Saudi Arabia as well as from other countries, Thailand, Myanmar (Southeast Asia) and Uganda (East Africa). We constructed haplotype networks to infer the evolutionary relationships of these gene regions. RESULTS: The three known kdr mutations, S989P, V1016G (IIS6) and F1534C (IIIS6), were detected in all five regions of Saudi Arabia. Interestingly, the triple homozygous wild genotype was reported for the first time in two individuals from the highlands of the Jazan region and one from the Al-Quoz, Sahil region. Overall, nine genotypes comprising four haplotypes were observed in southwestern Saudi Arabia. The median-joining haplotype networks of eight populations from Saudi Arabia, Southeast Asia and East Africa for both the IIS6 and IIIS6 domains revealed that haplotype diversity was highest in Uganda and in the Jazan and Sahil regions of Saudi Arabia, whereas haplotype diversity was low in the Jeddah, Makkah and Madinah regions. Median-joining haplotype networks of both domains indicated selection acting on the kdr-mutation containing haplotypes in Saudi Arabia. CONCLUSIONS: The presence of wild type haplotypes without any of the three kdr mutations, i.e. that are fully susceptible, in Saudi Arabia indicates that further consideration should be given to insecticide resistance management strategies that could restore pyrethroid sensitivity to the populations of Ae. aegypti in Saudi Arabia as part of an integrative vector control strategy.

Effects of blood meal sources on the biological characteristics of Aedes aegypti and Culex pipiens (Diptera: Culicidae).

Mosquitoes transmit many diseases to humans and animals e.g., malaria, yellow fever, dengue, filariasis and encephalitis. The fundamental target of this search was to study the effect of three different blood meal sources (human; rabbit and pigeon) on some biological and behavioral properties of Aedes aegypti and Culex pipiens. The results have assured that the females of the mosquito Ae. aegypti that were fed on human blood meal has registered the highest feeding activity from feeding on the blood meal whereas the females of the other mosquito Cx. pipiens have shown the highest feeding activity after being fed on pigeons when compared with its feeding on other factors. The results have shown non-significant variation in the average time necessary to digest the blood meal on both mosquito species Ae. aegypti and Cx. pipiens that were fed on vertebrate hosts under laboratory conditions. Furthermore, results assured that the difference in blood meal sources has yielded distinct variation in the reproductive capacity and efficiency of both female mosquitoes under investigation where both species Ae. aegypti and Cx. pipiens already fed on human blood meal have yielded a pronounced distinctive increase in egg production (oviposition) when compared with females that were fed on pigeon or rabbit blood meal respectively. Moreover, feeding of the female mosquitoes under lab conditions on different blood meal sources did not affect the level of the hatching eggs that were laid by both mosquito females.

Seagrasses extracts as potential mosquito larvicides in Saudi Arabia.

The present study aimed to evaluate the toxic and biological effects of some extracts of seagrasses (Cymodocea rotundata; Halophila ovata& Thalassia hemprichii) against Aedes aegypti, which transmits dengue fever, and Culex pipiens, which is the dominant species of mosquitoes in the Kingdom of Saudi Arabia, as a safe method for its control. The cumulative death rate during larval development into pupae and adults was used as a criterion for evaluating tested seaweed extracts against Ae. aegypti, Cx. Pipiens. According to the obtained IC50 values ​​(the concentration that inhibits the exit of 50 % of adult mosquitoes), the results showed that C. rotundata extract (70.78 & 77.47 ppm) was more effective against A. aegypti and Cx. pipiens in comparison with H. ovata (86,98 & 95,87 ppm) and T. hemprichii (83,94 & 88,82) extracts by (1.186, 1.229, 1.146 & 1.237) fold, respectively. The results showed that the treatment with marine plant extracts against mosquito larvae of Cx. Pipiens and Ae. Aegypti gave different biological effects similar to those of other insect growth regulators (IGRs). The results also revealed the presence of morphological abnormalities in larvae that were treated with all seaweed extracts and these effects extended to all stages of growth, which caused damage to the insect without completing its life cycle. Generally, the results indicate the importance of carrying out bio-assessment tests for the pesticides that are used against mosquitoes and establishing a database to be referenced when planning control programs and making the right decision about the pesticide used.

Molecular characterization and phylogenetic relationships among Rhynchophorus sp. haplotypes in Makkah Al-Mukarramah Region-KSA.

The study aims at detecting and characterizing haplotypes of red palm weevil (RPW) Rhynchophorus sp. in the Western region of Saudi Arabia based on the cytochrome oxidase subunit I (COI) gene sequence. The results indicated the occurrence of 17 nucleotide substitutions, of which three were nonsynonymous (NS). These three NS substitutions resulted in the variation in amino acid sequence in three positions, out of 133. These amino acids are isoleucine/valine, glycine/serine, and arginine/histidine. Based on the chemical properties of the cytochrome C oxidase (COX) enzyme, it is likely that the change at the first position has no effect, while changes at the other two positions can affect the chemical properties of the enzyme. At the three-dimensional (3D) level, the first two positions exist at the border or inside loop 3-4 of the enzyme, while the third position exists inside loop 4-5. These two loops influence the folding pattern of the enzyme, thus, likely affecting the function of the enzyme. However, it is unlikely that variations in the three positions will affect the binding ability of the heme group, which promotes the action of the COX enzyme in the electron transport chain. Variations in chemical properties and 3D structure of COX enzyme might be an evolutionary process (positive selection) that promotes in-time and in-site adaptation to the insect. In conclusion, this study can be helpful in pest management programs and in tracing RPW geographic spread and migration in Saudi Arabia.

Single Cell Oil (SCO)-Based Bioactive Compounds: I-Enzymatic Synthesis of Fatty Acid Amides Using SCOs as Acyl Group Donors and Their Biological Activities.

Fatty acid amides (FAAs) are of great interest due to their broad industrial applications. They can be synthesized enzymatically with many advantages over chemical synthesis. In this study, the fatty acid moieties of lipids of Cunninghamella echinulata ATHUM 4411, Umbelopsis isabellina ATHUM 2935, Nannochloropsis gaditana CCAP 849/5, olive oil, and an eicosapentaenoic acid (EPA) concentrate were converted into their fatty acid methyl esters and used in the FAA (i.e., ethylene diamine amides) enzymatic synthesis, using lipases as biocatalysts. The FAA synthesis, monitored using in situ NMR, FT-IR, and thin-layer chromatography, was catalyzed efficiently by the immobilized Candida rugosa lipase. The synthesized FAAs exhibited a significant antimicrobial activity, especially those containing oleic acid in high proportions (i.e., derived from olive oil and U. isabellina oil), against several human pathogenic microorganisms, insecticidal activity against yellow fever mosquito, especially those of C. echinulata containing gamma-linolenic acid, and anticancer properties against SKOV-3 ovarian cancer cell line, especially those containing EPA in their structures (i.e., EPA concentrate and N. gaditana oil). We conclude that FAAs can be efficiently synthesized using microbial oils of different fatty acid composition and used in specific biological applications.

Amidrazone modified acrylic fabric activated with cyanuric chloride: A novel and efficient support for horseradish peroxidase immobilization and phenol removal.

In this study, hydrazine treated acrylic fabrics (polyacrylonitrile, PAN) activated with cyanuric chloride was developed as supporting material for horseradish peroxidase (HRP) immobilization. The immobilization of HRP onto the modified supporting material was achieved after being end-over-end incubated for 12 h. Field emission scanning electron microscopy and Fourier-transform infrared spectroscopy techniques were used to confirm the successful immobilization. Reusability experiment was performed to estimate the ability of the immobilized HRP to recover the reaction medium, in which it was observed to retain 78% of its original activity after 10 cycles. Relative to the soluble HRP, the optimum pH and temperature for the immobilized HRP were shifted to 7-7.5 and 50 °C, respectively. The kinetic parameters of guaiacol and H2O2 for the immobilized HRP were determined to be Km/Vmax = 57.61, 11.35 and Kcat/Km = 1.87, 1.86, respectively, while the values for the free form were Km/Vmax = 41.49, 6.23 and Kcat/Km = 1.87, 1.86, respectively. Compared to the soluble form, the immobilized HRP exhibited higher resistance toward metal ions and some organic solvents. For an application perspective. The immobilization of HRP using this procedure has the potential to be used for industrial application and wastewater treatment.

Public health and veterinary important flies (order: Diptera) prevalent in Jeddah Saudi Arabia with their dominant characteristics and identification key.

Order Diptera of class insecta is of immense importance for the public and animal health and hygiene. Many dipteran flies are potential vectors of dreadful diseases. Therefore, it is required to have a simple characterization method and identification key for the field workers and entomologists. The present study fulfill the need and focus on the identification to generate a base line data with the help of original photographs. Nine families with 16 species of dipteran flies (other than mosquitoes) from Jeddah region of Saudi Arabia are included in this work. Major families which are more prevalent and common in this region are Muscidae, Calliphoridae, Sarcophagidae and Phoridae.

The desert wormwood (Artemisia herba-alba) - From Arabian folk medicine to a source of green and effective nanoinsecticides against mosquito vectors.

The development of eco-friendly and effective insecticides is crucial for public health worldwide. Herein, we focused on the desert wormwood (Artemisia herba-alba), a plant widely used in Arabian traditional medicine, as a source of green nanoinsecticides against mosquito vectors, as well as growth inhibitors to be employed against microbial pathogens. Ag nanoparticles (AgNPs) fabricated with the A. herba-alba extract were tested on Indian and Saudi Arabian strains of Anopheles, Aedes and Culex mosquitoes. The chemical profile of the A. herba-alba extract was determined by LC-DAD-MS and 1H NMR studies. Then, AgNPs were studied using UV-vis spectroscopy, XRD, FTIR spectroscopy, TEM, and EDX analyses. Artemisia herba-alba-synthesized AgNPs showed high larvicidal toxicity against mosquitoes from both Indian and Saudi Arabian strains. LC50 of AgNPs against Indian strains was 9.76 µg/ml for An. stephensi, 10.70 µg/ml for Ae. aegypti and 11.43 µg/ml for Cx. quinquefasciatus, whereas against Saudi Arabian strains it was 33.58 µg/ml for Ae. aegypti and 38.06 µg/ml for Cx. pipiens. In adulticidal experiments, A. herba-alba extract showed LC50 ranging from 293.02 to 450 µg/ml, while AgNP LC50 ranged from 8.22 to 27.39 µg/ml. Further, low doses of the AgNPs inhibited the growth of selected microbial pathogens. Overall, A. herba-alba can be further considered as a source of phytochemicals, with special reference to saponins, for effective and prompt fabrication of AgNPs with relevant insecticidal and bactericidal activity against species of high public health importance.

Promoting health education and public awareness about dengue and its mosquito vector in Saudi Arabia.

Currently, dengue fever is considered as the main health problem in several parts (Mekkah, Jeddah, Jazan and Najran) of Kingdom of Saudi Arabia (KSA) with dramatically increase in the number of cases reported every year. This is associated with obvious ineffectiveness in the recent control and management programs for the mosquito vector (Aedes aegypti). Here, we suggested promoting the health education and public awareness among Saudi people to improve the control of dengue mosquito vector. Several suggestions and recommendations were highlighted here to ensure effectiveness in the future control and management programs of dengue mosquito vector in KSA.

An update on the incidence of dengue gaining strength in Saudi Arabia and current control approaches for its vector mosquito.

BACKGROUND: The cases of dengue reported earlier in the late 1990s from the Kingdom of Saudi Arabia (KSA) occurred in the cities of Jeddah and Makkah. Although the kingdom has ample financial resources to establish effective control measures for the dengue vector, numerous cases of dengue occur and fluctuate in numbers from year to year. This necessitates a serious review of the current vector control strategies being practiced in order to identify the existing shortcomings. This short report provides an update on epidemiology of dengue in KSA (specifically in cities of Jeddah and Makkah) with a critical look at the current vector control strategies. FINDINGS: In 2013, 4411 cases of dengue were reported, with 8 cases of mortality. This number of dengue incidence was four times higher compared to 2012. In 2013, the highest number of 1272 dengue cases was reported in May, while the lowest number (37) of cases was reported in September. CONCLUSIONS: It is evident that the control strategies of the dengue vector presently employed are inadequate. There seems to be serious deficiencies in following proper scientific procedures during field application(s) of control materials against the vector as is evident by the increases in the number of dengue cases as well as frequent outbreaks of the vector mosquito populations. In this review, some specific suggestions are made to draw attention to the relevant KSA authorities of the possible reasons behind unsuccessful control results and as to how to improve the strategy of dengue vector control in the kingdom.

Household survey of container-breeding mosquitoes and climatic factors influencing the prevalence of Aedes aegypti (Diptera: Culicidae) in Makkah City, Saudi Arabia.

OBJECTIVE: To investigate the prevalence of container breeding mosquitoes with emphasis on the seasonality and larval habitats of Aedes aegypti (Ae. aegypti) in Makkah City, adjoining an environmental monitoring and dengue incidence. METHODS: Monthly visits were performed between April 2008 and March 2009 to randomly selected houses. During each visit, mosquito larvae were collected from indoors and outdoors containers by either dipping or pipetting. Mosquitoes were morphologically identified. Data on temperature, relative humidity, rain/precipitations during the survey period was retrieved from governmental sources and analyzed. RESULTS: The city was warmer in dry season (DS) than wet season (WS). No rain occurred at all during DS and even precipitations did fall, wetting events were much greater during WS. Larval survey revealed the co-breeding of Aedes, Culex and Anopheles in a variety of artificial containers in and around homes. 32 109 larvae representing 1st , 2nd, 3rd, and 4th stages were collected from 22 618 container habitats. Culicines was far the commonest and Aedes genus was as numerous as the Culex population. Ae. aegypti larval abundance exhibited marked temporal variations, overall, being usually more abundant during WS. Ten types of artificial containers were found with developing larvae. 70% of these habitats were located indoors. 71.42% of indoor containers were permanent and 28.58% was semi-permanent during WS. Cement tanks was the only container type permanent during DS. Ae. aegypti larval indices (CI, HI, BI) recorded were greater during WS. CONCLUSIONS: Taken together, these results indicate a high risk of dengue transmission in the holy city.