Aedes aegypti is inherently susceptible to arboviruses. The geographical expansion of this vector host species has led to the persistence of Dengue, Zika, and Chikungunya human infections. These viruses take advantage of the mosquito’s cell to create an environment conducive for their growth. Arboviral infection triggers transcriptomic and protein dysregulation in Ae. aegypti and in effect, host antiviral mechanisms are compromised. Currently, there are no existing vaccines able to protect human hosts from these infections and thus, vector control strategies such as Wolbachia mass release program is regarded as a viable option. Considerable evidence demonstrates how the presence of Wolbachia interferes with arboviruses by decreasing host cytoskeletal proteins and lipids essential for arboviral infection. Also, Wolbachia strengthens host immunity, cellular regeneration and causes the expression of microRNAs which could potentially be involved in virus inhibition. However, variation in the magnitude of Wolbachia’s pathogen blocking effect that is not due to the endosymbiont’s density has been recently reported. Furthermore, the cellular mechanisms involved in this phenotype differs depending on Wolbachia strain and host species. This prompts the need to explore the cellular interactions between Ae. aegypti-arboviruses-Wolbachia and how different Wolbachia strains overall affect the mosquito’s cell. Understanding what happens at the cellular and molecular level will provide evidence on the sustainability of Wolbachia vector control.

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Wolbachia are maternally transmitted bacteria that are utilized for arboviral disease prevention. Cytoplasmic incompatibility (CI) and viral blocking, two characteristics that make Wolbachia suitable for vector control, both depend on bacterial infection prevalence and density. Although numerous mosquito species naturally harbor Wolbachia, the prevalence of Wolbachia in Aedes aegypti varies from minimal to absent. The absence of natural Wolbachia in Ae. aegypti is considered advantageous for novel Wolbachia transinfection because interference caused by incompatible Wolbachia strains will not occur. In this study, we screened for Wolbachia density in 429 individual Ae. aegypti collected from Metropolitan Manila, Philippines using locally designed primers to determine whether primer compatibility improves Wolbachia detection in naturally infected Ae. aegypti. We also investigated the effects of host sex and Wolbachia strain on Wolbachia density. We found a high Wolbachia prevalence for 16S rRNA (40%) and wsp (62%) markers. Relative Wolbachia densities ranged from −3.840 to 2.710 for 16S rRNA and from −4.020 to 1.810 for wsp, and these relative densities were higher among male mosquitoes than female mosquitoes. Phylogenetic analysis revealed that most of the Ae. aegypti were clustered into supergroup B, with 63% (142/226) of these sequences representing a unique strain referred to here as "wAegML," which exhibited a significantly lower density in Ae. aegypti than the other two Wolbachia strains detected (wAegB/wAlbB and wAlbA) as well as a higher density in male Ae. aegypti than in females. Overall, locally designed primers improved Wolbachia detection in naturally infected Ae. aegypti. The unique strain wAegML occurs at a low density and is related to the use of wAlbB in mass-release programs; therefore, it is unlikely to interfere with such programs. Further studies on natural Wolbachia infection in Ae. aegypti combining methodological and biological factors are warranted to help resolve current conflicts in the field.

The hallmark of Acute Lung Injury/Acute Respiratory Distress Syndrome (ALI/ARDS) is inflammation-induced alveolar-vascular barrier destruction and neutrophilic infiltration that leads to the formation of cytokines and oxygen radicals. The objective of the study is to investigate the protective and toxicological effects of Antidesma bunius (L.) Spreng [Bignay] in murine model of Lipopolysaccharide E. coli (LPS)-induced ALI and compared with Fluticasone Propionate (FP), a synthetic corticosteroid. We showed that extracted Bignay fruits have high amount of phenols, steroids and flavonoids but insignificant amount of heavy metals and aflatoxins. BALB/c mice of either sex were divided into 4 groups in the ALI mouse model; Group 1: vehicle control; Group 2: LPS alone; Group 3: Bignay + LPS; and Group 4: FP + LPS. Bignay and FP were administered via intraperitoneal injection while LPS was given intra-tracheally. Biomarkers of ALI such as total lung inflammatory cell count, total lung protein content, lung edema and interleukin-6  (IL-6) secretion were measured 24 hrs after vehicle control or LPS treatment. Compared to vehicle controls, LPS caused significant increased in all measured biomarkers of ALI in samples collected from bronchoalveolar lavage fluid and were significantly attenuated by Bignay fruit extract or FP. Pulmonary vascular leakage caused by LPS was also evaluated after injection with Evans blue dye, an indication of lung injury. Extracted Bignay fruits or FP when given to mice 2 hrs after LPS administration substantially decreased the pulmonary vascular leak. Our findings are the first evidence demonstrating the preventive and non-toxic effects of extracted Bignay fruits in a murine model of LPS-induced ALI. The results could be attributed to the presence of active secondary metabolites such as flavonoids, phenols and steroids. It is also evident that extracted Bignay fruits are as effective as FP, well-established steroid, in blocking the biomarkers of ALI caused by LPS.

Aedes aegypti is inherently susceptible to arboviruses. The geographical expansion of this vector host species has led to the persistence of Dengue, Zika and Chikungunya human infections. These viruses take advantage of the mosquito’s cell to create an environment conducive for their growth. Arboviral infection triggers transcriptomic and protein dysregulation in Ae. aegypti and in effect, host antiviral mechanisms are compromised. Currently, there are no existing vaccines able to protect human hosts from these infections and thus, vector control strategies such as Wolbachia mass release program is regarded as a viable option. Considerable evidence demonstrates how the presence of Wolbachia interferes with arboviruses by decreasing cellular components vital for the pathogen and strengthening antiviral host responses. However, variation in the magnitude of Wolbachia’s viral inhibition that is neither due to strain nor density has been observed. Furthermore, the cellular mechanisms involved in the endosymbiont’s pathogen-blocking differs among hosts. This prompts the need to explore the cellular interactions between Ae. aegypti-arboviruses-Wolbachia and how these interactions overall affect the mosquito’s cell. Understanding what happens at the cellular and molecular level will provide evidence on the sustainability of Wolbachia vector control.