Abstract
Strains of Serratia marcescens, originally isolated from the gut lumen of adult female Anopheles stephensi mosquitoes, established persistent infection at high rates in adult A. stephensi whether fed to larvae or in the sugar meal to adults. By contrast, the congener S. fonticola originating from Aedes triseriatus had lower infection in A. stephensi, suggesting co-adaptation of Serratia strains in different species of host mosquitoes. Coinfection at high infection rate in adult A. stephensi resulted after feeding S. marcescens and Elizabethkingia anophelis in the sugar meal, but when fed together to larvae, infection rates with E. anophelis were much higher than were S. marcescens in adult A. stephensi, suggesting a suppression effect of coinfection across life stages. A primary isolate of S. marcescens was resistant to all tested antibiotics, showed high survival in the mosquito gut, and produced alpha-hemolysins which contributed to lysis of erythrocytes ingested with the blood meal. Genomes of two primary isolates from A. stephensi, designated S. marcescens ano1 and ano2, were sequenced and compared to other Serratia symbionts associated with insects, nematodes and plants. Serratia marcescens ano1 and ano2 had predicted virulence factors possibly involved in attacking parasites and/or causing opportunistic infection in mosquito hosts. S. marcescens ano1 and ano2 possessed multiple mechanisms for antagonism against other microorganisms, including production of bacteriocins and multi-antibiotic resistance determinants. These genes contributing to potential anti-malaria activity including serralysins, hemolysins and chitinases are only found in some Serratia species. It is interesting that genome sequences in S. marcescens ano1 and ano2 are distinctly different from those in Serratia sp. Ag1 and Ag2 which were isolated from Anopheles gambiae. Compared to Serratia sp. Ag1 and Ag2, S. marcescens ano1 and ano2 have more rRNAs and many important genes involved in commensal and anti-parasite traits.
Highlights
Bacteria of the genus Serratia are Gram-negative, rod-shaped, facultative anaerobes (Grimont and Grimont, 2006)
When S. marcescens ano1 was fed to 3rd instar A. stephensi larvae, the infection persisted to the adult stage for 62.5% (15/24) of adults at 4 days after adult emergence (Table 1), as evidenced by positive luciferase assay relative to negative controls
17.1% (6/35) and 32.5% (13/40) of the guts of A. stephensi adults were positive for S. fonticola MSU001 when fed to larval and adult stages, respectively, demonstrating that S. fonticola isolated from A. triseriatus had relatively poorer colonization and persistence than did S. marcescens in A. stephensi
Summary
Bacteria of the genus Serratia are Gram-negative, rod-shaped, facultative anaerobes (Grimont and Grimont, 2006). These bacteria are ubiquitously distributed in soil, sediments, water, plant roots, on surfaces of animals, as well as in the gastrointestinal tract of animals. Serratia marcescens or S. marcescens-like bacteria have been reported living symbiotically with, and sometimes causing disease in, economically important insects and nematodes (Benoit et al, 1990; Stanley et al, 2012; Wang et al, 2014; Aggarwal et al, 2015; Pei et al, 2015; Chen et al, 2016). Pathology in insects infected by S. marcescens appears to be related to establishment of disseminated infection in the hemolymph, whilst infection in the gut lumen is tolerated (Nehme et al, 2007)
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