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Identification and characterization of novel cecropins from the Oxysternon conspicillatum neotropic dung beetle.

ABSTRACT: Dung beetles are exposed to a complex microbiological ecosystem during their life cycle. Characterization of novel host-defense peptides (HDP) is essential to understanding the host innate immune response in insects. It constitutes a promising alternative to look for new therapeutic agents against pathogenic microbes. We identified four new HDP, Oxysterlins 1, 2, 3, and 4 from the transcriptome of the Oxysternon conspicillatum dung beetle. These HDP display a highly conserved signal peptide and a mature peptide, characterized by an overall positive charge (cationic) (pI: 10.23-11.49), a hydrophobic ratio (ΦH: 35-41), and amphipathicity. Oxysterlins 1, 2, and 3 have a linear α-helix structure, whilst Oxysterlin 4 has a mixture of both α-helix and β-sheet structures without disulfide bonds through bioinformatics prediction and circular dichroism. Oxysterlins are part of the cecropin family group in an exclusive clade related to beetle cecropins. They have predominant antimicrobial activity against Gram-negative bacteria, including multidrug resistant strains (3.12-50 μg/mL) measured by plate microdilution. Their kinetics, in a time-killing curve showed concentration-dependent bactericidal activity. Furthermore, these HDP have low toxicity against human erythrocytes (62.5-500 μg/mL) and Vero cells (250-500 μg/mL). This article describes new HDP of the cecropin family from the Oxysternon conspicillatum dung beetle, with antimicrobial activity against multidrug resistant bacteria and low toxicity.

SUBMITTER: Toro Segovia LJ

PROVIDER: S-EPMC5706684 | biostudies-other | 2017

REPOSITORIES: biostudies-other

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Identification and characterization of novel cecropins from the Oxysternon conspicillatum neotropic dung beetle.

Toro Segovia Lily Johanna LJ Téllez Ramírez Germán Alberto GA Henao Arias Diana Carolina DC Rivera Duran Juan David JD Bedoya Juan Pablo JP Castaño Osorio Jhon Carlos JC

PloS one 20171129 11

Dung beetles are exposed to a complex microbiological ecosystem during their life cycle. Characterization of novel host-defense peptides (HDP) is essential to understanding the host innate immune response in insects. It constitutes a promising alternative to look for new therapeutic agents against pathogenic microbes. We identified four new HDP, Oxysterlins 1, 2, 3, and 4 from the transcriptome of the Oxysternon conspicillatum dung beetle. These HDP display a highly conserved signal peptide and ...[more]

PMID: 29186139

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Project description:Abstract An important service in many ecosystems is the turnover and degradation of dung deposited by cattle. Dung beetles are the primary group of insects responsible for dung turnover, and factors affecting their abundance and distribution thus impact dung degradation. Lands lost to grazing due to dung buildup and pasture contamination total millions of acres per year in US pastures. We evaluated the structural differences in dung beetle assemblages in natural grasslands versus a managed agroecosystem in subtropical southeastern Florida (USA). We measured the direct effect of dung longevity when dung beetle fauna normally inhabiting dung pats were excluded. Our results indicate dung beetle abundance, functional diversity, and species richness have a substantial impact on the rate of dung turnover in subtropical pastoral lands with ~70% of dung removed from the soil surface after three months. Functional diversity and evenness did not have a significant positive effect on dung removal in managed, versus natural grasslands demonstrating a strong relationship between dung beetle assemblage composition and delivery of a key ecological process, dung degradation. We suggest the importance of trees, which provide a thermal refuge for beetles, should be dispersed within matrixes of open pasture areas and within proximity to adjacent closed‐canopy hammocks to facilitate the exchange of dung beetles between habitats and therefore maintain the provisioning of dung degradation services by dung beetle assemblages. An important service in many ecosystems is the turnover and degradation of dung deposited by cattle. Dung beetles are the primary group of insects responsible for dung turnover, and factors affecting their abundance and distribution thus impact dung degradation. Our results indicate dung beetle abundance, functional diversity, and species richness has a substantial impact on the rate of dung turnover in subtropical pastoral lands with ~70% of dung removed from the soil surface after three months.

Project description:BackgroundDung beetles provide many important ecosystem services, including dung decomposition, pathogen control, soil aeration, and secondary seed dispersal. Yet, the biology of most dung beetles remains unknown. Natural diets are poorly studied, partly because previous research has focused on choice or attraction experiments using few, easily accessible dung types from zoo animals, farm animals, or humans. This way, many links within natural food webs have certainly been missed. In this work, we aimed to establish a protocol to analyze the natural diets of dung beetles using DNA gut barcoding.MethodsFirst, the feasibility of gut-content DNA extraction and amplification of 12s rDNA from six different mammal dung types was tested in the laboratory. We then applied the method to beetles caught in pitfall traps in Ecuador and Germany by using 12s rDNA primers. For a subset of the dung beetles caught in the Ecuador sampling, we also used 16s rDNA primers to see if these would improve the number of species we could identify. We predicted the likelihood of amplifying DNA using gut fullness, DNA concentration, PCR primer, collection method, and beetle species as predictor variables in a dominance analysis. Based on the gut barcodes, we generated a dung beetle-mammal network for both field sites (Ecuador and Germany) and analyzed the levels of network specificity.ResultsWe successfully amplified mammal DNA from dung beetle gut contents for 128 specimens, which included such prominent species as Panthera onca (jaguar) and Puma concolor (puma). The overall success rate of DNA amplification was 53%. The best predictors for amplification success were gut fullness and DNA concentration, suggesting the success rate can be increased by focusing on beetles with a full gut. The mammal dung-dung beetle networks differed from purely random network models and showed a moderate degree of network specialization (H2': Ecuador = 0.49; Germany = 0.41).ConclusionWe here present a reliable method of extracting and amplifying gut-content DNA from dung beetles. Identifying mammal dung via DNA reference libraries, we created mammal dung-dung beetle trophic networks. This has benefits over previous methods because we inventoried the natural mammal dung resources of dung beetles instead of using artificial mammal baits. Our results revealed higher levels of specialization than expected and more rodent DNA than expected in Germany, suggesting that the presented method provides more detailed insights into mammal dung-dung beetle networks. In addition, the method could have applications for mammal monitoring in many ecosystems.

Project description:BackgroundConserving biodiversity in tropical landscapes is a major challenge to scientists and conservationists. Current rates of deforestation, fragmentation, and land use intensification are producing variegated landscapes with undetermined values for the conservation of biological communities and ecosystem functioning. Here, we investigate the importance of tropical variegated landscapes to biodiversity conservation, using dung beetle as focal taxa.MethodsThe study was carried out in 12 variegated landscapes where dung beetles were sampled using six pitfall traps, 30 m apart from each other, along a transect in each studied landscape use and cover classes-LUCC (forest fragment and corridor, coffee plantation, and pasture). We baited each pitfall trap with 30 g of human feces and left open for a 48 h period. We also measured three environmental variables reflecting structural differences among the studied classes: canopy cover, local vegetation heterogeneity and soil sand content.ResultsWe collected 52 species and 2,695 individuals of dung beetles. We observed significant differences in the mean species richness, abundance and biomass among classes, with forest fragments presenting the highest values, forest corridors and coffee plantations presenting intermediate values, and pastures the lowest values. Regarding community structure, we also found significant differences among classes. Canopy cover was the only variable explaining variation in dung beetle species richness, abundance, biomass, and community structure. The relative importance of spatial turnover was greater than nestedness-resultant component in all studied landscapes.DiscussionThis study evaluated the ecological patterns of dung beetle communities in variegated tropical landscapes highlighting the importance of these landscapes for conservation of tropical biodiversity. However, we encourage variegation for the management of landscapes that have already been fragmented or as a complementary initiative of current conservation practices (e.g., protection of natural habitats and establishment of reserves).

Project description:BackgroundDung beetle glycosaminoglycan is known to possess anti-aging activities. However, its anti-cancer mechanisms are not fully elucidated yet. The objective of this study was to evaluate the anti-cancer effect of insect-derived polymer dung beetle glycosaminoglycan (GAG) after intraperitoneally injecting it to melanoma mice induced by B16F10 cells.MethodsTo determine molecular mechanism involved in the anti-cancer effect of dung beetle GAG, its origin N-glycan under 3KD Dalton was assayed for melanoma cell cytotoxicity. Quantitative comparisons of adhesive molecule on extracellular matrix and activities of tissue inhibitor of metalloprotease 2 (TIMP-2) were also investigated. In vivo anti-cancer effect of dung beetle GAG on solid tumor size, survival time and gene-expression profiles was also assayed using B10F10 melanoma mice model. Mice with induced melanoma were then treated with Catharsius molossus (dung beetle) GAG (CaG) at 5?mg/kg for 8?weeks to investigate its anti-cancer effects compared to bumblebee (Bombus ignitus) queen glycosaminoglycan (IQG) and Huechys sanguinea glycosaminoglycan (HEG).ResultsThese N-glycans derived from these GAG were composed of many linear heparinoid polysaccharides, polymers with hexose and N-acetylhexose. Adminstration with these GAGs increased survival time and decreased melanoma sizes in mice, in accordance with their inhibitory effects on cell growth ratio of melanoma B16F10. In addition, treatment with N-glycans derived from theses glycosaminoglycan increased activities of TIMP-2 in HMVEC cells pretreated with TNF-alpha and in melanoma cells, suggesting that they had anti-inflammatory and anticancer activities. In DNA microarray results, compared to control, CaG treated mouse group showed upregulation of 192 genes including collagen,typeI,alpha1 (Col1a1), consistent with the highly increased in vitro extracellular matrix (ECM) adhesion on collagen 1 and up-regulation of heparanase (Hpse). After treatment with CaG, a total of 152 genes were down-regulated, including nuclear RNA export factor (Nxf3) and hyaluronan proteoglycan link protein1 (Hapln1).ConclusionsGlycosaminoglycan, CaG can strengthen ECM by increasing activity of TIMP-2 and adhesion activity on collagen known to inhibit changes of ECM, leading to tumor cell invasion and progression.

Dataset Information

Identification and characterization of novel cecropins from the Oxysternon conspicillatum neotropic dung beetle.

Publications

Identification and characterization of novel cecropins from the Oxysternon conspicillatum neotropic dung beetle.

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