Spatial characterization of proteolytic enzyme activity in the foregut region of the adult necrophagous fly, Protophormia terraenovae

Rivers, David; Acca, Gillian M.; Fink, Marc Y.; Brogan, Rebecca S.; Schoeffield, Andrew

doi:10.1016/j.jinsphys.2014.06.006

Cited by 10 publications

(14 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, the food is modified by the addition of salivary enzymes that are mixed into the gut fluids within the crop. In adult Protophormia terraenovae , enzymes consistent with trypsin, chymotrypsin and pepsin have been detected in the crop and exogenously deposited regurgitate [26]. Presumably a similar enzyme profile would be expected in other carrion-inhabiting calliphorids, although these same proteases have not been observed in Phormia regina [21].…”

Section: Insect Artifactsmentioning

confidence: 99%

“…In this respect, very little research has been done to analyze the chemical composition of fly artifacts, aside from DNA analysis for the purpose of identifying the source of blood, or inferences made based on the results of presumptive blood tests [6,9,10,13]. Rivers et al [26] demonstrated that regurgitate stains deposited by P. terraenovae possess at least three (trypsin-like, chymotrypsin-like, and pepsin-like) digestive enzymes that were also found in the crop of the adult fly, independent of the food source. These observations point to further research into the potential of fly digestive enzymes as the bases for new chemical methods of detection of insect artifacts.…”

Section: New Researchmentioning

confidence: 99%

See 1 more Smart Citation

Insect Artifacts Are More than Just Altered Bloodstains

Rivers

Geiman

2017

Insects

View full text Add to dashboard Cite

The bases for forensic entomology are that insects and their arthropod relatives can serve as evidence in criminal, medical and civil legal matters. However, some of the very same species that provide utility to legal investigations can also complicate crime scenes by distorting existing body fluid evidence (e.g., bloodstains, semen, saliva) and/or depositing artifacts derived from the insect alimentary canal at primary or secondary crime scenes. The insect contaminants are referred to as insect stains, artifacts, specks or spots, and are most commonly associated with human bloodstains. This review will discuss the different types of insect artifacts that have been described from crime scenes and laboratory experiments, as well as examine insect contaminates (non-blood based artifacts, transfer patterns, meconium, and larval fluids) that have received little research or case attention. Methods currently used for distinguishing insect stains from human body fluids will also be discussed and compared to presumptive tests used for identification of human body fluids. Since all available methods have severe limitations, areas of new research will be identified for the purpose of development of diagnostic techniques for detection of insect artifacts.

show abstract

Section: Insect Artifactsmentioning

confidence: 99%

Section: New Researchmentioning

confidence: 99%

Insect Artifacts Are More than Just Altered Bloodstains

Rivers

Geiman

2017

Insects

View full text Add to dashboard Cite

show abstract

“…On the other hand, in some species trypsin distribution along the digestive tract is more related to feeding habit. For example, high trypsin activity in the salivary glands of zoophytophagous Heteroptera (Zeng & Cohen, 2000;Zeng et al, 2002b;Swart et al, 2006) and the foregut of necrophagous Diptera (Rivers et al, 2014) is involved in extraoral digestion of dry food, whereas trypsin in foregut of xylophagous Isoptera (Sethi et al, 2011) has a role in trophallaxis, i.e., feeding of colony mates.…”

Section: Introductionmentioning

confidence: 99%

Dietary and phylogenetic correlates of digestive trypsin activity in insect pests

Lazarević

Janković-Tomanić

2015

Entomologia Exp Applicata

View full text Add to dashboard Cite

Because food proteins are crucial for insect survival, growth, and fecundity, enzymes involved in their digestion have attracted the attention of fundamental entomologists studying the mechanisms and patterns of dietary specialization, and applied entomologists searching for more efficient modes of pest control. Most insects digest proteins using trypsin, an endopeptidase that cleaves polypeptide chains on the carboxyl side of arginine and lysine, two basic amino acids. As the most ancestral proteinase, trypsin is wide-spread in the digestive tract of insects from various orders and with various feeding habits. The present review focuses on biochemical and molecular characteristics, mechanisms of regulation, and adaptive/non-adaptive changes of trypsin activity in response to heterogeneous food environments in a phylogenetic context. Within-and among-species variations in trypsin structure and regulation that contribute to better matching with specific food types are emphasized. We also discuss the relevance of these data for choosing an appropriate strategy for control of insect pests. Pest control strategies to reduce trypsin activity by interfering with substrate binding or trypsin synthesis and secretion have been suggested. Successful application of these procedures requires a multidisciplinary approach and knowledge about digestive physiology of the pest, as well as the structural characteristics of trypsins that determine their interaction with proteinaceous inhibitors and other food compounds, about patterns of developmental changes in trypsin gene expression, adaptive responses of insects to food composition, and various environmental effects on trypsin activity.

show abstract

“…This forensically important fly was selected for use in this study because it is one of the largest calliphorids in North America and the digestive physiology of adults, as well as for a sister species, Phormia regina , have been well characterized. Both attributes were essential to previous enzymatic studies conducted with P. terraenovae characterizing protease activity in the foregut and to the isolation of gut fluids and tissues during this study. Using sequence data for cathepsin D‐like proteinase from larvae of M. domestica (ppCAD3, Accession Number ), we constructed three synthetic peptides with predicted high antigenic probabilities, which in turn were used to generate polyclonal antisera.…”

mentioning

confidence: 98%

“…One avenue that has not been explored with respect to insect artifacts is the development of confirmatory tests based on the chemical composition of insect‐derived stains. Rivers et al demonstrated that regurgitate stains deposited by Protophormia terraenovae Robineau‐Desvoidy (Diptera: Calliphoridae) possessed at least three digestive enzymes (trypsin‐like, chymotrypsin‐like, and pepsin‐like) that were also found in the crop of the adult fly, independent of the food source. The pepsin‐like enzyme appears to be a cathepsin D‐like proteinase adapted for functionality in the strongly acidic midgut environment of larvae and adult flies .…”

mentioning

confidence: 99%

Distinction of Fly Artifacts from Human Blood using Immunodetection

Rivers

Acca

Fink

et al. 2018

Journal of Forensic Sciences

Self Cite

View full text Add to dashboard Cite

Insect stains produced by necrophagous flies are indistinguishable morphologically from human bloodstains. At present, no diagnostic tests exist to overcome this deficiency. As the first step toward developing a chemical test to recognize fly artifacts, polyclonal antisera were generated in rats against three distinct antigenic sequences of fly cathepsin D‐like proteinase, an enzyme that is structurally distinct in cyclorrhaphous Diptera from other animals. The resulting rat antisera bound to artifacts produced by Protophormia terraenovae and synthetic peptides used to generate the polyclonal antisera, but not with any type of mammalian blood tested in immunoassays. Among the three antisera, anti‐md3 serum displayed the highest reactivity for fly stains, demonstrated cross‐reactivity for all synthetic peptides representing antigenic sequences of the mature fly enzyme, and bound artifacts originating from the fly digestive tract. Further work is needed to determine whether the antisera are suitable for non‐laboratory conditions.

show abstract

Spatial characterization of proteolytic enzyme activity in the foregut region of the adult necrophagous fly, Protophormia terraenovae

Cited by 10 publications

References 38 publications

Insect Artifacts Are More than Just Altered Bloodstains

Insect Artifacts Are More than Just Altered Bloodstains

Dietary and phylogenetic correlates of digestive trypsin activity in insect pests

Distinction of Fly Artifacts from Human Blood using Immunodetection

Contact Info

Product

Resources

About