The properties of these CALs, except for their molecular

masses that apparently are underestimated due to interactions on the chromatographic column, are similar to those described before ( Terra and Ferreira, 2012). Predatory hemipterans are usually thought to rely on pre-oral digestion Entinostat concentration carried out by salivary enzymes. Although trypsin is usually described in salivary glands and considered to be responsible for prey tissue digestion, there is a lack of comparative work dealing with actual salivary hydrolase activities vis a vis midgut ones. Thus, unless this is done, one can not discount the possibility that prey tissues are pre-orally disrupted, but true digestion occurs only inside the midgut. For example, previous work on P. nigrispinus ( Oliveira et al., 2006) and B. tabidus ( Azevedo et al., 2007) implied a salivary trypsin on pre-oral digestion. Nevetheless, they did not rule out the possibility that they were assaying a cathepsin L instead of trypsin, nor evaluated the activity of this enzyme vis a vis the other proteinases to estimate its significance. Prey digestive enzymes are sometimes considered to play a role in digestion by predators,

although there is no experimental support for this. For instance, Pascual-Ruiz et al. (2009) suggested that P. maculiventris may well take advantage of prey proteolytic Bortezomib enzymes for digestion. Their conclusion is based on the increase of trypsin and chymotrypsin activity observed in P. maculiventris feeding on lepidopteran larvae in comparison to those feeding on beetles or dipteran pupae. As their proteolytic assays were done at pH 10, which favor lepidopteran enzymes ( Terra and Ferreira, 1994 and Terra and Ferreira, 2012) and maintain inactive hemipteran proteinases (this paper), their conclusions need to be re-evaluated. Our findings showed that prey muscle fibers are observed inside P. nigrispinus midguts and that they are no longer visible at

the posterior midgut. Flavopiridol (Alvocidib) This suggests that pre-oral digestion is restricted to tissue disruption. Midgut proteinases are found only in middle and posterior midgut, what discount the possibility that these enzymes are injected into prey. The only salivary proteinase with significant activity in comparison with midgut enzymes is collagenase. Thus, it is probable that collagenase-containing saliva is injected into the prey. This enzyme acting on the extracellular matrix disrupts tissues. Isolated cells or cell aggregates, like the observed muscle fibers, are then ingested by the bugs. True protein digestion then occurs inside the midgut under the action of cathepsin L-like enzymes and aminopeptidase. Although carboxypeptidases and dipeptidases have not been assayed, it is highly probable that they are also involved in protein digestion ( Terra and Ferreira, 1994 and Terra and Ferreira, 2012).