Competition, cooperation, and parental effects in larval aggregations formed on carrion by communally breeding beetles Necrodes littoralis (Staphylinidae: Silphinae).

Aggregations of juveniles are dominant forms of social life in some insect groups. Larval societies are shaped by competitive and cooperative interactions of the larvae, in parallel with parental effects. Colonies of necrophagous larvae are excellent systems to study these relationships. Necrodes littoralis (Staphylinidae: Silphinae), a carrion beetle that colonizes cadavers of large vertebrates, forms massive juvenile aggregations. By spreading over carrion anal and oral exudates, the beetles form the feeding matrix, in which the heat is produced and by which adults presumably affect the fitness of the larvae. We predict that exploitative competition shapes the behavior of N. littoralis larvae in their aggregations. However, cooperative interactions may also operate in these systems due mainly to the benefits of collective exodigestion. Moreover, indirect parental effects (i.e., formation of the feeding matrix) probably modulate larval interactions within the aggregations. By manipulating parental effects (present/absent) and larval density (0.02-1.9 larvae/g of meat), we found a strong negative group-size effect on fitness components of N. littoralis, in colonies with parental effects over almost the entire density range, and in colonies without parental effects for densities larger than 0.5 larva/g. This was accompanied by positive group-size effects in terms of development time (it shortened with larval density) and thermogenesis (it increased with larval density). A pronounced positive group-size effect on juvenile fitness was found only in colonies without parental effects and only in the low-density range. These results support the hypothesis that larval societies of N. littoralis are shaped by exploitation competition.

Field validation of post-mortem interval estimation based on insect development. Part 1: Accuracy gains from the laboratory rearing of insect evidence.

There is a consensus that forensic methods must be valid. The high quality of the method may be fully demonstrated only through validation. Unfortunately, there are very few experimental or casework validations of entomological methods for estimating post-mortem interval (PMI). Here, we present the first part of the results from the field validation of minimum PMI (PMImin) estimates based on insect development. From eight pig carcasses (24-46.4 kg) exposed in a forest habitat of Western Poland, one every one or two months, we collected insect evidence with standard entomological techniques. Using weather station temperatures and the thermal summation method, PMImin was estimated based on insect life stages that were reared under controlled laboratory conditions. Through rearing an insect until the next developmental landmark (or until eclosion), its age at the time of collection (and eventually PMImin) may be estimated by subtracting thermal accumulation in the laboratory from the corresponding thermal constant. We hypothesized that rearing insect evidence significantly improves the accuracy of PMImin compared to the estimation based on non-reared insect evidence. The results clearly supported this hypothesis. However, the accuracy significantly increased only in the case of these insects that developed normally during rearing. When their development in the laboratory was prolonged, PMImin was significantly less accurate. For the normal development sample the accuracy improved in all species and life stages. The largest accuracy gains from rearing were recorded for Calliphora vomitoria (Diptera: Calliphoridae) and Stearibia nigriceps (Diptera: Piophilidae). Moreover, when puparia or third instar larvae were reared, gains were larger than in the case of earlier life stages. In conclusion, this study demonstrated that the method of rearing insect evidence to improve the accuracy of PMImin is valid. However, it needs to be used with caution, since substantial part of the evidence may die or slow down their development in the laboratory, which violates assumptions of the method.

Calibrating insect age at eclosion by size in a gregarious carrion beetle Thanatophilus sinuatus (Staphylinidae: Silphinae).

Recent discoveries have shown that the physiological age at eclosion of forensically useful beetles differs between males and females and between beetles of various sizes. Accordingly, it was postulated that the size and sex of the beetles at eclosion may be used to calibrate their age, which may improve the accuracy of age (and post-mortem interval) estimates in forensic entomology. In this study, we derived thermal summation models for the eclosion for the Central European population of carrion beetles Thanatophilus sinuatus (Fabricius, 1775), (Staphylinidae: Silphinae), and tested the usefulness of sex and size for the calibration of beetle age at eclosion. Although in previous developmental studies, the beetles were reared individually, we reared them in larval aggregations, since in natural conditions T. sinuatus beetles are gregarious. Weak (r2 between 5% and 13%) negative correlations were observed between the size and age of T. sinuatus males or females at eclosion, demonstrating that calibration of age by beetle size and sex may bring only minimal benefits regarding the accuracy of age estimation in this species. However, it may still be worthwhile in the case of extremely large or small beetles. Moreover, the total development times recorded in this study were much shorter than in the previous T. sinuatus study, at 14°C by about 15 days and at 26°C by about 2 days. These differences emphasise the importance of gregariousness for the development of carrion beetles, and at the same time highlight the need for the ecologically-relevant protocols of development studies in forensic entomology.

Competition of insect decomposers over large vertebrate carrion: Necrodes beetles (Silphidae) vs. blow flies (Calliphoridae).

Large carrion is inhabited by highly variable and interactive communities of insects. Positive interactions in carrion insect communities have been recently the focus in carrion ecology. In contrast, competition between carrion insects is rather undervalued. Here we provide evidence that blow flies (Calliphoridae) and Necrodes beetles (Silphidae), dominant decomposers of large carcasses in terrestrial habitats, compete over carrion. By reanalyzing the results from 90 pig carcasses, we demonstrated that the contribution of the flies and the beetles to the decay was negatively related. The greater part of the large carrion pool was monopolized by blow flies, whereas Necrodes beetles abundantly colonized carcasses, on which blow flies were less effective as decomposers. In behavioral assays, we found that adult beetles killed 4 times more frequently feeding than postfeeding third instar larvae of the flies, with the large decrease in the killing frequency after the larvae reached the age of early third instar. Therefore, adult Necrodes beetles preferentially killed the larvae that were before or in their peak feeding. The study provides evidence that the interaction between blow flies and Necrodes beetles is a combination of indirect exploitative effects of the flies and direct interference effects of the beetles (the mixed competition).

Erratum to: Competition of insect decomposers over large vertebrate carrion: Necrodes beetles (Silphidae) vs. blow flies (Calliphoridae).

[This corrects the article DOI: 10.1093/cz/zoab100.].

Heat production in a feeding matrix formed on carrion by communally breeding beetles.

Insects regulate their body temperature mostly behaviourally, by changing posture or microhabitat. Usually they use heat that is already present in the environment. Sometimes, however, they may manipulate the environment to affect, focus or benefit from thermogenesis. Carrion beetles create a feeding matrix by applying to cadaver surface anal or oral exudates. We tested the hypothesis that the matrix, which is formed on carrion by communally breeding beetle Necrodes littoralis L. (Silphidae), produces heat that enhances insect fitness. Using thermal imaging we demonstrate that heat produced in the matrix formed on meat by adult or larval beetles is larger than in meat decomposing without insects. Larval beetles regularly warmed up in the matrix. Moreover, by comparing matrix temperature and larval fitness in colonies with and without preparation of meat by adult beetles, we provide evidence that formation of the matrix by adult beetles has deferred thermal effects for larval microhabitat. We found an increase in heat production of the matrix and a decrease in development time and mortality of larvae after adult beetles applied their exudates on meat in the pre-larval phase. Our findings indicate that spreading of exudates over carrion by Necrodes larvae, apart from other likely functions (e.g. digesting carrion or promoting growth of beneficial microbes), facilitates thermoregulation. In case of adult beetles, this behaviour brings distinct thermal benefits for their offspring and therefore may be viewed as a new form of indirect parental care with an important thermal component.

Post-mortem interval estimation based on insect evidence in a quasi-indoor habitat.

Insects collected on indoor cadavers are frequently used for post-mortem interval (PMI) estimation. Buildings encountered during crime investigations vary according to temperatures inside, the extent of insect access restriction or sanitary conditions. This article reports the PMI oriented analyses of insect evidence sampled from the human cadaver in the atypical indoor habitat. The body was found in the uninhabited house, on the floor covered with rubbish, in the room with no doors and windows. Thermal conditions in the room were less variable than in the local weather station, however still much more variable compared to the typical indoor habitat, indicating the need for retrospective correction of temperature records from the station. Cadaver entomofauna was surprisingly diverse and abundant. We recorded several taxa usually not occurring on indoor cadavers, e.g. immature stages of Necrodes littoralis (Coleoptera: Silphidae) or Stearibia nigriceps (Diptera: Piophilidae). PMI was based on the age and the pre-appearance interval estimated for live puparium of S. nigriceps, giving the total interval of 37 (±7.4) days plus 4-20 days resulting from the absence of first colonizing specimens of the species. This estimate was corroborated with the age estimate for empty puparia of Sarcophaga argyrostoma (Diptera: Sarcophagidae) with traces of Nasonia sp. (Hymenoptera: Pteromalidae) eclosion. Other insects indicated shorter but consistent PMI. Difficulties and limitations of insect-based PMI estimations in unusual indoor habitats are discussed.

Blowfly puparia in a hermetic container: survival under decreasing oxygen conditions.

Despite widely accepted standards for sampling and preservation of insect evidence, unrepresentative samples or improperly preserved evidence are encountered frequently in forensic investigations. Here, we report the results of laboratory studies on the survival of Lucilia sericata and Calliphora vomitoria (Diptera: Calliphoridae) intra-puparial forms in hermetic containers, which were stimulated by a recent case. It is demonstrated that the survival of blowfly intra-puparial forms inside airtight containers is dependent on container volume, number of puparia inside, and their age. The survival in both species was found to increase with an increase in the volume of air per 1 mg of puparium per day of development in a hermetic container. Below 0.05 ml of air, no insect survived, and above 0.2 ml of air per 1 mg of puparium per day, survival reached its maximum. These results suggest that blowflies reveal a single, general pattern of survival under decreasing oxygen conditions and that this pattern is a product of number of developing insects, their age and the initial amount of available air. Implications for forensic entomology are discussed.

Sex- and Size-Related Patterns of Carrion Visitation in Necrodes littoralis (Coleoptera: Silphidae) and Creophilus maxillosus (Coleoptera: Staphylinidae).

The estimation of postmortem interval (PMI) based on successional patterns of adult insects is largely limited, due to the lack of potential PMI markers. Sex and size of adult insects could be easily used for such estimation. In this study, sex- and size-related patterns of carrion attendance by adult insects were analyzed in Necrodes littoralis (Coleoptera: Silphidae) and Creophilus maxillosus (Coleoptera: Staphylinidae). For both species, abundance of males and females changed similarly during decomposition. A slightly female-biased sex ratio was recorded in N. littoralis. Females of N. littoralis started visiting carcasses, on average, one day earlier than males. There was a rise in size of males of N. littoralis at the end of decomposition, whereas for females of both species and males of C. maxillosus, no size-related patterns of carrion visitation were found. Current results demonstrate that size and sex of adult carrion beetles are poor indicators of PMI.

Validation of temperature methods for the estimation of pre-appearance interval in carrion insects.

The pre-appearance interval (PAI) is an interval preceding appearance of an insect taxon on a cadaver. It decreases with an increase in temperature in several forensically-relevant insects. Therefore, forensic entomologists developed temperature methods for the estimation of PAI. In the current study these methods were tested in the case of adult and larval Necrodes littoralis (Coleoptera: Silphidae), adult and larval Creophilus maxillosus (Coleoptera: Staphylinidae), adult Necrobia rufipes (Coleoptera: Cleridae), adult Saprinus semistriatus (Coleoptera: Histeridae) and adult Stearibia nigriceps (Diptera: Piophilidae). Moreover, factors affecting accuracy of estimation and techniques for the approximation and correction of predictor temperature were studied using results of a multi-year pig carcass study. It was demonstrated that temperature methods outperform conventional methods. The accuracy of estimation was strongly related to the quality of the temperature model for PAI and the quality of temperature data used for the estimation. Models for larval stage performed better than models for adult stage. Mean temperature for the average seasonal PAI was a good initial approximation of predictor temperature. Moreover, iterative estimation of PAI was found to effectively correct predictor temperature, although some pitfalls were identified in this respect. Implications for the estimation of PAI are discussed.