An illustrated identification key to early instar larvae of forensically important Muscidae (Diptera) of the western Palaearctic region.

There is a significant gap in the availability of comprehensive identification keys for the early larval stages of forensically important fly species. While well-documented identification keys exist for the third instar larvae, particularly for the Calliphoridae, Muscidae and Sarcophagidae families, there is a notable scarcity of keys for the first, except Calliphoridae, and the second instar larvae, with no such resources available for muscid species. The second instar larvae suffer the most from the lack of morphological descriptions and available identification keys. The Muscidae is one of the most frequently reported dipteran families of forensic importance colonising animal cadavers and human corpses. Nevertheless, descriptions of the morphology of their early instars remain scarce and limited to only a few species, thus their larval identification is challenging or impossible. Considering the numerous challenges associated with studying small-sized entomological material, we tested whether it is feasible to identify muscid flies to the species or at least genus level based predominantly on the details of the cephaloskeleton. To overcome the obstacle of observing details of small sclerites, especially their shapes and interconnections, we effectively employed confocal laser scanning microscopy (CLSM) as a supplementary method for light microscopy (LM). This study provides an identification key for first and second instar larvae of forensically important muscid species from the western Palaearctic (Europe, North Africa, Middle East). The proposed key primarily utilises details of the cephaloskeleton with only addition of external morphology.

The genome sequence of a muscid fly, Hydrotaea cyrtoneurina (Zetterstedt, 1845).

We present a genome assembly from an individual female Hydrotaea cyrtoneurina (muscid fly; Arthropoda; Insecta; Diptera; Muscidae). The genome sequence is 575.2 megabases in span. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules. The mitochondrial genome has also been assembled and is 19.6 kilobases in length.

Larval morphology of the avian parasitic genus Passeromyia: playing hide and seek with a parastomal bar.

The enigmatic larvae of the Old World genus Passeromyia Rodhain & Villeneuve, 1915 (Diptera: Muscidae) inhabit the nests of birds as saprophages or as haematophagous agents of myiasis among nestlings. Using light microscopy, confocal laser scanning microscopy and scanning electron microscopy, we provide the first morphological descriptions of the first, second and third instar of P. longicornis (Macquart, 1851) (Diptera: Muscidae), the first and third instar of P. indecora (Walker, 1858) (Diptera: Muscidae), and we revise the larval morphology of P. heterochaeta (Villenueve, 1915) (Diptera: Muscidae) and P. steini Pont, 1970 (Diptera: Muscidae). We provide a key to the third instar of examined species (excluding P. steini and P. veitchi Bezzi, 1928 (Diptera: Muscidae)). Examination of the cephaloskeleton revealed paired rod-like sclerites, named 'rami', between the lateral arms of the intermediate sclerite in the second and third instar larva. We reveal parastomal bars fused apically with the intermediate sclerite, the absence of which has so far been considered as apomorphic for second and third instar muscid larvae. Examination of additional material suggests that modified parastomal bars are not exclusive features of Passeromyia but occur widespread in the Muscidae, and rami may occur widespread in the Cyclorrhapha.

Linking bacteria, volatiles and insects on carrion: the role of temporal and spatial factors regulating inter-kingdom communication via volatiles.

Multi-kingdom community complexity and the chemically mediated dynamics between bacteria and insects have recently received increased attention in carrion research. However, the strength of these inter-kingdom interactions and the factors that regulate them are poorly studied. We used 75 piglet cadavers across three forest regions to survey the relationship between three actors (epinecrotic bacteria, volatile organic compounds (VOCs) and flies) during the first 4 days of decomposition and the factors that regulate this interdependence. The results showed a dynamic bacterial change during decomposition (temperature-time index) and across the forest management gradient, but not between regions. Similarly, VOC emission was dynamic across a temperature-time index and the forest management gradient but did not differ between regions. However, fly occurrence was dynamic across both space and time. The strong interdependence between the three actors was mainly regulated by the temperature-time index and the study regions, thereby revealing regulation at temporal and spatial scales. Additionally, the actor interdependence was stable across a gradient of forest management intensity. By combining different actors of decomposition, we have expanded our knowledge of the holistic mechanisms regulating carrion community dynamics and inter-kingdom interactions, an important precondition for better describing food web dynamics and entire ecosystem functions.

Larval morphology and temperature-dependent development models of Fannia pusio (Wiedemann): A forensic indicator with expanding distribution.

Fannia pusio (Wiedemann) is originally from tropical and subtropical regions of the Americas, where it has repeatedly been collected from both animal carrion and human cadavers. This species is expanding its distributional range, and it has been introduced to Africa, Asia, Australia, Oceania, and Europe. Newly introduced species may be confused with native species of local arthropod necrophagous assemblages, which from a forensic entomology perspective may impact the accuracy of post-mortem interval (PMI) estimations based on insect evidence. In this work we aim to raise awareness among forensic entomologists of the potential benefits and risks associated with the expanding range of F. pusio. Morphology of all larval instars is documented with a combination of light, confocal laser scanning and scanning electron microscopy. Characters allowing identification from other forensically important Fanniidae are listed. Thermal requirements for the development of immature stages of F. pusio were examined under nine ambient temperatures. Models describing changes in larval body length over time were produced for eight different temperatures. The thermal summation constant (k) and developmental zero (Tmin) were calculated for six developmental events: hatching, first ecdysis, second ecdysis, wandering, pupariation, and eclosion.

Diptera (Dryomyzidae, Fanniidae, Muscidae, Piophilidae) Associated With Pig Carcasses in a Forest Habitat of Poland: Sex-Related Patterns of Visitation and Effectiveness of Sampling Methods.

Our knowledge on many necrophilous dipterans was limited for a long time mainly due to taxonomic issues. Therefore, findings on the sex-related associations with pig carcasses in Dryomyzidae, Fanniidae, Muscidae, and Piophilidae are presented. Moreover, the effectiveness of pitfall traps and sweep nets in fly sampling were compared, since the correct method of collecting insects is critical for the development and application of entomological tools for forensic purposes. The trapped numbers of all species differed significantly in respect to both sexes. The sex ratios were strongly female-biased in most species of Muscidae and in Fanniidae. More females were also collected in Piophilidae, but the proportion of males was higher when compared to muscids. Within Dryomyzidae, the sex ratio differed according to species. Only Dryomyza flaveola (Fabricius, 1794) (Diptera: Dryomyzidae) and Muscina pascuorum (Meigen, 1826) (Diptera: Muscidae) had male-biased sex ratios. Differences in residency patterns between sexes in Stearibia nigriceps (Meigen, 1826) (Diptera: Piophilidae), Liopiophila varipes (Meigen, 1830) (Diptera: Piophilidae), and D. flaveola were statistically insignificant. All studied species were recorded for each sampling method. The effect of the sampling method on the number of individuals was statistically significant for all dipterans but Piophilidae. Pitfall traps outcompeted the aerial sweep net in representing Dryomyzidae and Muscidae, whereas both methods were almost equally effective in the collection of Piophilidae. A varied effectiveness of sampling methods was observed in Fanniidae. The sampling method significantly affected the determination of the appearance time of flies on carcasses, but the effect of species and sampling method was insignificant. Implications for forensic entomology are discussed.

Thermal requirements of immature stages of Chrysomya albiceps (Diptera: Calliphoridae) as a common forensically important fly.

Entomological material may be used to estimate the time since death occurred (postmortem interval, PMI) in forensically obscure cases. The method that is commonly used to calculate minimum post-mortem interval (mPMI, i.e., the least amount of time since one can be confident death occurred) is based on the relationship between insect development and ambient temerature. Isomegalen and isomorphen diagrams are among methods allowing to calculate the age of necorphagous insects, yet thermal summation models provide the most precise and acurate estimations. The digrams are prepared based on the length or the developmental stages of the larvae as a function of time and mean ambient temperature. A knowledge of thermal requirements, in particular lower temperature threshold (Dz) at which development of a species terminates, is of essential importance to calculate ADD (Accumulated Degree Days). In this study different temperature regimes were used to construct the isomorphen diagram, examinate changes in larval body length at different ambient temperatures and to estimate the thermal requirements for developemnt of Chrysomya albiceps, the most common dipteran species reported on human and animal cadavers in Iran. Six development events including hatching, 1st ecdysis, 2nd ecdysis, wandering, pupariation and eclosion were studied under eleven constant temperature regims (17-37 0C). The development rate of Ch. albiceps increased as temperature increased. The larval length peaked at the end of third stage and then decreased at wandering stage. The maximum larval length occurred at 72 h post oviposition at either 31, 33, or 35 °C. At 17 °C, larvae did not hatch from eggs and at 37 °C wandering larvae did not proceed to pupariation, and thus larval development were analysed at the nine left over temperatures. The development stages required at least (Dz ± SE) 13.04 ± 0.37, 14.29 ± 0.45, 15.69 ± 0.56, 15.18 ± 0.56, 14.94 ± 0.48, and 11.23 ± 0.41 °C to reach one of the successive developmentl events, respectively. The estimated thermal summation constant (k) for those the six events were 10.43 ± 0.27, 19.31 ± 0.32, 27.87 ± 1.3, 55.94 ± 1.82, 66.69 ± 3.5, and 143.52 ± 5.61 ADD accordingly.

DNA Barcoding Identifies Unknown Females and Larvae of Fannia R.-D. (Diptera: Fanniidae) from Carrion Succession Experiment and Case Report.

Application of available keys to European Fanniidae did not facilitate unequivocal species identification for third instar larvae and females of Fannia Robineau-Desvoidy, 1830 collected during a study of arthropod succession on pig carrion. To link these samples to known species, we took the advantage of molecular identification methods and compared newly obtained cytochrome oxidase subunit I (COI) barcode sequences against sequences deposited in reference databases. As an outcome of the results obtained, we describe for the first time a third instar larva of Fannia nigra Malloch, 1910 and Fannia pallitibia (Rondani, 1866) and a female of Fannia collini d'Assis-Fonseca, 1966. We provide combinations of characters allowing for discrimination of described insects from other Fanniidae. We provide an update for the key by Rozkosný et al. 1997, which allows differentiation between females of F. collini and other species of Fanniidae. Additionally, we provide a case of a human cadaver discovered in Southern Poland and insect fauna associated with it as the first report of F. nigra larvae developing on a human body.

Correction to: Development and validation of forensically useful growth models for central European population of Creophilus maxillosus L. (Coleoptera: Staphylinidae).

This article was published with an error in Figs. 5 and 6. The figures have multiple mistakes with lines extending beyond the chart and lack of grid lines in some places.

Development and validation of forensically useful growth models for Central European population of Creophilus maxillosus L. (Coleoptera: Staphylinidae).

The hairy rove beetle, Creophilus maxillosus (Linnaeus) (Staphylinidae), is recognized for its use in forensic entomology. However, insufficient developmental data exist for the Central European population of this species. Accordingly, we studied the development of C. maxillosus at ten constant temperatures (10-32.5 °C). Based on these results, linear and nonlinear developmental models were created and validated. We also studied the effect of different homogenous diets (third-instar larvae or puparia of Calliphora sp. Robineau-Desvoidy or Lucilia sp. Robineau-Desvoidy (Diptera: Calliphoridae) or mix of first- and second-instar larvae of Necrodes littoralis (Linnaeus) (Coleoptera: Silphidae)) on the development and mortality of C. maxillosus. Average total development times ranged between 122.21 days at 15 °C and 22.18 days at 30 °C. Beetles reached the adult stage in seven out of ten temperatures (15-30 °C). No beetles reached the adult stage when fed with larvae of N. littoralis; their development times at first and second larval stage were also significantly longer than in other food conditions. When C. maxillosus larvae were fed with blowfly larvae, the highest mortality was observed at the pupal stage, as compared when they were fed with blowfly puparia-at the first larval stage. While validating thermal summation models, the highest age estimation errors were found for beetles bred at 10 and 12.5 °C (between 21 and 43% for all developmental events). Age estimation errors were on average higher for pupation and eclosion than hatching and first and second ecdyses. While validating the models with specimens fed with different diets, the highest errors were recorded for beetles fed with N. littoralis larvae (22% for the first ecdysis and 33% for the second ecdysis) and Lucilia sp. puparia (32% for pupation and 22% for eclosion). Implications for C. maxillosus use in forensic entomology are discussed.

A comparative study of the entomofauna (Coleoptera, Diptera) associated with hanging and ground pig carcasses in a forest habitat of Poland.

Suicide by hanging is one of the most common causes of death, worldwide. Therefore, studies of insects associated with hanging cadavers are important for developing entomological tools capable of estimating the postmortem intervals in such cases. We conducted a two-year experiment, in the spring, summer, and autumn of 2012 and 2013, studying the entomofauna of 12 hanging and 12 ground, decomposing pig carcasses in a hornbeam-oak forest, in western Poland. Both carcass treatments showed colonization by similar insect species. Among the 224 taxa collected, 61 and 52 were minimally abundant (≥10 individuals) during the adult stage on at least one hanging and at least one ground carcass, respectively. During the spring and autumn, more adult flies were attracted to hanging carcasses, whereas during the summer, their numbers were similar on both hanging and ground carcasses. Adult beetles were higher in number on hanging carcasses during the spring, but during the summer and autumn, they were slightly more abundant on ground pig carcasses. Irrespective of the carcass treatment, Dryomyzidae, Muscidae, and Piophilidae were the most abundant flies, whereas representatives of Geotrupidae, Histeridae, and Silphidae were the dominant families in the studied beetle community. Only Anoplotrupes stercorosus (Scriba) and Hydrotaea similis Meade adults and Calliphora vomitoria (Linnaeus) larvae reached minimal abundance on all carcasses during all seasons. Carcass treatment only affected the numbers of minimally abundant fly species at the larval stage due to their higher numbers on ground pig carcasses. Although differences in the residency patterns of insects were observed between carcass treatments, they were not significant. In general, adult insects tended to be present for slightly longer times on hanging carcasses than on ground carcasses, whereas beetle larvae had longer presences on ground pig carcasses. Fly larvae were present on ground carcasses for the same amount of time as on hanging carcasses. More broken residency patterns were observed among the adult insects and beetle larvae that colonized hanging carcasses. Fly larvae that colonized ground carcasses had more breaks within the presence periods than fly larvae colonizing hanging carcasses. Neither treatment significantly affected the time of insect appearance on carrion. Most species appeared at the same time or at within one- or two-day intervals on both treatments. Seasonal differences in insect appearance were found, with most species colonizing quickly during the summer. The implications of these findings for forensic entomology are discussed.

Thermal requirements for the development of immature stages of Fannia canicularis (Linnaeus) (Diptera: Fanniidae).

Entomological material may be used to estimate the time since death occurred (minimum post-mortem interval, mPMI) in forensically obscure cases. The method commonly used to calculate mPMI is the application of a thermal summation model. Little is known about the thermal requirements of Fanniidae. To fill this gap, the influence of temperature on the development time of Fannia canicularis (Linnaeus), a cosmopolitan species with the potential to be used in forensic entomology, was studied. An experiment was performed under nine ambient temperatures. The thermal summation constants (k) and developmental zeros (Tmin) of this species were calculated for six developmental events: hatching, first ecdysis, second ecdysis, wandering, pupariation, and eclosion. Data were primarily analysed with two linear models, and then, since the relationship between temperature and development rate is curvilinear close to the lower and higher developmental thresholds, the relationship was also modelled using three nonlinear models. A fourth nonlinear model was also used to estimate the intrinsic optimum temperature of F. canicularis, at which the population size is maximal and the mortality is the lowest. At 33 °C, larvae did not hatch from eggs, and thus changes in larval body length were analysed at the eight remaining temperatures. Fannia canicularis development from egg to eclosion required a mean ± SE of 481.73 ± 9.89 degree-days above a lower threshold temperature of 4.64 ± 0.19 °C. At the lowest experimental temperature tested, 10 °C, F. canicularis successfully completed all developmental events, although ca. 30% of wandering larvae proceeded to the inactive stage instead of pupariation. Results obtained from nonlinear models were not satisfactory and did not allow us to consider them as reliable from a biological point of view.

Characterization and Identification of Puparia of Hydrotaea Robineau-Desvoidy, 1830 (Diptera: Muscidae) From Forensic and Archaeological Contexts.

Flies and beetles are the main components of the entomofauna colonizing a body after death. Following the recognition of constant and predictable colonization patterns and the knowledge about the dependence of the insect development to temperature, a new discipline, forensic entomology, has provided information useful to reconstruct criminal events. Funerary archaeoentomology has also applied the same rationale in archaeological contexts. Puparia represent a large fraction of the insect remains that can be found associated with a cadaver, especially when the body is mummified or in the advanced stages of decomposition. Puparium identification is still a problematic topic due to the lack of identification keys and, in several cases, a lack of diagnostic feature descriptions. Here, we focus the attention on some Hydrotaea Robineau-Desvoidy (Diptera, Muscidae) puparia from forensic and archaeological contexts. Puparia of Hydrotaea capensis (Wiedemann), Hydrotaea ignava (Harris), Hydrotaea aenescens (Wiedemann), Hydrotaea similis Meade, Hydrotaea pilipes Stein, and Hydrotaea dentipes (Fabricius) are here detailed and illustrated. Posterior spiracles, anal plate, and intersegmental spines have been considered as good diagnostic characters for the identification of these puparia.

DNA barcoding allows identification of European Fanniidae (Diptera) of forensic interest.

In forensic entomology practice, species identification is a prerequisite for any further analysis of collected material. Although morphology-based taxonomy may be hindered by a range of factors, these are not obstacles for a molecular identification approach, so-called DNA barcoding. The Fanniidae are a dipteran family that is attracted to and breeds in decomposing animal carrion and dead human bodies. However, morphological identification of fanniids, both at adult and immature stages, is considered to be difficult, particularly for non-experts. We investigated the usefulness of molecular taxonomy methods as an alternative/supplement for morphology-based identification in European Fanniidae of forensic interest. The material used in this study was collected from various regions in Asia, Europe and North America. We sequenced a barcode region of the mitochondrial cytochrome c oxidase subunit I (COI) in 27 species. For 13 species, including some taxa breeding in dead bodies, this study describes COI sequences for the first time. Our analysis revealed that both mini-barcode and full-length COI barcode sequences give very high specimen identification success. Despite the large number of COI barcode sequences referring to Fanniidae in the BOLD and GenBank databases, previous identification of forensically relevant Fanniidae was hindered by uneven taxonomic sampling. The majority of available sequences refer to species that are not of medico-legal interest, and many species of forensic interest are unrepresented or represented only by a single sequence. Because of erroneous data that are present in depository databases, DNA barcoding must be used with caution and cannot be considered to be the sole alternative to other identification methods. Wolbachia infections in the examined material did not disrupt specimen identification. The obtained results will facilitate precise identification of European Fanniidae of forensic interest, badly preserved material with degraded DNA, as well as matching of unidentified females and immature stages to already described specimens.

Identification of Muscidae (Diptera) of medico-legal importance by means of wing measurements.

Cadavers attract numerous species and genera of Muscidae, both regular elements of carrion insect assemblages, and accidental visitors. Identification of adult Muscidae may be considered difficult, particularly by non-experts. Since species identification is a vital first step in the analysis of entomological material in any forensic entomology orientated experiment and real cases, various alternative methods of species identification have been proposed. We investigated possibility of semiautomated identification by means of wing measurements as an alternative for classic morphology and DNA-based approaches. We examined genus-level identification success for 790 specimens representing 13 genera of the most common European cadavers visiting Muscidae. We found 99.8% of examined specimens correctly identified to the genus-level. Without error, the following were identified: Azelia, Eudasyphora, Graphomya, Hydrotaea, Musca, Muscina, Mydaea, Neomyia, Polietes, Stomoxys and Thricops. Genus-level misidentifications were found only in Helina and Phaonia. Discrimination of examined material on the species level within Hydrotaea (318 specimens representing eight species) and Muscina (163 specimens representing four species) showed lower, yet still high average identification success, 97.2 and 98.8%, respectively. Our results revealed relatively high success in both genus and species identification of Muscidae of medico-legal importance. Semiautomated identification by means of wing measurements can be used by non-experts and does not require sophisticated equipment. This method will facilitate the identification of forensically relevant muscids in comparison to more difficult and more time-consuming identification approaches based on taxonomic keys or DNA-based methods. However, for unambiguous identification of some taxa, we recommend complementary use of identification keys.

Muscidae (Diptera) of forensic importance-an identification key to third instar larvae of the western Palaearctic region and a catalogue of the muscid carrion community.

The Muscidae is one of the main dipteran families recognized as important for medico-legal purposes. Although an association of adult flies with decomposing human and animal bodies is documented for about 200 taxa worldwide, cadavers and carrion represents a breeding habitat for considerably fewer species. Species that do colonize dead human bodies can do so under diverse environmental conditions and, under certain circumstances, Muscidae may be the only colonizers of a body. Because of difficulties in identification, many studies have identified immature and/or adult muscids only to the genus or family level. This lack of detailed species-level identifications hinders detailed investigation of their medico-legal usefulness in carrion succession-oriented experiments. Identification to species level of third instars of Muscidae of forensic importance and the utility of larval morphological characters for taxonomic purposes were subjected to an in-depth revision. A combination of characters allowing for the discrimination of third instar muscids from other forensically important dipterans is proposed. An identification key for third instar larvae, which covers the full set of cadaver-colonising species of Muscidae from the western Palaearctic (Europe, North Africa, Middle East), is provided. This key will facilitate more detailed and species-specific knowledge of the occurrence of Muscidae in forensic entomology experiments and real cases. The carrion-visiting Muscidae worldwide are catalogued, and those species breeding in animal carrion and dead human bodies are briefly discussed with regard to their forensic importance.

Effect of body mass and clothing on carrion entomofauna.

Carcass mass largely affects pattern and rate of carrion decomposition. Supposedly, it is similarly important for carrion entomofauna; however, most of its likely effects have not been tested experimentally. Here, simultaneous effects of carcass mass and clothing are analyzed. A factorial block experiment with four levels of carcass mass (small carcasses 5-15 kg, medium carcasses 15.1-30 kg, medium/large carcasses 35-50 kg, large carcasses 55-70 kg) and two levels of carcass clothing (clothed and unclothed) was made in a grassland habitat of Western Poland. Pig carcasses (N = 24) were grouped into spring, early summer, and late summer blocks. Insects were sampled manually and with pitfall traps. Results demonstrate that insect assemblages are more complex, abundant, and long-lasting on larger carcasses, whereas clothing is of minor importance in this respect. Only large or medium/large carcasses were colonized by all guilds of carrion insects, while small or medium carcasses revealed high underrepresentation of late-colonizing insects (e.g., Cleridae or Nitidulidae). This finding indicates that carcasses weighing about 23 kg-a standard in forensic decomposition studies-give an incomplete picture of carrion entomofauna. Residencies of all forensically relevant insects were distinctly prolonged on larger carcasses, indicating that cadaver mass is a factor of great importance in this respect. The pre-appearance interval of most taxa was found to be unrelated to mass or clothing of a carcass. Moreover, current results suggest that rate of larval development is higher on smaller carcasses. In conclusion, this study demonstrates that carcass mass is a factor of crucial importance for carrion entomofauna, whereas the importance of clothing is small.

Confocal laser scanning microscopy as a valuable tool in Diptera larval morphology studies.

Larval morphology of flies is traditionally studied using light microscopy, yet in the case of fine structures compound light microscopy is limited due to problems of resolution, illumination and depth of field, not allowing for precise recognition of sclerites' edges and interactions. Using larval instars of cyclorrhaphan Diptera, we show the usefulness of confocal laser scanning microscopy (CLSM) for studying the morphological characters of immature stages by taking advantage of the autofluorescent properties of cephaloskeleton structures. We compare data obtained from killed but unprepared larvae with those from larvae prepared by clearing according to two commonly used methods, either with potassium hydroxide or with Hoyer's medium. We also evaluated the CLSM application for examining already slide-mounted larvae stored in museum collections and those freshly prepared. Our results indicate that CLSM and 3D reconstruction are excellent for visualizing small, compound structures of cylrorrhaphan larvae cephaloskeleton, if appropriate clearing techniques, i.e. the application of KOH, are used. Maximum intensity projection of confocal data sets obtained from material freshly prepared and that stored in museum collection does not differ. Because of this and the fact that KOH is commonly used as a clearing method to examine the cephaloskeleton of Diptera larvae, it is possible, and highly recommended, to use slides already prepared with this method for re-examination by CLSM. We conclude that CLSM application can be an invaluable source of data for studies of larval morphology of Cyclorrhapha by way of taxonomic diagnoses, character identification and improvement in characters homologization.

Hydrotaea similis Meade (Diptera: Muscidae) newly reported from a human cadaver: a case report and larval morphology.

A human cadaver is an attractive breeding habitat for necrophagous insects and for those species that are predators of necrophages. One of the important tasks of forensic entomologists is to provide a reliable list of species associated with decomposing bodies both in the adult and larval stages. We present here a case of a human cadaver discovered in Central Sweden and the insect fauna associated with it. This is the first report of Hydrotaea similis larvae developing in a dead human body. The larvae of H. similis have only recently been studied and, therefore, even if previously they had been collected during forensic casework or experiments, the larvae could not have been properly identified, except if reared to the adult stage. To facilitate future studies and casework, detailed descriptions are provided here of the morphology of third instar larvae of both H. dentipes and H. similis using combined SEM and light microscopy techniques. We describe characters that allow for easy species identification and discrimination of these two forensically important and closely related species.

Larval morphology of Atherigona orientalis (Schiner) (Diptera: Muscidae) -a species of sanitary and forensic importance.

Larval morphology is documented using both light and scanning electron microscopy for all three instars of the muscid fly Atherigona orientalis (Schiner), which is a species of known sanitary and forensic importance found in tropical and subtropical areas of all biogeographic regions. The unpaired sclerite in a form of a spicule is reported herein in the second and the third instar larvae. Occurrence of this sclerite was hitherto unknown in the second instar larvae of Muscidae and was only known from the third instar of several species, however not in a form of a spicule. Our study is the first report of the occurrence of the "sensory organ X" in all three larval instars of a species representing the family Muscidae. The bubble membrane, previously known only from third instar cyclorrhaphan larvae, is reported herein for the first time in the second instar. Characters allowing for discrimination of A. orientalis larvae from other forensically important Muscidae are summarised.