Cephalopharyngeal Skeleton Research Articles

Development of a multi-entry identification key for economically important fruit fly larvae (Diptera, Tephritidae, Dacinae).

Identification of fruit fly larvae is difficult due to the limited morphological characteristics present. However, this is the stage at which fruit flies are intercepted at ports of entry through horticultural imports. Molecular tools are useful but are time-consuming and expensive compared to morphological identifications. This project aims to use available information from the literature and our own research to build a multi-entry identification key for thirteen tephritid species and species groups that are of economic concern for the European Union. Third-instar larvae were obtained from different regions and hosts. Thirteen species or representatives of species groups were obtained, including Ceratitis, Dacus, Bactrocera and Zeugodacus spp. The cephalopharyngeal skeletons were dissected out, cleared in a 10% NaOH solution, dehydrated and mounted in Euparal on glass slides. Images of at least 20 larvae/species were captured using a compound microscope fitted with a camera. Measurements were taken of the mounted mandibles and the number of tubules and their position in the anterior spiracles in relation to the cephalic skeleton were noted. Differences between morphometric parameters were tested via ANOVA and verified using discriminant function analysis. A matrix was compiled including nine characters for which significant inter-specific differentiation was preliminarily detected. The key was converted into a mobile application by LucID.

Bolwig Organ and Its Role in the Photoperiodic Response of Sarcophaga similis Larvae

Flesh-fly Sarcophaga similis larvae exhibit a photoperiodic response, in which short days induce pupal diapause for seasonal adaptation. Although the spectral sensitivity of photoperiodic photoreception is known, the photoreceptor organ remains unclear. We morphologically identified the Bolwig organ, a larval-photoreceptor identified in several other fly species, and examined the effects of its removal on the photoperiodic response in S. similis. Backfill-staining and embryonic-lethal-abnormal-vision (ELAV) immunohistochemical-staining identified ~34 and 38 cells, respectively, in a spherical body at the ocular depression of the cephalopharyngeal skeleton, suggesting that the spherical body is the Bolwig organ in S. similis. Forward-fill and immunohistochemistry revealed that Bolwig-organ neurons terminate in the vicinity of the dendritic fibres of pigment-dispersing factor-immunoreactive and potential circadian-clock neurons in the brain. After surgical removal of the Bolwig-organ regions, diapause incidence was not significantly different between short and long days, and was similar to that in the insects with an intact organ, under constant darkness. However, diapause incidence was not significantly different between the control and Bolwig-organ-removed insects for each photoperiod. These results suggest that the Bolwig organ contributes partially to photoperiodic photoreception, and that other photoreceptors may also be involved.

Presence of the European bone-skipper fly Thyreophora cynophila (Panzer, 1798) (Diptera: Piophilidae) larvae in the cerebral cavity of large mammals and report of different insect larvae living inside mammal bones

Summary The iconic fly Thyreophora cynophila has long attracted the interest of entomologists but, despite its re-discovery in Spain and France, its biology is still poorly known. The larvae and puparium have been described in detail but their feeding habits on large carcasses remain largely unknown. Here, we took advantage of a carrion pile containing the remains of wild boar (Sus scrofa) and red deer (Cervus elaphus) as well as of two cow (Bos taurus) carcasses to search for T. cynophila larvae inside the bones and skulls. We document the occurrence of T. cynophila inside the cerebral cavity of dead animals, containing the brain. From the carrion pile, out of seven skulls, T. cynophila larvae could be found in five of them, and identified on the basis of their size, the shape and arrangement of the ventral tubercles, prothoracic spiracle and cephalopharyngeal skeleton. In contrast, no T. cynophila larvae could be found inside the nine bones we inspected. From the cow carcasses, no T. cynophila larvae could be found in the bones but one larva was found inside the cerebral cavity of one cow. In addition, other Diptera and Coleoptera larvae were found inside the bones.

The true identity of Periscelis winnertzii and description of P. laszloi sp. nov. from Europe (Diptera: Periscelididae)

The only preserved (damaged, sex unknown) type specimen of Periscelis (Periscelis) winnertzii Egger, 1862 has been revised and designated as a lectotype. Its revision revealed that P. winnertzii has recently been misinterpreted and is in fact identical with P. (P.) fugax Roháček & Andrade, 2017, syn. nov. Therefore, true P. winnertzii is re-diagnosed, with all its synonomies listed. Periscelis winnertzii auctt. (not Egger, 1862) is described as a new species, P. laszloi sp. nov., based on a series of specimens from Hungary (holotype), Portugal, Switzerland and Slovakia. An intersex specimen of P. laszloi sp. nov., female puparium and cephalopharyngeal skeleton of 3rd instar larva (ex puparium) of P. winnertzii (all from Hungary) are described and illustrated. Both species are also characterized by means of the barcoding region of COI, illustrated in detail (including structures of male and female terminalia) and their biology and distribution reviewed with new original data.

Distinguishing Four Calliphoridae Species (Diptera) from Jamaica Using the Cephalopharyngeal Skeleton: Application to Forensic Investigations.

Flies of the family Calliphoridae, commonly called blow flies, are important in the decomposition process. Knowledge on their succession pattern on corpses, species identification and the duration of their life cycle stages can be useful in forensic investigations especially when estimating the post-mortem interval. We performed linear-based morphometrics on the cephalopharyngeal skeleton of four blow fly species found in Jamaica to distinguish species and determine larval development stage. We collected eggs from pigs' heads used as bait in the field and conducted rearing exercises in the laboratory. We used the internal skeletonized structure, the cephalopharyngeal skeleton, to develop a practical and efficient method for species identification. For the first instar, we found species can be differentiated using all the measurements analysed in the study. We found that the mouth hook length may be useful in distinguishing larvae in the second instar. For the larvae in the third instar, the whole length of the skeleton, from mouth hook to length of the dorsal cornue, may be useful for separating species. We provide information on the cephalopharyngeal skeleton of Lucilia lucigerens (James), a blow fly species endemic to Jamaica, for the first time. Our work provides relevant information that could be utilized for species identification and life stage determination if fly evidence is to be incorporated in forensic investigations in Jamaica.

Elephantoloemus indicus Austen, 1930 (Diptera: Calliphoridae) as the cause of cutaneous myiasis in captive Indian elephants from Assam, India.

Elephantoloemus indicus Austen, 1930, a dipteran calliphorid fly is known to cause by its larval stage obligatory cutaneous myiasis in Indian subspecies of Asian elephants (Elephas maximus indicus Cuvier, 1798) in Myanmar and Thailand. The present study was undertaken on morphological identification of some specimens of fly larvae which were recovered from the warbles detected on the skin of captive Indian elephants at the Nameri National Park and Kaziranga National Park both situated in the state of Assam, India. The larval specimens were whitish to creamy white in colour and body conformation varied from cylindrical to barrel shaped depending on their measured size (Av 6.12±0.28×2.35±0.12mm). Microscopic examination of processed larvae revealed presence of numerous single pointed spines uniformly distributed on entire body surface, well developed mouth hooks and cephalopharyngeal skeleton at the anterior end and posterior spiracles each with lightly sclerotized peritreme enclosing three short and straight respiratory slits. Based on geographical distribution of the fly, host relation, larval parasitism and morphological characters, the larvae were determined as of the genus Elephantoloemus which is represented by E. indicus as the only species described so far. This finding seems to be the first record in India after its report from Myanmar and Thailand.

Case report of hook worm Grammocephalus hybridatus and stomach bot Cobboldia elephantis infections in a free-ranging Asian Elephant Elephas maximus in Tamil Nadu, India

Elephants in the wild are susceptible to many gastrointestinal parasites. In the present study, necropsy was conducted on a free-ranging Asian Elephant Elephas maximus female aged about 15 years which died at Coimbatore forest range, Tamil Nadu state, India. The necropsy revealed that the liver was infected with round worms and the stomach was heavily infested with dipteran larvae. These round worms and larvae were collected and processed by dehydrating in ascending grades of alcohol and then cleared in carbolic acid. The cleared samples were mounted and examined under light microscopy for species identification. Faecal samples collected from the rectum were analysed by sedimentation for the presence of helminth eggs. On microscopic examination, the head end of the round worms showed a buccal capsule which possessed a pair of semilunar ventral cutting plates. Male worms showed well-developed bursa at the posterior end. The anterior end of the dipteran larvae showed two powerful oral hooks with cephalopharyngeal skeleton. Anterior spiracle appeared as a short club-shaped tube with 12 lobes. The abdominal segments of the larvae had a row of belt-like triangular spines. The posterior spiracles of the larvae had three longitudinal parallel slits in each spiracle with closed peritreme. Based on the above morphological characters, the round worms and larvae were identified as Grammocephalus hybridatus and Cobboldia elephantis, respectively. Strongyle eggs were identified in the faecal sample based on the morphology of thin shell and segmented yolk. This appears to be the first report of G. hybridatus infection in a free-ranging elephant in Tamil Nadu state, India.

PM 7/53 (2) Liriomyza spp.

PM 7/53 (2) <i>Liriomyza</i> spp.

A PRELIMINARY EVALUATION ON BIVARIATE ALLOMETRY IN ACTIVE-FEEDING Chrysomya megacephala (FABRICIUS, 1794) (DIPTERA: CALLIPHORIDAE) LARVAE

The potential use of larval cephalopharyngeal skeleton as an alternative growth indicator in forensic entomology practice was assessed based on its developmental pattern, growth performance and allometric relationship with larval body. Chrysomya megacephala (Fabricius, 1794), was used as an experimental species and larval development were studied at ambient temperatures and relative humidity. Larval body size was measured from furthest part of the head to the last abdominal segment. Cephalopharyngeal skeleton was extracted from the body and measured from the tip of dorsal bridge to the left face of dorsal cornu. Daily progression of larval body length and cephalopharyngeal skeleton length showed the latter significantly had slower growth rates. The allometry of larval body and cephalopharyngeal skeleton showed they were correlated only because both variables increased across the three larval instars, which suggesting a spurious correlation. Separate bivariate correlations between the two variables showed only the first instar larvae had a significant (p&lt;0.01) but weak correlation, r=0.33. However, removing the effect of larval instars still produced statistically significant correlation (p&lt;0.001) albeit with moderate strength, r=0.57. From this study, there was not enough evidence to support cephalopharyngeal skeleton as an equal or a better alternative to larval body as growth indicator.

Morphological and phylogenetic characterization of Oestrus ovis larvae in sheep: Al-Qadisiyah province, Iraq

The fly larvae infect the nasal cavities and sinuses (frontal and maxillary) of sheep, goats, and a range of wild ruminants, forming a disease called oestrosis (Nasal myiasis or nasal bot). The disease is one of the significantly diseases for the Iraqi small ruminant industry that causes detrimental economic losses. The current work was carried out to morphologically- and molecularly-characterize O. ovis larvae collected from sheep in a slaughterhouse in Al-Qadisiyah province, Iraq. The study depended on collecting 20 larvae (at different stages) from 20 sheep from 15 October till 17 December 2020. The morphological examination was done using a stereomicroscope and relying on larval characteristic features, including the posterior end, spiracles, and cephalopharyngeal skeleton. The molecular characterization was performed utilizing polymerase chain reaction (PCR) and partial gene sequencing (PGS) methods of the cytochrome c oxidase subunit I (cox1) gene at 700-bp and 300-bp regions. Morphologically, the first-stage larvae (L1) showed characteristic mouth hooks, while the second-stage larvae (L2) revealed clear terminal stigmas. For the third-stage larvae (L3), the color of body segments and their spines' were the most important features for this larval stage. The PCR showed amplification at both regions 700bp and 300bp, in 8 and 7 isolates, respectively. The PGS revealed 15 different local isolates in genetic level aligned with isolates from Kyrgyzstan, Italy, Spain, and Turkey. This study shows the important strain differences of O. ovis that infect the local sheep in Al-Qadisiyah province, Iraq.

Development of forensically important scuttle fly, Megaselia spiracularis Schmitz (Diptera: Phoridae) at ambient temperatures with distinguishing features of the larval instars

Megaselia spiracularis Schmitz (Diptera: Phoridae) is a scuttle fly species of forensic importance and has been recorded colonizing human corpses in Malaysia, Korea and China. The necrophagous activity of M. spiracularis larvae on human corpses can be referred to estimate minimum postmortem interval (PMImin) in death investigation. However, developmental data of M. spiracularis derived from ambient fluctuating temperature is scarce and taxonomic descriptions to larval instars for species identification are still lacking. This research was conducted to address these limitations by studying the development of M. spiracularis at fluctuating ambient temperatures. From each sampling occasion, larval body lengths were obtained and samples were mounted on slides to determine larval instars based on taxonomic characteristics of the cephalopharyngeal skeleton, anterior spiracles and posterior spiracles. At fluctuating temperatures of 25–37 °C and relative humidity of 47–93%, developmental duration of egg stage was 7–9.5 h; first instar larva, 9.5–17 h; second instar, 6–12 h, feeding third instar, 18–36 h; post-feeding third instar, 6–12 h; and pupa, 168–204 h. Accumulated degree hours for each developmental stage based on fluctuating ambient temperatures were also determined. The size of M. spiracularis larvae varied from 0.64–0.78 mm after eclosion to 5.20–5.95 mm before entering post feeding phase. The first, second and third instar larva could be distinguished mainly by the morphology of cephalopharyngeal skeletons, anterior spiracles and posterior spiracles. Developmental durations of M. spiracularis larvae and the larval instars can be used as references to estimate PMImin as this species is likely to be featured in future forensic cases.

Effects of Preservative Concentrations on Larval Cephalopharyngeal Skeleton of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae), an Alternative Indicator to Larval Body Length for mPMI Estimation

Introduction: In forensic entomology, dipterous larval specimens found feeding on decomposing corpses are usually preserved in a range of 70-95% ethanol before being subjected to minimum postmortem interval (mPMI) assessment. However, larval body size, which is commonly used to infer mPMI, can be affected by preservatives resulting miscalculation of estimation. This study compared the effect of ethanol concentrations on larval body and cephalopharyngeal skeleton, a potential substitute to estimate larval age for mPMI estimation. Methods: Experiments were conducted on homogenised colony of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) third instar larvae reared in similar conditions. They were fixed with hot water (»80ºC) and preserved in 70% and 90% ethanol. After seven days in preservatives, larvae were measured for total body length from furthest part of the head to the last abdominal segment. Cephalopharyngeal skeleton was subsequently extracted from the body and measured based on morphometric landmarks on the pharyngeal sclerite. Centroid size of the cephalopharyngeal skeleton was also calculated based on the configuration of five morphometric landmarks. Results: In all four study replicates, pairwise comparisons with the original size indicated that larval body length was significantly affected by ethanol concentration (p&lt;0.001) whilst only two of the study replicates showed cephalopharyngeal skeleton maintained its size when preserved in different ethanol concentration. Conclusion: Possible causes of variations are discussed herein with the results clearly indicated cephalopharyngeal skeleton should be considered as alternative growth parameter for mPMI estimation.

Accidental intestinal myiasis caused by Musca domestica L. (Diptera: Muscidae) larvae in broiler chickens: a field study

Broiler chicken’s production considered a very important sector of Egyptian poultry production. Diarrhea is a common problem that Threaten poultry production. There are many causes of broiler chicken’s diarrhea as; nutritional causes, infectious causes (viral, bacterial and parasitic). Larvae rarely infest and cause diarrhea in chickens but if present they irritate intestinal mucosa which causes a secondary bacterial diarrhea. In our investigation, intestinal myiasis was reported as uncommon cause of broiler chicken diarrhea in Egypt. The collected larvae were identified using scanning electron microscope study which revealed that; it is 3rd larval stage of M. domestica; creamy white in color; the length of the larvae ranged from 10-12 mm (10.5 ± 0.3); the anterior end contain the black colored cephalopharyngeal skeleton; the anterior spiracles present in the 2nd segment and had a number ranged from 6 to 8 rounded spiracular openings. The spines present in the group resembles the combs which present in the spinose area. The spinose comb contains from 5 to 9 spines which is small and triangular with slightly tapering ends. The posterior peritremes contain three snake or S- shaped or even tortoise openings; with central button; the peritreme was complete; the distance between the two peritreme ranged from 70- 100 μm in length.

Taxonomic notes on the genus Campiglossa Rondani (Diptera, Tephritidae, Tephritinae, Tephritini) in India, with description of three new species.

Three new species of Campiglossa Rondani are described from India: adults of both sexes and third instar larvae of C. ialong David, Salini & Hancock, sp. nov. and C. sherlyae David & Hancock, sp. nov., plus an adult female of C. shaktii David, Sachin & Hancock, sp. nov., are described and illustrated. Postabdominal structures, cephalopharyngeal skeleton, and anterior and posterior spiracles of C. gemma (Hering, 1939) and C. sororcula (Wiedemann, 1830) are illustrated. DNA barcode sequences of C. ialongsp. nov., C. sherlyaesp. nov., and C. gemma were obtained and reported. Records of C. absinthii (Fabricius, 1805) and C. iracunda (Hering, 1938) are regarded as misidentifications of C. lyncea (Bezzi, 1913) and C. shaktiisp. nov., respectively, and excluded from the Indian fauna. A key to the known species of Campiglossa from India is provided. Results of preliminary phylogenetic analysis using COI revealed that C. ialongsp. nov. is paraphyletic to the Campiglossa misella group and C. C. sherlyaesp. nov. is a sister species of C. deserta.

Perianal Traumatic Myiasis in a Goat Caused by Wohlfahrtia magnifica (Schiner 1862) (Diptera: Sarcophagidae) in Konya Province of Turkey

Introduction: The infestation of live humans and animals with dipterous larvae is called Myiasis. These larvae feed on the living or dead tissues, body fluids, and ingested foods of hosts at least for a certain period in myiasis cases. Case Presentation: During clinical examination, several larvae were detected in the perianal area of a goat at Selcuk University, Faculty of Veterinary Medicine, Internal Medicine Clinics. All visible larvae were cleaned by the clinician with the help of forceps. Ivermectin administered against larvae possibly penetrated deep into the wound. A topical antibiotic was also applied, and the wound was washed with an antiseptic solution. Several larvae were preserved in tubes including ethanol (70%) and transferred to parasitology laboratory. Firstly, the larvae were incised with a scalpel and cleared in potassium hydroxide (10%). Then, they were rinsed in distilled water, put into 70% ethanol for dehydration, and stored in 99% ethanol until dissection. The larvae were morphologically examined under a stereo microscope and dissected. The larvae were identified as third instar larvae of Wohlfahrtia magnifica according to the morphological characteristics of cephalo-pharyngeal skeleton, anterior spiracles, and peritremes. Conclusion: It should be kept in mind that in myiasis cases, early diagnosis is a very important factor in preventing deep tissue loss.

Larvae of the Genus Rainieria Rondani, 1843 (Diptera, Micropezidae)

Data on the life history and morphology of the larvae of the fly genus Rainieria are presented, viz. R. hennigi Krivosheina et Krivosheina, 1996 and R. latifrons (Loew, 1870). The larvae inhabit the bark of damaged and weakened trees. The main diagnostic characters of the larvae are listed. The larvae of this two species differ in the structure of the anterior and posterior spiracles and the cephalopharyngeal skeleton.

Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) development by landmark-based geometric morphometrics of cephalopharyngeal skeleton: a preliminary assessment for forensic entomology application

BackgroundConsidering the practicality of geometric morphometrics which could discriminate insect species, this application was extended to the analysis of blow fly larval growth based on cephalopharyngeal skeleton. In forensic entomology, cephalopharyngeal skeleton plays a crucial role in species identification but the morphometric information of this part is scarce. In this study, Chrysomya megacephala (Fabricius, 1794) was reared in two study replicates in natural conditions and samplings were conducted at fixed daily intervals. Cephalopharyngeal skeletons were removed from larvae and mounted on glass slides. Images were obtained from the specimens; digitized and geometric morphometric analysis on C. megacephala cephalopharyngeal skeletons was performed with MorphoJ software based on the ordination of five landmarks. The assessments of this analysis were based on centroid size measurements, visualization on the landmarks displacements, classification of the relative landmarks by using canonical variate analysis, and ontogenetic allometry determination.FindingsCentroid size was strongly correlated with developmental time (p < 0.05) and significantly different between daily intervals (p < 0.05). Ontogenetic allometric effect based on multivariate regression on Procrustes coordinates and centroid size was significant (p < 0.0001), indicating that shape was influenced by growth (60.3%). Disposition occurred on all landmarks during development and was further discriminated based on age groups.ConclusionsOther than discriminating between species, geometric morphometrics was found to be practical to visualize larval growth based on cephalopharyngeal skeletons which can be useful in forensic entomology.

Non‐destructive DNA extractions from fly larvae (Diptera: Muscidae) enable molecular identification of species and enhance morphological features

AbstractInsects preserved as reference specimens are important in a wide range of fields, including health, pest management and forensics. The aim of the present study was to test a non‐destructive DNA extraction method on samples of soft‐bodied insects, fly larvae, which are otherwise hard to identify morphologically. This not only provides DNA enabling molecular identification but also retains morphological reference specimens for samples belonging to collections and museums that cannot be destroyed. In this work, fly larvae identified as belonging to the family Muscidae were non‐destructively processed. DNA barcoding molecular identification allowed most of these specimens to be assigned to species. Furthermore, the visibility of seven important larval morphological characters – anterior and posterior spiracles, mouth hooks, cephalopharyngeal skeleton, locomotory welts, segmentation and colour – was assessed pre‐ and post‐DNA extraction. It was found that the morphology generally did not deteriorate post‐DNA extraction but actually improved through increased visibility of internal features. Therefore, this non‐destructive DNA extraction method not only allowed COI barcode sequences to be obtained, but also enabled a better morphological identification of the fly larvae retaining physical reference voucher specimens and avoiding the need for dissections.

Preliminary assessment of cephalopharyngeal skeleton length and body length of Hemipyrellia ligurriens (Wiedemann) (Diptera: Calliphoridae) larvae as potential parameters to estimate minimum post mortem interval

Entomological evidence can be utilized in forensic investigation to estimate the elapsed time after death or minimum post mortem interval estimation (PMImin). This estimation is based on the age of dipterous specimens feeding on decomposing human tissues, which commonly refers to their larval body length. The objective of this research was to look for alternatives to larval body length which could be impaired by specimen handling and subsequently causing inaccuracy in PMImin estimation. In this research, development of forensic blow fly, Hemipyrellia ligurriens (Wiedemann) (Diptera: Calliphoridae), based on larval cephalopharyngeal skeleton length, was described for the first time. Development of H. ligurriens was represented by body length and cephalopharyngeal skeleton consisting anterodorsal process to dorsal cornu (ADP-DC), anterodorsal process to ventral cornu (ADP-VC) and dorsal cornu to ventral cornu (DC-VC). The cephalopharyngeal skeletons proportionally developed with larval body development, but periods of plateau existed between 0 and 10 h and 24–30 h, suggesting ecdysis periods of larvae. Data measurements were more consistent in cephalopharyngeal skeleton length than in larval body length, and there were strong positive significant correlations, r > 0.85 (p < 0.05). These findings warranted further investigations to examine cephalopharyngeal skeleton as an alternative growth parameter to larval body length.

European Chiropteromyzidae (Diptera): Taxonomic Revision, Nomenclature, Classification and Preimaginal Stages

European chiropteromyzid flies (Diptera: Acalyptrata: Heleomyzidae) are revised to clarify their nomenclature, taxonomic concepts and relationships. Two species are recognized which are considered to belong to the genus Neossos Malloch, 1927 viz., the widespread N. broersei (de Meijere, 1946) comb. nov. (= Chiropteromyza wegelii Frey, 1952) and N. nidicola (Frey, 1930) known only from type series from Finland. Consequently, the monotypic genera Ornitholeria Frey, 1930 (type species: O. nidicola Frey, 1930) and Chiropteromyza Frey, 1952 (type species: C. wegelii Frey, 1952) are treated as junior synonyms of Neossos Malloch, 1927. The above synonymies and generic affiliations are confirmed on the basis of comparative morphology of both European species and N. marylandicus Malloch, 1927, the type species of the genus Neossos. All studied taxa are redescribed and illustrated including detail of the male genitalia and/or female postabdomen, keyed and their full synonymies are presented. Preimaginal stages (3rd-instar larva, puparium) of N. broersei are studied for the first time. Research history, morphology of terminalia, relationships, biology and distribution of European species are reviewed and/or discussed. Lectotypes of Ornitholeria nidicola Frey, 1930 and Chiropteromyza wegelii Frey, 1952 are designated. Based on similarities in the external morphology, including the male and female terminalia, and the cephalopharyngeal skeleton of the larva, chiropteromyzids are considered to be a heleomyzoid lineage most closely related to the subfamily Heleomyzinae (of European authors) and, hence, not deserving family rank within Acalyptrata.