Ultrastructure of the larval salivary glands of Megaselia scalaris Loew (Diptera,Phoridae)

Each of the paired salivary glands of third instar larvae of the humpbacked fly Megaselia scalaris is a bag-like structure with a short neck region from which a single duct emerges. The two ducts form a common duct that empties into the ventral region of the pharynx near the mouthparts. The wall of the glands and ducts consists of a simple squamous epithelium that rests upon a connective tissue layer. Cells in the neck are less flattened than those found elsewhere. The basal surfaces of the cells are infolded most deeply in the neck and the least in the duct. The apical surfaces of the cells possess microvilli except in the duct where the apices of the cells are covered by a complex extracellular layer. This layer displays circularly arranged folds that accommodate a thread-like supportive structure resembling taenidial threads of tracheae. Elaborate junctional complexes are associated with the lateral surfaces of the cells. Elements of these complexes include a zonula adherens, a series of pleated septate desmosomes, and conventional desmosomes. The cytoplasm of the glandular cells is filled with RER and other organelles normally seen in cells that export proteins and mucosubstances. Secretory material found in the lumens of the glands reacts only moderately with the PAS procedure but more strongly with alcian blue and methods that demonstrate proteins. The nuclei of the glandular cells contain single large nucleoli and polytene chromosomes whose banding is rather indistinct. Treatment with EDTA produces detrimental effects on all of the foregoing ultrastructural features of the glands and ducts.     

Observations on first and second-instar larvae of Megaselia scalaris (Loew) (Diptera: Phoridae).

The ultrastructure of the first and second-instar larvae of Megaselia scalaris (Loew) was studied using scanning electron microscopy (SEM). Significant changes in morphological features were observed in the anterior and posterior spiracles, but only minimal changes in the labium and mouthhooks were seen. The ultrastructure of M. scalaris larvae not only provides chronological transformation of their larval instars, but it can also be used to explain their feeding behavior and mode of respiration. In addition, morphological structures useful for specific identification of first or second-instar larvae collected from human corpses may be used in forensic investigations.     

Ultrastructure of the spermatozoon of Megaselia scalaris Loew (Diptera:Brachycera:Cyclorrhapha:Phoridea:Phoridae)

When viewed by scanning electron microscopy (SEM), the spermatozoon of the phorid dipteran Megaselia scalaris appears threadlike, lacking distinct head and tail areas. These areas can be observed, however, in appropriately stained material. Measurements of Feulgen-stained material reveal average lengths of the head, tail, and total cell of 18.7, 128.7, and 147.4 μm, respectively. When tested for sulfhydryl and disulfide groups, the head displays only disulfide groups. Transmission electron microscopy (TEM) reveals 12 different regions: three (1–3) in the head, four (9–12) in the tail, and five (4–8) in a short zone of overlap between the head and tail. Most of the cell lies in regions 9 and 11 of the tail and 3 of the head, accounting for, respectively, 37.3%, 45.7%, and 11.2% of the total length. A tubelike acrosome indents the anterior end of the nucleus. The tail originates asymmetrically in relation to the long axis of the cell as a peglike structure associated with the dorsolateral region of the nucleus. No centriole is visible, and the nucleus has a notched appearance in longitudinal sections. Two mitochondrial derivatives and an axoneme displaying a 9+9+2 microtubule configuration and ATPase activity extend throughout most of the tail. In regions 9 and 10, an asymmetrically arranged accessory body is also present. Features having possible taxonomic utility include the asymmetrically arranged accessory body, the size and shape of the acrosome, and the notched appearance of the nucleus. The present report is apparently the first to describe the spermatozoon of a cyclorrhaphous dipteran which is not a member of the Schizophora.     

Surface ultrastructure of third-instar Megaselia scalaris (Diptera: Phoridae)

We describe some ultrastructure of the third-instar Megaselia scalaris (Diptera: Phoridae) using scanning electron microscopy, with the cephalic segment, anterior spiracle and posterior spiracle being emphasized. This study provides the taxonomic information of this larval species, which may be useful to differentiate from other closely-related species.     

蛆症异蚤蝇减数分裂染色体观察

以蛹精巢和卵巢组织为材料,采用空气干燥法制备染色体标本,Giemsa和硝酸银分别染色,对蛆症异蚤蝇Megaselia scalaris减数分裂染色体行为进行研究。结果表明:蛆症异蚤蝇的染色体数目n=3,由2条中着丝粒染色体和1条端着丝粒染色体组成;粗线期,第2条二价体具有较强的嗜银性,可能为性染色体;晚粗线期,第1条二价体的同源染色体之间出现一条细线,类似于联会复合体;终变期,第2条二价体形成环状结构;晚终变期,在3条二价体染色体臂上均产生条带,根据二价体着丝粒处是否成环可以将3条二价体分开。     

Competition between three types of sex determination in Megaselia scalaris Loew (Phoridae)

Summary Artificial populations of Megaselia scalaris were reared in population cages and in overcrowded glass cultures respectively. The three types of sex-determination according to the I- or II- or III- chromosome were set in competition with different frequencies starting the artificial populations. The genetic background was varied by using different strains. An equilibrium between the types of sex-determination was reached in all the populations. The II ^M -type proved a disadvantage in all examinations. In well ventilated cages the I ^M-type was strongly successful. In overcrowded glass cultures the III ^M-type proved an eminent advantage.     

First record of Megaselia scalaris (Loew) (Diptera: Phoridae) infesting laboratory colonies of Triatoma brasiliensis Neiva (Hemiptera: Reduviidae)

Megaselia scalaris (Loew) is a cosmopolitan and synanthropic scuttle fly, eclectic in its feeding habits and acts as detritivore, parasite, facultative parasite, and parasitoid. Here we report for the first time M. scalaris infesting laboratory colonies of Triatoma brasiliensis Neiva, the most important Chagas disease vector in semiarid areas of Brazil. M. scalaris larvae were found feeding inside bugs; pupae were found in the esophagus and intestinal regions of T. brasiliensis through dissection. Other relevant information about this finding is also described in this note, including some preventive measures to avoid laboratory colonies infestations.     

Estimation of population profiles of two strains of the fly Megaselia scalaris (Diptera: Phoridae) by bootstrap simulation

Based on experimental population profiles of strains of the fly Megaselia scalaris (Phoridae), the minimal number of sample profiles was determined that should be repeated by bootstrap simulation process in order to obtain a confident estimation of the mean population profile and present estimations of the standard error as a precise measure of the simulations made. The original data are from experimental populations founded with SR and R4 strains, with three replicates, which were kept for 33 weeks by serial transfer technique in a constant temperature room (25 +/- 1.0 degrees C). The variable used was population size and the model adopted for each profile was a stationary stochastic process. By these simulations, the three experimental population profiles were enlarged so as to determine minimum sample size. After sample size was determined, bootstrap simulations were made in order to calculate confidence intervals and to compare the mean population profiles of these two strains. The results show that with a minimum sample size of 50, stabilization of means begins.     

Reactions of seven basic fluorochromes with unfixed cells obtained from the salivary glands of the dipteran fly Megaselia scalaris Loew (Phoridae)

Seven basic fluorochromes with varying specificities were used to stain the large squamous epithelial cells isolated from the larval salivary glands of Megaselia scalaris (Phoridae). Although the EDTA-based method selected for isolating the cells produced permeabilization and a loss of viability of the cells, consistent results were obtained with the various fluorochromes. The "classical" pattern of green nuclear and red cytoplasmic fluorescence observed in cells stained with acridine orange could be changed to green cytoplasmic and red nuclear fluorescence by pretreatment with RNase. The predominantly cytoplasmic and nucleolar fluorescence obtained with pyronine Y could be changed to mainly nuclear fluorescence by RNase pretreatment. The other five fluorochromes tested were not affected appreciably by extraction with RNase. Quinacrine mustard, dicarbocyanine (DiOC3(3)), and rhodamine 123 produced primarily cytoplasmic and nucleolar fluorescence, while nile red revealed mainly cytoplasmic lipid droplets. Phosphine 3R initially stained lipid droplets but very rapidly redistributed throughout the cytoplasm and nucleus. Because of their large size, flatness, and content of histochemically demonstrable components, the cells of Megaselia are especially appropriate for use as "optical objects" or controls in various studies. New methods of isolating the cells, however, will be needed to prevent permeabilization and loss of viability of the cells.     

Reactions of seven basic fluorochromes with unfixed cells obtained from the salivary glands of the dipteran fly Megaselia scalaris Loew (Phoridae)

Summary Seven basic fluorochromes with varying specificities were used to stain the large squamous epithelial cells isolated from the larval salivary glands of Megaselia scalaris (Phoridae). Although the EDTA-based method selected for isolating the cells produced permeabilization and a loss of viability of the cells, consistent results were obtained with the various fluorochromes. The classical pattern of green nuclear and red cytoplasmic fluorescence observed in cells stained with acridine orange could be changed to green cytoplasmic and red nuclear fluorescence by pretreatment with RNase. The predominantly cytoplasmic and nucleolar fluorescence obtained with pyronine Y could be changed to mainly nuclear fluorescence by RNase pretreatment. The other five fluorochromes tested were not affected appreciably by extraction with RNase. Quinacrine mustard, dicarbocyanine (DiOC₆(3)), and rhodamine 123 produced primarily cytoplasmic and nucleolar fluorescence, while nile red revealed mainly cytoplasmic lipid droplets. Phosphine 3R initially stained lipid droplets but very rapidly redistributed throughout the cytoplasm and nucleus. Because of their large size, flatness, and content of histochemically demonstrable components, the cells of Megaselia are especially appropriate for use as optical objects or controls in various studies. New methods of isolating the cells, however, will be needed to prevent permeabilization and loss of viability of the cells.     

Alternative reproductive routines in a small fly, Puliciphora borinquenensis (Diptera: Phoridae)

Abstract. 1. Puliciphora borinquenensis (Wheeler), a very small phorid fly in which the female is apterous, breeds in small patches of decomposing organic matter such as dead insects. 2. Females leave the shelter of oviposition sites during the afternoon and ‘parade’ in exposed places making vigorous abdominal pumping movements which probably help to disperse a pheromone. 3. Males exhibit one of four reproductive routines. In the first they perform rapid sequences of stationary copulations with different parading females, achieving rates of 0.66 females min^-1 for periods of 30min (Al). In the second (A2) they wait on oviposition sites and grab non-parading females which have recently arrived. 4. In the third routine a male airlifts a parading female in copula to an oviposition site (Bl); in the final type of routine, females are airlifted and then deposited randomly when oviposition sites cannot be found (B2). 5. Males are able to learn the location of oviposition sites and this enables them to transport large numbers of females (up to thirty-three per male) to sites at a high rate (0.5 females min^-1). 6. Immediately a female is released after a B routine, the male follows her closely for up to 10s. This is thought to represent a phase of non-contact guarding, suggesting the occurrence of sperm competition. 7. Individual males persist with either A or B routines for long periods (c. 30 min), but they are able to switch between routines. 8. Young males (< 27 h after emergence) carry out A routines 18 times more commonly than B routines, whereas older males carry out A routines only 1.6 times more commonly than B routines. The choice between A and B routines in older flies does not depend on male or female density. 9. Males benefit from B routines because the eggs they fertilize are likely to reach oviposition sites, but they pay a considerable energy cost. A routines, on the other hand, are energetically cheaper, but mated females are less likely to reach oviposition sites unless they are subsequently transported by B males. An A male can then benefit provided his sperm are not completely displaced by the second male.     

Chronic exposure to elevated levels of manganese and nickel is not harmful to a cosmopolitan detritivore, Megaselia scalaris (Diptera: Phoridae)

Environmental contamination with metals such as manganese (Mn) and nickel (Ni) often results in elevated concentrations of these metals in plant tissues. At high concentrations, these metals are known to have detrimental effects on certain insect herbivores. Using laboratory bioassays and artificial diet, we investigated the development and survival of a cosmopolitan insect detritivore, Megaselia scalaris (Diptera: Phoridae), exposed to concentrations of Mn and Ni reaching 2600 mg Mn/kg and 5200 mg Ni/kg dry mass (dm) in artificial diet. Surprisingly, Ni and Mn at the concentrations tested did not harm this fly. Treatment groups from diets with 260–2600 mg Mn/kg dm and 1300–5200 mg Ni/kg dm had significantly shorter larval development times, overall times to adult emergence, and both pupariation and pupal eclosion times compared to a control group. Wing length of females, a correlate of adult fitness, was also greater in metal treatment groups. Other measures including rate of egg hatch, percentage of emerging flies that were female, and wing length of male flies, were not significantly different in metal treatment groups. We conclude that Megaselia scalaris is tolerant of exceptionally high levels of Mn and Ni.     

Structure,mitotic and meiotic behaviour,and stability of centromere-like elements devoid of chromosome arms in the fly Megaselia scalaris (Phoridae)

Minute elements detected in Megaselia scalaris (Phoridae, Diptera) lack chromosome arms but carry centromeres and possess kinetochore microtubules in mitosis as well as in meiosis. These centromere-like elements (CLEs) were present in two geographically independent strains of the fly. This indicates that their origin is not a recent event in the karyotype evolution of M. scalaris and that they are rather stable constituents of the karyotype. Most often, two CLEs were found in gonial and somatic mitosis. Spermatocytes contained one CLE. Two individuals examined deviated from this rule in that a metaphase spermatogonium showed three and an anaphase spermatogonium eight CLEs. These animals are believed to have been aneuploid relative to the CLEs. An analysis of spermatogonial division revealed that the CLEs behave like the centromeres of the regular chromosomes but seem to separate precociously, since they were closer to the spindle poles in late anaphase cells. Whereas the size of the CLEs was not significantly different between mitotic cells and secondary spermatocytes, the CLEs in primary spermatocytes were larger in volume by a factor of about 4.5 than those in mitosis and meiosis II. The additional material is interpreted as a glue that holds two CLEs together. This, in turn, is a prerequisite for orderly segregation. The function of the CLEs is not known. They are considered as B chromosomes reduced to the minimum required for segregation, the centromere.by J.B. RattnerAddress from April 1st 1991 until March 31st 1992: MRC, Human Genetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, Scotland     

Female reproductive system of the decapitating fly Pseudacteon wasmanni Schmitz (Diptera: Phoridae)

Pseudacteon wasmanni is a South American decapitating fly that parasitizes workers of Solenopsis fire ants. We used light microscopy (historesin serial-sectioning stained with Haematoxylin/Eosin) and scanning electron microscopy to show and analyze internal and whole external views of the female reproductive system. All specimens analyzed (n=9) by light microscopy showed post-vitellogenic oocytes inside the ovaries. The lack of typical follicles (oocyte-nurse cell complexes) in all specimens suggests that oogenesis occurs during the pupal stage. The total number of eggs found ranged from 31 to 280 (X=142±73, SD). The egg has a slugform or torpedo shape (about 130 by 20 μm) with a pointed apex at the posterior pole as defined by the fly; the micropyle appears to be in a depression or invagination at the anterior pole. An acute hypodermic-like ovipositor is evaginated from the hard sclerotized external genitalia during egg laying. The existence of a muscular bulb associated with the end of the common oviduct suggests that the egg is injected into the ant's body by a strong contraction of the bulb which probably is stimulated by bending of several ventral sensilla. During contraction, the abdomen extends out along a large fold between the sixth and seventh tergites in such a way that the sclerotized genitalia is rotated ventrally into a slightly anterior orientation in preparation for oviposition.