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。     

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.     

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 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.     

Karyotypes of four species of chironomids (Diptera, Chironomidae) from Northern Italy

Karyotypes of four chironomid species were studied: Cryptochironomus obreptans Walker, Cryptochironomus sp., Chironomus plumosus Linnaeus, and Stictochironomus rosenscholdi Zetterstedt. All these species belong to the subfamily Chironominae. Each species is characterized by a specific karyotype structure. The first species in the list has 2n = 4, while the three other species have 2n = 8.     

Internal reproductive organs of the female humpbacked fly,Megaselia scalaris Loew (Diptera : Phoridae)

The female reproductive system of the humpbacked fly Megaselia scalaris Loew (Diptera : Phoridae) was examined in whole mount preparations and serial sections. The system includes 2 ovaries, paired lateral oviducts, a common oviduct, and a genital chamber, opening externally through a gonopore, anteriad and ventrad to the anus. The ducts of the 2 accessory glands open independently into the dorsal region of the genital chamber. The terminal duct of a 2-armed spermatheca joins the right posterior and ventral wall of the genital chamber, immediately inside the gonopore. Passing dorally, the spermathecal duct lies immediately ventral to the duct of the right accessory gland. A short distance posteriad, it divides into two branches, each supplying an arm of the spermatheca. The genital chamber extends both anteriorly and posteriorly from its junction with the common oviduct, creating anterior and posterior compartments. In the right lateral wall of the genital chamber, a distinctive loop-shaped thickening (plate) resembles a darkened thread when it is observed through the integument. Features likely to have taxonomic utility include the posterior and ventral location of the terminal portion of the spermathecal duct; and the asymmetrically arranged, loop-shaped plate.     

Molecular differentiation of the homomorphic sex chromosomes inMegaselia scalaris (Diptera) detected by random DNA probes

Randomly cloned DNA fragments and a poly-(GATA) containing sequence were used as probes to identify sex chromosomal inheritance and to detect differences at the molecular level between the homomorphic X and Y in the phorid fly,Megaselia scalaris. Restriction fragment length differences between males and females and between two laboratory stocks of different geographic origin were used to differentiate between sex chromosomal and autosomal origin of the respective fragments. Five random probes detected X and Y chromosomal DNA loci and two others recognized autosomal DNA loci. One random probe and the poly(GATA) probe hybridized with both sex chromosomal and autosomal restriction fragments. Most of the Y chromosomal restriction fragments were conserved in length between the two stocks while most of the X chromosomal and autosomal fragments showed length polymorphism. It was concluded, therefore, that the Y chromosome contains a conserved segment in which crossover is suppressed and restriction site differences have accumulated relative to the X. These chromosomes, therefore, conform to a theoretically expected early stage of sex chromosome evolution.     

The complete mitochondrial genome of Nephrotoma scalaris parvinotata (Brunetti 1918) (Diptera: Tipulidae) and insights into the phylogeny of Tipulomorpha

Craneflies (Tipuloidea or Tipulidae sensu lato) are one of the most diverse groups of true flies (Insecta, Diptera). The Tipuloidea and perhaps the Trichoceridae formed the infraorder Tipulomorpha, which is traditionally considered the most basal group of Diptera. Relationships among Tipulomorpha and the phylogenetic position of this infraorder within the whole Diptera remain to be settled. A mitochondrial genome (mitogenome) phylogeny of Diptera was produced to test the relationships within Tipulomorpha and its phylogenetic position. A complete mitogenome of Nephrotoma scalaris parvinotata (Tipuloidea, Tipulidae) was firstly sequenced with a next-generation sequencing approach. Compared with the published mitogenomes of Tipuloidea, the new mitogenome had a larger genome size (17,862 nt), due to a longer non-coding control region. The gene order was identical to Drosophila yakuba. Phylogenetic reconstructions using different inference methods recovered Tipulomorpha as monophyletic. And the Tipulomorpha was retrieved in a relatively basal position in Diptera. Within Tipulomorpha, the Tipuloidea and the Trichoceridae were strongly supported as reciprocally monophyletic. Relationships within Tipuloidea were resolved as (Pedicidae + (Limoniidae + (Cylindrotomidae + Tipulidae))). Well supported relationships include: Pedicidae was the sister group of (Limoniidae + (Tipulidae + Cylindrotomidae)); Limoniidae was paraphyletic with respect to (Tipulidae + Cylindrotomidae); Cylindrotomidae was the sister group of Tipulidae. The newly sequenced N. scalaris parvinotata clustered with other two Nephrotoma species at a derived position in Tipuloidea.     

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.     

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.     

Conidium differentiation in Aspergillus nidulans wild-type and wet-white (wetA) mutant strains

Conidium (asexual spore) differentiation in wild-type and the wet-white (wetA) mutant of Aspergillus nidulans was compared in intact chains of successively older conidia. Carbohydrate cytochemistry helped define three stages (Stages I, II, and III) of wild-type conidium maturation on the basis of changes in the ultrastructure and composition of the conidium wall. Conidia of the wetA6 mutant strain formed normally but failed to mature during Stages II and III. Specifically, the inner wall layer of wetA6 conidia did not condense during Stage II and two wall layers that stained for carbohydrates did not form during the transition to Stage III. Concomitantly, wetA6 conidia formed large cytoplasmic vacuoles and underwent lysis. The wetA gene appears to have a conidium-specific function for the modification of the conidium wall during Stages II and III. These modifications of the conidium wall are essential for the stability of mature, dormant conidia.     

Karyotypes of Whytockia (Gesneriaceae)

The karyotypes of the genus Whytockia was investigated in order to reveal its sys tematic position in the tribe Klugieae and the evolution of karyotypes within this genus. The chromosome numbers are 2n＝18 with their length over 2.0 µm in all the investigated species of Whytockia, which represents the most primitive characteristics in the tribe Klugieae. This paper suggests that x＝9 be the primitive basic number of chromosomes in the Klugieae. The morphology of the resting nuclei in Whytockia belongs to the complex chromocenter type. The chromosomes at mitotic prophase are classified into the gradient type. The evolution of karyotypes in Whytockia is toward increase of the chromosomes with submedian centromeres and especially those with subterminal centromeres. The karyotype of the variety W. tsiangiana (Hand.-Mazz.)A. Weber var. wilsonii A. Weber is distinctly different from that of W. tsiangiana var. tsiangiana, which conforms well with the gross morphology. This paper suggests that this variety be separated from W. tsiangiana as an independent species with affinity to W. bijieensis.     

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.     

Quality of transgenic laboratory strains of Cochliomyia hominivorax (Diptera: Calliphoridae)

Genetically modified, mass reared insects present novel possibilities for the future of insect control. One concern about manipulation of insects is a possible loss of strain quality due to the introduction of a foreign gene of any sort into the insect genome. Eight transgenic strains of screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae), were compared with the wild-type parental laboratory strain in laboratory culture. Measurements of average fertility, fecundity, larval productivity, and longevity were analyzed. Two transgenic strains had significantly lower larval productivity than controls, one of which was explained by a homozygous lethal insertion of the transgene. Another strain produced significantly fewer eggs than controls. Overall strain characteristics, including measurements from egg, larva, pupa, and adult stages, were compared. Transgenic colonies did not consistently show significantly lower individual or aggregate strain quality characteristics than the control parental colony; hence, the presence of the transgene used to produce the strains tested did not incur a discrete cost to the colonies of laboratory-reared C. hominivorax.