Cytogenetic Features of Human Head and Body Lice (Phthiraptera: Pediculidae).

The genus Pediculus L. that parasitize humans comprise two subspecies: the head lice Pediculus humanus capitis De Geer and the body lice Pediculus humanus humanus De Geer. Despite the 200 yr of the first description of these two species, there is still a long debate about their taxonomic status. Some authors proposed that these organisms are separate species, conspecifics, or grouped in clades. The sequencing of both forms indicated that the difference between them is one gene absent in the head louse. However, their chromosomal number remains to be determined. In this study, we described the male and female karyotypes, and male meiosis of head and body lice, and examined the chromatin structure by means of C-banding. In P. h. humanus and P. h. capitis, the diploid chromosome complement was 2 n = 12 in both sexes. In oogonial prometaphase and metaphase and spermatogonial metaphase, it is evident that chromosomes lack of a primary constriction. No identifiable sex chromosomes or B chromosomes were observed in head and body lice. Neither chiasmata nor chromatin connections between homologous chromosomes were detected in male meiosis. The meiotic behaviour of the chromosomes showed that they are holokinetic. C-banding revealed the absence of constitutive heterochromatin. Our results provide relevant information to be used in mapping studies of genes associated with sex determination and environmental sensing and response.

Complementary sex determination in the parasitic wasp Diachasmimorpha longicaudata.

We studied the sex determination in Diachasmimorpha longicaudata, a parasitoid braconid wasp widely used as biological control agent of fruit pest tephritid flies. We tested the complementary sex determination hypothesis (CSD) known in at least 60 species of Hymenoptera. According to CSD, male or female development depends on the allelic composition of one sex locus (single-locus CSD) or multiple sex loci (multiple-locus CSD). Hemizygote individuals are normal haploid males, and heterozygotes for at least one sex locus are normal diploid females, but homozygotes for all the sex loci are diploid males. In order to force the occurrence of diploid males in D. longicaudata, we established highly inbred lines and examined their offspring using chromosome counting, flow cytometry, and sex ratio analysis. We found that when mother-son crosses were studied, this wasp produced about 20% of diploid males out of the total male progeny. Our results suggest that this parasitoid may represent the second genus with multiple-locus CSD in Hymenoptera. Knowledge about the sex determination system in D. longicaudata is relevant for the improvement of mass rearing protocols of this species. This information also provides the necessary background for further investigations on the underlying molecular mechanisms of sex determination in this species, and a better insight into the evolution of this pathway in Hymenoptera in particular and insects in general.

The significance of cytogenetics for the study of karyotype evolution and taxonomy of water bugs (Heteroptera, Belostomatidae) native to Argentina.

Male meiosis behaviour and heterochromatin characterization of three big water bug species were studied. Belostoma dentatum (Mayr, 1863), Belostoma elongatum Montandon, 1908 and Belostoma gestroi Montandon, 1903 possess 2n = 26 + X1X2Y (male). In these species, male meiosis is similar to that previously observed in Belostoma Latreille, 1807. In general, autosomal bivalents show a single chiasma terminally located and divide reductionally at anaphase I. On the other hand, sex chromosomes are achiasmatic, behave as univalents and segregate their chromatids equationally at anaphase I. The analysis of heterochromatin distribution and composition revealed a C-positive block at the terminal region of all autosomes in Belostoma dentatum, a C-positive block at the terminal region and C-positive interstitial dots on all autosomes in Belostoma elongatum, and a little C-positive band at the terminal region of autosomes in Belostoma gestroi. A C-positive band on one bivalent was DAPI negative/CMA3 positive in the three species. The CMA3-bright band, enriched in GC base pairs, was coincident with a NOR detected by FISH. The results obtained support the hypothesis that all species of Belostoma with multiple sex chromosome systems preserve NORs in autosomal bivalents. The karyotype analyses allow the cytogenetic characterization and identification of these species belonging to a difficult taxonomic group. Besides, the cytogenetic characterization will be useful in discussions about evolutionary trends of the genome organization and karyotype evolution in this genus.

Male meiosis, heterochromatin characterization and chromosomal location of rDNA in Microtomus lunifer (Berg, 1900) (Hemiptera: Reduviidae: Hammacerinae).

In the present work, we analysed the male meiosis, the content and distribution of heterochromatin and the number and location of nucleolus organizing regions in Microtomus lunifer (Berg, 1900) by means of standard technique, C- and fluorescent bandings, and fluorescent in situ hybridization with an 18S rDNA probe. This species is the second one cytogenetically analysed within the Hammacerinae. Its male diploid chromosome number is 31 (2n=28+X1X2Y), including a minute pair of m-chromosomes. The diploid autosomal number and the presence of m-chromosomes are similar to those reported in Microtomus conspicillaris (Drury, 1782) (2n=28+XY). However, Microtomus lunifer has a multiple sex chromosome system X1X2Y (male) that could have originated by fragmentation of the ancestral X chromosome. Taking into account that Microtomus conspicillaris and Microtomus lunifer are the only two species within Reduviidae that possess m-chromosomes, the presence of this pair could be a synapomorphy for the species of this genus. C- and fluorescent bandings showed that the amount of heterochromatin in Microtomus lunifer was small, and only a small CMA3 bright band was observed in the largest autosomal pair at one terminal region. FISH with the 18S rDNA probe demonstrated that ribosomal genes were terminally placed on the largest autosomal pair. Our present results led us to propose that the location of rDNA genes could be associated with variants of the sex chromosome systems in relation with a kind of the sex chromosome systems within this family. Furthermore, the terminal location of NOR in the largest autosomal pair allowed us to use it as a chromosome marker and, thus, to infer that the kinetic activity of both ends is not a random process, and there is an inversion of this activity.

Immature stages of development in the parasitoid wasp, Diachasmimorpha longicaudata.

The morphological changes experienced during the immature stages of the solitary parasitoid Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae: Opiinae) were studied. This natural enemy of several species of tephritid fruit flies is widely used in biological control strategies. Immature stages are poorly understood in endoparasitoids because they exist within the host. In the present work, developmental processes are described for this species, reared in Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) larvae under controlled environmental conditions. At 25 degrees C, 85% RH, and with an 18:6 L:D photoperiod, preimaginal development takes about 16 days. Five preimaginal stages can be described: egg, three larval instars, prepupa, pupa, and pharate adult. Superparasitism was found in 20% of the host pupae, and the number of oviposition scars was positively correlated with the number of parasitoid larvae per host puparium. The results are compared and discussed with previous studies on related species.

Cytogenetic analysis of three species of Pseudacteon (Diptera, Phoridae) parasitoids of the fire ants using standard and molecular techniques.

Pseudacteon flies, parasitoids of worker ants, are being intensively studied as potentially effective agents in the biological control of the invasive pest fire ant genus Solenopsis (Hymenoptera: Formicidae). This is the first attempt to describe the karyotype of P. curvatus Borgmeier, P. nocens Borgmeier and P. tricuspis Borgmeier. The three species possess 2n = 6; chromosomes I and II were metacentric in the three species, but chromosome pair III was subtelocentric in P. curvatus and P. tricuspis, and telocentric in P. nocens. All three species possess a C positive band in chromosome II, lack C positive heterochromatin on chromosome I, and are mostly differentiated with respect to chromosome III. P. curvatus and P. tricuspis possess a C positive band, but at different locations, whereas this band is absent in P. nocens. Heterochromatic bands are neither AT nor GC rich as revealed by fluorescent banding. In situ hybridization with an 18S rDNA probe revealed a signal on chromosome II in a similar location to the C positive band in the three species. The apparent lack of morphologically distinct sex chromosomes is consistent with proposals of environmental sex determination in the genus. Small differences detected in chromosome length and morphology suggests that chromosomes have been highly conserved during the evolutionary radiation of Pseudacteon. Possible mechanisms of karyotype evolution in the three species are suggested.

Cytogenetic analysis of three species of Pseudacteon (Diptera, Phoridae) parasitoids of the fire ants using standard and molecular techniques

Pseudacteon flies, parasitoids of worker ants, are being intensively studied as potentially effective agents in the biological control of the invasive pest fire ant genus Solenopsis (Hymenoptera: Formicidae). This is the first attempt to describe the karyotype of P. curvatus Borgmeier, P. nocens Borgmeier and P. tricuspis Borgmeier. The three species possess 2n = 6; chromosomes I and II were metacentric in the three species, but chromosome pair III was subtelocentric in P. curvatus and P. tricuspis, and telocentric in P. nocens. All three species possess a C positive band in chromosome II, lack C positive heterochromatin on chromosome I, and are mostly differentiated with respect to chromosome III. P. curvatus and P. tricuspis possess a C positive band, but at different locations, whereas this band is absent in P. nocens. Heterochromatic bands are neither AT nor GC rich as revealed by fluorescent banding. In situ hybridization with an 18S rDNA probe revealed a signal on chromosome II in a similar location to the C positive band in the three species. The apparent lack of morphologically distinct sex chromosomes is consistent with proposals of environmental sex determination in the genus. Small differences detected in chromosome length and morphology suggests that chromosomes have been highly conserved during the evolutionary radiation of Pseudacteon. Possible mechanisms of karyotype evolution in the three species are suggested.

Synapsis with and without recombination in the male meiosis of the leaf-footed bug Holhymenia rubiginosa (Coreidae, Heteroptera).

In organisms with chiasmatic meiosis two different relationships have been described between crossing over and synapsis: in one group of organisms synapsis depends on the initiation of meiotic recombination while in the other group it is independent of this initiation. These patterns have been observed mainly in organisms where all meiotic bivalents in the set have similar behaviors. In some heteropteran insects a pair of chromosomes named m chromosomes is known to behave differently from autosomes regarding synapsis and recombination. Here we used immunodetection of a synaptonemal complex component and acid-fixed squashes to investigate the conduct of the small m chromosome pair during the male meiosis in the coreid bug Holhymenia rubiginosa. We found that the m chromosomes form a synaptonemal complex during pachytene, but they are not attached by a chiasma in diakinesis. On the other hand, the autosomal bivalents synapse and recombine regularly. The co-existence of these variant chromosome behaviors during meiosis I add further evidence to the absence of unique patterns regarding the interdependence of synapsis and recombination.

Heterochromatin characterization in five species of Heteroptera.

The amount, composition and location of heterochromatin in Athaumastus haematicus (Stål, 1859), Leptoglossus impictus (Stål, 1859), Phthia picta (Drury, 1770) (Coreidae), Largus rufipennis Laporte, 1832 (Largidae) and Jadera sanguinolenta (Fabricius, 1775) (Rhopalidae) are analyzed by C-banding and DAPI/ CMA fluorescent banding. As the rule for Heteroptera the possession of holokinetic chromosomes and a pre-reductional type of meiosis cytogenetically characterize these five species. Besides, all of them (except L. rufipennis) present a pair of m chromosomes. C-banding technique reveals the absence of constitutive heterochromatin in A. haematicus, scarce C-positive blocks in L. impictus and J. sanguinolenta, and C-positive heterochromatin terminally located in P. picta and L. rufipennis. All C-bands are DAPI bright, except for a DAPI dull/CMA bright band at one telomeric end of the X chromosome in L. rufipennis, which probably corresponds to a nucleolar organizing region. The results of the banding techniques are analyzed in relation to the chiasma frequency and distribution in the five species, and it is concluded that there should exist some constraints to the acquisition and/ or accumulation of heterochromatin in their karyotypes.

Cytogenetic and nucleolar meiotic cycle analyses in Dysdercus imitator Blöte, 1931 (Pyrrhocoridae, Heteroptera) from Argentina.

So far, only seven and five species of Dysdercus from the Old and New Worlds, respectively, have been cytogenetically analysed. They all have holokinetic chromosomes and a pre-reductional type of meiosis. In the present study the chromosome complement, male meiosis and nucleolar meiotic cycle of Dysdercus imitator were analyzed. During male meiosis several cytogenetic features are remarkable, namely the presence of a long diffuse stage after pachytene, the finding of one or two ring bivalents per cell in almost all specimens, and the presence of several prenucleolar bodies lasting up to telophase II. The origin and function of these prenucleolar bodies could be related to a particular physiological cycle of the meiocytes.