Evaluation of the impact of Bacillus thuringiensis serovar israelensis and Temephos,used for the control of Simulium (Chirostilbia) pertinax Kollar, 1832 (Diptera,Simuliidae) on the associated entomofauna,Paraty, state of Rio de Janeiro,Brazil

The study was set up to evaluate the impact of two commercial larvicide formulations, Bacillus thuringiensis serovar israelensis base (Bti) at 15 ppm/1 min and temephos at 0.03 ppm of active ingredient, used to control Simulium pertinax populations, on associated non-target entomofauna occupying the same breeding sites. The experiments were carried out on the Pedra Branca and Muricana rivers, on the slopes of Serra do Mar massif, municipality of Paraty, state of Rio de Janeiro, Brazil. Bti was applied to the river Pedra Branca and temephosto theriver Muricana. On both rivers, treatment and control sections were labeled as such, each one with two observation posts: slow moving water and fast water regions respectively. Artificial substrata was used to evaluate the abundance of associated entomofauna. Attached immature stages of arthropods were removed from both of its surfaces fortnightly. Were collected, from the two rivers, 28 477 specimens of the entomofauna associated with S. pertinax. The families Hydropsychidae, Chironomidae, Bactidae, Simuliidae, Blephariceridae and Megapodagrionidae were represented. These was an impact of temephos on the entomofauna associated with S. pertinax only in Simuliidae and Chironomidae, and to Bti only in Simuliidae. However, the reduction in their numbers was not statistically significant.     

5S rDNA chromosomal mapping and COI sequence analysis reveal differentiation among distinct populations of a characid fish <Emphasis Type="Italic">Serrapinnus notomelas</Emphasis>

Cheirodontinae comprises a small-sized fish group with a wide geographical distribution throughout Central and South America, mainly in Brazilian Basins. The species Serrapinnus notomelas is widely distributed in the Upper Parana River Basin. In this work, chromosomal mapping of 5S ribosomal RNA (rRNA) gene was performed trough fluorescence in situ hybridization (FISH) in speciemens from three distinct localities belonging to two basins in the São Paulo State. All populations presented 5S clusters in two chromosome pairs. One additional pair was detected in samples from the Rio Paraitinguinha (Tietê Basin), and two additional pairs were detected in the population from Córrego Campo Novo (Tietê Basin). Analyses with partial sequences of COI were performed to verify the interelationship among the studied specimens, revealing Córrego Campo Novo population as a sister-group to the clade formed by the two other populations. The samples from Rio Paraitinguinha and Recanto dos Cambarás presented two distinct haplotypes each, while five haplotypes were observed in the Córrego Campo Novo population, sugesting that this could be older than the other populations. None of the nine haplotypes were shared among the three populations. The similarities and differences observed among the three populations using cytogenetic data and COI analysis are not related to geographic distances that separate the samples, suggesting that the origin of the Rio Paraitinguinha and Recanto dos Cambarás populations may be related to faunal exchanges. Additionally, the present data suggest that the analyzed populations of S. notomelas may be on independent evolutionary trajectories but in a very initial diversification stage. Moreover, the use of integrative information, such as molecular and chromosomal data in the analysis of population divergence and evolutionary trajectories in freshwater fishes is reinforced.     

Effects of Polydispyrenia simulii (Microspora; Duboscqiidae) on development of the gonads of Simulium pertinax (Diptera; Simuliidae).

The effects of the infection caused by the microsporidium Polydispyrenia simulii in gonads of the simuliid Simulium pertinax were evaluated. This blackfly species is the main target of control programmes in southeastern Brazil. Infected and noninfected blackflies were reared from larval stage to adults using a laboratory rearing system. There was no significant difference between the oogenic development and dimensions of ovarian follicles of healthy females compared to those reared from infected larvae. Otherwise, there was a significant difference between the fecundity of healthy females and those reared from infected larvae. In males, there was an evident reduction of motility of spermatozoids of males reared from infected larvae. The results obtained suggest that the microsporidium P. simulii have an important effect on the natural population dynamics of S. pertinax.     

A new black fly isolate of Bacillus thuringiensis autoagglutinating strain highly toxic to Simulium pertinax (Kollar) (Diptera, Simuliidae) larvae

Formulations containing the entomopathogenic Bacillus thuringiensis serovar israelensis strain IPS-82 has been widely applied for mosquito control around the world. Strain IPS-82 is highly active against Aedes aegypti but less active against other well-known vectors such as Culex quinquefasciatus and Simulium spp. larvae. Eighteen strains of B. thuringiensis were isolated from Simulium pertinax larvae naturally occurring in rivers of Southeast Brazil with one demonstrating special toxic effects. Simulated field tests against S. pertinax larvae showed that the native Brazilian autoagglutinating B. thuringiensis (LFB-FIOCRUZ 1035) has an LC50 at least 25 times lower than the standard IPS-82 strain. The same bacterial preparation was also tested against Ae. aegypti larvae in laboratory trials and the LC50 values obtained with LFB-FIOCRUZ 1035 were at least three times lower than the one for the IPS 82 strain. The results indicate that this strain is more toxic than the standard B. thuringiensis serovar israelensis (H14) in the two Dipteran species tested. It is noteworthy that differences between LC50 values were more pronounced in S. pertinax larvae, the source of the original isolation.     

The phylogeny of Simulium (Chirostilbia) (Diptera: Simuliidae) and perspectives on the systematics of the genus in the Neotropical Region

Recently, knowledge of Neotropical Simuliidae has been accumulating quickly. However, information about supra-specific relationships is scarce and diagnoses of Simulium subgenera are unsatisfactory. To investigate the relationships among Simulium (Chirostilbia) species and test the subgenus monophyly, we performed a cladistic analysis. The ingroup included all species of this subgenus and the outgroup included representatives of the 17 species groups of Neotropical Simulium and three Holarctic species. The study was based on a data matrix with 31 terminal taxa and 45 morphological characteristics of adult, pupa and larva. The phylogenetic analysis under equal weights resulted in eight most-parsimonious trees (length = 178, consistency index = 34, retention index = 67). The monophyly of the S. (Chirostilbia) was not supported in our analysis. The Simulium subpallidum species group was closer to Simulium (Psilopelmia) and Simulium (Ectemnaspis) than to the Simulium pertinax species group. Additionally, we describe the three-dimensional shape of the terminalia of male and female of Simulium (Chirostilbia) for the first time and provide comments about the taxonomic problems involving some species of the subgenus: Simulium acarayense, Simulium papaveroi, S. pertinax, Simulium serranum, Simulium striginotum and S. subpallidum.     

Chromosomal evidence of downstream dispersal of Astyanax fasciatus (Characiformes, Characidae) associated with river shed interconnection

In the 1960s, as a part of the construction of the Furnas Hydroelectric Power Dam, Minas Gerais, Brazil, the Rio Piumhi was diverted from the Rio Grande drainage to the S?o Francisco River basin, with the resulting introduction of species from one basin to the other. Chromosomal characteristics of various populations of Astyanax fasciatus sensu Eigenmann from the transect region were studied using cytogenetic techniques, with the goal to identify and map the dispersal of invasive species in the Rio S?o Francisco. Populations of the Rio Grande and Rio Piumhi are distinct from those of the S?o Francisco basin based on chromosome markers associated to the heterochromatin, Ag-NORs, 18S rDNA, 5S rDNA, and As-51 satellite DNA sites. Individuals with karyomorph originally present in the Piumhi drainage are now present in the S?o Francisco basin, downstream from the transposition channel. This expansion of the distribution corroborates a hypothesis of progressive substitution of native populations by the invasive Piumhi form.     

Nucleolar Organizing Regions, 18S and 5S rDNA in Astyanax Scabripinnis (Pisces, Characidae): Populations Distribution and Functional Diversity

Astyanax scabripinnis specimens from four distinct populations in Brazil were studied with respect to their karyotype macrostructure, nucleolar organizer regions, and 18S and 5S rRNA genes. The four populations showed a 2n = 50 chromosomes (3 M + 11 SM + 5 ST + 6 A pairs) and 1–2 B chromosomes. No chromosomal differentiations were observed between sexes. Although a karyotypic diversity has been characterized in this fish group, the populations now analyzed presented the same macrokaryotypic pattern. Chromosome mapping of 5S rDNA showed a total of eight sites located in four distinct chromosomal pairs, with no apparent differences among populations. A comparative study on 18S rDNA locations and Ag-NORs showed some secondary NOR sites that are not usually expressed in karyotypes and a probable differential NOR activity among populations. Correlations between these data, environmental conditions and B chromosomes are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Phylogenetic relationships and phylogeography of the Killifish species of the subgenus Chromaphyosemion (Radda, 1971) in West Africa, inferred from mitochondrial DNA sequences

We have analyzed the phylogenetic relationships of 160 specimens from 88 samples representing all defined species of the African Aplocheiloid subgenus Chromaphyosemion in order to examine the monophyly of this group, the species interrelationships, and to reveal trends in chromosomal evolution and formulate hypotheses about their evolutionary history. The data set comprised 1153 total nucleotides from the mitochondrial 12S rRNA, cytochrome oxidase I, and D-loop. The molecular-based topologies were analyzed by maximum parsimony, maximum likelihood, distance method and Bayesian inference support the monophyly of the subgenus Chromaphyosemion. All populations with ambiguous taxonomic status were assigned to an already described species except A. sp. Rio Muni which corresponds to a still undescribed species. Aphyosemion alpha and A. lugens were in basal position in the different trees that indicate a possible origin of the subgenus Chromaphyosemion in the South Cameroon-North Gabon region. Furthermore, the South Cameroon region (between 2 degrees and 3 degrees of North latitude) that accommodates half of the Chromaphyosemion species is considered to have been a refuge zone during the late quaternary dry events that Africa experienced. Phylogenetic relationships among the subgenus also revealed that chromosomal evolution is complex and should be studied at the intraspecific level.     

Relationships among native and introduced populations of the Argentine ant (Linepithema humile) and the source of introduced populations

The Argentine ant (Linepithema humile) is a damaging invasive species that has become established in many Mediterranean-type ecosystems worldwide. To identify likely sources of introduced populations we examined the relationships among native Linepithema populations from Argentina and Brazil and introduced populations of L. humile using mitochondrial cytochrome b sequence data and nuclear microsatellite allele frequencies. The mitochondrial phylogeny revealed that the populations in Brazil were only distantly related to both the introduced populations and the native populations in Argentina, and confirmed that populations in Brazil, previously identified as L. humile, are likely a different species. The microsatellite-based analysis provided resolution among native and introduced populations of L. humile that could not be resolved using the mitochondrial sequences. In the native range, colonies that were geographically close to one another tended to be genetically similar, whereas more distant colonies were genetically different. Most samples from the introduced range were genetically similar, although some exceptions were noted. Most introduced populations were similar to native populations from the southern Rio Parana and were particularly similar to a population from Rosario, Argentina. These findings implicate populations from the southern Rio Parana as the most likely source of introduced populations. Moreover, these data suggest that current efforts to identify natural enemies of the Argentine ant for biological control should focus on native populations in the southern Rio Parana watershed.     

Low courtship song variation in south and southeastern Brazilian populations of Drosophila meridionalis (Diptera, Drosophilidae)

Drosophila meridionalis is a cactus-breeding species with a wide distribution in South America. Most populations of this species are geographically isolated, what provides a promising scenario for studying evolution. Former studies of this species revealed a remarkable karyotypic variation among its populations. Up to six distinct metaphase chromosomes were described, showing that this species is polymorphic at least at the chromosomal level. In order to elucidate the taxonomic status of populations showing different metaphase chromosomes, we analyzed the courtship song of five populations of D. meridionalis in South and Southeastern Brazil. In addition, we analyzed the metaphase chromosomes of each population. Our results show that, despite the two karyotype observed, most courtship song parameters did not vary among the populations. Altogether, our results suggest that D. meridionalis from South and Southeastern Brazil represents one species with an inter-population chromosomal variability.     

Isoenzimatic analysis of four Anopheles (Kerteszia) cruzii (Diptera: Culicidae) populations of Brazil

Anopheles cruzii is a small sylvatic mosquito and primary human Plasmodium vector in Southern Brazil. The distribution of this bromeliad-breeding mosquito follows the Atlantic forest coastal distribution, where bromeliads are abundant. Morphological, genetic, and molecular polymorphisms among different populations have been reported and it has recently been suggested that An. cruzii is a complex of cryptic species. The aim of this work is to analyze the gene flow between different populations of An. cruzii collected in four localities within the geographic distribution range of the species, and to examine if An. cruzii is a complex of cryptic species. The genetic distances show that populations of the states of Santa Catarina, Sao Paulo, and Rio de Janeiro are genetically closer (0.032 to 0.083) than populations of Bahia (0.364 to 0.853) based on profiles from 10 distinct isoenzyme loci. The Fst was lower (0.077) when the Bahia population was excluded than when it was included (0.300) in the analyses. The inferred number of migrants per generation was 2.99 individuals among populations from the states of Santa Catarina, Sao Paulo, and Rio de Janeiro and 0.58 migrants per generation among all populations. Results suggest that An. cruzii is a complex of species and that the specimens of state of Bahia can be considered as belonging to a species that is distinct from other three closely-related populations studied.     

GEOGRAPHIC GENETIC STRUCTURE IN THE FRANCISCANA DOLPHIN (PONTOPORIA BLAINVILLEI)

The franciscana, Pontoporia blainvillei , is endemic to the South Atlantic coast of South America. The species is of conservation concern because it suffers elevated mortality rates due to incidental captures in fishing nets, and perhaps it is one of the most threatened small cetaceans in this region. Previous morphological and genetic studies have suggested the existence of at least two distinct stocks to the north and south of Santa Catarina Island in Brazil. Fixed differences were found between a sample from Rio de Janeiro and one from Rio Grande do Sul, in southern Brazil. Using 94 sequences of the control region of mitochondrial DNA, we examined the genetic structure of the species. We found no shared haplotypes between Rio de Janeiro and samples from the southern range of the species distribution. However, a phylogenetic analysis suggests that the former population is diphyletic with respect to the southern samples. This suggests that the populations have not been isolated long enough to reach reciprocal monophyly. Furthermore, genetic differentiation is broadly consistent with a simple model of isolation by distance, that therefore appears as an alternative to a model of strict isolation of two stocks. The estimated levels of gene flow are higher among neighboring populations, and decrease as more distant localities are compared. Finally, the molecular data suggest that franciscanas have expanded in Rio de Janeiro.     

Conserved 5S and variable 45S rDNA chromosomal localisation revealed by FISH in <Emphasis Type="Italic">Astyanax scabripinnis</Emphasis> (Pisces,Characidae)

Fluorescence in situhybridisation (FISH) and double FISH experiments were carried out to ascertain the chromosomal distribution pattern of the 45S and 5S ribosomal (r) DNAs in four populations of the characid fish Astyanax scabripinnis – a group considered to be a species complex for its wide karyotypical and morphological diversity. The results regarding the 45S rDNA agreed with this hypothesis, since these sites showed intra- and inter-populational, numerical and positional variations. However, the data obtained with the 5S rDNA probe revealed a highly conserved chromosomal distribution pattern of these sequences among individuals of each population, as well as among the populations analysed. We consider this contrasting situation as a functional divergence between 45S and 5S ribosomal DNAs, which may reflect the localisation of these sequences in distinct nuclear compartments, leading them to undergo differentiated evolutionary processes.     

Variations of 18S rDNA Loci Among Six Populations of Paeonia obovata Maxim. （Paeoniaceae） Revealed by Fluorescence In Situ Hybridization

The localization of 18S ribosomal RNA genes （rDNA） by fluorescence in situ hybridization （FISH） had been performed for some species of Paeonla. However, the pattern of 18S rDNA loci among populations Is Indistinct. In the present study, we localized 18S rDNA loci on meiotic or mitotic chromosomes of six populations of Paeonla obovata Maxim. （Paeonlaceae）. Different numbers of rDNA loci were found with different diploid （2n=10） populations, namely eight （Lushl and Mt. JIuhua populations）, 10 （Mt. Talbal population）, and seven （Mt. Guandl population）, whereas tetraplold （2n=20） populations were all found with 16 loci. Aii rDNA loci were mapped near teiomeres of mitotic chromosomes and there was no chromosome with two loci. The present results show that molecular cytological polymorphlsm exists among P. obovata diploid populations, Indicating that structural variations occurred frequently during the evolutionary history of this species, accompanied with differentiation among populations.     

Faunistic analysis of fruit flies (Diptera: Tephritidae) in the Northern and Northwestern Regions of Rio de Janeiro State, Brazil

This study aimed to characterize the fruit fly populations in three municipalities of the Northern region and two municipalities of the Northwestern region of Rio de Janeiro State, Brazil, and to evaluate the similarity among their populations. A faunistic analysis was performed from the fruit fly specimens captured in plastic McPhail traps with an aqueous solution of hydrolyzed protein to 5% placed in orchards of guava (Psidium guajava L.) and/or other fruits during 26 months. The total of 3,952 females of 15 species of Anastrepha Schiner and 277 females of Ceratitis capitata (Wied.) was captured. The species richness differed among the municipalities, with the highest value in S?o Francisco do Itabapoana (S = 14), resulting on the highest Shannon-Wiener index (H = 1.27). The equitability was low in the five municipalities due to the dominance of one unique fruit fly species. The predominant species (more frequent, constant and dominant) were Anastrepha obliqua (Macquart) in Campos dos Goytacazes and S?o Francisco do Itabapoana, Anastrepha fraterculus (Wied.) in Cambuci and Itaocara, and Anastrepha sororcula Zucchi in S?o Jo?o da Barra. Fruit fly populations had low diversity index of Margalef (alpha = 0.58 to 1.82). Regarding to fruit fly species composition, the populations in S?o Jo?o da Barra and Cambuci were more similar between each other, composing a distinct group from the populations in Campos dos Goytacazes and Itaocara. These two groups differed quite a lot from the population in S?o Francisco do Itabapoana.     

Wide but not impermeable: Testing the riverine barrier hypothesis for an Amazonian plant species

Wallace's riverine barrier hypothesis postulates that large rivers, such as the Amazon and its tributaries, reduce or prevent gene flow between populations on opposite banks, leading to allopatry and areas of species endemism occupying interfluvial regions. Several studies have shown that two major tributaries, Rio Branco and Rio Negro, are important barriers to gene flow for birds, amphibians and primates. No botanical studies have considered the potential role of the Rio Branco as a barrier, while a single botanical study has evaluated the Rio Negro as a barrier. We studied an Amazon shrub, Amphirrhox longifolia (A. St.‐Hil.) Spreng (Violaceae), as a model to test the riverine barrier hypothesis. Twenty‐six populations of A. longifolia were sampled on both banks of the Rio Branco and Rio Negro in the core Amazon Basin. Double‐digest RADseq was used to identify 8,010 unlinked SNP markers from the nuclear genome of 156 individuals. Data relating to population structure support the hypothesis that the Rio Negro acted as a significant genetic barrier for A. longifolia. On the other hand, no genetic differentiation was detected among populations spanning the narrower Rio Branco, which is a tributary of the Rio Negro. This study shows that the strength of riverine barriers for Amazon plants is dependent on the width of the river separating populations and species‐specific dispersal traits. Future studies of plants with contrasting life history traits will further improve our understanding of the landscape genetics and allopatric speciation history of Amazon plant diversity.     

Comparison of imaginal diets of various Syrphidae (Diptera) species

The full amount and species composition of pollen from intestines of 11 species of Syrphidae (Arctophila fulva, Eristalis arbustorum, E. nemorum, E. pertinax, Helophilus pendulus, Myiatropa florea, Rhingia campestris, Sericomyia silentis, Syrphus baltheatus, S. ribessii and S. vitripennis), collected at the some place (Torma, Jogeva distr., Estonia) and at the some time (18-20 July 1989) have been studied. Maximum number of pollen grains is different fly species varied from 67,800 (Rh. campestris) up to 240,700 (S. silentis) grains, and average number from 25,560 (Rh. campestris) up to 115,880 (E. pertinax) grains. Maximum volume of pollen in different fly species varied from 1.5 (S. ribesii) up to 23.6 (S. baltheatus) mm3, and average volume from 0.36 up to 7.0 mm3 (the same species). The difference in a imaginal diets of Syrphidae are found, and the degree of difference does not correlate with a degree taxonomic affinity of species. The difference in strategy of a feeding behavior of two Syrphus species, that have similar diet, are marked: in intestines of 80% specimens of S. ribesii we found pollen grains of less that 7 plant species, whereas intestines of more than 88% specimens of S. vitripennis contained more than 8 species of grains. Distinction in alimentary preferences of different species of files can not be explained neither particularities of their morphology, nor their color preferences.     

Speciation and chromosomal evolution of the Baikalian endemic chironomids of the genus Sergentia Kief. (Diptera, Chironomidae): karyotype divergence and chromosomal polymorphism in the populations of eurybathic species Sergentia flavodentata Tshern. and littoral species Sergentia baicalensis Tshern

Specific karyotype structure and chromosomal polymorphism was investigated in the populations of the Baikalian endemic chironomid species, eurybathic Sergentia flavodentata Tshernovskij, 1949 and littoral Sergentia baicalensis Tshernovskij, 1949. In addition to two fluctuating homozygous inversions in arms A and E, both species were characterized by species-specific karyotype features, namely, nucleolar organizer in the region 1 of chromosome IV in S. flavodentata, and inversion in arm F in S. baicalensis. Moreover, S. baicalensis demonstrated the presence of intraspecific population polymorphism. The populations differing in the presence of secondary overlapping inversion in arm A were found. The highest number of chromosomal rearrangements (7) was detected in S. flavodentata. Most of these rearrangements (six) were found in the population from the underwater thermal spring at a depth of 420 to 430 m (Northern Baikal, Frolikha Bay). In the populations from Middle and Southern Baikal, a rare pericentric inversion in chromosome I was described. In S. baicalensis, in addition to two common heterozygous inversions in arms C and F, disturbance of homologous pairing in different regions of the remaining arms were often detected. Stable chromosomal polymorphism preserved during 13 years in the population of S. flavodentata from the region of hydrothermal venting, serves as an evidence of high genetic plasticity of the species, which favors successful colonization of different Baikal depths and biotopes.     

Karyotypic and molecular identification of laboratory stocks of the South American fruit fly Anastrepha fraterculus (Wied) (Diptera: Tephritidae)

The taxonomic status of the tephritid pest Anastrepha fraterculus (Wied.) is a controversial subject, mainly because of misinterpretation of the observed genetic variation. In this work, the different karyotypes and DNA polymorphism of a geographically defined population from Northeastern Argentina were studied, using derived stocks maintained in the laboratory during 25 generations. The karyotypes were analyzed using C-banding and N-banding techniques, while DNA analysis was performed through the DNA polymerase chain reaction (PCR). The variants isolated from both the wild Montecarlo population and the derived laboratory stocks were fully compatible and are present in other wild populations from South Brazil (lat 31 degrees 30' S) to Mid-Argentina (lat 34 degrees 30' S). Single-pair crosses among stocks carrying different chromosomal variants demonstrated the absence of isolation barriers. The polymorphic fragments isolated by RAPDs/PCR showed polymorphisms among stocks whereas the analysis of rDNA ITS1 exhibit a unique ITS1 length. Our results seem to indicate that all the examined variants belong to a single species with extended polymorphism and therefore do not support the hypothesis that the extended chromosomal polymorphism in A. fraterculus implies the existence of a complex of cryptic species.     

Terena Amerindian group autosomal STR data: comparison studies with other Brazilian populations

Allele frequencies for the high polymorphic short tandem repeats (STR) loci PentaE, PentaD, D18S51, D21S11, TH01, D3S1358, FGA, D16S539, D7820, D13S317, vWA and D81179 were analysed in an native Amerindian population from Mato Grosso do Sul state named Terena. Deviations from Hardy–Weinberg expectations were evaluated and the results showed no differences from equilibrium in all loci. The combined power of discrimination and the combined power of exclusion for the 12 tested STR loci were 0.99999999 and 0.999999 respectively. The Terena population data were compared to other from 11 Brazilian populations (Amazônia, Pernambuco, Mato Grosso do Sul, São Paulo, Rio Grande do Sul, Alagoas, Sergipe, Rio Grande do Norte, Santa Catarina, Rondônia and Rio de Janeiro) representing the major Brazilian geographic regions. The F_ST comparative analysis showed no significant differences between all those populations except when comparing Terena with the remained ones.