Taxonomy and cladistic analysis of the subgenus Xenomorellia Malloch (Diptera: Muscidae: Morellia Robineau-Desvoidy) with description of two new species

Xenomorellia Malloch, a subgenus of Morellia Robineau-Desvoidy, is revised to include two new species, Morellia (Xenomorellia) inca Nihei and Carvalho sp. nov. from South America, and M. (X.) maia Carvalho and Nihei sp. nov. from Costa Rica and Mexico. Diagnoses for M. (X.) holti (Malloch) and M. (X.) montanhesa (Albuquerque) are provided, as well as an identification key to the four species of the subgenus. A cladistic analysis was performed to test the monophyly of Xenomorellia and to recover the phylogenetic relationships among its species. Tree searches resulted in one single most-parsimonious cladogram, wherein the monophyly of Xenomorellia is supported, as well as a sister-group relationship with the Neotropical subgenus Trichomorellia Stein. Xenomorellia was divided into two clades: one with Caribbean-Andean species (maia + inca), and another with species from southeastern South America (holti + montanhesa).

The Muscini flies of the world (Diptera, Muscidae): identification key and generic diagnoses

Flies of the tribe Muscini (Diptera, Muscidae) are worldwide in distribution and are represented by some 350 species in 18 genera. The present study provides an identification key and diagnoses for all the genera of world Muscini: Biopyrellia Townsend, Curranosia Paterson, Dasyphora Robineau-Desvoidy, Deltotus Seguy, Hennigmyia Peris, Mesembrina Meigen, Mitroplatia Enderlein, Morellia Robineau-Desvoidy, Musca Linnaeus, Myiophaea Enderlein, Neomyia Walker, Neorypellia Pont, Polietes Rondani, Polietina Schnabl & Dziedzicki, Pyrellia Robineau-Desvoidy, Pyrellina Malloch, Sarcopromusca Townsend, Ziminellia Nihei & de Carvalho. Most infrageneric taxa are also represented, namely, the sub-genera of Dasyphora and Morellia. Comments on phylogeny support (whenever pertinent) and the major references containing revisions and regional identification keys to species are provided for each genus and subgenus.

Immunocytochemical identification of adenohypophyseal cells in the pirarucu (Arapaima gigas), an Amazonian basal teleost

The adenohypophysis (AH) of juvenile pirarucu (Arapaima gigas), a representative species of the Osteoglossomorpha (bonytongue fishes, one of the oldest living groups of the teleosts), was studied using histochemical and immunocytochemical methods. The AH is comprised of the pars distalis (PD), without a clear distinction between rostral pars distalis (RPD) and proximal pars distalis (PPD), and the pars intermedia (PI). The neurohypophysis (NH) is positioned on top of the PD and penetrates and branches into the PI. In the most rostral dorsal portion of the PD, adrenocorticotropic cells and fusiform gonadotropic cells were found. In the central PD, scarce prolactin-producing cells and growth-hormone-producing cells were located mainly in the dorsal part, whereas round gonadotropic cells were abundant in the ventral portion of this region. Human thyrotropin immunoreactive cells were not found in the entire AH. In the PI, melanotropic, some adrenocorticotropic, and somatolactin-producing cells were located intermingled surrounding the neurohypophyseal branches. Our results showed that the A. gigas pituitary has some basal characteristics between the ancient Actinopterygii and the more derived teleosts.

Identification and lineage genotyping of South American trypanosomes using fluorescent fragment length barcoding

Trypanosoma cruzi and Trypanosoma rangeli are human-infective blood parasites, largely restricted to Central and South America. They also infect a wide range of wild and domestic mammals and are transmitted by a numerous species of triatomine bugs. There are significant overlaps in the host and geographical ranges of both species. The two species consist of a number of distinct phylogenetic lineages. A range of PCR-based techniques have been developed to differentiate between these species and to assign their isolates into lineages. However, the existence of at least six and five lineages within T. cruzi and T. rangeli, respectively, makes identification of the full range of isolates difficult and time consuming. Here we have applied fluorescent fragment length barcoding (FFLB) to the problem of identifying and genotyping T. cruzi, T. rangeli and other South American trypanosomes. This technique discriminates species on the basis of length polymorphism of regions of the rDNA locus. FFLB was able to differentiate many trypanosome species known from South American mammals: T. cruzi cruzi. T. cruzi marinkellei, T. dionisii-like, T. evansi, T. lewisi, T. rangeli, T. theileri and T. vivax. Furthermore, all five T. rangeli lineages and many T. cruzi lineages could be identified, except the hybrid lineages TcV and TcVI that could not be distinguished from lineages III and II respectively. This method also allowed identification of mixed infections of T. cruzi and T. rangeli lineages in naturally infected triatomine bugs. The ability of FFLB to genotype multiple lineages of T. cruzi and T. rangeli together with other trypanosome species, using the same primer sets is an advantage over other currently available techniques. Overall, these results demonstrate that FFLB is a useful method for species diagnosis, genotyping and understanding the epidemiology of American trypanosomes. (C) 2010 Elsevier B.V. All rights reserved.

Phylogenetic Validation of the Genera Angomonas and Strigomonas of Trypanosomatids Harboring Bacterial Endosymbionts with the Description of New Species of Trypanosomatids and of Proteobacterial Symbionts

We comparatively examined the nutritional, molecular and optical and electron microscopical characteristics of reference species and new isolates of trypanosomatids harboring bacterial endosymbionts. Sequencing of the V7V8 region of the small subunit of the ribosomal RNA (SSU rRNA) gene distinguished six major genotypes among the 13 isolates examined. The entire sequences of the SSU rRNA and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) genes were obtained for phylogenetic analyses. In the resulting phylogenetic trees, the symbiont-harboring species clustered as a major clade comprising two subclades that corresponded to the proposed genera Angomonas and Strigomonas. The genus Angomonas comprised 10 flagellates including former Crithidia deanei and C. desouzai plus a new species. The genus Strigomonas included former Crithidia oncopelti and Blastocrithidia cuiicis plus a new species. Sequences from the internal transcribed spacer of ribosomal DNA (ITS rDNA) and size polymorphism of kinetoplast DNA (kDNA) minicircles revealed considerable genetic heterogeneity within the genera Angomonas and Strigomonas. Phylogenetic analyses based on 16S rDNA and ITS rDNA sequences demonstrated that all of the endosymbionts belonged to the Betaproteobacteria and revealed three new species. The congruence of the phylogenetic trees of trypanosomatids and their symbionts support a co-divergent host-symbiont evolutionary history. (C) 2011 Elsevier GmbH. All rights reserved.

Species diversity of Leishmania (Viannia) parasites circulating in an endemic area for cutaneous leishmaniasis located in the Atlantic rainforest region of northeastern Brazil

OBJECTIVES To identify the aetiological agents of cutaneous leishmaniasis and to investigate the genetic polymorphism of Leishmania (Viannia) parasites circulating in an area with endemic cutaneous leishmaniasis (CL) in the Atlantic rainforest region of northeastern Brazil. METHODS Leishmania spp. isolates came from three sources: (i) patients diagnosed clinically and parasitologically with CL based on primary lesions, secondary lesions, clinical recidiva, mucocutaneous leishmaniasis and scars; (ii) sentinel hamsters, sylvatic or synanthropic small rodents; and (iii) the sand fly species Lutzomyia whitmani. Isolates were characterised using monoclonal antibodies, multilocus enzyme electrophoresis (MLEE) and polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region rDNA locus. RESULTS Seventy-seven isolates were obtained and characterised. All isolates were identified as Leishmania (Viannia) braziliensis serodeme 1 based on reactivity to monoclonal antibodies. MLEE identified 10 zymodemes circulating in the study region. Most isolates were classified as zymodemes closely related to L. (V.) braziliensis, but five isolates were classified as Leishmania (Viannia) shawi. All but three of the identified zymodemes have so far been observed only in the study region. Enzootic transmission and multiclonal infection were observed. CONCLUSIONS Our results confirm that transmission cycle complexity and the co-existence of two or more species in the same area can affect the level of genetic polymorphism in a natural Leishmania population. Although it is not possible to make inferences as to the modes of genetic exchange, one can speculate that some of the zymodemes specific to the region are hybrids of L. (V.) braziliensis and L. (V.) shawi.

Plant Leaf Identification Using Gabor Wavelets

This article presents a novel method of plant classification using Gabor wavelet filters to extract texture filters in a foliar surface. The aim of this promising method is to add to the results obtained by other leaf attributes (such as shape, contour, color, among others), increasing, therefore, the percentage of classification of plant species. To corroborate the efficiency of the technique, an experiment using 20 species from Brazilian flora was done and discussed. The results are also compared with texture Fourier descriptors and cooccurrence matrices. (C) 2009 Wiley Periodicals, Inc. Int J Imaging Syst Technol, 19, 236-243, 2009; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ima.20201

Fractal dimension applied to plant identification

This article discusses methods to identify plants by analysing leaf complexity based on estimating their fractal dimension. Leaves were analyzed according to the complexity of their internal and external shapes. A computational program was developed to process, analyze and extract the features of leaf images, thereby allowing for automatic plant identification. Results are presented from two experiments, the first to identify plant species from the Brazilian Atlantic forest and Brazilian Cerrado scrublands, using fifty leaf samples from ten different species, and the second to identify four different species from genus Passiflora, using twenty leaf samples for each class. A comparison is made of two methods to estimate fractal dimension (box-counting and multiscale Minkowski). The results are discussed to determine the best approach to analyze shape complexity based on the performance of the technique, when estimating fractal dimension and identifying plants. (C) 2008 Elsevier Inc. All rights reserved.

Characterization and Identification of Proteolytic Bacteria From the Gut of the Velvetbean Caterpillar (Lepidoptera: Noctuidae)

The characterization and identification of proteolytic bacteria from the gut of the velvetbean caterpillar (Anticarsia gemmatalis) were the objectives of this study. Twelve aerobic and anaerobic isolates of proteolytic bacteria were obtained from the caterpillar gut in calcium caseinate agar. The number of colony forming units (CFUs) of proteolytic bacteria was higher when the bacteria were extracted from caterpillars reared on artificial diet rather than on soybean leaves (1.73 +/- 0.35 X 10(3) and 0.55 +/- 0.22 X 10(3) CFU/mg gut, respectively). The isolated bacteria were divided into five distinct groups, according to their polymerase chain reaction restriction fragment-length polymorphism profiles. After molecular analysis, biochemical tests and fatty acid profile determination, the bacteria were identified as Bacillus subtilis, Bacillus cereus, Enterococcus gallinarum, Enterococcus mundtii, and Staphylococcus xylosus. Bacterial proteolytic activity was assessed through in vitro colorimetric assays for (general) proteases, serine proteases, and cysteine proteases. The isolated bacteria were able of hydrolyzing all tested substrates, except Staphylococcus xylosus, which did not exhibit serine protease activity. This study provides support for the hypothesis that gut proteases from velvetbean caterpillar are not exclusively secreted by the insect cells but also by their symbiotic gut bacteria. The proteolytic activity from gut symbionts of the velvetbean caterpillar is suggestive of their potential role minimizing the potentially harmful consequences of protease inhibitors from some of this insect host plants, such as soybean, with implications for the management of this insect pest species.

Effectiveness of different solid-phase microextraction fibres for differentiation of selected Madeira island fruits based on their volatile metabolite profile: identification of novel compounds

A headspace solid-phase microextraction (HS-SPME) procedure based on five commercialised fibres (85 μm polyacrylate – PA, 100 μm polydimethylsiloxane – PDMS, 65 μm polydimethylsiloxane/divinylbenzene – PDMS/DVB, 70 μm carbowax/divinylbenzene – CW/DVB and 85 μm carboxen/polydimethylsiloxane – CAR/PDMS) is presented for the characterization of the volatile metabolite profile of four selected Madeira island fruit species, lemon (Citrus limon), kiwi (Actinidia deliciosa), papaya (Carica papaya L.) and Chickasaw plum (Prunus angustifolia). The isolation of metabolites was followed by thermal desorption gas chromatography–quadrupole mass spectrometry (GC–qMS) methodology. The performance of the target fibres was evaluated and compared. The SPME fibre coated with CW/DVB afforded the highest extraction efficiency in kiwi and papaya pulps, while in lemon and plum the same was achieved with PMDS/DVB fibre. This procedure allowed for the identification of 80 compounds, 41 in kiwi, 24 in plums, 23 in papaya and 20 in lemon. Considering the best extraction conditions, the most abundant volatiles identified in kiwi were the intense aldehydes and ethyl esters such as (E)-2-hexenal and ethyl butyrate, while in Chicasaw plum predominate 2-hexenal, 2-methyl-4-pentenal, hexanal, (Z)-3-hexenol and cyclohexylene oxide. The major compounds identified in the papaya pulp were benzyl isothiocyanate, linalool oxide, furfural, hydroxypropanone, linalool and acetic acid. Finally, lemon was shown to be the most divergent of the four fruits, being its aroma profile composed almost exclusively by terpens, namely limonene, γ-terpinene, o-cymene and α-terpinolene. Thirty two volatiles were identified for the first time in the fruit or close related species analysed and 14 volatiles are reported as novel volatile metabolites in fruits. This includes 5 new compounds in kiwi (2-cyclohexene-1,4-dione, furyl hydroxymethyl ketone, 4-hydroxydihydro-2(3H)-furanone, 5-acetoxymethyl-2-furaldehyde and ethanedioic acid), 4 in plum (4-hydroxydihydro-2(3H)-furanone, 5-methyl-2-pyrazinylmethanol, cyclohexylene oxide and 1-methylcyclohexene), 4 in papaya (octaethyleneglycol, 1,2-cyclopentanedione, 3-methyl-1,2-cyclopentanedione and 2-furyl methyl ketone) and 2 in lemon (geranyl farnesate and safranal). It is noteworthy that among the 15 volatile metabolites identified in papaya, 3-methyl-1,2-cyclopentanedione was previously described as a novel PPARγ (peroxisome proliferator-activated receptor γ) agonist, having a potential to minimize inflammation.

Morphologic and molecular characterization of Myrciaria spp species

A jabuticabeira é considerada uma das fruteiras mais típicas do Brasil. Entretanto, há poucos estudos sobre esta planta na literatura, e mesmo sua classificação botânica é muito controvertida. Este trabalho faz comparações entre as espécies de jabuticabeiras, usando as técnicas de marcadores morfológicos (organografia) e moleculares RAPD. As características morfológicas das plantas, usadas como marcadores morfológicos, foram comparadas com espécimes presentes nos herbários dos Estados de São Paulo e Minas Gerais e com as descrições obtidas em revisão de literatura especializada. As diferenças moleculares entre as espécies foram determinadas por meio do uso de marcadores RAPD. O experimento foi realizado nas cidades de Piracicaba, Jaboticabal e Ituverava do Estado de São Paulo, Brasil. Diferenças morfológicas e moleculares entre as plantas estudadas foram identificadas, e quatro grupos distintos de espécies foram definidos: Myrciaria cauliflora (Mart.) O. Berg, M. coronata Mattos, M. jaboticaba (Vell.) O. Berg. e M. phytrantha (Kiaersk.) Mattos. A técnica de marcadores moleculares, aliada à técnica de marcadores morfológicos, mostrou ser uma ferramenta importante na identificação de espécies de jabuticabeiras.

A revision of the genus Pulaeus Den Heyer, with descriptions of a new genus and four new Brazilian species (Acari: Prostigmata: Cunaxidae)

A new genus, Lupaeus gen. nov., is created to contain part of the described species in the genus Pulaeus Den Heyer. A key to the genera of the tribe Pulaeini, to which these two genera belong, and one auxiliary key to the species of Brazil and South Africa, are provided. Four new species, viz. Pulaeus myrtaceus sp. nov., P. quadrisolenidius sp. nov., Lupaeus lectus sp. nov. and L. lobidorsalis sp. nov. are described and figured. P. martini Den Heyer, 1981 is designated as type species for the new genus. New nomenclatural combinations are reported.

Morphological characterisation of five brazilian species of Fidicinoides (Hemiptera: Cicadidae)

The objective of this study was to describe and illustrate the morphological characteristics of some species of Brazilian Fidicinoides to aid in their identification. Descriptions and illustrations are given of the head, thorax, abdomen, right forewing and the last urostemite of the male of the Fidicinoides besti, Fidicinoides brunnea, Fidicinoides duckensis, Fidicinoides pseudethelae and Fidicinoides sucinalae species. The main diagnostic characteristics of these species are markings on pronotum and/or mesonotum and male stemite VIII characteristics.

Molecular Identification and Characterization of Ileal and Cecal Fungus Communities in Broilers Given Probiotics, Specific Essential Oil Blends, and Under Mixed Eimeria Infection

Broiler digestive tract fungal communities have gained far less scrutiny than that given corresponding bacterial communities. Attention given poultry-associated fungi have focused primarily on feed-associated toxin-producers, yeast, and yeast products. The current project focused on the use of pyrosequencing and denaturing gradient gel electrophoresis (DGGE) to identify and monitor broiler digestive fungal communities. Eight different treatments were included. Four controls were an Uninfected-Unmedicated Control, an Unmedicated-Infected Control, the antibiotic bacitracin methylene disalicylate plus the ionophore monensin as Positive Control, and the ionophore monensin alone as a Negative Control. Four treatments were two probiotics (BC-30 and Calsporin) and two specific essential oil blends (Crina Poultry Plus and Crina Poultry AF). All chickens except the Unmedicated-Uninfected Control were given, at 15 days of age, a standard oral Eimeria inoculum of sporulated oocysts. Ileal and cecal digesta were collected at pre-Eimeria infection at 14 days of age and at 7 days post-Eimeria infection at 22 days of age. Extracted cecal DNA was analyzed by pyrosequencing to examine the impact of diet supplements and Eimeria infection on individual constituents in the fungal community, while DGGE was used to compare more qualitative changes in ileal and cecal communities. Pyrosequencing identified three phyla, seven classes, eight orders, 13 families, 17 genera, and 23 fungal species. Ileal and cecal DGGE patterns showed fungal communities were clustered mainly into pre- and post-infection patterns. Post-infection Unmedicated-Uninfected patterns were clustered with pre-infection groups demonstrating a strong effect of Eimeria infection on digestive fungal populations. These combined techniques offered added versatility towards unraveling the effects of enteropathogen infection and performance enhancing feed additives on broiler digestive microflora.

Evaluation of four molecular typing Methodologies as tools for determining taxonomy relations and for identifying. species among Yersinia isolates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)