Reproductive behavior, development and eye regression in the cave armored catfish, Ancistrus cryptophthalmus Reis, 1987 (Siluriformes: Loricariidae), breed in laboratory

The troglobitic armored catfish, Ancistrus cryptophthalmus (Loricariidae, Ancistrinae) is known from four caves in the São Domingos karst area, upper rio Tocantins basin, Central Brazil. These populations differ in general body shape and degree of reduction of eyes and of pigmentation. The small Passa Três population (around 1,000 individuals) presents the most reduced eyes, which are not externally visible in adults. A small group of Passa Três catfish, one male and three females, reproduced spontaneously thrice in laboratory, at the end of summertime in 2000, 2003 and 2004. Herein we describe the reproductive behavior during the 2003 event, as well as the early development of the 2003 and 2004 offsprings, with focus on body growth and ontogenetic regression of eyes. The parental care by the male, which includes defense of the rock shelter where the egg clutch is laid, cleaning and oxygenation of eggs, is typical of many loricariids. On the other hand, the slow development, including delayed eye degeneration, low body growth rates and high estimated longevity (15 years or more) are characteristic of precocial, or K-selected, life cycles. In the absence of comparable data for close epigean relatives (Ancistrus spp.), it is not possible to establish whether these features are an autapomorphic specialization of the troglobitic A. cryptophthalmus or a plesiomorphic trait already present in the epigean ancestor, possibly favoring the adoption of the life in the food-poor cave environment. We briefly discuss the current hypotheses on eye regression in troglobitic vertebrates.

A cave population of Isbrueckerichthys alipionis (Gosline, 1947) in the Upper Ribeira karst area, southeastern Brazil (Siluriformes: Loricariidae)

A cave population of the armored catfish Isbrueckerichthys alipionis is reported from the Santana Cave, in the rio Betari watershed, Upper Ribeira karst area, Iporanga, São Paulo State, southeastern Brazil. The cave population was compared to an epigean population of I. alipionis and no significant differences where found in morphology or degree of pigmentation. As the cave population is known for at least 30 years and is apparently isolated from epigean streams, it is classified as troglophilic. The discovery of this troglophilic species in the Santana Cave is an additional strong argument for the conservation of that cave.

Chetoneura shennonggongensis, a new species of cave-dwelling Keroplatini from China (Diptera : Keroplatidae), with a discussion of the position of Chetoneura

Chetoneura shennonggongensis Amorim & Niu, sp. n., inhabiting caves in East China, is described based on the adult male and female, plus larva. Chetoneura Colless, previously associated with Orfelia and known from a single species from caves in Malaysia, is formally transferred to the Keroplatini. This is the first description of the larva and of the general biology of the genus. The larva of this new species is suspended in a roughly horizontal hammock of silken threads amidst mucous coated snare threads from the roof of caves, but is not bioluminescent as some species of other genera in Keroplatidae ( e. g., Arachnocampa, Keroplatus, and Orfelia). A key to separate both species of the genus is provided. The position of the genus within the Keroplatini is considered. Chetoneura is considered to be closely related to a clade containing Heteropterna and Ctenoceridion.

Chromosome homologies of the highly rearranged karyotypes of four Akodon species (Rodentia, Cricetidae) resolved by reciprocal chromosome painting: the evolution of the lowest diploid number in rodents

Traditionally comparative cytogenetic studies are based mainly on banding patterns. Nevertheless, when dealing with species with highly rearranged genomes, as in Akodon species, or with other highly divergent species, cytogenetic comparisons of banding patterns prove inadequate. Hence, comparative chromosome painting has become the method of choice for genome comparisons at the cytogenetic level since it allows complete chromosome probes of a species to be hybridized in situ onto chromosomes of other species, detecting homologous genomic regions between them. In the present study, we have explored the highly rearranged complements of the Akodon species using reciprocal chromosome painting through species-specific chromosome probes obtained by chromosome sorting. The results revealed complete homology among the complements of Akodon sp. n. (ASP), 2n = 10; Akodon cursor (ACU), 2n = 15; Akodon montensis (AMO), 2n = 24; and Akodon paranaensis (APA), 2n = 44, and extensive chromosome rearrangements have been detected within the species with high precision. Robertsonian and tandem rearrangements, pericentric inversions and/or centromere repositioning, paracentric inversion, translocations, insertions, and breakpoints, where chromosomal rearrangements, seen to be favorable, were observed. Chromosome painting using the APA set of 21 autosomes plus X and Y revealed eight syntenic segments that are shared with A. montensis, A. cursor, and ASP, and one syntenic segment shared by A. montensis and A. cursor plus five exclusive chromosome associations for A. cursor and six for ASP chromosome X, except for the heterochromatin region of ASP X, and even chromosome Y shared complete homology among the species. These data indicate that all those closely related species have experienced a recent extensive process of autosomal rearrangement in which, except for ASP, there is still complete conservation of sex chromosomes homologies.

Evaluation of iris color stability in ocular prosthesis

Accurate iris reproduction in the fabrication of ocular prosthesis in order to match the remaining eye is a key factor to mask the loss and achieve an esthetic outcome for anophthalmic patients. This study evaluated the stability of acrylic paints used for replicating iris color in ocular prostheses by the analysis of two factors: the temperature of the acrylic resin polymerization cycle during prosthesis fabrication and the incidence of sun light, which is the main photodegrading agent undermining the longevity of ocular prostheses. An accelerated aging assay was used for both analyses. Specimens simulating the prosthetic iris in the colors blue, yellow, black, brown and green were fabricated, and were submitted to a colorimetric reading before and after undergoing the thermal conditions of acrylic resin polymerization. Next, the specimens were submitted to an artificial accelerated aging assay with ultraviolet radiation A and weekly colorimetric readings during a 3-week period. The color change (??*) values for the four specimens painted with the same color paint were averaged and the resulting values were considered for statistical analysis. Levine's test and Student's t-test were used to analyze the influence of the temperature of the polymerization cycle during prosthesis fabrication on the color stability of each acrylic resin paint. Friedman's test for three dependent samples was used for analysis of color photodegradation as function of time. Significance level was set at 0.05 for all analyses. It was observed that, after the action of the temperature of the polymerization cycle, alteration above clinically acceptable level of ??*> 3.3 was observed only for the yellow color. After the accelerated aging assay, there were statistically significant differences (p<0.05) as a function of time in the green, brown, black and blue colors. Changes were clinically acceptable for the brown and black colors; slightly above the clinically acceptable limit for the green color; and significantly high and impracticable from a clinical standpoint for the blue color. There was no statistically significant differences (p>0.05) for the yellow color, which presented color change only a little above the clinically acceptable limit. In conclusion: 1. Only the yellow color presented alterations above the clinically acceptable levels after the polymerization cycle; 2. After accelerated aging, there was no changes in the yellow color above the clinically acceptable levels; 3. For the green color, degradation was significant and slightly above the clinically acceptable levels; 4. The black, brown and blue colors presented significant alterations as function of time; the alterations of the brown and black colors were within acceptable clinical levels, while the blue color presented a more accentuated degradation over time.

Mechanisms of pigmentation loss in subterranean fishes

Troglobitic (exclusively subterranean) organisms usually present, among their apomorphies related to the subterranean life (troglomorphisms), the regression of eyes and melanic pigmentation. The degree of regression varies among species, from a slight reduction to the complete loss of eyes and dark pigmentation, without a taxonomic correlation. While mechanisms of eye reduction have been intensively investigated in some troglobites such as the Mexican blind tetra characins, genus Astyanax, and the European salamander, Proteus anguinus, few studies have focused on pigmentation. The Brazilian subterranean ichthyofauna distinguishes not only by the species richness (23 troglobitic fishes so far known) but also by the variation in the degree of reduction of eyes and pigmentation. This study focused on Brazilian fishes completely devoid of melanic pigmentation: the characiform Stygichthys typhlops (Characidae) and the siluriforms Ancistrus formoso (Loricariidae), Rhamdiopsis sp.1 (Heptapteridae; from caves in the Chapada Diamantina, Bahia) and Rhamdiopsis sp. 2 (cave in Campo Formoso, Bahia). In order to investigate if such depigmentation is the result of blockage in some step in the melanogenesis, in vitro tests of administration of L-DOPA were done, using caudal-fin fragments extracted from living fish. Except for Rhamdiopsis sp. 2, all the studied species were DOPA(+), i.e., melanin was synthesized after L-DOPA administration. This indicates these fish do have melanophores but they are unable to convert L-tyrosine to L-DOPA. On the other hand, Rhamdiopsis sp. 2, like the albino specimens of Trichomycterus itacarambiensis previously studied (which correspond to one third of the population), are DOPA(-), either because the block of melanin synthesis occurs downstream in melanogenesis, which is probably the case with T. itacarambiensis (monogenic system in view of the phenotypic discontinuity), or because the so-called albinos do no possess melanophores. The physiological loss in the ability to synthesize melanin, apparently caused by different genetic processes in DOPA(+) and in DOPA(-) fishes, may co-exist in subterranean populations with a decrease in the density of melanophores, as observed in the pigmented two thirds of T. itacarambiensis population, a morphological reduction apparently controlled by polygenic systems producing a continuous phenotypic variation.

Use of chromosome microdissection in fish molecular cytogenetics

Chromosome microdissection is a technique in which whole chromosomes or chromosomal segments are dissected under an inverted microscope yielding chromosome-specific sequences. Several protocol modifications introduced during the past 15 years reduced the number of chromosomes required for most applications. This is of particular interest to fish molecular cytogenetics, since most species present highly uniform karyotypes which make impossible the collection of multiple copies of the same chromosome. Probes developed in this manner can be used to investigate chromosome homologies in closely related species. Here we describe a protocol recently used in the gymnotiform species group Eigenmannia and review the major steps involved in the generation of these markers focusing on protocol modifications aiming to reduce the number of required chromosomes.

Trichomycterus dali: a new highly troglomorphic catfish (Silurifomes: Trichomycteridae) from Serra da Bodoquena, Mato Grosso do Sul State, Central Brazil

Trichomycterus dali, new species, is described from flooded limestone caves in Serra da Bodoquena karst area, State of Mato Grosso do Sul, Central Brazil. The new species is diagnosed by a unique character in the genus, the presence of conspicuous, ridge-like adipose folds lining dorsally throughout the body. Trichomycterus dali can be further distinguished readily from epigean congeners by the reduction of eyes and skin pigmentation (except for T. gorgona), and from remaining congeners (i.e., all hypogean plus T. gorgona) by the total loss of eyes, not visible externally (except for T. sandovali and T. spelaeus). Other diagnostic features includes very long barbels, especially the nasal (99.3-143.5% HL) and the maxillary (97.0-131.3% HL), pectoral-fin ray count reaching I,9 and a unique cranial fontanel with a conspicuous constriction on the meeting point of supraoccipital and the two frontal bones. The troglobitic status of the species is suggested by the presence of troglomorphisms on an advanced degree, especially the reduction of skin pigmentation, the total loss of eyes and the enlarged barbels. In addition, the presence of a well developed adipose fold in adults may indicate a distinctive adaptation acquired by neoteny to withstand the food scarce conditions of its hypogean habitat.

Ituglanis mambai, a new subterranean catfish from a karst area of Central Brazil, rio Tocantins basin (Siluriformes: Trichomycteridae)

Ituglanis mambai, new species, is described from a cave in the Mambaí karst area, State of Goiás, Central Brazil. The new species distinguishes from epigean and cave congeners by the combination of the following characteristics: posterior supraoccipital fontanel absent; pectoral-fin rays usually i,7; six pleural ribs; total vertebrae 37-38 behind Weberian apparatus; shorter predorsal length (65.1-70.8% SL); shorter caudal peduncle length (8.4-11.9% SL); shorter dorsal-fin base length (7.7-11.3% SL); wider interobital width (29.2-36.5% HL); larger mouth width (43.4-64.0% HL); intermediate between epigean and other cave Ituglanis species as regards to both eyes (diameter varying from 0.5 to1.0 mm in adults, 7.8-10.1 % HL) and pigmentation, composed by irregular light brown spots along the body. The latter indicate the troglobitic status for I. mambai. In addition, this species has the maxillae with a discrete medial-posterior projection; fronto-lacrimal one half-length of the maxillae and pointed backwards; posterior process of palatine half its length, with a tenuous medial concavity; 14 dorsal and 12 ventral procurrent rays. In the natural habitat, I. mambai displayed cryptobiotic habits, trying to hide in the graveled bottom or under boulders when disturbed, apparently showing a negative response to light. It was observed a preference to slow-moving waters. Recent flood marks were observed in the stream conduit in March/April 2007 (end of the rainy season) when less individuals were observed on 300 m of the subterranean stream compared to September 2004 (end of the dry season).

Epigean and subterranean ichthyofauna in Cordisburgo karst area, eastern Brazil

After an ichthyofaunistic survey conducted in May 2007 on surface (epigean) water bodies of Cordisburgo karst area, State of Minas Gerais, 13 species were recorded, mostly characiforms; in addition three non-troglomorphic (normally eyed and pigmented) and one troglomorphic catfish (siluriforms) species were recorded in two caves surveyed at different occasions, totaling 17 fish species for the area. All the nominal species herein reported for Cordisburgo area have been previously reported for the Rio das Velhas basin. None of the species observed in caves were found in epigean habitats and vice-versa. The four cave species are distributed throughout subterranean stream reaches, with individuals at different size/age classes. This, associated to the lack of conspicuous morphological differences in relation to epigean congeners, indicate that Trichomycterus brasiliensis, Gymnotus cf. carapo and Pimelodella cf. vittata are troglophiles (species encompassing individuals able to live and complete their life cycle either in the surface or in the subterranean environment) in the Morena Cave; the latter forms a large population and may be at the beginning of a differentiation process due to isolation in the subterranean habitat, as indicated by a slight reduction in eye size. Topographic isolation may be the cause for the incipient, but unmistakable troglomorphism of the Rhamdiopsis population found in the Salitre Cave, allowing for its classification as troglobite (exclusively subterranean species). The Cordisburgo area is subject to significant anthropic pressure, mainly represented by deforestation for agriculture, cattle raising and timbering. Tourism is an additional important threat for cave communities, calling for urgent protection measures.

Study of mural paintings by Fulvio Pennacchi in São Paulo City by mineralogical techniques

The present research deals with two mural paintings made in 1947 with the fresco technique by Fulvio Pennacchi in the Catholic Chapel of the Hospital das Clínicas (São Paulo City, Brazil), namely the Virgin Annunciation and the Supper at Emmaus. This study regards the materials and painting techniques used by the artist, based on historical research,on in situ observations and laboratory analytical techniques (stereomicroscopy,scanning electron microscopy with an energy dispersive spectrometer, X-ray diffractometry, electron microprobe, images obtained with UV-light), aiming to improve the methods of characterization of objects of our cultural heritage, and to enhance its preservation accordingly. The results lead to the identification of the plaster components and of distinct layers in the frescoes, besides further information on grain size, impurities and textures, composition of pigments, and features of deterioration, such as efflorescences. The degree of degradation of the murals painting was assessed by this way. Our data suggest that a single layer of plaster was used by Pennacchi, as a common mortar with fine- and medium-grained aggregates. Differences in texture were obtained by adding gypsum to the plaster.

A new species of Glaphyropoma: the first subterranean copionodontine catfish and the first occurrence of opercular odontodes in the subfamily (Siluriformes: Trichomycteridae)

A new species of the rare copionodontine genus Glaphyropoma is described from subterranean waters in the Diamantina Plateau, Bahia State, central northeastern Brazil. This is the first troglomorphic species in the subfamily Copionodontinae. It is distinguished from all other copionodontines by the presence of opercular odontodes, and further distinguished from its only congener, G. rodriguesi, by the reduction of dark integumentary pigmentation. The new species shares the single synapomorphy previously proposed for Glaphyropoma, the marked narrowing of the first hypobranchial and indirect character evidence also supports its inclusion in the genus. The presence of opercular odontodes in the new species, in combination with a reviewed hypothesis of sister group relationship between Copionodontinae and Trichogeninae, indicate that the absence of opercular odontodes in previously-known copionodontines is secondary, rather than primitive.

Phlebotomines (Diptera, Psychodidae) in the Ribeira Valley Speleological Province - 1. Parque Estadual Intervales, state of São Paulo, Brazil

The identification of the sandfly fauna and investigation of some ecological aspects of its populations in areas frequented by tourists of the PEI, an Atlantic forest reserve with many caves, were the objective of this study. Captures were undertaken monthly from January 2001 to December 2002, with automatic light traps installed in 13 ecotopes, including caves, forests, domiciliary and peridomiciliary environments, and by aspiration in armadillo burrows. Additionally, although not at regular intervals, Shannon traps were installed in forests and anthropic environments, aspirations were made on cave walls, among roots and fallen leaves, and some insects were captured while biting researchers. A total of 891 sandflies belonging to 21 species were captured. Six hundred specimens representing 19 species were captured with light traps, 215 in anthropic (2.24 insects/trap) and 385 in extra-domiciliary (1.46 insects/trap) environments. Brumptomyia troglodytes was the most abundant species (the Standardised Index of Species Abundance = 0.705). Pintomyia monticola predominated in the Shannon traps and showed anthropophilic and diurnal activity. Psathyromyia pascalei predominated in the aspirations; the largest number being in armadillo burrows. Eleven species were captured in caves; although some might be troglophiles, the majority used these ecotopes as resting places. Nyssomyia intermedia, Nyssomyia neivai and Migonemyia migonei, implicated in the transmission of cutaneous leishmaniasis in the Southeastern Brazilian region, were all found, though in such low densities as to suggest minimal risk of the disease in the PEI.

Lutzomyia longipalpis (Diptera, Psychodidae) em Cuesta Basáltica, na bacia hidrográfica do Rio Corumbataí, Região Centro-leste do Estado de São Paulo

A incidência das leishmanioses tegumentar e visceral americanas, em especial esta última (LVA), em hospedeiros caninos e humanos, encontra-se em crescente processo de expansão no Estado de São Paulo. Para a vigilância epidemiológica dessas endemias, torna-se fundamental o conhecimento da distribuição e da ecologia das diferentes espécies da fauna flebotomínea vetoras. Assim, a divulgação de novos encontros de seus vetores, sobretudo da Lutzomyia longipalpis, o principal vetor da LVA, é fundamental para apontar novas áreas de risco para a transmissão dessas doenças. Neste estudo, capturas de flebotomíneos foram realizadas em ambiente domiciliar, peridomiciliar e de mata, em diferentes localidades rurais dos municípios de Ipeúna e Itirapina, entre outubro de 2001 e fevereiro de 2004. Foram utilizadas armadilhas luminosas automáticas do tipo CDC, das 18h às 8h, em 14 noites, resultando 420 horas de exposição. Foram capturados 177 flebotomíneos pertencentes a doze espécies. A espécie mais abundante, Nyssomyia neivai, apontada como a principal vetora de LTA no Estado, contribuiu com 85,4% dos espécimes capturados em Ipeúna. O encontro de Lutzomyia longipalpis em uma caverna em Itirapina, aponta para o risco de estabelecimento da LVA na área e a necessidade de mais estudos locais sobre sua ecologia, sobretudo em relação à ocupação de ambientes antrópicos.

New reports of Antricola guglielmonei and Antricola delacruzi in Brazil, and a description of a new argasid species (Acari)

Adults of 3 tick species (Acari: Argasidae) identified as Antricola guglielmonei, Antricola delacruzi, and Carios rondoniensis n. sp. were collected oil bill guano in a cave in the state of Rondonia, western Amazon. Brazil. Adults of C. rondoniensis Possess a unique combination of characters that distinguish them front all described adults in the Argasidae. i.e.. a large spiracular plate densely filled with small goblets, a well-developed flap covering the female genital opening, and palpi containing several tufts of long setae oil articles 2 and 3. Unlike Ornithodoros or other Carios species. adults of C rondoniensis have a scooplike hypostome devoid of denticles, as in Antricola spp. Conversely, the presence of a pair of long posthypostomal setae, and a slitlike transverse fissure at the Capsule opening of the Haller's organ, are characters of C. rondonensis that are also found ill species of Carios and Ornithodoros, but not in Antricola species. Molecular analyses inferred from a portion of the 16S rRNA mitochondrial gene indicate that C. rondoniensis is phylogenetically closest to species of Carios. followed by species of Antricola. and then Ornithodoros. Because the highest bootstrap value linking C. rondoniensis to Carios spp. was 62%, further phylogenetic studies are needed to better evaluate the taxonomic Status of the former species.