Circadian Pattern of Wheel-Running Activity of a South American Subterranean Rodent (Ctenomys cf knightii)

Circadian rhythms are regarded as essentially ubiquitous features of animal behavior and are thought to confer important adaptive advantages. However, although circadian systems of rodents have been among the most extensively studied, most comparative biology is restricted to a few related species. In this study, the circadian organization of locomotor activity was studied in the subterranean, solitary north Argentinean rodent, Ctenomys knightii. The genus, Ctenomys, commonly known as Tuco-tucos, comprises more than 50 known species over a range that extends from 12S latitude into Patagonia, and includes at least one social species. The genus, therefore, is ideal for comparative and ecological studies of circadian rhythms. Ctenomys knightii is the first of these to be studied for its circadian behavior. All animals were wild caught but adapted quickly to laboratory conditions, with clear and precise activity-rest rhythms in a light-dark (LD) cycle and strongly nocturnal wheel running behavior. In constant dark (DD), the rhythm expression persisted with free-running periods always longer than 24h. Upon reinstatement of the LD cycle, rhythms resynchronized rapidly with large phase advances in 7/8 animals. In constant light (LL), six animals had free-running periods shorter than in DD, and 4/8 showed evidence of splitting. We conclude that under laboratory conditions, in wheel-running cages, this species shows a clear nocturnal rhythmic organization controlled by an endogenous circadian oscillator that is entrained to 24h LD cycles, predominantly by light-induced advances, and shows the same interindividual variable responses to constant light as reported in other non-subterranean species. These data are the first step toward understanding the chronobiology of the largest genus of subterranean rodents.

Field and Laboratory Studies Provide Insights into the Meaning of Day-Time Activity in a Subterranean Rodent (Ctenomys aff. knighti), the Tuco-Tuco

South American subterranean rodents (Ctenomys aff. knighti), commonly known as tuco-tucos, display nocturnal, wheel-running behavior under light-dark (LD) conditions, and free-running periods >24 h in constant darkness (DD). However, several reports in the field suggested that a substantial amount of activity occurs during daylight hours, leading us to question whether circadian entrainment in the laboratory accurately reflects behavior in natural conditions. We compared circadian patterns of locomotor activity in DD of animals previously entrained to full laboratory LD cycles (LD12:12) with those of animals that were trapped directly from the field. In both cases, activity onsets in DD immediately reflected the previous dark onset or sundown. Furthermore, freerunning periods upon release into DD were close to 24 h indicating aftereffects of prior entrainment, similarly in both conditions. No difference was detected in the phase of activity measured with and without access to a running wheel. However, when individuals were observed continuously during daylight hours in a semi-natural enclosure, they emerged above-ground on a daily basis. These day-time activities consisted of foraging and burrow maintenance, suggesting that the designation of this species as nocturnal might be inaccurate in the field. Our study of a solitary subterranean species suggests that the circadian clock is entrained similarly under field and laboratory conditions and that day-time activity expressed only in the field is required for foraging and may not be time-dictated by the circadian pacemaker.

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.

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.

Surface and subterranean ichthyofauna in the Serra do Ramalho karst area, northeastern Brazil, with updated lists of Brazilian troglobitic and troglophilic fishes

After an ichthyofaunistic survey in several epigean (surface) water bodies of the Serra do Ramalho, southern Bahia, conducted in May 2007, 44 species were recorded; in addition, three non-troglomorphic (normally eyed and pigmented) and two troglomorphic species were recorded only in caves, totaling 49 species of fishes for the area, which represents a little more than one fourth of the total registered in the literature for the entire Rio São Francisco basin. In these caves, which have been studied since 2005, eight non-troglomorphic species were sampled and their presence in both epigean and subterranean habitats, associated to the lack of morphological differences, indicate that they may be either troglophiles (species encompassing individuals able to live and complete their life cycle either in the surface or in the subterranean environment), trogloxenes (individuals regularly found in subterranean habitats, but which must return periodically to the surface in order to complete their life cycle) or even accidental in caves. In addition, two troglomorphic species (with reduced eyes and melanic pigmentation when compared to close epigean relatives), belonging respectively to the genera Rhamdia and Trichomycterus, were recorded exclusively in caves, thus classified as troglobites. Interestingly, no epigean representative of the genus Trichomycterus was collected. The new data are integrated into updated lists of Brazilian troglobitic and troglophilic fishes, based on published data and new records recently confirmed.

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.

Environmental conditions and rodent infestation in Campo Limpo district, Sao Paulo municipality, Brazil

Rodents are involved in the transmission to human beings of several diseases, including liptospirosis, which shows high lethality rates in Sao Paulo municipality. Despite this, few studies have assessed the relationship existing between urban environmental conditions and building rodent infestation. With the purpose of clarifying this relationship, an analysis has been conducted in order to quantify the influence of environmental factors upon rodent infestation on a low-income district. Diagnosis of the environmental situation has been performed to evaluate the frequency according to which harborage, food and access sources occur, and a survey on infestation rates in 2175 dwellings in the area studied. The logistic regression analysis showed that among the environmental variables, the one that showed the closest association with rodent infestation was access; followed by harborage, and food. It was concluded that poor socioeconomic and environmental conditions in the area propitiate the occurrence of high rodent infestation rates

Survival and Neuronal Differentiation of Mesenchymal Stem Cells Transplanted into the Rodent Brain Are Dependent upon Microenvironment

Introduction: The successful integration of stem cells in adult brain has become a central issue in modern neuroscience. In this study we sought to test the hypothesis that survival and neurodifferentiation of mesenchymal stem cells (MSCs) may be dependent upon microenvironmental conditions according to the site of implant in the brain. Methods: MSCs were isolated from adult rats and labeled with enhanced-green fluorescent protein (eGFP) lentivirus. A cell suspension was implanted stereotactically into the brain of 50 young rats, into one neurogenic area (hippocampus), and into another nonneurogenic area (striatum). Animals were sacrificed 6 or 12 weeks after surgery, and brains were stained for mature neuronal markers. Cells coexpressing NeuN (neuronal specific nuclear protein) and GFP (green fluorescent protein) were counted stereologically at both targets. Results: The isolated cell population was able to generate neurons positive for microtubule-associated protein 2 (MAP2), neuronal-specific nuclear protein (NeuN), and neurofilament 200 (NF200) in vitro. Electrophysiology confirmed expression of voltage-gated ionic channels. Once implanted into the hippocampus, cells survived for up to 12 weeks, migrated away from the graft, and gave rise to mature neurons able to synthesize neurotransmitters. By contrast, massive cell degeneration was seen in the striatum, with no significant migration. Induction of neuronal differentiation with increased cyclic adenosine monophosphate in the culture medium before implantation favored differentiation in vivo. Conclusions: Our data demonstrated that survival and differentiation of MSCs is strongly dependent upon a permissive microenvironment. Identification of the pro-neurogenic factors present in the hippocampus could subsequently allow for the integration of stem cells into nonpermissive areas of the central nervous system.

Placentation in Sigmodontinae: a rodent taxon native to South America

Background: Sigmodontinae, known as ""New World rats and mice,"" is a large subfamily of Cricetidae for which we herein provide the first comprehensive investigation of the placenta. Methods: Placentas of various gestational ages ranging from early pregnancy to near term were obtained for five genera, i.e. Necromys, Euryoryzomys, Cerradomys, Hylaeamys, and Oligoryzomys. They were investigated by means of histology, immunohistochemistry, a proliferation marker, DBA-lectin staining and transmission electron microscopy. Results: The chorioallantoic placenta was organized in a labyrinthine zone, spongy zone and decidua and an inverted yolk sac persisted until term. The chorioallantoic placenta was hemotrichorial. The interhemal barrier comprised fetal capillary endothelium and three layers of trophoblast, an outermost, cellular layer and two syncytial ones, with interspersed trophoblast giant cells (TGC). In addition, accumulations of TGC occurred below Reichert's membrane. The junctional zone contained syncytial trophoblast, proliferative cellular trophoblast, glycogen cells and TGC that were situated near to the maternal blood channels. In three of the genera, TGC were also accumulated in distinct areas at the placental periphery. PAS-positive glycogen cells derived from the junctional zone invaded the decidua. Abundant maternal uNK cells with positive response to PAS, vimentin and DBA-lectin were found in the decidua. The visceral yolk sac was completely inverted and villous. Conclusion: The general aspect of the fetal membranes in Sigmodontinae resembled that found in other cricetid rodents. Compared to murid rodents there were larger numbers of giant cells and in some genera these were seen to congregate at the periphery of the placental disk. Glycogen cells were found to invade the decidua but we did not identify trophoblast in the walls of the deeper decidual arteries. In contrast these vessels were surrounded by large numbers of uNK cells. This survey of wild-trapped specimens from five genera is a useful starting point for the study of placentation in an important subfamily of South American rodents. We note, however, that some of these rodents can be captive bred and recommend that future studies focus on the study of time dated pregnancies.

Comparative study on free-running locomotor activity circadian rhythms in Brazilian subterranean fishes with different degrees of specialization to the hypogean life (Teleostei: Siluriformes; Characiformes)

In this paper we report findings on the presence of circadian rhythms in six species of cave-dwelling fishes from Brazil. Locomotor activity of islolated individuals was automatically recorded for 10 consecutive days under constant darkness. The species tested show varied degrees of specialization to subterranean life and we found varying degrees of the circadian components of locomotor activity as measured by the periodogram algorhythm of Lomb-Scargle. Both the presence and robustness of the circadian components seem to vary according to the degree of specialization to subterranean life, the more specialized, the less circadian rhythmicity was detected.

A new genus and species of rodent from the Brazilian Atlantic Forest (Rodentia: Cricetidae: Sigmodontinae: Oryzomyini), with comments on oryzomyine biogeography

We describe in this paper a new genus and species of cricetid rodent from the Atlantic Forest of Brazil, one of the most endangered eco-regions of the world. The new form displays some but not all synapomorphies of the tribe Oryzomyini, but a suite of unique characteristics is also observed. This new forest rat possesses anatomical characteristics of arboreal taxa, such as very developed plantar pads, but was collected almost exclusively in pitfall traps. Phylogenetic analyses of morphological (integument, soft tissue, cranial, and dental characters) and molecular [nuclear - Interphotoreceptor retinoid binding protein (Irbp) - and mitochondrial - cytochrome b - genes] datasets using maximum likelihood and cladistic parsimony approaches corroborate the inclusion of the new taxon within oryzomyines. The analyses also place the new form as sister species to Eremoryzomys polius, an Andean rat endemic to the Maranon valley. This biogeographical pattern is unusual amongst small terrestrial vertebrates, as a review of the literature points to few other similar examples of Andean-Atlantic Forest pairings, in hylid frogs, Pionus parrots, and other sigmodontine rodents. (C) 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 161, 357-390. doi:10.1111/j.1096-3642.2010.00643.x

Association between decay fungi and subterranean termites in the wood biodeterioration process of Tipuana tipu (Benth.) O. Kuntze trees of Sao Paulo city, SP

In the present paper the process of wood biodeterioration of tipuana trees planted in 7 regions of the city of Sao Paulo, SP was evaluated. On the sidewalks, 1109 trees were analyzed taking into consideration the occurrence and association of the xylophagous organisms (decay fungi and subterranean termites), the wood deterioration and the BHD (breast height diameter). The percentage of wood internal deterioration (%) was obtained by non destructive analysis, using a penetrometer. The results had shown that 75% of the tipuana trees presented BHD superior to 50 cm, characterizing them as adult. Decay fungi in the roots and/or trunk had been observed in 338 trees (30.5%). Subterranean termites of Heterotermes sp. and Coptotermes gestroi species had occurred in 307 trees (27.7%), the latter in high infestation level. The association between the fungi and termites was observed, as well as its relation with the BHD, where a greater value of BHD meant higher wood biodeterioration intensity. For tipuana trees, the BHD was considered an indicative attribute of the internal deterioration intensity, caused by these xylophagous organisms.

Driving mechanisms for groundwater flow and salt transport in a subterranean estuary

This paper presents field measurements and numerical simulations of groundwater dynamics in the intertidal zone of a sandy meso-tidal beach. The study, focusing on vertical hydraulic gradients and pore water salinities, reveals that tides and waves provide important forcing mechanisms for flow and salt transport in the nearshore aquifer. Such forcing, interacting with the beach morphology, enhances the exchange between the aquifer and ocean. The spatial and temporal variations of vertical hydraulic gradients demonstrate the complexity and dynamic nature of the processes and the extent of mixing between fresh groundwater and seawater in a subterranean estuary''. These results provide evidence of a potentially important reaction zone in the nearshore aquifer driven by oceanic oscillations. Land-derived contaminants may undergo important biogeochemical transformations in this zone prior to discharge.

Effects of (-)-RO363 at human atrial beta-adrenoceptor subtypes, the human cloned beta(3)-adrenoceptor and rodent intestinal beta(3)-adrenoceptors

1 Chronic treatment of patients with beta-blockers causes atrial inotropic hyperresponsiveness through beta(2)-adrenoceptors, 5-HT4 receptors and H-2-receptors but apparently not through beta(1)-adrenoceptors despite data claiming an increased beta(1)-adrenoceptor density from homogenate binding studies. We have addressed the question of beta(1)-adrenoceptor sensitivity by determining the inotropic potency and intrinsic activity of the beta(1)-adrenoceptor selective partial agonist (-)-RO363 and by carrying out both homogenate binding and quantitative beta-adrenoceptor autoradiography in atria obtained from patients treated or not treated with beta-blockers. In the course of the experiments it became apparent that (-)-RO363 also may cause agonistic effects through the third atrial beta-adrenoceptor. To assess whether (-)-RO363 also caused agonistic effects through beta(3)-adrenoceptors we studied its relaxant effects in rat colon and guinea-pig ileum, as well as receptor binding and adenylyl cyclase stimulation of chinese hamster ovary (CHO) cells expressing human beta(3)-adrenoceptors. 2 beta-Adrenoceptors were labelled with (-)-[I-125]-cyanopindolol. The density of both beta(1)- and beta(2)-adrenoceptors was unchanged in the 2 groups, as assessed with both quantitative receptor autoradiography and homogenate binding. The affinities of (-)-RO363 for beta(1)-adrenoceptors (pK(i) = 8.0-7.7) and beta(2)-adrenoceptors (pK(i) = 6.1-5.8) were not significantly different in the two groups. 3 (-)-RO363 increased atrial force with a pEC(50) of 8.2 (beta-blocker treated) and 8.0 (non-beta-blocker treated) and intrinsic activity with respect to (-)-isoprenaline of 0.80 (beta-blocker treated) and 0.54 (non-beta-blocker treated) (P<0.001) and with respect to Ca2+ (7 mM) of 0.65 (beta-blocker treated) and 0.45 (non-beta-blocker treated) (P<0.01). The effects of (-)-RO363 were resistant to antagonism by the beta(2)-adrenoceptor antagonist, ICI 118,551 (50 nM). The effects of 0.3-10 nM (-)-RO363 were antagonized by 3-10 nM of the beta(1)-adrenoceptor selective antagonist CGP 20712A. The effects of 20-1000 nM (-)-RO363 were partially resistant to antagonism by 30-300 nM CGP 20712A. 4 (-)-RO363 relaxed the rat colon, partially precontracted by 30 mM KCl, with an intrinsic activity of 0.97 compared to (-)-isoprenaline. The concentration-effect curve to (-)-RO363 revealed 2 components, one antagonized by (-)-propranolol (200 nM) with pEC(50)=8.5 and fraction 0.66, the other resistant to (-)-propranolol (200 nM) with pEC(50)=5.6 and fraction 0.34 of maximal relaxation. 5 (-)-RO363 relaxed the longitudinal muscle of guinea-pig ileum, precontracted by 0.5 mu M histamine, with intrinsic activity of 1.0 compared to (-)-isoprenaline and through 2 components, one antagonized by (-)-propranolol (200 nM) with pEC(50)=8.7 and fraction 0.67, the other resistant to (-)-propranolol with pEC(50)=4.9 and fraction 0.33 of maximal relaxation. 6 (-)-RO363 stimulated the adenylyl cyclase of CHO cells expressing human beta(3)-adrenoceptors with pEC(50)=5.5 and intrinsic activity 0.74 with respect to (-)-isoprenaline (pEC(50)=5.9). (-)-RO363 competed for binding with [I-125]cyanopindolol at human beta(3)-adrenoceptors transfected into CHO cells with pK(i)=4.5. (-)-Isoprenaline (pk(i)=5.2) and (-)-CGP 12177A (pK(i)=6.1) also competed for binding at human beta(2)-adrenoceptors. 7 We conclude that under conditions used in this study, (-)-RO363 is a potent partial agonist for human beta(1)- and beta(3)-adrenoceptors and appears also to activate the third human atrial beta-adrenoceptor. (-)-RO363 relaxes mammalian gut through both beta(1)- and beta(3)-adrenoceptors. (-)-RO363, used as a beta(1)-adrenoceptor selective tool, confirms previous findings with (-)-noradrenaline that beta(1)-adrenoceptor mediated atrial effects are only slightly enhanced by chronic treatment of patients with beta-blockers. Chronic treatment with beta(1)-adrenoceptor-selective blockers does not significantly increase the density of human atrial beta(1)- and beta(2)-adrenoceptors.