Trypanosoma cruzi: distinct patterns of infection in the sibling caviomorph rodent species Thrichomys apereoides laurentius and Thrichomys pachyurus (Rodentia, Echimyidae)

Thrichomys apereoides, a caviomorph rodent species common in a highly endemic area for Chagas disease in Brazil, may act as reservoir of the parasite. However, no information is available concerning its sibling species Thrichomys pachyurus, found in the Pantanal region, where Trypanosoma cruzi is found only in the enzootic cycle. We followed up the cross infection of these cryptic species with two isolates derived from naturally infected T. pachyurus and Thrichomys apereoides laurentius. No regional co-adaptation between Thrichomys species and the regional isolates were noticed. However, significant differences in the outcome of the infection were observed. T. a. laurentius was more resistant than T. pachyurus, as expressed by lower parasitemia and less histopathological damage. The routine biochemical markers used for laboratory rodents were unsuitable for follow up of infection in Thrichomys spp, since they did not correlate with the histopathological findings or allowed the kinetic follow-up of tissue colonization by the parasite.     

Differences in ingestive balance of two populations of neotropical Thrichomys apereoides (Rodentia, Echimyidae)

Thrichomys apereoides is widely distributed in the Caatinga, a semi-arid region in Brazil, but is presumed to lack capabilities for water conservation. In the present study, we compared two populations of adult individuals living under different precipitation conditions (700 and 450 mm year(-1)). Animals from the less dry area were twice as heavy as those from the drier locality. Under ad libitum water regimen, there were differences between populations in relative food intake as well as in water intake and urine concentration, but not in normalized body mass water intake. Under short-term water deprivation, both populations presented similar body mass loss. Whereas individuals from the more arid locality maintained food consumption, urine volume and urine osmolality, Thrichomys from the less dry locality reduced food consumption and urine volume. The occurrence of anuria in 75% of animals from this population indicates that the limits of their ability to deal with water shortage had been reached. The morphological and physiological difference and the non-allometric similarities found between the two populations of T. apereoides fulfill the criteria for physiological adaptations to differences in annual rainfall. Our data challenge the hypothesis that the irregularity of annual rainfall in the Caatinga precludes the evolution of adaptations to this semi-arid climate.     

Karyologic evidence of diversification of the genus Thrichomys (Rodentia,Echimyidae)

Karyologic analysis of Thrichomys specimens from different Brazilian localities, in Pantanal, Cerrado and Caatinga biomes, shows different chromosome complements. The 2n = 34, FN = 64 karyotype is found in Mato Grosso do Sul state; the 2n = 30, FN = 54 karyotype in Bahia, Pernambuco, Piauí and Ceará states; the 2n = 30, FN = 56 karyotype in Goiás and Tocantins states, the 2n = 28, FN = 50 karyotype in Minas Gerais state, the 2n = 28, FN = 52 and 2n = 26, FN = 48 karyotypes in Bahia state. Comparisons of G-band patterns allowed the identification of homologies shared by all karyotypes and show that the two karyotypes with the lowest diploid number (2n = 26 and 2n = 28) belong to two different evolutionary lineages. The most proper names for each karyomorphic population are: Thrichomys pachyurus for 2n = 34; Thrichomys apereoides apereoides for 2n = 28, FN = 50; Thrichomys apereoides laurentius for 2n = 30, FN = 54 and Thrichomys inermis for 2n = 26. Two karyotypes (2n = 28, FN = 52 and 2n = 30, FN = 56) could not be attributed to any subspecies. These different karyomorphotypes are allopatric and/or parapatric.     

Maintenance and breeding of Thrichomys (Trouessart, 1880) (Rodentia: Echimyidae) in captivity

South American histricognath rodents Thrichomys apereoides laurentius and Thrichomys pachyurus are natural hosts of Trypanosoma cruzi, agent of Chagas disease. We established breeding colonies of these species to serve as experimental models in various parasitological studies. Both species of Thrichomys have all the requirements necessary to become excellent laboratory models: they can be easily maintained in the standard laboratory conditions and breed throughout the year and they do not have any special dietary demands and can be fed by standard food pellets designed for laboratory mice. Both species produce precocious offspring that have their eyes and ears open, teeth erupted, fur well developed, and can eat solid food in the first week of life. T. a. laurentius has larger litter sizes and lower body masses at birth and weaning than T. pachyurus. Moreover, females of T. a. laurentius reach puberty earlier and with lower body mass than T. pachyurus.     

Trypanosoma cruzi in a caviomorph rodent: parasitological and pathological features of the experimental infection of Trichomys apereoides (Rodentia, Echimyidae)

To understand the interaction of Trypanosoma cruzi with caviomorph rodents, which supposedly have an ancient co-evolutionary history with this parasite, experimental infection of laboratory reared Trichomys apereoides with several isolates of both genotypes of the parasite was studied. Parasitemia, pattern of hematic cells, specific humoral immune response, histopathological features and parasite clearance were appraised. T. apereoides maintained stable infections independent of the T. cruzi genotype as demonstrated by positive PCR results in analyses of several tissues after a 5 months follow-up. The acute phase was characterized by abundant and disseminated presence of amastigotes, vacuolization and/or myocytolysis. Lymphocytosis was a common feature. The chronic phase was characterized mainly by lymphomacroeosinophilic infiltrates independent of the inoculated T. cruzi isolate. T. cruzi of different genotypes did not show any tissular preference in T. apereoides.     

Variation in the skull morphometry of four taxonomic units of Thrichomys (Rodentia: Echimyidae), from different Neotropical biomes

The echimyid rodents of the genus Thrichomys vary considerably in their behavior and feeding ecology, reflecting their occurrence in environments as different as the Caatinga, Cerrado, Pantanal, and Chaco biomes. While the genus was originally classified as monospecific, a number of Thrichomys species have been recognized in recent decades, based on morphometric, cytogenetic, and molecular analyses. While Thrichomys is well studied, the variation found in its cranial morphology is poorly understood, given the taxonomic and ecological complexities of the genus. Using a geometric morphometric approach, we characterized the differences found in the cranial morphology of four Thrichomys taxonomic units, including three established species, Thrichomys apereoides, Thrichomys fosteri, and Thrichomys laurentius, and one operational taxonomic unit (OTU), Thrichomys aff. laurentius. No significant differences were found among these units in cranium size, but significant variation was found in skull shape. The Procrustes distances provided a quantification of the differences in the shape of the skull, with the largest distances being found between T. aff. laurentius and T. fosteri in the dorsal view, and between T. aff. laurentius and T. apereoides in the ventral view. A Discriminant Function Analysis (DFA) with cross-validation determined that the pairings with the highest correct classification were T. aff. laurentius vs. T. apereoides and T. aff. laurentius vs. T. fosteri, in both views. The principal variation in skull shape was found in the posterior region and the zygomatic arch, which may be related to differences in diet.     

Reproductive isolation of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) in southern Spain

The phenology, reproductive cycle and cone crop of two sympatric subspecies ofJuniperus phoenicea (Cupressaceae) were studied during three consecutive years in southern Spain. The flowering pattern of each subspecies was constant during this period: inJ. phoenicea subsp.turbinata flowering was always in October–November, whileJ. phoenicea subsp.phoenicea flowered in February–March. There was no overlap between the flowering periods of the two taxa. The reproductive cycle of the two subspecies was similar, though there were some important differences because of their flowering times. Male cone development in both subspecies occurred in autumn, but the male cones ofJ. phoenicea subsp.phoenicea did not shed pollen until the end of February. InJ. phoenicea subsp.turbinata, the gap between pollination and fertilisation was seven months while inJ. phoenicea subsp.phoenicea it was only three months, fertilisation taking place at the same time in the two taxa. Post-fertilisation events were similar in the two subspecies, leading to a cycle of two years inJ. phoenicea subsp.turbinata and a shorter one of about 20 months inJ. phoenicea subsp.phoenicea. Alternating good and slight seed cone crops were found in both taxa during the three years studied, the good crops of one subspecies coinciding with the slight crops of the other one. Although the proximity of the two subspecies and method of pollination could make hybridisation between the taxa possible, the different flowering seasons prevent hybridisation isolating them reproductively.     

Molecular and karyologic variation in the genus Isothrix (Rodentia, Echimyidae)

Phylogenetic relationships among available samples of the genus Isothrix using mitocondrial gene cytochrome b were carried out and a new karyotype of Isothrix specimens referred to Isothrix negrensis from the mid course of Rio Negro, in the Brazilian Amazon, was described. Molecular, karyological and morphological data of Isothrix negrensis was compared to that of I. bistriata and I. pagurus. Cytochrome b DNA sequence analyses clearly separate I. negrensis from I. bistriata and genetic distances estimates between then are greater than between species of the related genus Mesomys. The median-joining analysis postulated the presence of two median vectors between I. bistriata haplotypes from adjacent localities, suggesting that genetic isolation between them is unrelated to geographic distance. These results confirm previous molecular differences suggesting that I.bistriata is a composite of several taxa. The karyotype of Isothrix negrensis also differs from those reported for I. bistriata and I. pagurus. In relation to the external morphology I. negrensis differs from I. bistriata by the overall darker dorsal coloration, head darker than dorsum, presence of a longitudinal line of blackish hairs in the proximal part of the tail extending until near to its base, dorsal surface of hind feet covered with yellowish hairs and stripe of light hairs in the head reduced to a small patch.     

Physical and ecological isolation contribute to maintain genetic differentiation between fire salamander subspecies

Landscape features shape patterns of gene flow among populations, ultimately determining where taxa lay along the continuum between panmixia to complete reproductive isolation. Gene flow can be restricted, leading to population differentiation in two non-exclusive ways: “physical isolation”, in which geographic distance in combination with the landscape features restricts movement of individuals promoting genetic drift, and “ecological isolation”, in which adaptive mechanisms constrain gene flow between different environments via divergent natural selection. In central Iberia, two fire salamander subspecies occur in parapatry across elevation gradients along the Iberian Central System mountains, while in the adjacent Montes de Toledo Region only one of them occurs. By integrating population and landscape genetic analyses, we show a ubiquitous role of physical isolation between and within mountain ranges, with unsuitable landscapes increasing differentiation between populations. However, across the Iberian Central System, we found strong support for a significant contribution of ecological isolation, with low genetic differentiation in environmentally homogeneous areas, but high differentiation across sharp transitions in precipitation seasonality. These patterns are consistent with a significant contribution of ecological isolation in restricting gene flow among subspecies. Overall, our results suggest that ecological divergence contributes to reduce genetic admixture, creating an opportunity for lineages to follow distinct evolutionary trajectories.Subject terms: Evolutionary ecology, Speciation     

Chorioallantoic and yolk sac placentation in Thrichomys laurentinus (Echimyidae) and the evolution of hystricognath rodents

The evolutionary history of Hystricognathi is associated with major transformations in their placental system. Data so far indicate that key characters are independent from size dimensions in medium to very large species. To better understand the situation in smaller species, we analyzed placental development in a spiny rat, Thrichomys laurentinus. Fourteen individuals ranging from early implantation to near term were investigated by histology, immunohistochemistry, proliferation activity and electron microscopy. Placentation in Thrichomys revealed major parallels to the guinea pig and other hystricognath rodents with respect to the early and invasive implantation, the process of trophoblast invasion, the internal organization of the labyrinth and the trophospongium as well as the establishment of the complete inverted yolk sac placenta. In contrast to systematically related small-sized species, the placental regionalization in Thrichomys was characterized by a remarkable lobulated structure and associated growing processes. Reverse to former perspectives, these conditions represented ancient character states of hystricognaths. The subplacenta was temporarily supplied by both the maternal and fetal blood systems, a rare condition among hystricognaths. The extraplacental trophoblast originating from the subplacenta was partly proliferative in mid gestation. In conclusion, the presented results indicated that only minor variations occurred in small-sized hystricognath species, independent of their systematic interrelationships. Previous views were supported that placentation in hystricognaths followed an extraordinary stable pattern, although the group had distinct habitats in South America and Africa that were separated 30-40 million years ago.     

Chromosome differentiation of spiny rats of the genus Proechimys (Rodentia, Echimyidae)]

Karyological analysis of spiny rats of the genus Proechimys revealed four chromosome forms: 2n = 30; 2n = 29; 2n = 28; 2n = 24. 30-chromosome spiny rats are characterized by unique karyotype constitution. 27-chromosome spiny rats from the territory near Iquitos differ from the same of Pucalpa (Ucayali Department) by Y-chromosome structure and banding of long arms with two pairs of autosomes. 24-chromosome rats from the regions mentioned above do not differ in G-banded chromosomes. Diploid chromosome set equal to 29, has a family originated from the parents which had 2n = 30 and 2n = 28 under the laboratory conditions. The absence of clear morphological differentiation, similarity of constituent parts of the karyotype and hybridization possibility point to relatively recent origination of the chromosome forms under study.     

Thrichomys laurentius (Rodentia; Echimyidae) as a Putative Reservoir of Leishmania infantum and L. braziliensis: Patterns of Experimental Infection

The importance of the genus Thrichomys in the retention of infection and transmission of Leishmania species is supported by previous studies that describe an ancient interaction between caviomorphs and trypanosomatids and report the natural infection of Thrichomys spp. Moreover, these rodents are widely dispersed in Brazil and recognized as important hosts of other tripanosomatids. Our main purpose was to evaluate the putative role of Thrichomys laurentius in the retention of infection and amplification of the transmission cycle of Leishmania infantum and L. braziliensis. Male and female T. laurentius (n = 24) born in captivity were evaluated for the retention of infection with these Leishmania species and followed up by parasitological, serological, hematological, biochemical, histological, and molecular assays for 3, 6, 9, or 12 months post infection (mpi). T. laurentius showed its competence as maintenance host for the two inoculated Leishmania species. Four aspects should be highlighted: (i) re-isolation of parasites 12 mpi; (ii) the low parasitic burden displayed by T. laurentius tissues; (iii) the early onset and maintenance of humoral response, and (iv) the similar pattern of infection by the two Leishmania species. Both Leishmania species demonstrated the ability to invade and maintain itself in viscera and skin of T. laurentius, and no rodent displayed any lesion, histological changes, or clinical evidence of infection. We also wish to point out the irrelevance of the adjective dermotropic or viscerotropic to qualify L. braziliensis and L. infantum, respectively, when these species are hosted by nonhuman hosts. Our data suggest that T. laurentius may act at least as a maintenance host of both tested Leishmania species since it maintained long-lasting infections. Moreover, it cannot be discarded that Leishmania spp. infection in free-ranging T. laurentius could result in higher parasite burden due the more stressing conditions in the wild. Therefore the tissular parasitism of the skin, infectiveness to the vector, and amplification of the transmission cycle of both Leishmania species could be expected.     

Disruption of genetic interaction between two autosomal regions and the X chromosome causes reproductive isolation between mouse strains derived from different subspecies

Reproductive isolation that initiates speciation is likely caused by incompatibility among multiple loci in organisms belonging to genetically diverging populations. Laboratory C57BL/6J mice, which predominantly originated from Mus musculus domesticus, and a MSM/Ms strain derived from Japanese wild mice (M. m. molossinus, genetically close to M. m. musculus) are reproductively isolated. Their F1 hybrids are fertile, but successive intercrosses result in sterility. A consomic strain, C57BL/6J-ChrX(MSM), which carries the X chromosome of MSM/Ms in the C57BL/6J background, shows male sterility, suggesting a genetic incompatibility of the MSM/Ms X chromosome and other C57BL/6J chromosome(s). In this study, we conducted genomewide linkage analysis and subsequent QTL analysis using the sperm shape anomaly that is the major cause of the sterility of the C57BL/6J-ChrX(MSM) males. These analyses successfully detected significant QTL on chromosomes 1 and 11 that interact with the X chromosome. The introduction of MSM/Ms chromosomes 1 and 11 into the C57BL/6J-ChrX(MSM) background failed to restore the sperm-head shape, but did partially restore fertility. This result suggests that this genetic interaction may play a crucial role in the reproductive isolation between the two strains. A detailed analysis of the male sterility by intracytoplasmic sperm injection and zona-free in vitro fertilization demonstrated that the C57BL/6J-ChrX(MSM) spermatozoa have a defect in penetration through the zona pellucida of eggs.     

Tooth morphology of Echimyidae (Rodentia,Caviomorpha): homology assessments,fossils, and evolution

Echimyidae constitute the most important radiation of caviomorph rodents in the Neotropical region, represented by 20 extant genera and several extinct species. Both in extant and fossil forms, this diversity is reflected by a significant morphological variation found in crown structures of the cheek teeth. Different hypotheses of primary homology have been proposed for these structures, which, in turn, support diverse dental evolutionary hypotheses. In this contribution we inspect the main structures (cusps and lophids) of the lower deciduous teeth and molars in extinct and extant Echimyidae, and establish their topological correspondences. Comparisons with cusps and lophids of Erethizontidae are emphasized. We explore the testing of alternative primary hypotheses of lophid correspondences in a cladistic context. Following a ‘dynamic’ approach, we select the hypothesis of primary homology, which produced the more parsimonious results, and evaluate the evolutionary transformations of the dental characters analysed. In this context, the phylogenetic relationships of living Myocastor coypus (Molina, 1782) with the extinct Tramyocastor and Paramyocastor are tested. Our results indicate that pentalophodonty is the derived condition for the lower molars in Echimyidae, that trilophodonty evolved independently at least three times during the evolutionary history of these rodents, and that tetralophodonty represents the plesiomorphic condition. This study shows that dental evolution in echimyids can be better understood when occlusal structures are expressed as reliably comparable characters, and when fossils are taken into account. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 451–480.     

Morphological evolution in the mandible of spiny rats, genus Trinomys (Rodentia: Echimyidae)

The development and evolution of the rodent mandible have been studied in depth in recent years. The mandible is a complex structure because it consists of six morphogenetic components formed by different condensations of mesenchymal cells. Using recent techniques for the geometric analysis of shape, we have combined developmental information with a powerful quantification of shape variation and an independent estimate of phylogeny (molecular data) to assess the evolutionary patterns of shape change in mandibles of the rodent genus Trinomys . In general, the major trends in shape variation did not agree with the expected phylogenetic pattern. However, for small-scale morphological differences, one species ( T. yonenagae ) was responsible for the lack of association between morphology and molecular divergence. This species is genetically similar to but morphologically different from other Trinomys . The coronoid process was considered to be the most conservative morphogenetic component in the mandible.     

Reproductive and habitat isolation between two populations of the striated spined loach

Synopsis This study deals with ecological differences influencing reproductive isolation between two sympatric populations, the small and middle forms, of the striated spined loach. As natural hybrids are rare, isolation between these populations must be complete. There are some differences in their reproduction and early life history. Both reproduce in temporary waters flooded in summer, but their spawning sites are isolated from each other. The small form spawns in shallow areas of still water around irrigation creeks or fens. On the other hand, the middle form spawns in slow-flowing ditches. Spawning site segregation between them is an important isolating mechanism, and is a by-product of adaptation in their early life history.     

Karyological analysis of Proechimys cuvieri and Proechimys guyannensis (Rodentia, Echimyidae) from central Amazon

The aim was to characterize the karyotype of rodents of the genus Proechimys from three localities in the central Brazilian Amazon, in the search for new markers that might shed light on our understanding of the taxonomy and evolutionary history of this taxon. Two karyotypes were found, viz., 2n = 28, FN = 46 in individuals from the NRSP (Cuieiras River) and REMAN (Manaus), and 2n = 46, FN = 50 in individuals from the Balbina Hydroelectric Plant. While individuals with the karyotype with 2n = 28 chromosomes were morphologically associated with Proechimys cuvieri, their karyotype shared similarities with those of the same diploid number in two other regions. Although three karyotypes are described for Proechimys cuvieri, no geographic distribution pattern that defined a cline could be identified. Based on the morphological examination of voucher specimens and additional results from molecular analysis, the karyotype with 2n = 46 and FN = 50 could be associated with P. guyannensis.     

Molecular characterization and mitochondrial sequence variation in two sympatric species of Proechimys (Rodentia: Echimyidae) in French Guiana

Variations in mitochondrial DNA characters were used to characterize two morphologically similar and sympatric species of Neotropical terrestrial rodents of the genus Proechimys (Mammalia: Echimyidae). We sequenced both cytochrome b (1140 pb) and part of the control region (445 pb) from four individuals of P. cuvieri and five of P. cayennensis from French Guiana, which allowed us to depict intra- and inter-specific patterns of variation. The phylogenetic relationships between the nine sequences evidence the monophyly of each species, and illustrate that more polymorphism might exist within P. cuvieri than within P. cayennensis. By developing species-specific primers to amplify a fragment of the cytochrome b gene, we were able to identify 50 individuals of Proechimys spp. caught in two localities of French Guiana. In both sites of primary rainforests, we showed that the two species live in syntopy, and this observation emphasizes the need to document ecological differences which should exist in order to diminish inter-specific competition.