Epithalamus of the nine-banded armadillo, Dasypus novemcinctus

The epithalamus of embryonic, neonatal and adult nine-banded armadillo (Dasypus novemcinctus) was examined for evidence of pineal-like tissue. The evagination of the diencephalic roof (the anlage of the epiphysis) was not found in any embryonic specimens. In the adult brain, the epithalamus is dominated by the sub-commissural organ (SCO) which is surrounded entirely by the posterior commissure. At the most caudal aspect of the SCO, previous investigators have observed pineal-like tissue. Using pineal-specific staining techniques however, we found no evidence of this tissue. Because the armadillo produces melatonin in a rhythmic manner, exhibits exacting circadian rhythms, and shows altered rhythms when exposed to exogenous melatonin, we believe other organs, perhaps the retina or Harderian gland, must be involved in maintaining the coordinated melatonin titer.     

The chromosomes of the nine-banded armadillo,Dasypus novemcinctus

Summary The chromosomes of the nine-banded armadillo, Dasypus novemcinctus, were studied using cultured peripheral leukocytes, tissue cells and direct preparation of male germ cells.The diploid number was found to be 64 in the blood and tissue cells, the haploid number was 32 in the testicular preparations.The autosomes can be divided into six groups, according to their size and the position of the centromere. Group A contains two pairs of large metacentrics, Groups B and C contain together 18 pairs of acrocentric chromosomes, the distinguishing feature being a gradual decrease in size. Group D is composed of six pairs of small metacentric chromosomes with almost medial centromeres. One prominentld satellited chromosome constitutes a ready marker element of this groud and of the karyotype. It is not entirely certain whether this satellitey chromosome has a homologue or not. Five pairs of small acrocentrics constitute Group E. These are almost indistinguishable from one another.Buccal smears showed a typical sex chromatin body in interphase nuclei of epithelial cells from female animals similar to that observed in some other mammals. A sex difference was also noted on examination of peripheral blood films, typical drumsticks being present on polymorphonuclear leukocytes from female armadillos.     

Helminth fauna of the nine-banded armadillo (Dasypus novemcinctus) in north-central Florida

The gastrointestinal tracts of 32 free-ranging, 9-banded armadillos (Dasypus novemcinctus) from north-central Florida were examined for the presence of helminths during July 1991 to November 1993. Aspidodera sp. (Nematoda: Aspidoderidae), most closely resembling Aspidodera sogandaresi, were recovered from 20 of 32 armadillos examined. Total numbers of A. sogandaresi ranged from 1 to 1,021 per infected animal, and followed an inverse correlation to body condition index for those animals. The cystacanth stage of 1 acanthocephalan, Macracanthorhynchus ingens, was present in 1 armadillo, and is the first report of M. ingens in the 9-banded armadillo. The present study contributes to the known natural history of the 9-banded armadillo, an important animal research model.     

Determination of reproductive maturity in the female nine-banded armadillo (Dasypus novemcinctus)

At 3-month intervals from birth to 27 months of age, 3 female armadillos were killed. The number and size of follicles greater than 202 micron were determined, plasma progesterone concentration was measured, and values were correlated with age. Blood samples were taken monthly by femoral vein puncture and plasma was analysed by radioimmunoassay for progesterone concentration. At necropsy, both ovaries were visually inspected for a corpus luteum, weighed and then processed for routine histology. The number of normal, antral follicles greater than 202 micron were counted. These follicles were arbitrarily categorized into 15 different size groups (every 77 micron). Total number of follicles greater than 202 micron varied from 15.5 +/- 1.3 at 15 months of age to 26.3 +/- 1.9 at 21 months. Follicles of a size (greater than 978 micron) most likely to ovulate were present only at greater than or equal to 9 months of age. Progesterone values remained below the adult concentration (5 ng/ml) until 15 months of age. A concentration of progesterone indicative of ovulation (approximately 10 ng/ml) occurred between 17 and 20 months of age. The findings of the present study demonstrate that the female armadillo is reproductively mature after 15 months of age.     

Cold-induced fever and peak metabolic rate in the nine-banded armadillo (Dasypus novemcinctus)

In response to cold exposure, some mammals, including the nine-banded armadillo (Dasypus novemcinctus), exhibit an increase in core temperature. This response, which can be qualified as a cold-induced fever, could increase cold tolerance by increasing peak metabolic rates because of the Q(10) effects. This hypothesis, however, is not compatible with the observation that peak core temperature can occur up to 100 min before peak metabolic rate in nine-banded armadillos during acute exposure to cold heliox (79% He; 21% O(2)). This temporal separation between the timing of peak metabolic rate and core temperature could be the result of regional heterothermy, of the confounding effects of activity, or of using heliox as a respiratory gas. We tested these potential sources of error by exposing nine-banded armadillos to cold air while simultaneously monitoring behavior, metabolic rates (V dot o2 and V dot co2), and four core temperatures. Cold air exposure resulted in a smaller but significant temporal separation, with peak core temperature occurring on average 10 min before peak metabolic rate. Animals exhibited low activity levels, and the four core temperatures changed according to the same temporal pattern, thus eliminating the possibility that activity or regional heterothermy caused the temporal separation. Using a conceptual model, we propose that the temporal separation resulted from a rate of cooling that was too fast.     

The nine-banded armadillo (Dasypus novemcinctus) is naturally infected with Sarcocystis neurona

Sarcocysts were dissected from the tongue of a nine-banded armadillo (Dasypus novemcinctus). DNA was extracted and characterised by PCR amplification followed by restriction fragment length polymorphism analysis and nucleotide sequencing. A total of 1879 nucleotides were compared; the sarcocyst DNA sequence was identical to that reported for Sarcocystis neurona. DNA was extracted from the sarcocysts of five more nine-banded armadillos. A 254-nucleotide sequence was determined for each and found to be identical to S. neurona. Western blot techniques for detection of anti-S. neurona antibody were developed for use with armadillo plasma and samples from 19 wild-caught and 17 captive-raised armadillos were examined. Whereas all of the 19 wild-caught armadillos had antibodies to S. neurona, only one of 17 captive-raised armadillos did. These results suggest that the nine-banded armadillo are naturally infected with S. neurona.     

The nine-banded armadillo (Dasypus novemcinctus) is an intermediate host for Sarcocystis neurona

The nine-banded armadillo (Dasypus novemcinctus) is an intermediate host of at least three species of Sarcocystis, Sarcocystis dasypi, Sarcocystis diminuta, and an unidentified species; however, life cycles of these species have not been determined. Following feeding of armadillo muscles containing sarcocysts to the Virginia opossum (Didelphis virginiana), the opossums shed sporulated Sarcocystis sporocysts in their faeces. Mean dimensions for sporocysts were 11.0x7.5 microm and each contained four sporozoites and a residual body. Sporocysts were identified as Sarcocystis neurona using PCR and DNA sequencing. A 2-month-old foal that was negative for S. neurona antibodies in the CSF was orally inoculated with 5x10(5) sporocysts. At 4 weeks post-infection, the foal had a 'low positive' result by immunoblot for CSF antibodies to S. neurona and by week 6 had a 'strong positive' CSF result and developed an abnormal gait with proprioceptive deficits and ataxia in all four limbs. Based on the results of this study, the nine-banded armadillo is an intermediate host of S. neurona.     

Expression of nine-banded armadillo (Dasypus novemcinctus) interleukin-2 in E. coli

The nine-banded armadillo (Dasypus novemcinctus) is the only immunologically intact animal that regularly develops lepromatous-type leprosy when inoculated with Mycobacterium leprae. However, the ability to exploit this model for understanding the pathogenesis of leprosy has been limited by a lack of suitable immunological reagents. Recently, efforts began to sequence the entire armadillo genome, and this sequence information will help make possible the development of a wide array of new immunological reagents suitable for use with armadillos. Using the available sequence data, a region of high homology to interleukin-2 of other mammals was identified. Primers were designed to amplify the coding region corresponding to the mature peptide and its exact sequence was confirmed. cDNA was made from ConA-stimulated armadillo PBMC. The amplified coding region was sub-cloned into a pET expression vector and transformed into Escherichia coli for over-expression. The subsequent product was characterized by SDS-PAGE and bioassays. Tritiated thymidine incorporation by CTLL-2 and armadillo lymphoblasts confirmed functionality of the recombinant product. The advent of the D. novemcinctus genome sequence and subsequent generation of immunological tools will assist in advancing the armadillo as a translational model for leprosy.     

A novel abundant family of retroposed elements (DAS-SINEs) in the nine-banded armadillo (Dasypus novemcinctus)

About half of the mammalian genome is composed of retroposons. Long interspersed elements (LINEs) and short interspersed elements (SINEs) are the most abundant repetitive elements and account for about 21% and 13% of the human genome, respectively. SINEs have been detected in all major mammalian lineages, except for the South American order Xenarthra, also termed Edentata (armadillos, anteaters, and sloths). Investigating this order, we discovered a novel high-copy-number family of tRNA derived SINEs in the nine-banded armadillo Dasypus novemcinctus, a species that successfully crossed the Central American land bridge to North America in the Pliocene. A specific computer algorithm was developed, and we detected and extracted 687 specific SINEs from databases. Termed DAS-SINEs, we further divided them into six distinct subfamilies. We extracted tRNA(Ala)-derived monomers, two types of dimers, and three subfamilies of chimeric fusion products of a tRNA(Ala) domain and an approximately 180-nt sequence of thus far unidentified origin. Comparisons of secondary structures of the DAS-SINEs' tRNA domains suggest selective pressure to maintain a tRNA-like D-arm structure in the respective founder RNAs, as shown by compensatory mutations. By analysis of subfamily-specific genetic variability, comparison of the proportion of direct repeats, and analysis of self-integrations as well as key events of dimerization and deletions or insertions, we were able to delineate the evolutionary history of the DAS-SINE subfamilies.     

Recent range expansion and distributional limits of the nine-banded armadillo (Dasypus novemcinctus) in the United States

Abstract. Prior to the mid 1850s armadillo dispersal may have been inhibited by native subsistence hunters, the Rio Grande acting as a physical hindrance, and natural habitat barriers posed by fire-maintained grasslands. European colonists settled south Texas in mass during the latter half of the 19th century and largely removed existing impediments to armadillo range expansion, effectively releasing the species into the United States. The speed of the armadillo's natural range expansion after 1850 was probably accelerated by human travel and commerce into and out of its historic range in south Texas on a proliferation of roadways and railroads. This translocation process will likely continue and, combined with natural dispersal, will provide the armadillo the opportunity to ultimately become established in any habitats in the United States in which it can survive.
Limits to future distribution will likely be determined by climatic factors, and may be bounded by regions receiving at least 38 cm annual precipitation and having mean January temperatures >−2°C, or fewer than 24 total annual freeze days. Armadillos may have already approached a precipitation-defined boundary to the west and a distribution on the Great Plains limited by winter temperature minima. They may be expected to continue their advance up the East Coast to about the region of 41° North Latitude, decreasing to 39° North Latitude across the midwestern states. Future introductions on the West Coast may result in the establishment of armadillo populations in suitable habitats from California to Washington, an event that has already taken place with another tropical mammal emigrant, the opossum.     

The convergent evolution of the secondary structure of mitochondrial cysteine tRNA in the nine-banded armadillo Dasypus novemcinctus

The case of the convergent loss of the D-hairpin in mitochondrial cysteine tRNA of the nine-banded armadillo Dasypus novemcinctus is described. This evolutionary event sheds light on the molecular structure-function relationship and on the effect of this relationship on the processes of evolution of biopolymers and macromolecules.     

Pathogenicities and GP43kDa gene of three Paracoccidioides brasiliensis isolates originated from a nine-banded armadillo Dasypus novemcinctus )

We studied three different isolates of Paracoccidioides brasiliensis obtained from the mesenteric lymph node (D3LY1), the spleen (D3S1) and the liver (D3LIV1) of the same armadillo ( Dasypus novemcinctus ).Pulmonal inflammatory area was evaluated by intravenous inoculation of 10⁶ yeast cells of each isolates in young, male, ddY mice. Moreover, the partial sequence of GP43kDa gene of P. brasiliensis was analyzed. The lung inflammatory area was greater in animals inoculated with isolate D3S1. The partial sequence of GP43kDa gene indicated that isolate D3S1 is different from isolates D3LY1 and D3LIV1. This study suggested that the same armadillo might be susceptible to multiple P. brasiliensis isolates simultaneously. This revised version was published online in August 2006 with corrections to the Cover Date.     

Expression and characterization of recombinant interferon gamma (IFN-gamma) from the nine-banded armadillo (Dasypus novemcinctus) and its effect on Mycobacterium leprae-infected macrophages

Armadillos (Dasypus novemcinctus) manifest the full histopathological spectrum of leprosy, and are hosts of choice for in vivo propagation of Mycobacterium leprae. Though potentially useful as a model of leprosy pathogenesis, few armadillo-specific reagents exist. We have identified a region of high homology to the interferon gamma (IFN-γ) of other mammals within the recently published armadillo whole genomic sequence. cDNA was made from ConA-stimulated armadillo peripheral blood mononuclear cells (PBMC), amplified, and cloned into a pET expression vector for transformation and over-expression in Escherichia coli. The recombinant protein (rDnIFN-γ) was characterized by western blot and its biological function confirmed with bioassays including intracellular killing of Toxoplasma gondii and induction of indoleamine 2, 3-dioxygenase activity. In using rIFN-γ to activate macrophages from mice, humans or armadillos, similar to humans, rIFN-γ-activated armadillo MΦ did not produce nitrite and or inhibit the viability of M. leprae in vitro. Conversely, murine rIFN-γ-activated mouse MΦ produced high levels of nitrite and killed intracellular M. leprae in vitro. These data indicate that the response of armadillo MΦ to rDnIFN-γ is similar to that which occurs in humans, and demonstrates a potentially important value of the armadillo as a model in leprosy research.     

Stress-induced adrenal changes and their relation to reproductive failure in captive nine-banded armadillos (Dasypus novemcinctus)

Histological features of adrenal glands from captive and unstressed free-ranging nine-banded armadillos were compared to determine if undefined stress factors associated with captivity were capable of causing changes in adrenal morphology. Animals in captivity less than 3 mo showed histological features identical to unstressed free-ranging animals. Animals in captivity 6 mo or longer, however, showed adrenal changes like those previously described in other mammals exposed to various types of stress, except that there was no accompanying increase in adrenal weight. Similar adrenal changes were found in free-ranging animals exposed to extremely harsh winter weather. Females from both the chronic captive population and this stressed free-ranging population were seen to suffer reproductive failure. Reproductive function in males was not affected. There was no detectable sex difference in gross or histological adrenal changes; however, previous reports indicate there is a sex difference in the secretory product from the adrenals of captive nine-banded armadillos. A possible relationship between reproductive failure in the female and the sex difference in adrenal secretory products is discussed.     

Isolation of Paracoccidioides brasiliensis from the nine-banded armadillo Dasypus novemcinctus, in an endemic area for paracoccidioidomycosis in Colombia

The microniche of the dimorphic fungus Paracoccidiodes brasiliensisremains undefined in spite of the many attempts to isolate it from natural sources. Until recently, knowledge was also scanty concerning the presence of natural infections in animals; however, in the last decade, the fungus has been repeatedly isolated from the nine-banded armadillo, Dasypus novemcinctusin Brazil. A study aimed at determining the presence of infected armadillos in one of the paracoccidioidomycosis endemic areas of Colombia (Manizales, Department of Caldas) was undertaken. Based on the records of paracoccidioidomycosis patients available in the regional hospital, we selected a locality corresponding to a permanent resident, and found that it also had armadillo's burrows. Counting with the proper authorization, two animals were captured, sacrificed under prolonged anaesthesia and various internal organs cultured in mycological media. PCR with specific P. brasiliensis'primers was also done. The fungus was isolated from the mesenteric lymph node of one of the animals; fungal DNA amplification was positive in the same specimen as well as in the liver. The isolate from the Colombian armadillo indicates that these animals are regular hosts to P. brasiliensis in at least two endemic countries. Due to the restricted life pattern of these mammals they represent an important link with the natural habitat of the fungus. Consequently, a study of their movements and habits could prove rewarding in the search for this habitat.     

Survey for leprosy in nine-banded armadillos (Dasypus novemcinctus) from the southeastern United States

Ears from 853 nine-banded armadillos (Dasypus novemcinctus) from Alabama, Arkansas, Florida, Georgia and Mississippi were examined microscopically for evidence of leprosy. All were negative for both acid-fast bacteria (Mycobacterium leprae) and lesions compatible with leprosy.