Identification of bloodmeal sources of triatomines captured in the Paraguayan Chaco region of South America by means of molecular biology analysis.

The Paraguayan Chaco is an isolated environment with its own unique ecosystem. In this region, Chagas disease remains a health problem. Chagas disease is caused by the parasite Trypanosoma cruzi, and it is primarily transmitted by triatomines. In order to identify the blood meal sources of triatomines, specimens of the vector were collected in domestic and peridomestic areas and the PCR-RFLP method was implemented. Cytochrome b was amplified from the samples and later subjected to digestion with two restriction enzymes: Hae III and Xho I.It was possible to generate distinct restriction patterns on the amplified material to identify several blood meal sources for the vectors. We employed the blood from several species as positive controls: human, chicken, canine, feline, and armadillo blood. However, we identified only 3 sources for the blood meals of the insect vectors: human, chicken and canine blood. In total, 76 triatomines were captured. T. cruzi was not found in any of them. In 61% of the captured specimens, the blood meal sources for the vectors could be identified. In 30% of these cases, the presence of DNA from more than one vertebrate was detected in the same triatomine. The most common blood meal source found was chicken blood. The presence of human and chicken blood in triatomines captured in domestic and peridomestic areas strongly suggests that the parasite can freely move amongst both areas regardless of food availability. Free vector movement in these areas constitutes an epidemiological threat for the inhabitants of the community under study.

Survey of Leptospira spp. and Brucella abortus in Free-Ranging Armadillos from Pantanal, Brazil.

Out of the 20 recognized species of armadillos in the world, 11 are found in Brazil, and five of them are found in Pantanal, one of the world's largest wetlands. Beef cattle (Bos taurus) farming is the main economic activity in this region, which promotes intense wildlife-livestock contact and increases the likelihood of pathogen exposure, including to agents with zoonotic and economic relevance. Previous studies demonstrated that several wildlife species in Pantanal have been exposed to Brucella abortus and Leptospira spp.; however, little is known regarding the exposure and/or prevalence of zoonotic pathogens in armadillos. We used conventional PCR, the rose Bengal test (RBT), and the microscopic agglutination test (MAT) to investigate the exposure to and infection by Brucella spp. and Leptospira spp. using blood samples from four species of armadillos: nine-banded armadillo (Dasypus novemcinctus, n=2), southern naked-tailed armadillo (Cabassous unicinctus, n=8), yellow armadillo (Euphractus sexcinctus, n=16), and giant armadillo (Priodontes maximus, n=22), captured in Nhecolândia, Pantanal, Brazil. Samples were PCR- and RBT-negative for Brucella spp. infection and exposure. However, MAT revealed a Leptospira spp. seroprevalence of 31% (5/16; 95% confidence interval [CI]=0.11-0.58) in yellow armadillo and 18% (4/22; 95% CI=0.05-0.40) in giant armadillo specimens to serogroups Autumnalis, Cynopteri, and Pomona, with titers ranging from 200 to 1,600. Our results contribute to the understanding of zoonotic pathogens in armadillos in Pantanal and reinforce the importance of wildlife health surveillance in this area.

Antibodies to Toxoplasma gondii, Leishmania spp., and Leptospira spp. in Free-Ranging Six-Banded Armadillos (Euphractus sexcinctus) from Northeastern Brazil.

Antibodies to pathogens of public health importance were investigated in 33 free-ranging six-banded armadillos (Euphractus sexcinctus) from Brazil. The frequency of seropositive animals for Toxoplasma gondii, Leishmania spp., and Leptospira spp. were two, three, and two, respectively.

Cytogenetic damage in peripheral blood cultures of Chaetophractus villosus exposed in vivo to a glyphosate formulation (Roundup).

Different concentrations of a glyphosate formulation, Roundup® Full II (66.2% glyphosate) were tested in culture peripheral blood of armadillo Chaetophractus villosus with cytogenetic biomarkers like mitotic index (MI), chromosomal aberrations (CA), sister chromatid exchange (SCE) and cell proliferation kinetics (CPK) by means of replication index. Adults animals of both sexes were exposed to RU at four concentrations ranging from 0.026 mL RU solution to 0.379 mL RU daily in oral treatment with the same volume (0.2 mL) during 7 days. We analyzed the induced damage at different times considering T0 as control value, one (T1), seven (T7) and 30 days (T30). One day after, only the higher concentration shows MI significant differences (p < 0.05), at T7 the frequency increases and at T30 it decreases reaching T0 values. The analysis of CA frequencies shows that only 0.106 mL RU/day exhibit significant differences vs T0 values. A great variability is expressed in the values of standard deviation (SD) and in the wide confidence intervals of the media. One day after treatments (T1) all four concentrations shows significant differences in SCE vs T0 values. Replication Index (RI) does not show significant differences. The dose-response behavior was not observed in either CA or SCE. The consistency of the findings obtained with the same biomarkers in vitro support the idea of expanding studies in order to characterize the risk doses for these mammals.

Presence of antibodies against Leptospira serovars in Chaetophractus villosus (Mammalia, Dasypodidae), La Pampa province, Argentina.

Leptospirosis is a zoonosis of worldwide distribution. The aim of this study was to examine the presence of antibodies against 21 Leptospira reactive serovars in Chaetophractus villosus in La Pampa province, Argentina, using the microscopic agglutination test (MAT). Pathologic changes compatible with leptospirosis and in situ detection of the agent by immunohistochemistry were studied in 24 and 3 individuals respectively. Only 35/150 (23.3%) serum samples had antibodies against Leptospira sp. Six percent of the samples reacted with serovar Canicola, 4.7% with serovar Castellonis, 1.3% with serovar Icterohemorrhagieae and 0.7% with serovar Hardjo. Sixteen (10.6%) serum samples agglutinated with Castellonis-Icterohemorrhagiae and Canicola-Castellonis serovars, both with 4.7%, and Canicola-Hardjo and Castellonis-Canicola-Icterohemorrhagiae both with 0.6%. Fourteen animals had variable degrees of lesions, which were more severe in animals with higher serological titers (3200), and Leptospira sp. was detected in 3 animals by immunohistochemistry. These results represent the first record of the presence of Leptospira in C. villosus in La Pampa.

Valores hematológicos del armadillo peludo (Chaetophractus nationi) en cautiverio en un parque zoológico de Lima, Perú / Hematological values of the hairy armadillo (Chaetophractus nationi) in captivity at one Zoological Park Lima, Perú

El objetivo del presente estudio fue determinar los valores hematológicos del armadillo peludo (Chaetophractus nationi) en cautiverio en un parque zoológico. Su distribución geográfica se ubica entre los 3 500 y 4 300 m.s.n.m., en los arenales de las mesetas andinas de Perú, Argentina, Bolivia y Chile. En la actualidad, su situación es vulnerable según la Unión Internacional para la Conservación de la Naturaleza (UICN). Siete armadillos adultos aparentemente sanos fueron sometidos a un estudio hematológico con el fin de determinar parámetros referenciales en esta población específica. El diseño estadístico incluyó la determinación del promedio y desviación estándar. Los valores obtenidos en promedio fueron: eritrocitos = 4 738 571,43 /ul; leucocitos = 6 064,29 /ul; hemoglobina = 13,30 g/dl; hematocrito = 36,43 %; neutrófilos = 60,86 %; linfocitos = 23,71 %; monocitos = 0,86 %; eosinófilos = 10,00 %; basófilos = 2,14 % y recuento de plaquetas = 267 285,71 /ul.

The purpose of the present study was to determine the hematological values of the hairy armadillo (Chaetophractus nationi) in captivity at the one Zoological Park. Its geographic distribution ranges between 3 500 to 4 300 m. a .s. l. in the Andean sand plates of Peru, Argentina, Bolivia y Chile. Actually its situation is vulnerable according to the International Union for Conservation of Nature (IUCN). Seven adult hairy armadillos apparently healthy were used for a hematological study in order to determinate the referential parameters for this species. The average and the standard deviation were determined Erythrocytes = 4 738 571,43 /ul; leucocytes = 6 064,29 /ul; hemoglobin = 13,30 g/dl; hematocrit = 36,43 %; neutrophils = 60,86 %; lymphocytes = 23,71 %; monocytes = 0,86 %; eosinophils = 10,00 %; basophiles = 2,14 % and platelets = 267 285,71 /ul.

Health assessment of free-ranging three-banded (Tolypeutes matacus) and nine-banded (Dasypus novemcinctus) armadillos in the Gran Chaco, Bolivia.

The Gran Chaco, Bolivia, has a total of seven species of armadillos with the three-banded (Tolypeutes matacus) and nine-banded (Dasypus novemcinctus) the most commonly hunted by the local Isoseño-Guarani people. Armadillos are known carriers of zoonotic pathogens, including Mycobacterium leprae, Toxoplasma gondii, and Trypanosoma cruzi; thus human handling and consumption of these species may have a significant public health impact. A health assessment that included physical examinations, hematology, plasma biochemical analyses, levels of exposure to selected infectious agents, and endoparasite and ectoparasite identification was performed on nine-banded and three-banded armadillos in the Gran Chaco, Bolivia. Based on clinical findings, the general health of these armadillos was rated as good. However, many of the nine-banded armadillos (64%) had abrasions and wounds, probably related to the capture method. The blood value results from a subset of these armadillos are presented as baseline values for free-ranging populations of both these species in Bolivia. Serologic antibody tests for M. leprae were negative in three-banded (n = 8) and nine-banded (n = 2) armadillos. Three-banded armadillos were antibody positive for Eastern equine encephalitis virus (8/8; 100%) and Saint Louis encephalitis virus (5/8; 62.5%). Two of 12 (16.7%) three-banded armadillos tested were antigen positive for Dirofilaria immitis. Nine-banded armadillos were antibody positive for T. gondii (3/9; 33.3%), Eastern equine encephalitis virus (5/8; 62.5%), and T. cruzi (2/9; 22.2%). Two of eight (25%) nine-banded armadillos were antigen positive for D. immitis. A number of endo- and ectoparasites were identified in/on both species of armadillos. Results from this study support the possibility that the handling and consumption of these species by the local Isoseño-Guarani people may have a public health impact.

Health evaluation of free-ranging and captive pichis (Zaedyus pichiy; Mammalia, Dasypodidae), in Mendoza Province, Argentina.

The health of free-ranging and captive pichis (Zaedyus pichiy) was assessed in Mendoza Province, Argentina, between November 2001 and December 2006. Postmortem examinations of 150 confiscated and vehicle-killed pichis and clinical examinations of 139 wild-caught individuals suggest that the wild populations are currently in good health. Lesions and scars were observed in a large proportion of wild-caught pichis. The most common lesions were associated with parasitism or parasite larva migration. Sarcocystis cysts were relatively common in the skeletal muscle, and Besnoitia cysts were observed in the lungs of 24 evaluated animals. Elevated ambient humidity levels often caused moist dermatitis with epidermal detachment in captive pichis. This report constitutes the first health evaluation of free-ranging and captive Z. pichiy. It will be a starting point for future health studies and will be beneficial for the captive management of this species.

Evaluation of endocytic capacity and NADPH-oxidase activity from armadillo (Dasypus novemcinctus) eosinophils infected with microfilariae.

Endocytic activity of phagocytic cells from armadillos infected with viruses, parasites or bacteria is unknown. This report shows that eosinophils from armadillos infected with microfilaria act against these helmintic parasites but have deficiencies in their oxygen-dependent bacteriocidal mechanisms and also in endocytic capacity against yeast.

Hematology and serum chemistry values in captive and wild pichis, Zaedyus pichiy (Mammalia, Dasypodidae).

As part of an ongoing study on the health status of pichis, Zaedyus pichiy (Mammalia, Dasypodidae), blood was collected under manual restraint from 72 free-ranging pichis captured in Mendoza Province, Argentina, between November 2001 and December 2006, and from 22 captive-kept pichis in January 2007. Reference values were established for hematology and serum chemistry. Pichis had lower leukocyte counts and higher mean corpuscular volumes than most other mammals. Blood values were similar for captive and wild pichis, and only a few significant differences were found among genders or age classes.

Lepra en el armadillo salvaje / No disponible

Los armadillos salvajes de nueve bandas (Dasypus novemcinctus) de la zona central y sur de Estados Unidos son huéspedes naturales y endémicos de Mycobacterium leprae. Los estudios llevados a cabo durante los últimos 30 años o más en alrededor de 5.000 animales confirman que la infección está presente en Arkansas, Louisiana, Mississippi y Texas. Los índices de prevalencia más elevada se encuentran entre los animales de las zonas aluviales de menor altura y áreas costeras, principalmente en Lousiana y Texas. Tanto la densidad poblacional de animales como los factores locales pueden contribuir a la que se detecte lepra en el armadillo de cualquiera de estas zonas de los EE.UU. y sólo hay pocos casos que describan la infección entre animales de América del Sur o Central. Sin embargo, este tema ha suscitado escaso interés en otros países. Los armadillos son relativamente recientes en los EE.UU. y la lepra ya estaba presente en Texas y Lousiana antes de su llegada. No está muy clara la relación ecológica entre los humanos y los armadillos. Sin embargo, los armadillos infectados son un gran reservorio de M. leprae y pueden ser fuente de infección para algunos individuos de este país y quizás en otras partes comprendidas dentro del alcance de estos animales

Wild nine-banded armadillos (Dasypus novemcictus) in the south central United States are highly endemic natural hosts of Mycobacterium leprae. Surveys conducted over the lass 30 years on more that 5.000 animals confirm that the infection is present among armadillos in Arkansas, Louisiana, Mississippi and Texas. Highest prevalence rates are found among the animals in low-lying alluvial and coastal areas, primarily in Lousiana and Texas. Both animal density and local factors may contribute to the detectability of armadillo leprosy in those regions. Little evidence for M.leprae infection is found among armadillos elsewhere in the US range, and only a few reports relate finding the infection among animals in Central or South America. However, the issue has received only scant attention in other countries. Armadillos only recently expanded their range into the Us, and leprosy was present in Texas and Lousiana prior to the arrival of armadillos. The ecological relationship between humans and armadillos with M. leprae in this region remains unclear. However, infected armadillos constitute a large reservoir of M. leprae and they may be a source of infection for some humans in this country, and perhaps in other locations across the animal´s range

A study of platelets in the armadillo chaetophractus villosus (xenarthra, dasypodidae).

Nothing is known about haemostasis in Xenarthra, a widely distributed Order of American mammalians. Chaetophractus villosus, a member of the Dasypodidae family of this group, which is easily adapted to captivity, is of growing interest for biomedical research. In this work, we studied platelet number, MPV, ultrastructure of the platelets by SEM and TEM, and aggregation responses to ADP and ristocetin in this species. Blood samples were obtained by cardiac puncture in 20 anaesthetised animals. Platelet count and MPV were evaluated at the beginning and at the end of a 3-year experimental period, to detect possible variations related to time of captivity. SEM and TEM were done by routine methods adapted to the material, and aggregation response to ADP and ristocetin were evaluated by the Born method. The parameters studied did not show any sex-related differences, nor did the platelet count change during captivity. Nevertheless, MPV decreased during this period. Platelets were ultrastructurally similar to those of other mammals and human beings and responded to proven agonists. Data provided in this study will contribute to the understanding of the haemostatic process in this species.

Major plasma lipids and fatty acids in four HDL mammals.

Lipid classes and their fatty acids were compared in plasma from four mammals: a laboratory rodent, the mouse; two domestic animals, the cat and dog; and a wild animal, the South American armadillo, Chaetophractus villosus. In all, the most abundant lipoprotein was high-density lipoprotein (HDL). In the total lipid of plasma, phospholipids (PL) predominated in all four species, in correlation with the proportion of HDL, both being largest in dogs. The major PL was phosphatidylcholine (PC), followed by sphingomyelin (SM) and lysophosphatidylcholine. The total plasma lipid from the four species contained long-chain n-6 polyunsaturated fatty acids as the predominant acyl groups, followed by comparable proportions of total saturated and monoenoic fatty acids and small percentages of n-3 PUFA. The percentages of these four major groups of fatty acids in PC, SM, triacylglycerols and cholesterol esters were similar among species, but showed significant differences in the ratios between major individual fatty acids composing these groups.

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.

Reactivity of a population of armadillo lymphocytes with an antibody to human gamma,delta T-cells.

Reactivity of lymphocytes from the nine-banded armadillo (Dasypus novemcinctus) was examined by flow cytometry using a panel of 16 commercially available fluorochrome-conjugated monoclonal antibodies raised against human or murine leukocyte antigens. The only reactivity observed was with antibody TCRdelta1, directed against a common determinant on the delta chain of the human gamma,delta T-cell receptor. Using this antibody, a distinct, bright population of lymphocytes was seen in the peripheral blood in all of 47 animals examined, accounting for 2.0-47.1% of lymphocytes (median, 10.6%). The gamma,delta-reactive lymphocyte population comprised a greater percentage of intraepithelial lymphocytes in the small intestine than in the blood; variable percentages of gamma,delta-reactive cells were also observed in the spleen, thymus, lymph nodes, and bone marrow, and in cutaneous lepromas. In armadillos with disseminated Mycobacterium leprae infection, a significantly greater percentage of circulating lymphocytes reacted with the anti-gamma,delta antibody. This is the first described reactivity of armadillo lymphocytes with a monoclonal antibody to a lymphocyte antigen, and it may offer a useful tool in disease models involving the armadillo.

Plasma and erythrocyte cholinesterase values for the common long-nosed armadillo, Dasypus novemcinctus.

Plasma and erythrocyte cholinesterase activities were determined for 40 free-living and 12 captive common long-nosed armadillos (Dasypus novemcinctus) in order to establish normal values for monitoring pesticide exposure. Plasma cholinesterase activity ranged from 105 to 549 U/liter with no sexual or seasonal differences. Plasma values from captive animals were significantly lower than those from wild armadillos. Erythrocyte cholinesterase activity ranged from 2,915 to 15,126 U/liter with no differences detected between captive and wild animals or between sexes. However, erythrocyte cholinesterase values varied seasonally. Erythrocyte and plasma cholinesterase activities were not significantly correlated. Packed cell volume ranged from 24 to 51% and did not vary significantly between captive and wild samples, between sexes or among seasons. However, both whole blood and erythrocyte cholinesterase activities showed significant negative correlations with packed cell volume. Controlled experiments are needed to find the factors responsible for the statistically significant difference between plasma cholinesterase activities of captive and wild armadillos. The seasonal variation in erythrocyte cholinesterase activity and the negative correlation between erythrocyte cholinesterase activity and packed cell volume can be explained by an hypothesis that relates the variation in erythrocyte cholinesterase activity to variation in erythrocyte turnover rate. Future work should involve experiments to test this hypothesis.