Neuronal vacuolation in raccoons from Oregon.

During a 2-year period (1995-1997), vacuoles were detected in neurons of 21/50 (42% prevalence) raccoons (Procyon lotor) in Oregon. Age or sex predisposition was not apparent. Twenty of these raccoons were from within a radius of 40 km of Corvallis in western Oregon. Microscopically, the vacuoles were variable in size, were in the perikarya, and were consistently present in pontine nuclei. Brain tissues were negative for rabies virus antigen by fluorescent antibody test and for the protease-resistant protein prion by immunohistochemistry. Electron microscopic examination of the brain stem of selected animals revealed accumulation of electron-dense material within neuronal perikarya. Light and electron microscopic examination indicated that the accumulated intracellular material had a high lipid content. These lesions suggest a form of neuronal storage condition. Further research is required to identify the composition of the intracellular lipid material, to elucidate the mechanism of neuronal vacuolation in raccoons, and to understand the basis for the apparent geographic restriction of this lesion.     

Chronic dilatative myocarditis caused by Trypanosoma cruzi in two dogs

Trypanosoma cruzi was believed responsible for causing chronic dilatative myocarditis in 2 female hunting dogs. Clinical signs included ascites, respiratory distress, thoracic effusion, cyanosis, and weak pulse with ventricular arrhythmias. Electrocardiography indicated first-degree heart block, chamber enlargement, and ventricular-based arrhythmias unresponsive to treatment. M-mode echocardiography of 1 dog confirmed bilateral cardiac enlargement and septal and left ventricular free wall thinning. Multifocal infiltrates of plasma cells, lymphocytes, and histiocytes, cardiocyte degeneration, and multifocal fibrosis were the predominant histologic lesions. Trypanosoma cruzi pseudocysts were infrequently found.     

Dehydroepiandrosterone increases resistance to experimental infection by Trypanosoma cruzi

Dehydroepiandrosterone (DHEA) enhances immune responses against a wide range of viral, bacterial, and parasitic pathogens. In a previous study, we reported that administration of DHEA significantly decreased the numbers of blood parasites in Trypanosoma cruzi experimental infection. The present study was undertaken to determine the effectiveness of DHEA in reducing the severity of acute phase T. cruzi infection of male and female Wistar rats. Animals were treated subcutaneously with 40 mg/kg body weight/day of DHEA. The concentration of nitric oxide (NO) was determined in spleen peritoneal cavity. Interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were determined in the sera of uninfected and infected animals. DHEA treatment augments NO production for both sexes after in vitro LPS treatment for uninfected animals. Infection triggered enhanced NO levels although not significant. IL-2 and IFN-gamma were detectable in higher concentrations in treated and infected rats of both genders when compared to untreated controls. These data suggest that DHEA may have a potent immunoregulatory function that can affect the course of T. cruzi infection.     

Epidemiological survey of Trypanosoma cruzi infection in domestic owned cats from the tropical southeast of Mexico

American trypanosomiasis is an infectious disease of importance for public health and caused by the protozoa Trypanosoma cruzi mainly transmitted by triatomine bugs. The precise role of cats in the peridomestic transmission of T. cruzi and the mechanism by which cats become infected remain uncertain. The objective of this work was to determine the prevalence of T. cruzi infection in domestic cats from an urban area of tropical Mexico by serological and molecular methods and evaluate associated risk factors. A total of 220 domestic cats from Merida Yucatan, Mexico, were studied. Animals older than 3 months were blood sampled. Serum and DNA were obtained. Specific T. cruzi IgG antibodies were detected using a commercial indirect ELISA with an anti-cat antibody HRP labelled. Positive cases were confirmed by Western blot (WB). Polymerase chain reaction (PCR) was also performed using the primers TC1 and TC2. From the 220 cats, 8.6% had antibodies against T. cruzi using ELISA test and later confirmed by WB. In 75 cats (34%), the sequence of ADNk of T. cruzi was amplified. The bad-regular body condition was the only risk factor associated with PCR positive to T.cruzi (P < 0.001). In Mexico, there are no previous epidemiological reports that demonstrate the importance of the cat as a reservoir of T. cruzi. Few individuals were identified with a serological response because they were probably at an early stage of infection or antibodies were not detected because they could be immunocompromised (FIV, FeLV or others). It is necessary to monitor PCR-positive patients and conduct further studies for better understanding of the epidemiology and pathogenesis of Chagas disease in domestic cats.     

Haematological and histopathological findings after ovariectomy in Trypanosoma cruzi infected mice

The aim of this study was to investigate the influences of ovariectomy on histopathological and hematological parameters during the course of Trypanosoma cruzi infection. Hematological and immunological homeostasis is influenced by gonadal steroid hormones. Ovariectomy exerts profound influences on parasitic diseases including T. cruzi infection through modulation of the host's immune response. Three groups of female Mus musculus were infected with 4000 blood trypomastigotes of the Y strain of T. cruzi. One group was subjected to ovariectomy, another to simulated surgery before the infection, and a third group of unoperated animals were used as controls. Marked differences were detected in the responses of blood and tissue parasites. On day 9, post-infection parasitism was significantly higher in ovariectomized animals (P<0.05). These results were confirmed by histopathological studies, in which ovariectomized animals displayed hearts with higher number of amastigote burdens, increased inflammatory infiltrate, enhanced tissue fibers disorganization and decreased lytic antibody percentage, when compared to their counterparts. On day 9 the hematological changes were more apparent, with a decrease in erythrocytes, platelets and leucocytes for ovariectomized infected animals. Simulated surgery, as a stressful agent, did not cause any imbalance in parasitism or in the hemogram profile. The results confirm the importance of the female steroids in resistance against T. cruzi infection.     

An atypical case of Trypanosoma cruzi infection in a young English Mastiff

A case of Trypanosoma cruzi infection in a young English Mastiff from Texas is presented. Clinical signs and laboratory findings included subcutaneous edema, lymphadenopathy, weight loss, and hypoalbuminemia. Cytology of a lymph node revealed numerous amastigotes. No trypomastigotes were observed in buffy coat preparation of peripheral blood, and on histologic evaluation, most organs contained numerous interstitial pseudocysts. Initial serology was positive for both T. cruzi and Leishmania, and immunohistochemistry supported a diagnosis of Leishmania. However, additional serology supported a T. cruzi infection, and cultivation of organisms isolated from a lymph node revealed morphology consistent with T. cruzi. In addition, PCR analysis resulted in a 504 bp fragment with 99% homology to a flagellar protein of T. cruzi. Although uncommon, autochthonous cases of both T. cruzi and Leishmania have been reported in the United States. Clinical signs observed with both diseases can show many similarities, cytology may be indistinguishable, as in this case, and serological cross-reactivity is common. This case demonstrates an unusual presentation of T. cruzi and the use of multiple testing strategies to support its diagnosis.     

Isolation, mouse pathogenicity, and genotyping of Trypanosoma cruzi from an English Cocker Spaniel from Virginia, USA

Trypanosoma cruzi was demonstrated in blood smears and heart tissue from a 5-year old, female, English Cocker Spaniel that had never been outside of the state of Virginia, USA. Plasma from the dog was positive in a commercially available immunochromatographic dipstick assay for T. cruzi and negative in an immunochromatographic dipstick assay for visceral Leishmania spp. The plasma from the dog had an indirect immunofluorescent antibody titer of 1:800 against epimastigotes of T. cruzi while the titer was 1:50 against promastigotes of L. infantum. The parasite was isolated from the blood in vitro from the dog (TcVT-1 isolate) and used to experimentally infect female C3H and ICR mice. The parasite was nonpathogenic for experimentally inoculated mice. DNA was isolated from parasites grown in vitro and used to determine that the genotype of T. cruzi present in the dog was genotype TcIV. This genotype is common in raccoons, Procyon lotor, in North America and suggests that raccoons may serve as reservoirs for canine infection.     

An endemic focus of Trypanosoma cruzi infection in a subhuman primate research colony.

Leishmanial forms of Trypanosoma cruzi with multifocal pseudocysts were identified in a deceased adult female rhesus monkey (Macaca mulatta) that had been maintained in an outdoor primate colony in southern Texas. Subsequent clinical investigation of the remainder of the colony identified 20 additional monkeys seropositive for T cruzi. Concurrent epizootiologic surveys confirmed the presence of sylvatic reservoirs and triatomine vectors of T cruzi infection and identified a natural T cruzi transmission cycle. Veterinarians associated with the research colony were informed of the potential for further T cruzi transmission and appropriate insect vector and reservoir animal control programs were implemented..     

Neuroaxonal dystrophy in raccoons (Procyon lotor) from Iowa.

During a 12-month period (1998-1999), microscopic evidence of neuroaxonal dystrophy (NAD) in medullae oblongata of raccoons (Procyon lotor) was observed in 17/39 (47% prevalence in adults) from Iowa, USA. Three of the animals were kits (<3 months), 26 were between 1 and 2 years, and 10 were over 7 years. Lesions were not seen in the medullae of the 3 kits. In young adults, the lesions were mild and were seen in 7 animals. More severe lesions were present in the 10 older raccoons. Grossly, the brains were unremarkable. Microscopically, NAD was confined to the dorsal caudal medulla, where certain nuclei (predominantly gracilis and cuneate) were bilaterally affected. Severely affected animals had vacuolar degeneration of neurons or neuronal loss and extensive areas of spongiosis. Tests for the presence of PrP(res) in the brain were negative. Spongiotic areas often contained axonal spheroids. Degenerate neurons and axons occasionally contained amphophilic periodic acid-Schiff-positive granular material. There was a paucity of inflammatory cells in the affected areas. Since lesions were not present in kits, were either absent or mild in young adults, and were severe in older raccoons, the findings may be related to advancing age. Neuroaxonal dystrophy has not been previously reported in raccoons. Retrospective examination of raccoon brains from the eastern and northwestern areas of the country revealed very low prevalence of NAD. Because of the apparently high prevalence of this condition at this geographic location, factors other than age (genetic, nutritional, and/or environmental) may influence this degenerative process in the brains of raccoons in Iowa.     

Specific humoral and cellular immunity induced by Trypanosoma cruzi DNA immunization in a canine model

Chagas disease has a high incidence in Mexico and other Latin American countries. Because one of the most important known methods of prevention is vector control, which has been effective only in certain areas of South America, the development of a vaccine to protect people at risk has been proposed. In this study, we assessed the cellular and humoral immune response generated following immunization with pBCSP and pBCSSP4 plasmids containing the genes encoding a trans-sialidase protein (present in all three forms of T. cruzi) and an amastigote specific glycoprotein, respectively, in a canine model. Thirty-five beagle dogs were divided randomly into 5 groups (n = 7) and were immunized twice intramuscularly with 500 μg of pBCSSP4, pBCSP, pBk-CMV (empty plasmid) or saline solution. Fifteen days after the last immunization the 4 groups were infected intraperitoneally with 500 000 metacyclic trypomastigotes. The fifth group was unimmunized/infected. The parasitaemia in the immunized/infected dogs was for a shorter period (14 vs. 29 days) and the parasite load was lower. The concentration of IgG1 (0.612 ± 0.019 O.D.) and IgG2 (1.167 ± 0.097 O.D.) subclasses was measured (absorbance) 15 days after the last immunization with both recombinant plasmids, the majority of which were IgG2. The treatment of parasites using the serum from dogs immunized with pBCSP and pBCSSP4 plasmids produced 54% (± 11.8) and 68% (± 21.4) complement-mediated lysis, respectively. At 12 h post immunization, an increase in cytokines was not observed; however, vaccination with pBCSSP4 significantly increased the levels of IFN-γ and IL-10 at 9 months post-infection. The recombinant plasmid immunization stimulated the spleen cell proliferation showing a positive stimulatory index above 2.0. In conclusion, immunization using both genes effectively induces a humoral and cellular immune response.     

Pythiosis of the digestive tract in dogs from Oklahoma

Enteric pythiosis was diagnosed in nine dogs in Oklahoma. Eight dogs had anorexia and weight loss. Two of these dogs had diarrhea; two dogs exhibited vomiting and diarrhea; and one dog had vomiting. One dog presented with dysphagia. Seven dogs had either a palpable or radiographically visible abdominal mass. These seven dogs had localized regions of mucosal ulceration and thickened gastric or intestinal walls with some involvement of the adjacent mesentery or omentum. Two dogs had enlarged regional mesenteric lymph nodes. One dog that presented with dysphagia had an oropharyngeal mass involving the larynx and cranial esophagus. Microscopically, there was transmural chronic sclerosing and granulomatous to pyogranulomatous inflammation with arteritis. Pythium spp. were identified in all specimens by immunohistochemistry.     

IgG isotype profile is correlated with cardiomegaly in Beagle dogs infected with distinct Trypanosoma cruzi strains

A systematic study following infection by various strains of the protozoan parasite, Trypanosoma cruzi, and the simultaneous monitoring of the humoral immune response together with the elicited cellular response, could add greatly to our understanding of differences between strains of this important human pathogen. In that sense, acute and chronic infections with distinct T. cruzi strains (Y, Berenice-78 and ABC) in Beagle dogs were studied through a longitudinal evaluation of immunoglobulin G (IgG), IgG1 and IgG2 isotypes (by ELISA and flow cytometry (FC)), as well as measurements of peripheral blood mononuclear cell (PBMC) proliferation over a 100-week period, and their correlation with cardiomegaly. Our results show that infected animals presenting cardiomegaly showed lower or absent levels of IgG1 during the chronic phase of the infection, when compared to those that did not show an increase in heart weight. In that manner, our results suggest that IgG1 could be used as a marker for cardiac pathogenicity in Chagas disease.     

Clinical, clinicopathologic, and parasitologic observations of trypanosomiasis in dogs infected with North American Trypanosoma cruzi isolates

Nineteen purebred Beagles of various ages (4, 5, 13, and 47 weeks) were inoculated with North American Trypanosoma cruzi isolates obtained from an opossum (Tc-O), an armadillo (Tc-A), or a dog (Tc-D). Dogs were grouped on the basis of clinical outcome of infection. During the acute stage of disease, dogs of group 1 (n = 7 inoculated with Tc-O or Tc-A) died or were euthanatized because of the severity of disease. Dogs of group 2 (n = 5 inoculated with Tc-O or Tc-A) developed acute disease, but survived to develop chronic disease. Dogs of group 3 (n = 7 Tc-D-inoculated dogs) developed neither acute nor chronic disease. Dogs of group 4 (n = 4--2 dogs 13 weeks old and 2 dogs 47 weeks old) served as noninoculated controls. Clinical signs associated with severe acute myocarditis developed in dogs of groups 1 and 2 between postinoculation day (PID) 15 and 28. Generalized lymphadenopathy and lymphocytosis were observed in all dogs of groups 1, 2, and 3 between PID 14 and 17. Serum alanine transaminase and aspartate transaminase activities and urea nitrogen concentration were high, and glucose concentration was low prior to death of dogs in group 1. Serum activities of isoenzymes of creatine kinase were significantly (P less than 0.05) high in only 1 dog (group 1), whereas serum lactate dehydrogenase isoenzyme activities were not significantly high in any dog.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevalence of Toxoplasma gondii infection in raccoons.

Serum samples from 427 raccoons (93 from Pennsylvania, 45 from New Jersey, 72 from South Carolina, 68 from Virginia, 30 from Iowa, and 119 from Ohio) were evaluated for Toxoplasma gondii antibodies in dilutions of 1:25, 1:50, and 1:500. The distribution of T gondii antibody titers was less than 1:25 for 212 raccoons (49.6%), 1:25 for 34 raccoons (7.9%), 1:50 for 117 raccoons (27.4%), and greater than or equal to 1:500 for 64 raccoons (14.9%). Tissue cysts were seen in the liver, and tachyzoites were in the brain of a raccoon with abnormal neurologic signs and concurrent infection with canine distemper virus. Organisms in the liver were stained with anti-T gondii serum, and the raccoon had a T gondii titer of 1:160 in the agglutination test.     

Growth of Chickasaw Plum in Oklahoma

Management of rangelands for wildlife and livestock entails understanding growth of clonal shrubs such as Chickasaw plum (Prunus angustifolia Marsh.). We studied growth of this species in one county in north-central (Payne) and two counties in northwestern Oklahoma (Ellis, Harper) during 2006 and 2007. We estimated age of stems and roots by growth rings and area of stands with the use of a handheld GPS unit. Based on zero-intercept regression models, stands grew at similar rates (overlapping 95% confidence intervals [CIs]) among counties with a pooled estimate of 31.0 m² · yr⁻¹ (95% CI = 26.5–35.6 m² · yr⁻¹; n = 95). This rate showed considerable variability within and among study sites (r = 0.52). Stem diameter increased (zero-intercept models) more rapidly in north-central Oklahoma (5.27 mm · yr⁻¹; 95% CI = 5.01–5.53 mm · yr⁻¹; r = 0.90; n = 53) than in northwestern Oklahoma (3.68 mm · yr⁻¹; 95% CI = 3.55–3.81 mm · yr⁻¹; r = 0.91; n = 102); data were pooled because of similar rates in Ellis and Harper counties. Stem height was a power function of stem age (y = 0.97x^0.28; r = 0.56), indicating rate of growth in height (m · yr⁻¹) declined with age according to dy/dx = 0.27x^−0.72. Knowledge of the area expansion rate of Chickasaw plum clones aids in management planning to increase or decrease canopy coverage by this shrub.     

Plasmid DNA immunization with Trypanosoma cruzi genes induces cardiac and clinical protection against Chagas disease in the canine model

The only existing preventive measure against American trypanosomosis, or Chagas disease, is the control of the transmitting insect, which has only been effective in a few South American regions. Currently, there is no vaccine available to prevent this disease. Here, we present the clinical and cardiac levels of protection induced by expression to Trypanosoma cruzi genes encoding the TcSP and TcSSP4 proteins in the canine model. Physical examination, diagnostic chagasic serology, and serial electrocardiograms were performed before and after immunization, as well as after experimental infection. We found that immunization with recombinant plasmids prevented hyperthermia in the acute phase of experimental infection and produced lymphadenomegaly as an immunological response against the parasite and additionally prevented heart rate elevation (tachycardia) in the acute and/or chronic stages of infection. Immunization with T. cruzi genes encoding the TcSP and TcSSP4 antigens diminished the quality and quantity of the electrocardiographic abnormalities, thereby avoiding progression to more severe developments such as right bundle branch block or ventricular premature complexes in a greater number of dogs.