Intracellular Parasites Toxoplasma gondii and Besnoitia besnoiti, Unveiled in Single Host Cells Using AP-SMALDI MS Imaging.

The obligate intracellular apicomplexan parasites Toxoplasma gondii and Besnoitia besnoiti are important causes of disease in both humans and cattle. To date, effective specific treatments are lacking for both infections. To counteract severe symptoms leading to, e.g., disabilities and even abortion in the case of human toxoplasmosis and bovine besnoitiosis, novel targets are required for development of drugs and vaccines. A promising emerging technique for molecular characterization of organisms is high-resolution atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization (AP-SMALDI) mass spectrometry imaging (MSI) which enables semiquantitative visualization of metabolite distributions. MSI was here used to trace and characterize lipid metabolites in primary bovine umbilical vein endothelial cells (BUVECs) upon infection with tachyzoites, an early and pathogenic fast-replicating life stage of T. gondii and B. besnoiti. A cell bulk, derived from noninfected controls and parasite-infected cell pellets, was analyzed by AP-SMALDI MSI in technical and biological triplicates. Multivariate statistical analysis including hierarchical clustering and principle component analysis revealed infection-specific metabolites in both positive- and negative-ion mode, identified by combining database search and LC-MS2 experiments. MSI analyses of host cell monolayers were conducted at 5 µm lateral resolution, allowing single apicomplexan-infected cells to be allocated. This is the first mass spectrometry imaging study on intracellular T. gondii and B. besnoiti infections and the first detailed metabolomic characterization of B. besnoiti tachyzoites. MSI was used here as an efficient tool to discriminate infected from noninfected cells at the single-cell level in vitro.

X-ray evaluation of intestinal dysmotility induced by Eimeria pragensis infection in C57BL/6 mice.

This study was conducted to elucidate the intestinal dysmotility during coccidiosis. C57BL/6 male mice at seven weeks of age were inoculated with Eimeria pragensis sporulated oocysts (100 to 1,000 oocysts). The intestinal motility was evaluated by observing discharging time of barium sulfate (Ba2SO4) after oral administration (WITT: the whole intestinal transit time). The exact location of the dysmotility was analyzed by intermittent barium gastrography. Upper intestinal dysmotility was evaluated by charcoal propulsion study. Additionally, the occurrence of dysmotility was observed at different post-infection times (4, 7, and 14 days post-infection (d.p.i.)) and in infection-dose dependent manner (100, 300, and 1,000 oocysts). As the E. pragensis infected mice had significantly lower feed intake compared to the control group, we designed a feed apprehension study to evaluate the effect of low feed intake on the intestinal dysmotility. The WITT of infected mice at 7 d.p.i. was significantly longer (6 hr) than the uninfected mice (2.5 hr). Intestinal dysmotility was observed in the small intestine, caecum, and colorectum in the infected mice. Charcoal propulsion was slower in infected group (reaching to 40.4% of the whole small intestine) compared to control group (68.0%). The dysmotility was observed at the beginning of the patent period (7 d.p.i.) and subsided as the patency ended (14 d.p.i.). Mice with lower feed intake appeared to have similar intestinal motility as control mice. In summary, this study revealed the evidence of intestinal hypomotility during E. pragensis infection.

Comparative ultrasonographic investigations of the gastrointestinal tract and the liver in healthy and diseased pigeons.

The use of ultrasound as a diagnostic tool in birds has been documented for cardiac, urogenital, and liver disease. However, its use in gastrointestinal tract disease is not defined. Therefore, the purpose of this study was to compare the ultrasonographic findings of the intestine and liver of six healthy racing pigeons with those of six racing pigeons with gastrointestinal disease. The echogenicity of the liver was significantly different between the two groups. Pigeons with gastrointestinal disease had less homogeneous liver echogenicity with focal heterogeneous areas and the hepatic blood vessels were visible and dilated. The duodenum was visualized and its mean diameter of 7.2 +/- 0.3 mm in the diseased pigeons was significantly wider (P < or = 0.001) than the 5.7 +/- 0.2 mm in healthy birds. The thickness of the duodenal wall in healthy and diseased pigeons was 1.6 +/- 0.1 and 2.4 +/- 0.1 mm, respectively, and they were significantly different (P < or = 0.001). We defined baseline measurements for the duodenal loop in pigeons and provided evidence that ultrasound can be a useful diagnostic tool for investigating intestinal disease in pigeons.

Comparative features of double-contrast barium studies in patients with isosporiasis and strongyloidiasis.

OBJECTIVE: To compare the gastrointestinal features of isosporiasis and strongyloidiasis. METHODS: Two patients with isosporiasis and three patients with strongyloidiasis were assessed by double-contrast radiography of the duodenum and small intestine, with reference to histology of the duodenal biopsy specimens. RESULTS: Both conditions affected the duodenum and the proximal jejunum, and showed similar radiographic changes as the diseases progressed. Thus, three patients with diarrhoea lasting 1 year or less showed only minimal or irregularly thickened mucosal folds, which seemed to result from mucosal inflammation. Two patients with long-standing disease periods (17 years and 30 years) presented a markedly granular mucosal appearance with effacement of the folds on radiography. These chronological differences in the radiographic features seemed to reflect the degree of villous atrophy. CONCLUSION: Isosporiasis has similar radiographic features of strongyloidiasis.

Cecal filling and defecation of chickens infected with Eimeria tenella.

White Leghorn cockerels, 11 to 22 days old, were inoculated each with a single oral dose of 4-5 X 10(4) sporulated oocysts of Eimeria tenella. Radiographic study of urinary backflow in infected chickens injected with sodium iothalamate subcutaneously indicated that retrograde movement of ceca was impaired particularly 7, 10, and 14 days after infection. No inflow was noted 7 days after infection when barium sulfate was inoculated into cloaca. Weight of cecal contents examined 7 days after infection was significantly smaller than uninfected control. Number of cecal feces was counted every 24 h beginning 4 through 14 days after infection. The counts in infected birds were significantly fewer than uninfected control 8, 9, and 10 days after infection. Outflow of cecal contents was studied in chickens surgically injected with barium sulfate into cecum 7 days after infection. Radiographic study indicated that most of uninfected control ceca excreted or evacuated the medium between 10 and 24 h after injection, while a few infected birds cleared ceca during the same period.

Cryptosporidiosis of the stomach and small intestine in patients with AIDS.

Cryptosporidiosis is a parasitic disease of the gastrointestinal tract that causes a choleralike diarrhea in patients with the acquired immunodeficiency syndrome (AIDS). Review of barium studies of the stomach and small intestine in 16 AIDS patients with cryptosporidiosis showed that the studies were abnormal in 13. Of these, five had moderate or marked prominence of the mucosal folds, and eight had slight prominence. The jejunum was predominantly involved in nine; the entire small intestine was uniformly affected in the other four. Three of the 16 patients had moderate or marked dilatation of the small intestine. One had marked dilution of the barium, and one had marked fragmentation and flocculation of the barium. There was narrowing and rigidity of the gastric antrum in two patients. These radiographic findings are nonspecific, but are indicative of cryptosporidiosis in a patient with AIDS and protracted diarrhea.

Hepatozoon canis infection in dogs: clinical, radiographic, and hematologic findings.

Hepatozoon canis gametocytes were identified in circulating neutrophils of 3 dogs from different parts of Texas. Clinical signs included intermittent fever, stiffness, and inappetence. The dogs had marked neutrophilic leukocytosis (35,000--51,000/mm3). Parasitemia varied from less than 1% to 60% of the circulating neutrophils. Two of the dogs had periosteal new born reaction on the vertebrae, ilia, ribs, and bones of the limbs proximal to the carpus and tarsus. There was no evidence that specific drug therapy was of value, although use of corticosteroids, aspirin, and antimicrobial agents was accompanied by clinical improvement.